diff --git a/README.md b/README.md index 3bbf953..002ecaf 100644 --- a/README.md +++ b/README.md @@ -4,25 +4,45 @@ [PubTator](https://www.ncbi.nlm.nih.gov/CBBresearch/Lu/Demo/PubTator/) and its 2.0 version ([PubTator Central](https://www.ncbi.nlm.nih.gov/CBBresearch/Lu/Demo/PubTatorCentral/)) uses text mining to tag PubMed abstracts/artciles with standardized concepts. This repository retrieves and processes PubTator annotations for use in [`greenelab/snorkeling`](https://github.com/greenelab/snorkeling) and elsewhere. -## Environment +# Get Started -Install the [conda](https://conda.io) environment specified in [`environment.yml`](environment.yml) by running: +## Set-up Environment + +### Conda + +1. Install the [conda](https://conda.io) environment. +2. Create the pubtator environmenmt by running: + +```sh +conda create --name Pubtator python=3.8 +``` +3. Install packages via pip by running the following: + +```sh +pip install -r requirements.txt +``` + +4. Activate with `conda activate pubtator`. + +### Pip + +1. Make sure you have python version **3.8** installed. +2. Install packages by running the following: ```sh -conda env create --file environment.yml +pip install -r requirements.txt ``` -Activate with `conda activate pubtator`. ## Execution -To download and extract Pubator Central's data (default) run the following: +To start processing Pubtator/Pubtator Central run the following command: ```sh -bash execute.sh {email address here} +python execute.py --config config_files/pubtator_central_config.json ``` -If the original Pubtator is desired run the above command with the following flag: --pubtator. You do not need to provide your email address when running the first version of Pubtator. +If the original Pubtator is desired replace `pubtator_central_config.json` with `pubtator_config.json`. The json file contains all the necessary parameters needed to run. More information for the json file can be found [here](config_files). ## License diff --git a/config_files/CONFIG.md b/config_files/CONFIG.md new file mode 100644 index 0000000..9008a35 --- /dev/null +++ b/config_files/CONFIG.md @@ -0,0 +1,96 @@ +# Configuration Files Explained + +This file explains the pipeline steps and parameters needed for each step. +**Note: any added parameter or step will be ignored unless `execute.py` is manually changed.** + +## Repository Download + +This is the first step of the Pubtator pipeline. +Basically this step downloads Pubtator Central's annotation file from their ftp server. + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| url | the url to download the file from | a string with a url path | +| download_folder | the folder to hold the downloaded file | a string name for the folder | +| skip | tell execute.py to ignore this step and contine | true or false | + +## Pubtator to XML + +This is the second step of the Pubtator pipeline. +This step converts Pubtator/Pubtator Central's annotation file into xml format. +**Note: This step may take awhile to complete** + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| documents | The file path pointing to the downloaded file from the previous step. | a string for the file path | +| output | The file path to save the xml file. Make sure to keep the xz extension. | a string for the file path | +| skip | Tell execute.py to ignore this step and contine | true or false | + +## Extract Tags + +This is the second step of the Pubtator pipeline. +This step extracts Pubtator/Pubtator Central's annotations from the xml file. +It outputs a tsv file that contains all extracted annotation. +**Note: This step may take awhile to complete** + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| input | The file path pointing to the xml file in previouss step. Make sure to keep the xz extension. | a string for the file path | +| output | The file path to save the tsv file. Make sure to keep the xz extension. | a string for the file path | +| skip | Tell execute.py to ignore this step and contine | true or false | + +## Hetnet ID Extractor + +This is the third step of the Pubtator pipeline. +This step filters out extracted annotations to only include tags within [Hetionet's Database](https://het.io/). + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| input | The file path pointing to the tsv file in previous step. Make sure to keep the xz extension. | a string for the file path | +| output | The file path to save the tsv file. Make sure to keep the xz extension. | a string for the file path | +| skip | Tell execute.py to ignore this step and contine | true or false | + +## Map PMIDS to PMCIDS + +This is the forth step of the Pubtator pipeline. +This step querys NCBI's pmid to pmcid mapper in order to grab PMCIDS. +**Note: To download full text you will need to have PMCIDS. PMIDS will not work.** + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| input | The file path pointing to the tsv file in extract tags step. Make sure to keep the xz extension. | a string for the file path | +| output | The file path to save the tsv file. | a string for the file path | +| debug | This is a flag for debugging purposes. Feel free to ignore and leave as false. | true or false | +| skip | Tell execute.py to ignore this step and contine | true or false | + +## Download Full Text + +This is the fifth step of the Pubtator pipeline. +This step queries Pubtator Central's api and downloads annotated full text if text is present. + +Following Parameters for this section: +| Param | Description | Accepted Values | +| --- | --- | --- | +| input | The file path pointing to the tsv file in previous step. | a string for the file path | +| output | The file path to save the xml file. | a string for the file path | +| temp_dir | The folder to hold temporary batch files for this step of the pipeline | a string for the folder path | +| log_file | A log file that keeps track of the IDs that have already been queried. It is used to monitor progress in case the process is interrupted. Make sure it has the tsv extension. | a file path for the file | +| skip | Tell execute.py to ignore this step and contine | true or false | + +## Extract Full Text Tags + +This is the sixth step of the Pubtator pipeline. +This step extracts tags from full text documents. +Please refer to [Extract Tags Section](#extract-tags) for parameter details. + +## Hetnet ID Extractor Full Text + +This is the last step of the Pubtator pipeline. +This step filters tags to only have Hetionet tags. +Please refer to [Hetnet ID Extractor Section](#hetnet-id-extractor) for parameter details. + diff --git a/config_files/README.md b/config_files/README.md new file mode 100644 index 0000000..64580ed --- /dev/null +++ b/config_files/README.md @@ -0,0 +1,37 @@ +# Configuration Files + +## File Description + +| File | Description | +| --- | --- | +| [pubtator central config](pubtator_central_config.json) | This is a configuration file for parsing Pubtator Central. | +| [pubtator config](pubtator_config.json) | This is a configuration file for parsing Pubtator (older version of Pubtator Central). | +| [tests config](tests_config.json) | This is a configuration file for testing the pubtator system. Feel free to ignore this file. | + +## Usage + +Each configuration file is in json format and contains parameters for each step within the pubtator pipeline. +All files are organized by order of operation, which means the very first step occurs at the top and the subsequent step comes right afterwards. +Every step can be skipped, which allows one to continue the pipeline at any step one chooses. +Please refer to [CONFIG.md](CONFIG.md) for more details on each pipeline step and their respective parameters. + +Example config file: +```json +{ + "pipeline step 1":{ + "param1":"param1_value", + "param2":"param2_value", + "skip":false + }, + "pipeline step 2":{ + "param1":"param1_value", + "param2":"param2_value", + "skip":false + }, + "pipeline step 3":{ + "param1":"param1_value", + "param2":"param2_value", + "skip":false + } +} +``` diff --git a/config_files/pubtator_central_config.json b/config_files/pubtator_central_config.json new file mode 100644 index 0000000..c6146a2 --- /dev/null +++ b/config_files/pubtator_central_config.json @@ -0,0 +1,54 @@ +{ + "repository_download":{ + "url":"ftp://ftp.ncbi.nlm.nih.gov/pub/lu/PubTatorCentral/bioconcepts2pubtatorcentral.offset.gz", + "download_folder":"download", + "skip":false + }, + + "pubtator_to_xml": { + "documents":"download/bioconcepts2pubtatorcentral.offset.gz", + "output":"data/pubtator-central-docs.xml.xz", + "skip":false + }, + + "extract_tags":{ + "input":"data/pubtator-central-docs.xml.xz", + "output":"data/pubtator-central-tags.tsv.xz", + "skip":false + }, + + "hetnet_id_extractor":{ + "input":"data/pubtator-central-tags.tsv.xz", + "output":"data/pubtator-central-hetnet-tags.tsv.xz", + "skip":false + }, + + "map_pmids_to_pmcids":{ + "input":"data/pubtator-central-tags.tsv.xz", + "output":"data/pubtator-pmids-to-pmcids.tsv", + "debug":false, + "skip":false + }, + + "download_full_text":{ + "input":"data/pubtator-pmids-to-pmcids.tsv", + "document_batch":100, + "output":" data/pubtator-central-full-text.xml", + "temp_dir":"data/temp", + "log_file":"batch_log.tsv", + "skip":false + }, + + "extract_full_text_tags":{ + "input":"data/pubtator-central-full-text.xml", + "output":"data/pubtator-central-full-text-tags.tsv.xz", + "skip":false + }, + + "hetnet_id_extractor_full_text":{ + "input":"data/pubtator-central-full-text-tags.tsv.xz", + "output":"data/pubtator-central-full-hetnet-tags.tsv.xz", + "skip":false + } + +} diff --git a/config_files/pubtator_config.json b/config_files/pubtator_config.json new file mode 100644 index 0000000..5693046 --- /dev/null +++ b/config_files/pubtator_config.json @@ -0,0 +1,25 @@ +{ + "repository_download":{ + "url":"ftp://ftp.ncbi.nlm.nih.gov/pub/lu/PubTator/bioconcepts2pubtator_offsets.gz", + "download_folder":"download", + "skip":false + }, + + "pubtator_to_xml": { + "documents":"download/bioconcepts2pubtator_offsets.gz", + "output":"data/pubtator-docs.xml.xz", + "skip":false + }, + + "extract_tags":{ + "input":"data/pubtator-docs.xml.xz", + "output":"data/pubtator-tags.tsv.xz", + "skip":false + }, + + "hetnet_id_extractor":{ + "input":"data/pubtator-tags.tsv.xz", + "output":"data/pubtator-hetnet-tags.tsv.xz", + "skip":false + } +} diff --git a/config_files/tests_config.json b/config_files/tests_config.json new file mode 100644 index 0000000..782281f --- /dev/null +++ b/config_files/tests_config.json @@ -0,0 +1,54 @@ +{ + "repository_download":{ + "url":"ftp://ftp.ncbi.nlm.nih.gov/pub/lu/PubTatorCentral/bioconcepts2pubtatorcentral.offset.gz", + "download_folder":"download", + "skip":true + }, + + "pubtator_to_xml": { + "documents":"data/example/1-sample-annotations.txt", + "output":"data/example/2-sample-docs.xml", + "skip":false + }, + + "extract_tags":{ + "input":"data/example/2-sample-docs.xml", + "output":"data/example/3-sample-tags.tsv", + "skip":false + }, + + "hetnet_id_extractor":{ + "input":"data/example/3-sample-tags.tsv", + "output":"data/example/4-hetnet-tags.tsv", + "skip":false + }, + + "map_pmids_to_pmcids":{ + "input":"data/example/3-sample-tags.tsv", + "output":"data/example/5-sample-pmids-to-pmcids.tsv", + "debug":true, + "skip":false + }, + + "download_full_text":{ + "input":"data/example/5-sample-pmids-to-pmcids.tsv", + "document_batch":100, + "output":"data/example/6-sample-full-text.xml", + "temp_dir":"data/temp", + "log_file":"batch_log.tsv", + "skip":false + }, + + "extract_full_text_tags":{ + "input":"data/example/6-sample-full-text.xml", + "output":"data/example/7-sample-full-text-tags.tsv", + "skip":false + }, + + "hetnet_id_extractor_full_text":{ + "input":"data/example/7-sample-full-text-tags.tsv", + "output":"data/example/8-hetnet-full-text-tags.tsv", + "skip":false + } + +} diff --git a/data/example/2-sample-docs.xml b/data/example/2-sample-docs.xml index cb448f6..5d1b442 100644 --- a/data/example/2-sample-docs.xml +++ b/data/example/2-sample-docs.xml @@ -1,7 +1,7 @@ Pubtator - 2020/03/02 + 2020/08/03 Pubtator.key 1560033 diff --git a/data/example/6-sample-full-text.xml b/data/example/6-sample-full-text.xml index 72e1e67..f64814d 100644 --- a/data/example/6-sample-full-text.xml +++ b/data/example/6-sample-full-text.xml @@ -1,7 +1,7 @@ PubTator -2020/03/02 +2020/08/03 BioC.key @@ -54,7 +54,7 @@ Cronobacter sakazakii - MESH:C489032 + MESH:D011392 Chemical proline @@ -95,89 +95,89 @@ paragraph 709 Survival of the foodbourne pathogen Cronobacter sakazakii in low water activity (aw) environments, e.g. powdered infant formula (PIF), is largely attributed to the accumulation of organic compounds termed osmolytes or compatible solutes. Synthesised de novo, or transported from the bathing solution, compatible solutes function to increase cell turgor thereby counterbalancing the external osmotic upshift and preventing water loss from the cell, which if left unchecked can lead to plasmolysis and ultimately cell death. - + 28141 Species Cronobacter sakazakii - + 9606 Species infant + + MESH:D014867 + Chemical + + water + + + MESH:D014867 + Chemical + + water + INTRO paragraph 1232 In Escherichia coli, a model organism for the study of bacterial osmoadaptation, the transmembrane protein ProP is perhaps the best characterised compatible solute uptake system; facilitating the uptake of both proline and glycine betaine. A member of the major facilitator superfamily (MFS), E. coli ProP is a 500-amino acid protein comprising of 12 transmembrane domains and a characteristic carboxy-terminal extension. In a previous in silico study we identified seven ProP homologues on the C. sakazakii BAA-894 genome; one of which, ESA_02131, encodes a protein exhibiting 90% identity to E. coli ProP. While the remaining six homologues encode proteins exhibiting features of classic secondary transporters, they are all 60-70 amino acids shorter than the E. coli ProP; lacking the extended carboxyl tail. Notwithstanding the lack of structural consistency, particularly at the C-terminal end, we have shown that six of the seven C. sakazakii proP homologues contribute to C. sakazakii osmotolerance, albeit to varying degrees. - + 562 Species Escherichia coli - + 562 Species E. coli - + 28141 Species C. sakazakii - + 562 Species E. coli - + 562 Species E. coli - + 28141 Species C. sakazakii - + 28141 Species C. sakazakii - - MESH:C015732 - Chemical - - carboxy - - - MESH:C489032 + + MESH:D011392 Chemical proline - - MESH:D005998 - Chemical - - glycine - - MESH:C041069 + MESH:D001622 Chemical - - carboxyl + + glycine betaine @@ -185,29 +185,23 @@ paragraph 2268 Culham et al. first described the E. coli ProP as harbouring unusual structural features which appeared unique within the transporter superfamily. This study predicted the formation of an alpha helical coiled coil resulting from the presence of the carboxyl terminal extension. Indeed, a synthetic polypeptide corresponding to the C-terminal extension of ProP formed a dimeric alpha helical coiled coil. Interestingly, when amino acid changes were introduced to the coiled coil, ProP required a larger osmotic upshift to become activated, suggesting that the C-terminal domain likely plays a role in osmosensing. Furthermore, a derivative of ProP which lacked the 26 amino acid C-terminal domain was expressed, but inactive. In contrast, despite the structural degeneracy observed between the homologues, C. sakazakii ProP homologues lacking the C-terminal extension do contribute to osmotolerance, albeit to a lesser extent than the extended ProP (which we designate Prop1) encoded by ESA_02131. - + 562 Species E. coli - + 28141 Species C. sakazakii - - MESH:C041069 - Chemical - - carboxyl - - - MESH:D000596 + + - Chemical - - amino acid C + + ProP @@ -215,30 +209,18 @@ paragraph 3265 While several studies have focused on elucidating the role of the carboxyl extension in E. coli, little is known about the role, if any, of the ProP1 carboxyl extension in the far more osmotolerant C. sakzakii. Herein, we investigate the role of the C-terminal coiled coil of ProP1 in contributing to C. sakazakii osmotolerance, by creating a chimeric protein (ProPc) in which the extended C-terminal domain of ProP1 (encoded by ESA_02131) is spliced onto the truncated C-terminal end of ProP2 (encoded by ESA_01706). - + 562 Species E. coli - + 28141 Species C. sakazakii - - MESH:C041069 - Chemical - - carboxyl - - - MESH:C041069 - Chemical - - carboxyl - METHODS @@ -257,6 +239,12 @@ table 3844 Ampr. This strain is resistant to ampicillian. + + - + Chemical + + ampicillian + TABLE @@ -273,30 +261,24 @@ Tab1.xml 3924 Strain or plasmid Relevant genotype or characteristics Source or reference Plasmids pUC18 Ampr, lacZ', pMB9 replicon pUC18: ESA_02131 pUC18 harboring ESA_02131 gene under control of the native promoter pUC18: ESA_01706 pUC18 harboring ESA_01706 gene under control the native promoter pUC18: ESA_01706CTE pUC18 harboring chimeric ESA_01706 with fused C-terminal extension (ESA_02131) under control of the native promoter This work Strains Cronobacter sakazakii BAA-894 C.sakazakii strain isolated from powdered formula associated with neonatal intensive care unit Escherichia coli DH5alpha Intermediate cloning host.supE44 DeltalacU169(80lacZDeltaM15)R17 recA1 endA1 gyrA96 thi-1 relA1 Invitrogen MKH13 MC4100Delta(putPA)101Delta(proP)2Delta(proU) MKH13 pUC18:ESA_02131+ Host strain harbouring pUC18: ESA_02131 plasmid. Ampr MKH13 pUC18:ESA_01706+ Host strain harbouring pUC18: ESA_01706 plasmid. Ampr MKH13 pUC18:ESA_01706CTE Host strain harbouring pUC18: ESA_01706CTE plasmid. Ampr This work - + 28141 Species Cronobacter sakazakii - + 28141 Species C.sakazakii - + 668369 Species Escherichia coli DH5alpha - - MESH:C538052 - Disease - - MC4100Delta(putPA)101Delta(proP)2Delta - METHODS @@ -325,59 +307,41 @@ Tab2.xml 5068 Primer name Primer sequence (5' to 3') Length Characteristics ESA_02131 F CATCGGCCGACAGGCCAGTCAATGAATGATGC 32 EagI cut site R CATTCTAGAGAGTACAACGGAATGCGGGG 29 XbaI cut site ESA_01706 F CATTCTAGAGTCGGGCGGCTCTTTATCTGG 30 XbaI cut site R CATGGATCCTTGACCAGATGACGCAGTCTTTC 32 BamHI cut site Chimeric-01706 F AATAAGCTTGTGGCTTTTTATGCCGGGCTGC 31 HindIII cut site R CAGGCCAGTAATCAGCGCCGCGCCCATGAC 30 3' SOEing overhang Chimeric-02131CTE F CGCGGCGCTGATTACTGGCCTGACGATGAAAG 32 5' SOEing overhang R AATGGATCCTTACTCGTTAATACGAGGATGCTGG 34 BamHI cut site pUC18 MCS Check F CATTAGCTCACTCATTAGGCACC 23 pUC18 insert check R CATTGTAAAACGACGGCCAGTG 22 pUC18 insert check - - MESH:C016800 - Chemical - - Chimeric-02131CTE F - - - MESH:C094836 - Chemical - - 02131 F - METHODS paragraph 5734 PCR primers (Table 2) were designed for each proP homologue based on C. sakazakii strain BAA-894 sequence data available from the NCBI database (NC_009778.1). The formation of the chimeric ProP protein (ProPc), which consists of the extended coiled coil region of ProP1 (amino acid position 422 to 505) fused to the C-terminus of ProP2 (encoded by ESA_01706), was performed using a modified SOEing (Splicing by overlap extension) technique. In silico comparative analysis of the native ProP1 and ProP2 sequences, revealed a point of amino acid homology within the twelfth predicted transmembrane domain, a leucine/isoleucine/threonine triplet (LIT) at position 422-424 and 437-439 respectively, which was selected as the splice site. Briefly, the fusion was performed using three separate PCR reactions: the first PCR (primer set Chimeric-01706) resulted in an ESA_01706 (proP2) amplicon lacking the C-terminal extension but containing a 15-bp 3'overhang corresponding to the LIT triplet of the ProP1 C- terminal extension. The second PCR (primer set Chimeric-02131CTE) formed an amplicon of 210-bp encoding ProP1 C-terminal extension with a 5'-overhang, also corresponding to the LIT triplet. The final PCR (primer set Chimeric-01706-F Chimeric-02131tail-R) was performed with the two previous amplicons as template; resulting in a final product of 1,623-bp, representing the ESA_01706 native promoter and modified coding region (encoding the fused ProP1 C-terminal extension after the LIT triplet). This product was digested with restriction enzymes BamHI and HindIII and ligated to a similarly digested pUC18 vector forming pUC18:ESA_01706CTE (C-Termini Extension). The integrity of the chimeric sequence was confirmed by sequencing (MWG Operon, Germany and GATC, Germany) and transformed into E. coli MKH13. - + 28141 Species C. sakazakii - + 562 Species E. coli - - MESH:C061951 + + MESH:D007930 Chemical - - threonine + + leucine - - MESH:C043801 + + MESH:D007532 Chemical isoleucine - - MESH:C038361 - Chemical - - leucine - - - MESH:D000922 + + MESH:D013912 Chemical - - Chimeric-01706-F Chimeric-02131tail + + threonine @@ -391,53 +355,59 @@ paragraph 7575 Overnight cultures of E. coli MKH13 clones expressing the wild-type and chimeric ProP proteins (ProP1, ProP2 and ProPC respectively) were grown at 37 C with shaking at 200 rpm in either 10 ml LB or M9 minimal media containing 0.5% glucose, 0.04% arginine, 0.04% isoleucine, 0.04% valine (Sigma-Aldrich Co.). Cells were pelleted by centrifugation at 5,000 g, washed and re-suspended in 200 mul 1/4 strength Ringer's solution. The cell suspension was added to the appropriate filter sterilized media with varying concentrations (0-10%) of added NaCl. Growth was monitored in the relevant media over a 48 hour period, with optical density (OD600) readings being taken every hour. Triplicate readings were taken and graphs were plotted using SigmaPlot version 11.0. E. coli MKH13 harbouring the empty pUC18 plasmid was used as a negative control. - + 562 Species E. coli - + 562 Species E. coli - + 1239833 - Strain + Species at 5 - + + MESH:D001120 + Chemical + + arginine + + MESH:D012965 Chemical NaCl - - MESH:D001127 - Chemical - - arginine - - - MESH:D005947 + + MESH:D014633 Chemical - - glucose + + valine - - MESH:C043801 + + MESH:D007532 Chemical isoleucine - - MESH:C521924 + + - Chemical - - valine + + MKH13 + + + MESH:D005947 + Chemical + + glucose @@ -451,7 +421,7 @@ title_2 8434 C. sakazakii ProP structures - + 28141 Species @@ -471,73 +441,49 @@ paragraph 8591 Based on sequence similarity to the E. coli ProP protein, we identified ProP1 (the product of ESA_02131) as the most likely ProP homolog in C. sakazakii; exhibiting 90% amino acid sequence identity and structural features characteristic of E. coli ProP. Indeed, further analysis using TMHMM and TexTopo software predicted ProP1 to be a membrane protein with 12-transmembrane domains, an extended central hydrophilic loop and carboxy terminal extension (Figure 1). While the remaining five ProP homologues on the C. sakazakii BAA-894 genome were also predicted to encode proteins with 12 transmembrane domains and an extended central hydrophilic loop, they each lacked the extended carboxy-terminal domain identified in ProP1, a feature which likely affects the final protein structure and function. - + 562 Species E. coli - + 28141 Species C. sakazakii - + 562 Species E. coli - + 28141 Species C. sakazakii - - MESH:C015732 - Chemical - - carboxy - - - MESH:C015732 - Chemical - - carboxy - RESULTS paragraph 9391 Figure 1B illustrates the tertiary structure for ProP1 (predicted using the I-TASSER server). Most notably the presence of a coiled coil domain is evident as a result of the extended carboxy-terminal identified by sequence analysis. The coiled coil domain likely protrudes into the intracellular cytoplasm of the organism where its function remains unclear. By contrast, the tertiary structure of ProP2 (Figure 1A), representative of the remaining 6 ProP homologues and exhibiting 40% identity to E. coli ProP and 49% identity to ProP1, lacks the coiled coil domain at the carboxy-terminal end. - + 562 Species E. coli - - MESH:C015732 - Chemical - - carboxy - - - MESH:C015732 - Chemical - - carboxy - RESULTS title_2 9988 Chimeric protein (ProPc) expression in E. coli MKH13 - + 562 Species @@ -549,25 +495,25 @@ paragraph 10041 The osmoprotective properties of ProP1, ProP2 and ProPc were measured and compared in E. coli MKH13; an osmosensitive mutant incapable of growth in high osmolality environments (>=4%). The pUC18 plasmid containing each gene of interest was transformed to E. coli MKH13. Transformation efficiencies of 60 CFU/mug DNA were achieved, with successful transformation being confirmed by colony PCR, followed by sequencing. Transformants were screened for osmotolerance on media (both LB and M9 plus 1 mM proline) containing between 4% and 10% added NaCl. - + 562 Species E. coli - + 562 Species E. coli - - MESH:C489032 + + MESH:D011392 Chemical proline - + MESH:D012965 Chemical @@ -585,25 +531,25 @@ paragraph 10621 To determine the effect of each ProP homologues, both native (ProP1 and ProP2) and chimeric (ProPc), on the osmotolerance of E. coli MKH13, each of the strains was grown in media containing varying concentrations of NaCl. Growth was monitored over a 48 hour period in minimal media supplemented with 1 mM proline and containing 0-10% added NaCl. - + 562 Species E. coli - - MESH:C489032 - Chemical - - proline - - + MESH:D012965 Chemical NaCl - + + MESH:D011392 + Chemical + + proline + + MESH:D012965 Chemical @@ -617,25 +563,25 @@ fig_caption 10969 Physiological analysis of E. coli MKH13 expressing ProP1, ProP2, ProPc and the empty pUC18 plasmid. A) Optical density was measured over a 48 hour period in media supplemented with 0-10% added NaCl. B) The growth rate of E. coli MKH13 expressing each of the proteins of interest was measured in media supplemented with 0-10% added NaCl. - + 562 Species E. coli - + 562 Species E. coli - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical @@ -647,60 +593,66 @@ paragraph 11312 Media containing 5% NaCl yielded the most discriminatory results. While E. coli MKH13 expressing the empty pUC18 vector showed no growth at 5%, each of the other three strains tested conferred some degree of osmotolerance on the host (Figure 2A). The strain expressing ProP1 was the most osmotolerant, with a maximum optical density (OD600) of 0.326 after 37 hours growth at 5% NaCl. The strain possessing ProPc grew to an OD significantly higher than the strains expressing either ProP1 or ProP2. E. coli MKH13 expressing ProP2 in 5% NaCl grew to a maximum OD of 0.111 after 48 hours, whereas E. coli MKH13 containing ProPc grew to a final OD of 0.189 at the same time point. Interestingly, growth of the stains expressing ProP2 and the chimeric protein continued to increase up to 48 hours, while growth of E. coli MKH13 expressing ProP1 reached maximum OD after only 37 hours. - + 562 Species E. coli - + 562 Species E. coli - + 562 Species E. coli - + 562 Species E. coli - + 1239833 - Strain + Species at 5 - + 1239833 - Strain + Species at 5 - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical NaCl + + - + Chemical + + ProP2 + TABLE @@ -723,55 +675,55 @@ paragraph 12826 Each E. coli MKH13 strain expressing a proP gene of interest conferred osmotolerance (Figure 2B). As expected, E. coli MKH13 demonstrated a significant reduction in growth rate as NaCl concentrations increased, with a final growth rate of 0.0004 hr-1 recorded in media supplemented with 4% NaCl and no subsequent growth recorded thereafter. E. coli MKH13 expressing ProP1 demonstrated the highest osmotolerance of all the strains tested with growth rates of 0.009 hr-1 to 0.004 hr-1 recorded in media supplemented with 5% to 10% NaCl respectively. The next most osmotolerant strain was that expressing ProPc. Growth rates of 0.004 hr-1 to 0.003 hr-1 were recorded in media supplemented with 5% to 10% NaCl respectively (Table 3). This was higher than the growth rates observed when E. coli MKH13 expressing the native ESA_01706 gene (ProP2) was grown in a high osmolality environment, suggesting an important role for the C. sakazakii ProP C-terminal extension in osmotolerance. - + 562 Species E. coli - + 562 Species E. coli - + 562 Species E. coli - + 562 Species E. coli - + 28141 Species C. sakazakii - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical NaCl - + MESH:D012965 Chemical @@ -789,73 +741,73 @@ paragraph 13833 A unique feature of the neonatal pathogen C. sakazakii is its ability to survive for prolonged periods in environments of low aW, such as powdered infant formula (PIF), making it a significant cause for concern. Indeed, up to 80% of infants infected with C. sakazakii die within days of birth, while survivors often suffer delayed neurological symptoms, brain abscesses or hydrocephalus. However, despite this, little is known about the molecular mechanisms that allow this organism to survive in environments such as PIF where it is subject to extreme hyper-osmotic stress. - + 28141 Species C. sakazakii - + 9606 Species infant - + 9606 Species infants - + 28141 Species C. sakazakii - - MESH:D054549 - Disease - - hyper-osmotic stress - - + MESH:D006849 Disease hydrocephalus - + MESH:D001922 Disease brain abscesses - + MESH:D009422 Disease neurological symptoms + + MESH:D004194 + Disease + + hyper-osmotic stress + DISCUSS paragraph 14408 In a previous in silico study we identified seven copies of an E. coli proP homolog on the BAA-894 genome. Physiological analysis confirmed that six of the proP homologues identified played a role in osmotolerance. The availability of osmolytes in the media also had an effect on the osmotolerance of the host, with growth rates varying depending on the type or variety of compatible solutes present. While all six ProP proteins exhibited features characteristic of classic secondary transporters, five of the proteins were between 60-70 amino acids shorter than ProP1; lacking the characteristic C-terminal cytoplasmic extension Previous studies in our lab have demonstrated that the six C. sakazakii ProP homologues lacking the C-terminal coiled coil are significantly less osmoprotective than ProP1, suggesting an important role for this domain in modulating C. sakazakii osmotolerance. - + 562 Species E. coli - + 28141 Species C. sakazakii - + 28141 Species @@ -867,119 +819,126 @@ paragraph 15300 In the current study, the C. sakazakii ProP2 (encoded by ESA_01706) was chosen as the prototypical ProP homologue to study the role of the alpha helical coiled coil in osmotolerance. Genetic splicing yielded a chimeric protein structure (ProPc) possessing the native ProP2 domains in addition to the C-terminal alpha helical coiled coil domain from ProP1 (Figure 1C). E. coli MKH13 expressing ProPc grew to a higher OD in minimal media supplemented with proline, when compared to the native protein ProP2 which lacked the extended coiled coil domain (Figure 2A). These data demonstrate that the addition of the coiled coil domain from ProP1 to ProP2 results in a protein with an increased osmoprotective effect on the usually osmotically sensitive E. coli MKH13. However, as the osmolality of the medium increased, this trend appeared to reverse with OD readings becoming similar at 9% NaCl and the chimeric protein growing to a higher OD than the native in 10% NaCl, suggesting that the extent of osmotic pressure also has a role to play in the activity of the proteins. - + 28141 Species C. sakazakii - + 562 Species E. coli - + 562 Species E. coli - + MESH:D012965 Chemical NaCl - - MESH:C489032 + + MESH:D011392 Chemical proline - + MESH:D012965 Chemical NaCl + + - + Chemical + + MKH13 + DISCUSS paragraph 16374 The role of the C-terminal domain of other osmolyte transporters, such as BetP (Corynebacterium glutamicum) and OpuA (Bacillus subtilis), was demonstrated to be important for the activation of these proteins during an increase in the osmolality of the surrounding medium. Furthermore, Culham et al. created a synthetic polypeptide corresponding to the C-terminal domain of E. coli ProP which formed a dimeric alpha helical coiled coil structure, similar to the coiled coil structure of ProP1 (illustrated in Figure 1A). In the same study ProP proteins from both E. coli and Agrobacterium tumefaciens, possessing the characteristic alpha helical coiled coil, were activated at a lower osmolality than orthologues lacking the coiled coil structure. C. glutamicum possesses a ProP protein which lacks the C-terminal alpha helical coiled coil domain and, presumably as a result of this, requires a higher osmolality for activation. E. coli ProP variants lacking the coiled coil or with an amino acid substitution disrupting the formation of the alpha helical coiled coil, also require a larger osmotic upshift than the wild-type transporter. This study demonstrates that the activity of these ProP orthologues is dependent on the osmolality of the surrounding medium, and the alpha helical coiled coil is believed to tune the transporter to osmoregulate the cell over a low osmolality range. These data may therefore offer an explanation for the increased growth observed in E. coli MKH13 expressing ProP2, which lacks the coiled coil domain, in media supplemented with 10% NaCl relative to either ProP1 or ProPc (Figure 2). It is likely that the coiled coil domain of C. sakazakii ProP1 has a similar tuning function. Furthermore, the presence of multiple ProP porters lacking the C-terminal coiled coil domain, and therefore only active at a higher osmolality, may well explain the extreme osmotolerance unique to C. sakazakii; allowing the pathogen to survive in environments like PIF. The ProP1 protein, on the other hand possessing the coiled coil, may be the only osmolyte transporter required to respond to low or moderate hyperosmotic challenge. - + 1718 Species Corynebacterium glutamicum - + 1423 Species Bacillus subtilis - + 562 Species E. coli - + 562 Species E. coli - + 358 Species Agrobacterium tumefaciens - + 1718 Species C. glutamicum - + 562 Species E. coli - + 562 Species E. coli - + 28141 Species C. sakazakii - + 28141 Species C. sakazakii - - Disease - - hyperosmotic challenge - - + MESH:D012965 Chemical NaCl + + - + Chemical + + BetP + CONCL @@ -992,19 +951,19 @@ paragraph 18537 The addition of the coiled coil domain from ProP1 to ProP2 resulted in a chimeric protein (ProPc) which demonstrated higher osmotolerance compared to the native ProP2 (under moderate osmotic stress conditions). Furthermore, the growth rate of E. coli MKH13 expressing ProP2 increased in minimal media supplemented with 10% NaCl; suggesting that, as is the case in E. coli, the coiled coil domain tunes ProP at low osmolality, whereas ProP transporters lacking the coiled coil domain are more active at a higher osmolality range. - + 562 Species E. coli - + 562 Species E. coli - + MESH:D012965 Chemical @@ -1040,25 +999,25 @@ footnote 19249 AF, CJ and AL carried out the the experimental work. AF drafted the manuscript together with together with CJ, AL and RDS. All authors read and approved the final manuscript. - + MESH:D001281 Disease AF - + MESH:D009101 Disease AL - + MESH:D009101 Disease AL - + MESH:D001281 Disease @@ -1089,6 +1048,12 @@ surname:Somero;given-names:GN 19435 Living with water stress: evolution of osmolyte systems + + MESH:D014867 + Chemical + + water + 2 @@ -1122,29 +1087,29 @@ 2577 19594 Identification and disruption of the proBA locus in Listeria monocytogenes: role of proline biosynthesis in salt tolerance and murine infection - + 1639 Species Listeria monocytogenes - + 10090 Species murine - - MESH:C489032 + + MESH:D011392 Chemical proline - - MESH:D007239 - Disease - - infection + + MESH:D012492 + Chemical + + salt @@ -1183,18 +1148,12 @@ surname:Steer;given-names:BA 19820 Isolation and sequencing of escherichia coli gene proP reveals unusual structural features of the osmoregulatory proline/betaine transporter, ProP - + 562 Species escherichia coli - - MESH:C489032 - Chemical - - proline - surname:Voegele;given-names:RT @@ -1215,12 +1174,6 @@ surname:Hodges;given-names:RS 19967 The role of the carboxyl terminal alpha-helical coiled-coil domain in osmosensing by transporter ProP of Escherichia coli - - MESH:C041069 - Chemical - - carboxyl - surname:O'Mahony;given-names:J @@ -1239,7 +1192,7 @@ surname:Coffey;given-names:A 20089 Analysis of the role of the Cronobacter sakazakii ProP homologues in osmotolerance - + 28141 Species @@ -1263,13 +1216,13 @@ 1046 20172 A possible role of ProP, ProU and CaiT in osmoprotection of Escherichia coli by carnitine - + 562 Species Escherichia coli - + MESH:D002331 Chemical @@ -1324,7 +1277,7 @@ surname:Porwollik;given-names:S 20379 Genome sequence of Cronobacter sakazakii BAA-894 and comparative genomic hybridization analysis with other Cronobacter species - + 28141 Species @@ -1346,11 +1299,17 @@ 16713 20506 OpuA, an osmotically regulated binding protein-dependent transport system for the osmoprotectant glycine betaine in bacillus subtilis - - MESH:D005998 + + - Chemical - - glycine + + OpuA + + + MESH:D001622 + Chemical + + glycine betaine @@ -1370,6 +1329,12 @@ 268 20640 Analysis of the role of betL in contributing to the growth and survival of Listeria monocytogenes LO28 + + - + Chemical + + betL + 9 @@ -1417,13 +1382,13 @@ 1237 20880 The human gut microbiome: the ghost in the machine - + 9606 Species human - + 749906 Species @@ -1449,13 +1414,13 @@ surname:Frye;given-names:JG 20931 Enterobacter sakazakii invades brain capillary endothelial cells, persists in human macrophages influencing cytokine secretion and induces severe brain pathology in the neonatal rat - + 28141 Species Enterobacter sakazakii - + 9606 Species @@ -1476,7 +1441,7 @@ 34 21113 Enterobacter sakazakii brain abscess in the neonate: the importance of neuroradiologic imaging - + 28141 Species @@ -1502,7 +1467,7 @@ surname:Boggs;given-names:JM 21208 Transmembrane Helix I and Periplasmic Loop 1 of Escherichia coli ProP are involved in osmosensing and osmoprotectant transport - + 562 Species @@ -1528,6 +1493,12 @@ surname:Dowhan;given-names:W 21335 The osmotic activation of transporter ProP is tuned by both its c-terminal coiled-coil and osmotically induced changes in phospholipid composition + + MESH:D010743 + Chemical + + phospholipid + @@ -1775,7 +1746,7 @@ Lynch syndrome - MESH:D016889 + MESH:D014591 Disease endometrial @@ -1942,21 +1913,15 @@ paragraph 5123 Bioinformatic analysis and in vitro assays previously showed that genetic mutations at exon-intron boundaries in the MLH1 gene generate aberrantly spliced transcripts (see Table 1). All mutations were classified as pathogenic (Class 5) according to the International Society for Gastrointestinal Hereditary Tumours Variant Interpretation Committee (InSiGHT VIC). - + 4292 Gene 208 MLH1 - - MESH:C059568 - Chemical - - InSiGHT VIC - - - MESH:D005770 + + MESH:D004067 Disease Gastrointestinal Hereditary Tumours @@ -1973,77 +1938,83 @@ paragraph 5500 LCLs were established from patient blood by transformation with Epstein-Barr Virus (as described previously), and cultured in RPMI with 10% fetal bovine serum (Gibco, Life Technologies) at 37 C in 5% CO2. Lymphocytes for transformation from five patients (c.588 + 1G > T, c.589-2A > G, c.791-1G > C, c.884G > A and c.1559-2A > T) were obtained from the Australasian Colorectal Cancer Family Registry (ACCFR). Lymphocytes from the patient with the c.790 + 1G > T mutation were obtained from the MCO collection. - + 9606 Species patient - + 9913 Species bovine - + 9606 Species patients - + 9606 Species patient - + c.790+1G>T;RS#:267607789 DNAMutation c.790 + 1G > T - + c.791-1G>C;RS#:267607795 DNAMutation c.791-1G > C - + c.589-2A>G;RS#:267607767 DNAMutation c.589-2A > G - - MESH:D002245 + + - Chemical CO2 - + c.588+1G>T;RS#:267607772 DNAMutation c.588 + 1G > T - + c.884G>A;RS#:63750144 DNAMutation c.884G > A + + c.1559-2A>T;RS#:267607836 + DNAMutation + + c.1559-2A > T + MESH:D015179 Disease - - Colorectal Cancer + + Australasian Colorectal Cancer - c.1559-2A>T;RS#:267607836 - DNAMutation - - c.1559-2A > T + - + Chemical + + RPMI @@ -2067,54 +2038,49 @@ Tab2.xml 6109 Region NOMe-seq primers Annealing ( C) c.588 + 1G > T F:TTGATATTTAGTGTGTGTTTTTGGYAAT F/R = 54 C R:CACATAATATCTTAAAAAATTCCAAAATAATA F/RN = 56 C RN:ATACCRACTAACARCATTTCCAAAAATAA c.589-2A > G F:TTAGGTATTTAGTATATAATGYAGG F/R = 51 C R:CACTATAAATATTTTCAAAACTAAAACCTTA F/RN = 52 C RN:CACAAAATCTAAAAAATTACATACACCTAA c.790 + 1G > T F:TAGGYATAGGAGGATTTTAAATGGATTAAGTT F/R = 52 C R:CAATTTCTTTAATAACAATRCCTATACCTAAA F/RN = 52 C RN:TTACTCRTAAAAACTCTAACACCATCAA c.791-1G > C F:GATGTGATGTGYATATTATTATAGAAATGTT F/R = 55 C R:TATCARCACCTCCTAATAAAATGAARCATA F/RN = 56 C RN:ATCCTTTTRCCAATAATATATAAAATTCACTCTA c.884G > A F:GGATGTGATGTGYATATTATTATAGAAATGTT F/R = 58 C R:CTATTATARCTTCCCAACTAACCCCARCAA F/RN = 58 C RN:CTACAARCTATCARCACCTCCTAATAA c.1559-2A > T F:TATTAGGAGGYTTAATTTAGGYTTTTTTGYTTAT F/R = 58 C R:ACCCTCACCACCTAATTCACAACATTTATAA F/RN = 57 C RN:ACTAARCAACTACCAAAAACTAARCTTCTTA HSPA5 F: GAGAAGAAAAAGTTTAGATTTTATA F/R = 56 C R: AAACACCCCAATAAATCAATC - + 3309 Gene 3908 HSPA5 - + c.1G>T DNAMutation 1G > T - + c.1G>T DNAMutation 1G > T - - MESH:C504520 - Chemical - - C F - - - c.884G>A;RS#:63750144 - DNAMutation - - c.884G > A - - - Chemical - - CACAAAATCTAAAAAATTACATACACCTAA - - + c.589-2A>G;RS#:267607767 DNAMutation c.589-2A > G - + c.1559-2A>T;RS#:267607836 DNAMutation c.1559-2A > T + + c.791-1G>C;RS#:267607795 + DNAMutation + + c.791-1G > C + + + c.884G>A;RS#:63750144 + DNAMutation + + c.884G > A + METHODS @@ -2155,16 +2121,16 @@ bisulfite - MESH:D012436 + - Chemical - - S-adenosylmethionine + + HCl - MESH:D002725 + MESH:D012436 Chemical - - phenol chloroform + + S-adenosylmethionine MESH:D004492 @@ -2179,10 +2145,10 @@ NaCl - MESH:C014843 + MESH:D002725 Chemical - - HCl + + chloroform MESH:C042345 @@ -2191,12 +2157,24 @@ bisulfite - MESH:C032259 + - + Chemical + + Tris + + + MESH:D012967 Chemical SDS - + + MESH:D019800 + Chemical + + phenol + + MESH:D000431 Chemical @@ -2216,49 +2194,49 @@ fig_caption 8510 Allele-specific positioning of nucleosomes at MLH1 variants. (A) The MLH1 gene with arrows showing the location of the splice site mutations investigated in the LCLs derived from the six patients. The numbers correspond to the NOMe-seq assays shown in panel B (exon, vertical bar; intron, horizontal bar with arrows). (B) Each panel shows the nucleosome occupancy on the wild-type allele (upper) and the allele harbouring the indicated splice site mutation (lower). Blue box = exon; blue arrows = intron and direction of transcription; yellow diamond = location of splice site mutation; vertical black bars below line = GpC sites; vertical black bars above line = CpG sites; the pink bar represents a single nucleosome (150 bp, drawn to scale); pink shading indicates the location of nucleosomes on individual DNA molecules as determined by GpC methyltransferase inaccessibility; black circles = methylated GpC/accessible to M.CviPI; white circles = unmethylated GpC/inaccessible to M.CviPI. CpG sites are depicted by triangles. Black triangle = methylated CpG; white triangle = unmethylated CpG. - + 4292 Gene 208 MLH1 - + 4292 Gene 208 MLH1 - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 9606 Species @@ -2272,63 +2250,63 @@ fig_caption 9644 Nucleosome occupancy and endogenous CpG methylation at the HSPA5 gene. (A) Schematic showing the location of the NOMe-seq assay (black bar) in relation to the HSPA5 promoter. The arrow represents the transcriptional start site. (B) NOMe-seq was used to map the position of nucleosomes on individual DNA molecules at the HSPA5 gene in LCLs derived from the six individuals with splice site mutations in MLH1. Vertical black bars below line = GpC sites; vertical black bars above line = CpG sites; pink bars = the location of nucleosomes on individual DNA molecules as determined by GpC methyltransferase inaccessibility; black circles = methylated GpC/accessible to M.CviPI; white circles = unmethylated GpC/inaccessible to M.CviPI. All endogenous CpG sites were unmethylated and triangles are not shown to provide better clarity of GpC sites. - + 3309 Gene 3908 HSPA5 - + 3309 Gene 3908 HSPA5 - + 3309 Gene 3908 HSPA5 - + 4292 Gene 208 MLH1 - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 2995 Gene 128415 GpC - + 2995 Gene 128415 @@ -2341,47 +2319,47 @@ paragraph 10508 We determined the positioning of nucleosomes across exons 7, 8, 9, 10 or 14 of the MLH1 gene in LCLs derived from six Lynch syndrome patients. Splice site mutations within or near these exons affected splice donor or acceptor sites (Table 1). NOMe-seq and single molecule sequencing allowed us to distinguish mutant from wild-type alleles using the splice site sequence mutation. At all sites analysed the majority of molecules were inaccessible at GpC sites and methylated at CpG sites (Figure 1). NOMe-seq analysis of the control gene HSPA5, a constitutively active gene that maintains a nucleosome-depleted region at the transcription start site, showed that the CpG island was unmethylated (data not shown) and confirmed that GpC inaccessibility was due to nucleosome occupancy rather than incomplete M.CviPI treatment (Figure 2). This shows that the majority of molecules at the sites analysed are occupied by nucleosomes in these cells. - + 4292 Gene 208 MLH1 - + 2995 Gene 128415 GpC - + 3309 Gene 3908 HSPA5 - + 2995 Gene 128415 GpC - + 9606 Species patients - + 9606 Species donor - + MESH:D003123 Disease @@ -2393,7 +2371,7 @@ paragraph 11454 Separation of alleles by presence or absence of the splice site mutation showed no overall difference in either nucleosome positioning or occupancy between wild-type and mutant alleles (Figure 1 and Table 1). In the six splice site mutations analysed, three were located within donor splice sites and three were located within acceptor splice sites (Table 1). Though some difference in precise positioning between mutant and wild-type alleles was observed across some exons (for example, exons 7 and 10, Figure 1B) this was confined to a small subset of molecules, rather than the majority of molecules as would be expected if the mutations affected nucleosome positioning. Our study therefore shows that nucleosome occupancy is unaltered at exon-intron boundaries containing these splice site mutations. - + 9606 Species @@ -2417,55 +2395,55 @@ paragraph 12560 NOMe-seq has been used to measure nucleosome occupancy at gene promoters, CTCF binding sites and regulatory elements but this is the first study that has utilised the technique to measure nucleosome occupancy at splice sites. NOMe-seq previously showed that the active CpG island (CGI) promoter of HSPA5 has a nucleosome-depleted region (NDR) upstream of the transcriptional start site. This region was included as a control in each assay, and in all cases the NDR was present. This indicated that the absence of GpC methylation observed in MLH1 was caused by nucleosome occlusion, not an artefact caused by a defective M.CviPI enzyme. In addition, CpG methylation was absent from the HSPA5 NDR which is a feature of active CGI promoters. Finally, gene body methylation is a feature of human cells and the majority of CpG sites within the MLH1 exons and introns were methylated. Together, these controls indicated that the NOMe-seq assays captured the true state of nucleosome positioning at MLH1 splice sites. - + 10664 Gene 4786 CTCF - + 3309 Gene 3908 HSPA5 - + 2995 Gene 128415 GpC - + 4292 Gene 208 MLH1 - + 4292 Gene 208 MLH1 - + 4292 Gene 208 MLH1 - + 9606 Species human - + 3309 Gene 3908 @@ -2478,21 +2456,21 @@ paragraph 13571 Alternative splicing of MLH1 occurs in a range of normal tissues, but several pieces of evidence indicate that the transcripts observed in our study are caused by the mutation, rather than being normal splicing events. First, the splicing aberrations were predicted with bioinformatic splicing software, and second, although one of the aberrant transcripts (Delta10) has been reported among naturally occurring MLH1 splice transcripts, the clinical data associated with these variants are also indicative of pathogenicity as demonstrated by the results of previous multifactorial likelihood analyses. - + 4292 Gene 208 MLH1 - + 4292 Gene 208 MLH1 - + c.del10 DNAMutation @@ -2504,7 +2482,7 @@ paragraph 14172 One strength of our study was the ability to determine the allele-specific position of nucleosomes by incorporating a heterozygous single nucleotide variant into each NOMe-seq assay. Small changes in positioning were observed but these molecules constituted approximately the same small proportion of total molecules on both alleles. Subtle changes in nucleosome positioning have been considered important in regulating the expression of cell cycle-dependent genes and enhancer accessibility, but in those studies the subtle changes were observed on a significant number of molecules that resulted in an overall change in nucleosome occupancy. There was no consistent alteration in nucleosome occupancy between the wild-type and mutant alleles at a variety of donor and acceptor sites, which would be expected if nucleosome positioning at these sites played a role in mediating the effects of splice site mutations. Together our data shows that alleles containing a splice site mutation show no differences in nucleosome occupancy to wild-type alleles. - + 9606 Species @@ -2516,43 +2494,43 @@ paragraph 15225 A limitation of the approach was that in most of the assays (assays 2 to 6) it was only possible to measure nucleosome positions at either the donor site or the acceptor site. Bisulfite conversion causes fragmentation of genomic DNA and it is technically challenging to amplify fragments greater than 500 bp in length. The donor and acceptor sites were separated by more than 2 kb of intronic sequence, making it impossible to concurrently determine the allele-specific position of nucleosomes at the donor and acceptor site on the same molecule. Although an independent NOMe-seq assay could be performed at the other donor or acceptor site, it would provide no information on allele-specificity. The data from one assay (c.588 + 1 G > T), however, in which both splice sites were present in the one amplicon, showed no significant change in nucleosome occupancy at the donor or acceptor site in intron 7. - + 9606 Species donor - + 9606 Species donor - + 9606 Species donor - + 9606 Species donor - + 9606 Species donor - + c.588+1G>T;RS#:267607772 DNAMutation c.588 + 1 G > T - + MESH:C042345 Chemical @@ -2564,20 +2542,20 @@ paragraph 16139 Previous studies have utilised genome-wide datasets of micrococcal nuclease (MNase) digested chromatin to investigate nucleosome occupancy. Here we used NOMe-seq to map the position of nucleosomes relative to exons and splice sites in a single gene at single molecule resolution. This approach provides the most accurate possible measurement of nucleosome positioning and enables allele-specific mapping of nucleosomes. In agreement with previous reports in humans, Caenorhabditis elegans and Drosophila melanogaster, we observed high nucleosome occupancy at exons. Thus, the findings from genome-wide studies were supported by our independent, single-molecule approach at the MLH1 gene. - + 4292 Gene 208 MLH1 - + 9606 Species humans - + Disease Caenorhabditis elegans and Drosophila melanogaster @@ -2588,41 +2566,42 @@ paragraph 16827 A key finding of our study was that nucleosome positioning was not significantly affected by mutations at the MLH1 splice sites investigated. Recent work with an in vitro IKBKAP mini-gene system found that alternative splicing changed chromatin organisation, with splice site strength and factors needed for splicing, such as U1 snSNP, playing a role in the regulation of nucleosome occupancy in exons. This and an earlier study indicate that splice site strength is an important determinant of nucleosome occupancy in exons. Exons with stronger polypyrimidine tracts (PPT; one of the conserved DNA elements located at 3' splice sites) have increased nucleosome occupancy compared to immediately adjacent introns, suggesting that nucleosomes act as a barrier that help to define the physical location of the 3' splice site. Although we investigated an equal number of donor and acceptor splice site mutations, nucleosome positioning was not significantly different between exons and the immediate 5' and 3' intronic sequence on the wild-type or mutant allele. The discrepancy between earlier studies and ours may in part relate to the strength of the MLH1 splice sites investigated. Alternatively, changes to chromatin organisation induced by splice mutations may be different to those caused by normal splicing events. Co-transcriptional splicing involves a complex interplay between RNAPII, the spliceosome and histone modifications and alterations in these factors, in addition to DNA sequence changes may be needed to alter nucleosome occupancy at exons. - + 4292 Gene 208 MLH1 - + 8518 Gene 2699 IKBKAP - + 6863 Gene 2394 PPT - + 4292 Gene 208 MLH1 - + 9606 Species donor - + + - Chemical polypyrimidine @@ -2633,27 +2612,27 @@ paragraph 18394 In summary this study used NOMe-seq to determine the allele-specific position of nucleosomes in relation to MLH1 splice site mutations. We conclude that splice site mutations that cause aberrant splicing of MLH1 do not alone significantly affect local nucleosome positioning in LCLs from Lynch syndrome patients. - + 4292 Gene 208 MLH1 - + 4292 Gene 208 MLH1 - + 9606 Species patients - + MESH:D003123 Disease @@ -2683,20 +2662,38 @@ footnote 18811 MAS and LBH designed NOM-seq assays. ACN and MAS performed NOMe-seq assays. BAT provided LCL samples. RLW and LBH conceived the study, participated in its coordination and helped to draft the manuscript. All authors read and approved the final manuscript. - + 81606 Gene 12687 LBH - + 81606 Gene 12687 LBH + + MESH:D005357 + Disease + + MAS + + + MESH:D005357 + Disease + + MAS + + + MESH:C084683 + Chemical + + ACN + REF @@ -2789,7 +2786,7 @@ 453 19311 Regulation of a mammalian gene bearing a CpG island promoter and a distal enhancer - + 9606 Species @@ -2845,7 +2842,7 @@ 1976 19576 The CTD role in cotranscriptional RNA processing and surveillance - + 1283 Gene @@ -2885,18 +2882,18 @@ 685 19716 Extracolonic cancer in hereditary nonpolyposis colorectal cancer - + + MESH:D009369 + Disease + + cancer + + MESH:D003123 Disease hereditary nonpolyposis colorectal cancer - - MESH:D009369 - Disease - - Extracolonic cancer - 36 @@ -2911,7 +2908,7 @@ 818 19781 Genetic susceptibility to non-polyposis colorectal cancer - + MESH:D015179 Disease @@ -2994,13 +2991,13 @@ surname:Walters;given-names:R 19989 A multifactorial likelihood model for MMR gene variant classification incorporating probabilities based on sequence bioinformatics and tumor characteristics: a report from the Colon Cancer Family Registry - + MESH:D009369 Disease tumor - + MESH:D015179 Disease @@ -3012,11 +3009,11 @@ ref 20194 Australasian Colorectal Cancer Family Registry [http://www.pedigree.org.au/pedigree-studies/accfr.aspx] - + MESH:D015179 Disease - - Colorectal Cancer + + Australasian Colorectal Cancer @@ -3024,13 +3021,13 @@ ref 20298 MCO study tumour collection [http://researchdata.ands.org.au/mco-study-tumour-collection] - + MESH:D009369 Disease tumour - + MESH:D009369 Disease @@ -3057,7 +3054,7 @@ surname:Nichols;given-names:PW 20388 Hypomethylation of a LINE-1 promoter activates an alternate transcript of the MET oncogene in bladders with cancer - + MESH:D009369 Disease @@ -3078,7 +3075,7 @@ 274 20503 Discrimination of pseudogene and parental gene DNA methylation using allelic bisulfite sequencing - + MESH:C042345 Chemical @@ -3104,13 +3101,18 @@ surname:Lin;given-names:JC 20601 H2A.Z maintenance during mitosis reveals nucleosome shifting on mitotically silenced genes - + 3015 Gene 80218 H2A.Z + + Disease + + mitosis + surname:Liang;given-names:G @@ -3136,7 +3138,7 @@ ref 20790 Taberlay PC, Statham AL, Kelly TK, Clark SJ, Jones PA: Reconfiguration of nucleosome depleted regions at distal regulatory elements accompanies DNA methylation of enhancers and insulators in cancer.Genome Res 2014, Epub ahead of print. - + MESH:D009369 Disease @@ -3179,7 +3181,7 @@ surname:LeProust;given-names:EM 21074 Targeted and genome-scale strategies reveal gene-body methylation signatures in human cells - + 9606 Species @@ -3223,12 +3225,6 @@ ref 21230 Thompson BA, Martins A, Spurdle AB: A review of mismatch repair gene transcripts: issues for interpretation of mRNA splicing assays.Clin Genet 2014. Epub ahead of print. - - MESH:D049290 - Disease - - Spurdle AB - surname:Frenkel;given-names:B @@ -3249,13 +3245,7 @@ surname:Jia;given-names:L 21400 Dynamic nucleosome-depleted regions at androgen receptor enhancers in the absence of ligand in prostate cancer cells - - MESH:D000728 - Chemical - - androgen - - + MESH:D011471 Disease @@ -3277,7 +3267,7 @@ 5 21517 Bisulfite genomic sequencing: systematic investigation of critical experimental parameters - + MESH:C042345 Chemical @@ -3326,89 +3316,77 @@ Aluru N, Karchner SI, Franks DG, Nacci D, Champlin D, Hahn ME 0 Targeted mutagenesis of aryl hydrocarbon receptor 2a and 2b genes in Atlantic killifish (Fundulus heteroclitus) - + 8078 Species Atlantic killifish - + 8078 Species Fundulus heteroclitus - - MESH:D006838 - Chemical - - aryl hydrocarbon - ABSTRACT abstract 112 Understanding molecular mechanisms of toxicity is facilitated by experimental manipulations, such as disruption of function by gene targeting, that are especially challenging in non-standard model species with limited genomic resources. While loss-of-function approaches have included gene knock-down using morpholino-modified oligonucleotides and random mutagenesis using mutagens or retroviruses, more recent approaches include targeted mutagenesis using zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology. These latter methods provide more accessible opportunities to explore gene function in non-traditional model species. To facilitate evaluations of toxic mechanisms for important categories of aryl hydrocarbon pollutants, whose actions are known to be receptor mediated, we used ZFN and CRISPR-Cas9 approaches to generate aryl hydrocarbon receptor 2a (AHR2a) and AHR2b gene mutations in Atlantic killifish (Fundulus heteroclitus) embryos. This killifish is a particularly valuble non-traditional model for this study, with multiple paralogs of AHR whose functions are not well characterized. In addition, some populations of this species have evolved resistance to toxicants such as halogenated aromatic hydrocarbons. AHR-null killifish will be valuable for characterizing the role of the individual AHR paralogs in evolved resistance, as well as in normal development. We first used five-finger ZFNs targeting exons 1 and 3 of AHR2a. Subsequently, CRISPR-Cas9 guide RNAs were designed to target regions in exon 2 and 3 of AHR2a and AHR2b. We successfully induced frameshift mutations in AHR2a exon 3 with ZFN and CRISPR-Cas9 guide RNAs, with mutation frequencies of 10% and 16%, respectively. In AHR2b, mutations were induced using CRISPR-Cas9 guide RNAs targeting sites in both exon 2 (17%) and exon 3 (63%). We screened AHR2b exon 2 CRISPR-Cas9-injected embryos for off-target effects in AHR paralogs. No mutations were observed in closely related AHR genes (AHR1a, AHR1b, AHR2a, AHRR) in the CRISPR-Cas9-injected embryos. Overall, our results demonstrate that targeted genome-editing methods are efficient in inducing mutations at specific loci in embryos of a non-traditional model species, without detectable off-target effects in paralogous genes. - + 105937229 Gene AHR2 - + 105920361 Gene AHRR - + 8078 Species Atlantic killifish - + 8078 Species Fundulus heteroclitus - + 8078 Species killifish - + 8078 Species killifish - - MESH:D006838 - Chemical - - aryl hydrocarbon - - - MESH:D006838 + + MESH:D009841 Chemical - - aryl hydrocarbon + + oligonucleotides - + MESH:D064420 Disease toxicity - - MESH:D006846 + + MESH:D006841 Chemical - - halogenated aromatic hydrocarbons + + aromatic hydrocarbons @@ -3422,215 +3400,209 @@ paragraph 2511 The Atlantic killifish (Fundulus heteroclitus) is one of the most ecologically and environmentally important estuarine fish distributed along the East coast of the United States. Their ability to tolerate wide changes in environmental conditions, including temperature, salinity, oxygen and pH, have made them an ideal model species to investigate the biochemical, physiological and evolutionary basis of environmental adaptation. Some populations of killifish are also valuable models for understanding the mechanisms of evolved resistance to toxicants. Populations of killifish inhabiting contaminated coastal waters along the North Atlantic U.S. coast have evolved resistance to some contaminants representing major categories of aryl hydrocarbon pollutants, such as polynuclear aromatic hydrocarbons (PAHs), and halogenated aromatic hydrocarbons such as polychlorinated biphenyls (PCBs), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 'dioxin') and other dioxin-like compounds (DLCs). This evolved resistance involves alterations in signaling through the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor through which some PAHs, PCBs, and TCDD alter gene expression and cause toxicity. Killifish express four AHR paralogs (AHR1a, AHR1b, AHR2a and AHR2b), the products of distinct loci. Although the respective functions of these AHRs are not well understood, AHR2 proteins appear to play a major role in mediating the developmental effects of PAHs and PCBs in fish. - + 105937229 Gene AHR2 - + 8078 Species Atlantic killifish - + 8078 Species Fundulus heteroclitus - + 8078 Species killifish - + 8078 Species killifish - + 8078 Species Killifish - + 29154 Species Atlantic - + 29278 Species expression - + MESH:D010100 Chemical oxygen - - MESH:D006838 + + MESH:D011084 Chemical - - aryl hydrocarbon + + PAHs - + MESH:D011078 Chemical polychlorinated biphenyls - - MESH:D006846 - Chemical - - halogenated aromatic hydrocarbons - - + MESH:D011078 Chemical PCBs - - MESH:D013749 + + MESH:D000072317 Chemical - + TCDD - - MESH:D011078 + + MESH:D011084 Chemical - - PCBs + + PAHs - + MESH:D006838 Chemical - - aryl hydrocarbon + + hydrocarbon - + + MESH:D011084 + Chemical + + PAHs + + + MESH:D011078 + Chemical + + PCBs + + MESH:D064420 Disease toxicity - - MESH:D006841 - Chemical - - aromatic hydrocarbons - - - MESH:D013749 + + MESH:D000072317 Chemical - + TCDD - - MESH:C087686 - Chemical - - PAHs - - - MESH:C087686 + + MESH:D006841 Chemical - - PAHs + + aromatic hydrocarbons - - MESH:D013749 + + MESH:D011084 Chemical - - 2,3,7,8-tetrachlorodibenzo-p-dioxin + + polynuclear aromatic hydrocarbons - + MESH:D004147 Chemical dioxin - + MESH:D004147 Chemical dioxin - + + MESH:D000072317 + Chemical + + 2,3,7,8-tetrachlorodibenzo-p-dioxin + + MESH:D011078 Chemical PCBs - - MESH:C087686 - Chemical - - PAHs - INTRO paragraph 4005 While there is a great deal of understanding about the physiological and biochemical basis of environmental adaptation in killifish, very little is known about its genetic basis, mainly due to the lack of genetic tools to evaluate gene function. Recently, the antisense morpholino oligonucleotide (MO)-based gene knockdown method was adapted to killifish to determine the role of aryl hydrocarbon receptors and cytochrome P4501A (CYP1A) in developmental toxicity, and to study the role of aquaporins in adult killifish. However, MOs are effective only for a limited period of time and do not completely eliminate gene expression. In order to conclusively demonstrate the function of any protein, gene knockouts are essential. - + 105930482 Gene cytochrome P4501A - + 105930482 Gene CYP1A - + 8078 Species killifish - + 8078 Species killifish - + 8078 Species killifish - + 29278 Species expression - + MESH:D064420 Disease developmental toxicity - - MESH:D006838 + + MESH:D060172 Chemical - - aryl hydrocarbon + + morpholino oligonucleotide @@ -3638,43 +3610,43 @@ paragraph 4731 In the past decade, several targeted genome-editing methods have been successfully employed to manipulate genes in a variety of model and non-model organisms. They include zinc-finger nucleases (ZFNs), transactivator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system (Fig. 1). ZFN and TALENs utilize engineered nucleases consisting of a sequence-specific DNA-binding domain fused to a nonspecific DNA cleavage module. The DNA-binding domains can be customized to recognize any sequence of interest. Distinct from the site-specific nucleases, CRISPR-Cas9 is an RNA-guided DNA nuclease (RGN), where a small guide RNA (sgRNA) complementary to a 20 base target DNA sequence guides Cas9, a bacterially derived nuclease, to induce a double-strand break (DSB) at the target site. Because the DSB repair process by non-homologous end-joining (NHEJ) is error prone, it leaves insertions or deletions (indels) at the target site. The relative ease of generating target-specific mutations using these techniques has opened up new avenues for conducting functional studies in any organism of interest. Several studies have demonstrated the efficacy of ZFNs, TALENs and RGNs in producing heritable mutations in non-model organisms such as rainbow trout, medaka, tilapia and yellow catfish (Table 1). Mutagenesis rates of up to 95% have been observed with CRISPR-Cas9. So far, there are no studies describing the generation of gene knock-outs in killifish or of targeted AHR mutants in any non-mammalian vertebrate. In this study, we utilized ZFN and CRISPR-Cas9 approaches to induce mutations in paralogous killifish AHR2a and AHR2b genes, and we observed indels in the injected embryos. We discuss strategies for the application of this method to knock out closely related paralogs in a species with a long generation time. - + 8022 Species rainbow trout - + 8090 Species medaka - + 1234273 Species yellow catfish - + 8078 Species killifish - + 9606 Species mammalian - + 8078 Species killifish - + Disease tilapia @@ -3691,7 +3663,7 @@ title_2 6638 2.1. Killifish adults and embryos - + 8078 Species @@ -3703,30 +3675,54 @@ paragraph 6672 Mature male and female Atlantic killifish were collected from Scorton Creek (Sandwich, MA) during full moon and new moon in Spring and Summer of each year (2011-2014) using minnow traps, as described previously. Although killifish populations are known to vary dramatically in their sensitivity to DLCs, this source population is known to be relatively sensitive to DLCs, characteristic of other populations of this species resident to similarly uncontaminated sites. Fish were maintained in tanks with continuous flow-through seawater (SW) at 18-20 C and 14h:10h light/dark photoperiod conditions. The animal husbandry practices were approved by the Animal Care and Use Committee of the Woods Hole Oceanographic Institution. Female fish were lightly anesthetized with Tricaine (MS222; buffered with sodium bicarbonate, Sigma-Aldrich, St. Louis, MO, USA) and oocytes were obtained for in vitro fertilization (IVF) by gently squeezing the abdomen. Oocytes were collected in glass petri dishes with filtered SW (25 parts per thousand; ppt). Milt was obtained by euthanizing mature males in MS222, dissecting out the gonads, and chopping them with a scalpel blade in seawater. A few drops of milt were added to the oocytes for fertilization. Each IVF experiment included a pool of oocytes stripped from at least 2-3 females and milt from 1 or 2 males. Approximately 20 minutes after the addition of milt, embryos were rinsed with filtered SW to remove any excess sperm. Fertilized embryos were maintained at 23 C until further use. - + 8078 Species Atlantic killifish - + 8078 Species killifish - + + MESH:C537182 + Disease + + IVF + + MESH:C003636 Chemical Tricaine + + - + Chemical + + 14h + + MESH:C537182 + Disease + + IVF + + MESH:D017693 Chemical sodium bicarbonate + + MESH:C003636 + Chemical + + MS222 + METHODS @@ -3739,13 +3735,13 @@ paragraph 8242 AHR2a exon 2 and 3 (Fig. 2) were each targeted with a five-finger ZFN pair using the CompoZr custom ZFN service (Sigma-Aldrich, St. Louis, MO, USA) (Table 2; Fig. 2). Left- and right-ZFN plasmids were in vitro transcribed and polyadenylated using the mMessage mMachine T7 kit (Life Technologies, Carlsbad, CA, USA). ZFN mRNA from each half-site was diluted to a concentration of 100-400 ng/microl and 2.5 nl of each was co-injected into 1 or 2-cell stage killifish embryos, as described below. - + 105937229 Gene AHR2a exon 2 and 3 - + 8078 Species @@ -3763,35 +3759,41 @@ paragraph 8787 Guide RNAs (gRNA) targeting AHR2a and AHR2b were designed using the ZiFiT Targeter website (http://zifit.partners.org/ZiFiT_Cas9). Exons 1, 2 and 3 of AHR2a and AHR2b were searched for target sites in order to obtain the shortest truncated mutant protein. A single target site was found in each exon. The AHR2a exon 1 target site did not begin with GG residues, which prevented it from being cloned into the gRNA expression vector. The AHR2b exon 1 target site was not optimal due to repetitive sequences. Therefore, the target sites for exons 2 and 3 of AHR2a and AHR2b were selected for gRNA synthesis (Table 2). PAGE-purified complementary oligonucleotides (Eurofins Genomics, Huntsville, AL, USA) were annealed and cloned into an expression vector following a published protocol. The gRNA oligonucleotide sequences are provided in Table 2. Expression vectors for guide RNA (pDR274) and Cas9 endonuclease (MLM3613) were obtained from Addgene (https://www.addgene.org/). The gRNA was transcribed using the MAXIscript T7 kit, and the Cas9 mRNA was transcribed and poly(A)-tailed with the mMESSAGE mMACHINE T7 ULTRA kit (Life Technologies, Carlsbad, CA, USA). gRNA and Cas9 mRNA were mixed at a ratio of 1:12 for the microinjections. - + 105937229 Gene AHR2 - + 29278 Species Expression vectors - + 29278 Species expression - + 29278 Species expression - - MESH:C017937 + + MESH:D009841 Chemical - - poly(A) + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotides @@ -3805,37 +3807,55 @@ paragraph 10074 Microinjection of killifish embryos was performed following a previously established protocol with some modifications. Briefly, prior to microinjection, 1-cell stage embryos were rolled on wet paper towels to prevent the filaments on the chorion from adhering to the microinjection needle. They were subsequently placed on custom-made agarose microinjection embryo trays and oriented so that the cell directly faces the microinjection needle. The microinjection setup consisted of a Zeiss Stemi 2000-C stereomicroscope, Narishige IM-300 microinjector and a micromanipulator (Narishige International, East Meadow, NY, USA). Aluminosilicate glass tubes (1.0 mm outer diameter, 0.64 mm inner diameter) were used to prepare microinjection needles. This type of needle was found to be superior to borosilicate needles for microinjection of killifish embryo, which have a thick chorion. A P-30 vertical micropipette puller (Sutter Instruments, Novato, CA, USA) at a heater output setting of 97 was used to pull aluminosilicate glass tubing into microinjection needles. The injection volume was calibrated by injecting the solution into mineral oil and measuring the diameter of the droplet using a stage micrometer. The microinjection volume was approximately 5 nL per embryo. To compensate for variations in needle opening size, injection time and pressure were altered during microinjection. Injected embryos were maintained in filtered SW at 23 C. Some of the injected embryos were transferred to the fish-rearing facility at the National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, Environmental Protection Agency, Narragansett, RI, to be raised to adulthood. The numbers of injected fish that are being raised for identifying founders are provided in Supplementary Table 1. - + 8078 Species killifish - + 8078 Species killifish - + 29154 Species Atlantic - - MESH:C049037 + + MESH:D012685 Chemical - - aluminosilicate + + agarose - - MESH:C415406 + + MESH:D008899 + Chemical + + mineral oil + + + - Chemical borosilicate - + + MESH:C049037 + Chemical + + aluminosilicate + + + MESH:D005597 + Disease + + opening + + MESH:C049037 Chemical @@ -3853,65 +3873,89 @@ paragraph 11913 ZFN-injected and uninjected embryos were sampled at 3, 6, 9 and 24 hours post-injection to determine the temporal profile of ZFN protein expression. At each time point, one pool of 10 embryos was sampled. Embryos were homogenized in 1x sample treatment buffer with beta-mercaptoethanol and boiled for 5 minutes to denature the protein. Samples were separated on an 8% polyacrylamide gel using the discontinuous buffer system. The proteins were transferred onto a nitrocellulose membrane with a semi-dry transfer unit (BioRad, Waltham, MA, USA), and the membrane was blocked with 5% milk in TBST (20 mM Tris pH 7.5, 300 mM NaCl and 0.1% (v/v) Tween 20) overnight. ZFN protein was detected using a mouse monoclonal anti-Flag antibody (Sigma, St. Louis, MO, USA) at 1:5000 dilution. The secondary antibody used was horseradish peroxidase (HRP)-conjugated goat anti-mouse antibody (Santa Cruz Biotechnology, Dallas, TX, USA) at 1:5000 dilution. The membranes were incubated in the primary antibody for 60 min at room temperature, washed with TBST twice (10 min each), incubated with secondary antibody for 60 min, and finally washed with TBST (15 min). Detection was done using the ECL Prime Western blotting reagent (GE Healthcare, Pittsburgh, PA, USA). - + 10090 Species mouse - + 3704 Species horseradish - + 9925 Species goat - + 10090 Species mouse - + 29278 Species expression - - MESH:C040388 + + - + Chemical + + Tris + + + MESH:C016679 Chemical polyacrylamide - + MESH:D012965 Chemical NaCl - - MESH:C053576 + + - Chemical - - Tris + + TBST - + + - + Chemical + + TBST + + + MESH:D011136 + Chemical + + Tween 20 + + + MESH:D015352 + Disease + + semi-dry transfer unit + + MESH:D008623 Chemical beta-mercaptoethanol - - MESH:D011136 + + - Chemical - - Tween + + TBST @@ -3925,7 +3969,13 @@ paragraph 13197 Somatic mutations were determined using the Surveyor mutation detection kit (Transgenomic Inc., Omaha, NE, USA) following the manufacturer's instructions. Embryos (pool of 5 embryos per replicate) were sampled at 48 hours post-injection and genomic DNA was isolated using the proteinase K digestion method. The AHR2a and AHR2b target regions were amplified by polymerase chain reaction (PCR) using Advantage polymerase (Clontech, Mountain View, CA, USA). The primer sequences are given in Table 2. PCR cycling conditions were 94 C for 1 min, followed by 35 cycles of [94 C for 5 s, 68 C for 1 min], followed by 72 C for 7 min. The PCR products were melted and re-annealed to form hetero-duplex DNA and digested with Surveyor nuclease for 20 min at 42 C. The resulting products were electrophoresed on a 15% TBE gel and visualized by ethidium bromide staining. - + + - + Chemical + + TBE + + MESH:D004996 Chemical @@ -3943,13 +3993,13 @@ paragraph 14086 PCR products from samples that were positive in the Surveyor endonuclease assay were cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) and sequenced in 96-well plates (Functional Biosciences, Madison, WI, USA). Insertions and deletions were identified by aligning the sequences with the wildtype AHR sequence (Sequencher 5.1, Gene Codes Corp., Ann Arbor, MI, USA). Off-target screening for AHR2b CRISPR-Cas9 exon2 was done by direct-sequencing of PCR products. The amplification reactions were purified using the GeneClean kit (MP Biomedicals, Santa Ana, CA, USA). Direct-sequencing of the amplicons for AHR1a, AHR2a, and AHRR was done with primers Fh2a-ex2rev, 2b-1aF, and RR-ex2fwd, respectively (Table 2). The chromatograms were visually inspected for regions of double peaks indicating insertions or deletions. - + 105937229 Gene AHR2 - + 105920361 Gene @@ -3973,42 +4023,48 @@ paragraph 14958 We tested different concentrations of ZFN mRNA on killifish embryos by injecting 500, 1000 and 2000 pg per embryo. No overt toxic effects or phenotypic abnormalities were observed at any of the concentrations as determined by microscopic observation of embryos during development. Hence, we selected 2000 pg to inject in subsequent experiments. ZFN protein expression was confirmed in the embryos injected with 2000 pg ZFN mRNA. Embryos were sampled at 3, 6, 9 and 24 hours post-microinjection. The maximum ZFN protein expression was observed at 6 hours post-microinjection (Fig. 3A). At 3 and 9 hours, ZFN protein expression was much lower, and at 24 hours, no expression was detected. These results suggest that the maximum efficiency of ZFNs in inducing DSBs in killifish is around 6 hours post-injection. - + 8078 Species killifish - + 8078 Species killifish - + 29278 Species expression - + 29278 Species expression - + 29278 Species expression - + 29278 Species expression + + MESH:C007563 + Chemical + + DSBs + RESULTS @@ -4039,7 +4095,7 @@ paragraph 17669 CRISPR-Cas9 guide RNAs targeting exon 2 and 3 of the AHR2b locus were both efficient at generating mutations in the microinjected embryos. Surveyor mutation detection assay showed the presence of mutations in all 4 pools of AHR2b exon 2 CRISPR-Cas-injected embryos analyzed (20 embryos total; 5 embryos per pool) (Fig. 5A). The PCR products from these 4 pools were combined and cloned into the pGEM-T Easy vector. Sequencing results revealed that 16 sequences out of 70 (23%) had indels at the target site, represented by 8 distinct mutations (5 deletions and 3 insertions) (Fig. 5B). - + 105937229 Gene @@ -4063,66 +4119,60 @@ paragraph 18766 In order to assess potential off-target effects of the CRISPR-Cas9 experiments, we chose one of the guide RNAs that gave positive results (exon 2 of AHR2b) to screen for possible mutations in other AHR paralogs and AHRR. The multiple AHR paralogs and the AHRR are closely related genes and share substantial sequence identity, especially in the N-terminus. Alignment of the AHR2b CRISPR-Cas9 target sequence with the other genes is shown in Fig. 6A. The numbers of mismatches between the 20 bp AHR2b target site and AHR1a, AHR2a, AHR1b, and AHRR are 4, 4, 6, and 3, respectively. These corresponding regions were amplified from the uninjected and AHR2b CRISPR-Cas9-injected embryo genomic DNA samples in which AHR2b mutations were observed. The PCR products were analyzed for mutations using the Surveyor endonuclease assay. The AHR1a, AHR2a, and AHRR samples contained small fragments indicating endonuclease digestion of the PCR products (Fig. 6B). These fragments were also present in the AHR2a and AHRR reactions from the uninjected embryos. These amplification reactions were purified and analyzed by direct-sequencing. The chromatograms of the AHR1a and AHR2a sequencing reactions contained double peaks at previously characterized SNP locations (#141 and #159 for AHR1a, and #156 for AHR2a). Surveyor endonuclease is sensitive to even a single nucleotide mismatch between the two DNA strands, therefore it will cleave DNA at the SNP sites. The sizes of the observed Surveyor endonuclease fragments in these samples (Fig. 6B) can be explained by these SNPs, thereby suggesting that they do not represent off-target mutations. On the other hand, we did not detect a SNP in the AHRR sequences that would explain the fragments observed in the Surveyor assay. However the presence of these fragments in both uninjected and injected samples suggest that they are rare SNPs that may not be detected by direct sequencing. - + 105920361 Gene AHRR - + 105920361 Gene AHRR - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 105920361 Gene AHRR - + 105920361 Gene AHRR - + 105920361 Gene AHRR - + 105920361 Gene AHRR - - MESH:D009584 - Chemical - - N - DISCUSS @@ -4135,25 +4185,25 @@ paragraph 20701 In this study, we demonstrated the feasibility of targeted mutagenesis approaches in inducing mutations in an environmental model species with limited genomic and genetic resources. The ease with which ZFN and CRISPR-Cas9 methods can be utilized to cleave specific genomic DNA sequences has led to the generation of gene knockouts in species not traditionally used for conducting genetic and functional studies (Table 1). Using these methods, we produced mutations in AHR2a and AHR2b loci in killifish embryos. These embryos are currently being raised to adulthood, and their progeny will be screened to identify potential founders. As killifish have a long generation time (2 years minimum) and lack established breeding techniques, the time it takes to generate homozygous mutants is relatively long. Here, we propose some strategies to expedite the process of generating homozygous mutants in killifish. AHR-null killifish will be valuable for characterizing the role of the individual AHR paralogs in evolved resistance, as well as in normal development. - + 8078 Species killifish - + 8078 Species killifish - + 8078 Species killifish - + 8078 Species @@ -4171,78 +4221,78 @@ paragraph 22459 In order to determine the translational efficiency of the injected ZFN mRNA, we measured ZFN protein expression in embryos using an anti-FLAG antibody. Our results demonstrated that maximal ZFN protein expression is observed at 6 hours post-injection, suggesting that ZFN protein binding to the target site and creating DSBs occurs around this time. In comparison, in zebrafish embryos injected with a ZFN targeting the aryl hydrocarbon receptor repressor (ahrra) gene, we observed maximal ZFN protein expression at 3 hours post-injection. These species-specific differences in ZFN protein expression could be linked to the developmental rate in the two species. The developmental period for zebrafish from fertilization to hatching is 2 days, whereas killifish hatch at 15 days post-fertilization. Another factor that could contribute to the differences in protein expression is the water temperature at which embryos are incubated. Zebrafish embryos are reared at a higher temperature (28 C) than killifish (23 C), which may affect the rate of transcription and translation. - + 641291 Gene 123929 ahrra - + 7955 Species zebrafish - + 7955 Species zebrafish - + 8078 Species killifish - + 7955 Species Zebrafish - + 8078 Species killifish - + 29278 Species expression - + 29278 Species expression - + 29278 Species expression - + 29278 Species expression - + 29278 Species expression - - MESH:D006838 + + MESH:D014867 Chemical - - aryl hydrocarbon + + water @@ -4256,13 +4306,13 @@ paragraph 24866 Genome-editing methods have become a powerful resource for generating mutants in non-model species, but there are some concerns with regard to their off-target effects. ZFNs, as well as CRISPR-Cas9, have been associated with cytotoxicity, presumably due to binding and cleavage at non-target sites, although toxic effects could also result from the target gene disruption. Screening the genome for off-target effects a priori by blast searches with the target sequence is not feasible for species that do not have sequenced genomes. However, paralogs and closely related genes that have been characterized can be screened for mutations, as these represent likely targets. We checked for off-target effects of CRISPR-Cas9 in all AHR paralogs, as well as AHRR, a gene closely related to AHR (Fig. 6). Based on the Surveyor assay and sequencing results, no off-target effects were detected, even though the number of sequence mismatches among these genes was as low as 3 residues. If genomic resources are available, several tools exist for in silico identification of potential off-target sites. Alternatively, potential off-target sites can be searched using the fuzznuc algorithm (http://embossgui.sourceforge.net/demo/manual/fuzznuc.html) in RNAseq data, if available. - + 105920361 Gene AHRR - + MESH:D064420 Disease @@ -4274,85 +4324,85 @@ paragraph 26136 In ZFN and CRISPR-Cas9 injected embryos we did not observe any developmental toxicity, including the highest concentration of ZFN (2000 pg of ZFN mRNA/embryo). These results are in contrast to studies in other fish species, in which overt developmental phenotypes were observed at these concentrations. One reason for the lack of toxicity could be the relative size of the killifish embryos in comparison to zebrafish embryos, in which most of these methods have been optimized. Freshly fertilized killifish embryos are 2 mm in diameter, and are larger than zebrafish (0.7 mm), where ZFNs above 100 pg per embryo have been shown to cause embryo toxicity. Similarly, in medaka with an egg diameter (1.2 mm), 160 pg of ZFN caused embryo toxicity. In addition to the size of the embryo, rapid turnover of the ZFN protein might be responsible for the lack of embryo toxicity in killifish. Our results suggest that the ZFN protein is expressed transiently around 6 hours post-injection and is degraded by 9 hours (Fig. 3A), suggesting faster degradation of the injected mRNA in killifish. We cannot rule out the possibility of additional factors, yet to be determined, contributing to the low toxicity of injected mRNA in killifish. - + 8078 Species killifish - + 7955 Species zebrafish - + 8078 Species killifish - + 7955 Species zebrafish - + 8090 Species medaka - + 8078 Species killifish - + 8078 Species killifish - + 8078 Species killifish - + MESH:D064420 Disease toxicity - + MESH:D064420 Disease toxicity - + MESH:D064420 Disease - - developmental toxicity + + toxicity - + MESH:D064420 Disease toxicity - + MESH:D064420 Disease toxicity - + MESH:D064420 Disease @@ -4364,49 +4414,49 @@ paragraph 27364 Our results demonstrate the utility of the ZFN and CRISPR-Cas9 approaches in generating mutations in an ecologically relevant fish species that have successfully adapted to environments contaminated with PCBs and PAHs. Generating AHR knockout mutants will provide a powerful tool to functionally characterize the role of these proteins in evolved resistance to PCBs and PAHs. We are currently raising the ZFN- and CRISPR-Cas9-injected fish to adults. The numbers of injected animals that are being raised and the breeding strategy that will be followed are provided in the supplemental information (Supp. Table 1 and Supp. Fig. 1). Killifish mature in 1 year; however, larger fish, producing sufficient offspring per pair for testing, require > 2 years of laboratory rearing. Most organisms in which these approaches are extensively used to generate mutants have short life history cycles (e.g. 2-3 months). However, in the past few years targeted mutagenesis approaches have been successfully used in species with longer life cycles such as yellow catfish, rainbow trout and Atlantic salmon. In order to expedite the process of obtaining homozygous mutants, several strategies can be followed. During certain steps of the homozygous mutant breeding, fin-clips from juvenile fish can be screened by direct-sequencing to identify mutants (Supp. Fig. 1). Direct-sequencing allows screening a single DNA sample per fish, as opposed to cloning the PCR product and screening multiple clones. Another possible approach to accelerate the identification of founders is to screen gametes stripped from juveniles. Once the founder animals are identified, it is possible to artificially induce precocious maturation of gametes by treating the founders with gonadotropins, a common practice in breeding aquaculture species. - + 8078 Species Killifish - + 1234273 Species yellow catfish - + 8022 Species rainbow trout - + 8030 Species Atlantic salmon - + MESH:D011078 Chemical PCBs - - MESH:C087686 + + MESH:D011084 Chemical - + PAHs - - MESH:C087686 + + MESH:D011084 Chemical - + PAHs - + MESH:D011078 Chemical @@ -4418,13 +4468,13 @@ paragraph 29178 Other strategies to consider while generating mutants in species with long life history cycles is to generate at least two mutant lines targeting different sites in each gene. As it is time consuming to raise embryos to reproductive maturity, targeting multiple regions of the same gene confirms the specificity of the observed phenotypes associated with the gene knockout. In addition, if a frameshift mutation is not produced in one of the targets, the other target will serve as an alternative. We followed these approaches and designed CRISPR-Cas9 guide RNAs against AHR2a, in addition to the ZFN method. Similarly, we designed two different gRNAs against AHR2b, and observed mutations at both target sites. Another important consideration in target site selection in species such as the Atlantic killifish is to avoid regions with known single nucleotide polymorphisms (SNPs). Killifish have high genetic diversity, and if the target site has multiple mismatches due to SNPs, the efficacy of the ZFN proteins or guide RNAs will be reduced. In addition, SNPs interfere with the mutation detection assays such as Surveyor endonuclease by creating cuts at polymorphic sites. An additional strategy to consider is optimizing injection conditions to maximize the chances for mutating both alleles at the target locus simultaneously. Such bi-allelic mutations in CRISPR-Cas9-injected embryos have been demonstrated previously. The presence of bi-allelic mutations in the germline will expedite the process of generating homozygous individuals. - + 8078 Species Atlantic killifish - + 8078 Species @@ -4448,13 +4498,13 @@ paragraph 31170 In this study we demonstrated the utility of targeted gene-editing methods for generating ahr mutants in Atlantic killifish. We observed higher frequency of mutations with CRISPR-Cas9 compared to ZFN mRNA injected embryos. We are currently raising the injected embryos to identify potential germ line mutants. Some of the challenges in generating homozygous mutants in killifish include long generation time and lack of established breeding methods. We have proposed some strategies to overcome these challenges and expedite the process of identifying the founders and generating ahr-null mutants. The strategies proposed here are applicable to any fish species with limited aquaculture resources, providing an opportunity to conduct functional studies. - + 8078 Species Atlantic killifish - + 8078 Species @@ -4496,18 +4546,12 @@ surname:Hahn;given-names:ME 32448 Understanding the physiological role of aryl hydrocarbon receptor repressor (AHRR) using gene knock-down and targeted mutagenesis in zebrafish - + 105920361 Gene AHRR - - MESH:D006838 - Chemical - - aryl hydrocarbon - surname:Schartl;given-names:M @@ -4547,7 +4591,7 @@ surname:Gou;given-names:Y 32731 Targeted disruption of exogenous EGFP gene in medaka using zinc-finger nucleases - + 8090 Species @@ -4572,7 +4616,7 @@ surname:Uemura;given-names:N 32812 Efficient targeted mutagenesis in medaka using custom-designed transcription activatorlike effector nucleases - + 8090 Species @@ -4591,6 +4635,11 @@ 168 32922 Stages in Normal Development of Fundulus heteroclitus + + Disease + + Fundulus heteroclitus + surname:Adelman;given-names:ZN @@ -4606,7 +4655,7 @@ ref 32976 Germline excision of transgenes in Aedes aegypti by homing endonucleases - + 7159 Species @@ -4663,13 +4712,13 @@ 91 33224 Acquired resistance to Ah receptor agonists in a population of Atlantic killifish (Fundulus heteroclitus) inhabiting a marine superfund site: In vivo and in vitro studies on the inducibility of xenobiotic metabolizing enzymes - + 8078 Species Atlantic killifish - + 8078 Species @@ -4706,6 +4755,12 @@ 1175 33511 Targeted chromosomal cleavage and mutagenesis in Drosophila using zinc-finger nucleases + + 7227 + Species + + Drosophila + surname:Cho;given-names:KW @@ -4722,7 +4777,7 @@ 834 33599 Biallelic genome modification in F(0) Xenopus tropicalis embryos using the CRISPR/Cas system - + 8364 Species @@ -4766,7 +4821,7 @@ surname:Cohen;given-names:S 33692 Fundulus as the premier teleost model in environmental biology: opportunities for new insights using genomics - + 8078 Species @@ -4787,29 +4842,29 @@ 200 33802 Effects of ovaprim, a commercial spawning inducer, on vasotocin and steroid hormone profiles in the catfish Heteropneustes fossilis: in vivo and in vitro studies - + 71179 Species catfish - + 93621 Species Heteropneustes fossilis - + MESH:D014668 Chemical vasotocin - + MESH:D013256 Chemical - - steroid + + steroid hormone @@ -4827,31 +4882,31 @@ 240 33964 AHR2 mediates cardiac teratogenesis of polycyclic aromatic hydrocarbons and PCB-126 in Atlantic killifish (Fundulus heteroclitus) - + 105937229 Gene AHR2 - + 8078 Species Atlantic killifish - + 8078 Species Fundulus heteroclitus - - MESH:C023035 + + MESH:D011078 Chemical - - PCB-126 + + PCB - + MESH:D011084 Chemical @@ -4881,24 +4936,18 @@ surname:Liang;given-names:D 34094 Heritable targeted inactivation of myostatin gene in yellow catfish (Pelteobagrus fulvidraco) using engineered zinc finger nucleases - + 1234273 Species yellow catfish - + 1234273 Species Pelteobagrus fulvidraco - - MESH:D055435 - Chemical - - myostatin - surname:Faraji;given-names:F @@ -4924,7 +4973,7 @@ surname:Ngo;given-names:C 34227 Heritable targeted gene disruption in zebrafish using designed zinc-finger nucleases - + 7955 Species @@ -4946,13 +4995,13 @@ surname:Wargelius;given-names:A 34312 Targeted Mutagenesis in Atlantic Salmon (Salmo salar L.) Using the CRISPR/Cas9 System Induces Complete Knockout Individuals in the F0 Generation - + 8030 Species Atlantic Salmon - + 8030 Species @@ -4974,19 +5023,19 @@ surname:Woltering;given-names:DS 34457 Altered CYP1A expression in Fundulus heteroclitus adults and larvae: a sign of pollutant resistance? - + 105930482 Gene CYP1A - + 8078 Species Fundulus heteroclitus - + 29278 Species @@ -5025,7 +5074,7 @@ surname:Crews;given-names:CM 34647 Highly efficient targeted mutagenesis in axolotl using Cas9 RNA-guided nuclease - + 8296 Species @@ -5062,8 +5111,8 @@ 370 34791 A simple "universal" DNA extraction procedure using SDS and proteinase K is compatible with direct PCR amplification - - MESH:C032259 + + MESH:D012967 Chemical SDS @@ -5122,7 +5171,7 @@ 361 35094 Induced ovulation in Atlantic croaker (Sciaenidae) using hCG and an LHRH analog: A preliminary study - + 29154 Species @@ -5140,7 +5189,7 @@ 443 35195 Mecahnisms of innate and acquired resistance to dioxin-like compounds - + MESH:D004147 Chemical @@ -5162,29 +5211,23 @@ 13748 35265 Molecular evolution of two vertebrate aryl hydrocarbon (dioxin) receptors (AHR1 and AHR2) and the PAS family - + 105937232 Gene AHR1 - + 105937229 Gene AHR2 - - MESH:D004147 - Chemical - - dioxin - - - MESH:D006838 + + MESH:D011478 Chemical - - aryl hydrocarbon + + PAS @@ -5207,13 +5250,13 @@ surname:Inoue;given-names:T 35374 Transcription activator-like effector nucleases efficiently disrupt the target gene in Iberian ribbed newts (Pleurodeles waltl), an experimental model animal for regeneration - + 8319 Species Iberian ribbed newts - + 8319 Species @@ -5235,7 +5278,7 @@ 97 35549 Targeted mutagenesis in sea urchin embryos using TALENs - + 7656 Species @@ -5262,7 +5305,7 @@ surname:Tsai;given-names:SQ 35605 Efficient genome editing in zebrafish using a CRISPR-Cas system - + 7955 Species @@ -5298,7 +5341,7 @@ 13909 35754 Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system - + 7955 Species @@ -5321,37 +5364,31 @@ surname:Stegeman;given-names:JJ 35840 Role of AHR2 in the expression of novel cytochrome P450 1 family genes, cell cycle genes, and morphological defects in developing zebra fish exposed to 3,3',4,4',5-pentachlorobiphenyl or 2,3,7,8-tetrachlorodibenzo-p-dioxin - + 105937229 Gene AHR2 - + 7955 Species zebra fish - + 29278 Species expression - - MESH:D065308 - Disease - - morphological defects - - - MESH:D013749 + + MESH:D000072317 Chemical 2,3,7,8-tetrachlorodibenzo-p-dioxin - + MESH:C023035 Chemical @@ -5372,29 +5409,22 @@ 33824 36063 Identification and functional characterization of two highly divergent aryl hydrocarbon receptors (AHR1 and AHR2) in the teleost Fundulus heteroclitus Evidence for a novel subfamily of ligand-binding basic helix loop helix-Per-ARNT-Sim (bHLH-PAS) factors - + 105937232 Gene AHR1 - + 105937229 Gene AHR2 - - 8078 - Species - - Fundulus heteroclitus - - - MESH:D006838 - Chemical - - aryl hydrocarbon + + Disease + + teleost Fundulus heteroclitus @@ -5415,7 +5445,7 @@ surname:Sawada;given-names:H 36318 Efficient targeted mutagenesis of the chordate Ciona intestinalis genome with zinc-finger nucleases - + 7719 Species @@ -5484,7 +5514,7 @@ surname:Deng;given-names:Y 36622 Efficient targeted gene disruption in Xenopus embryos using engineered transcription activator-like effector nucleases (TALENs) - + 8364 Species @@ -5536,29 +5566,23 @@ surname:Jiang;given-names:XL 36815 Antagonistic roles of Dmrt1 and Foxl2 in sex differentiation via estrogen production in tilapia as demonstrated by TALENs - + 105938153 Gene Dmrt1 - + 105926779 Gene Foxl2 - + Disease tilapia - - MESH:D004967 - Chemical - - estrogen - surname:Deshmukh;given-names:H @@ -5597,7 +5621,7 @@ 287 37054 Fundulus heteroclitus: ovarian reproductive physiology and the impact of environmental contaminants - + 8078 Species @@ -5621,7 +5645,7 @@ surname:Zhao;given-names:H 37154 A highly effective TALEN-mediated approach for targeted gene disruption in Xenopus tropicalis and zebrafish - + 8364 Species @@ -5668,7 +5692,7 @@ surname:Xu;given-names:H 37401 Highly efficient and specific genome editing in silkworm using custom TALENs - + 7091 Species @@ -5692,7 +5716,7 @@ surname:Hahn;given-names:ME 37478 Development of the morpholino gene knockdown technique in Fundulus heteroclitus: a tool for studying molecular mechanisms in an established environmental model - + 8078 Species @@ -5727,7 +5751,7 @@ 527 37723 Inducing high rates of targeted mutagenesis in zebrafish using zinc finger nucleases (ZFNs) - + 7955 Species @@ -5749,7 +5773,7 @@ 548 37815 Genetic diversity and structure of an estuarine fish (Fundulus heteroclitus) indigenous to sites associated with a highly contaminated urban harbor - + 8078 Species @@ -5772,7 +5796,7 @@ surname:Reppert;given-names:SM 37963 Efficient targeted mutagenesis in the monarch butterfly using zinc-finger nucleases - + 13037 Species @@ -5793,31 +5817,31 @@ 81 38047 Cytochrome P4501A (CYP1A) in killifish (Fundulus heteroclitus): Heritability of altered expression and relationship to survival in contaminated sediments - + 105930482 Gene CYP1A - + 105930482 Gene Cytochrome P4501A - + 8078 Species killifish - + 8078 Species Fundulus heteroclitus - + 29278 Species @@ -5843,7 +5867,7 @@ surname:McKinney;given-names:R 38201 Adaptations of wild populations of the estuarine fish Fundulus heteroclitus to persistent environmental contaminants - + 8078 Species @@ -5863,30 +5887,30 @@ 864 38318 Adaptation of the Estuarine Fish Fundulus heteroclitus (Atlantic Killifish) to Polychlorinated Biphenyls (PCBs) - - 8078 - Species - - Fundulus heteroclitus - - + 8078 Species Atlantic Killifish - + MESH:D011078 Chemical Polychlorinated Biphenyls - + MESH:D011078 Chemical PCBs + + MESH:D005393 + Disease + + Fish Fundulus heteroclitus + surname:Yaoita;given-names:Y @@ -5903,7 +5927,7 @@ 784 38430 Generation of albino Xenopus tropicalis using zinc-finger nucleases - + 8364 Species @@ -5957,17 +5981,17 @@ surname:Stanton;given-names:BA 38635 Morpholino gene knockdown in adult Fundulus heteroclitus: role of SGK1 in seawater acclimation - + 105930542 Gene SGK1 - - 8078 - Species - - Fundulus heteroclitus + + MESH:C538052 + Disease + + adult Fundulus heteroclitus @@ -5988,7 +6012,7 @@ surname:Shibata;given-names:T 38730 Targeted mutagenesis in the sea urchin embryo using zinc-finger nucleases - + 7656 Species @@ -6025,7 +6049,7 @@ 446 38847 Mammalian gene targeting with designed zinc finger nucleases - + 9606 Species @@ -6044,7 +6068,7 @@ ref 38908 Chimeric nucleases stimulate gene targeting in human cells - + 9606 Species @@ -6066,25 +6090,30 @@ 239 38967 Developmental and tissue-specific expression of AHR1, AHR2, and ARNT2 in dioxin-sensitive and -resistant populations of the marine fish Fundulus heteroclitus - + 105937232 Gene AHR1 - + 105937229 Gene AHR2 - + 29278 Species expression - + + Disease + + Fundulus heteroclitus + + MESH:D004147 Chemical @@ -6110,20 +6139,20 @@ surname:Hiraga;given-names:T 39125 Aryl hydrocarbon receptor 2 mediates 2,3,7,8-tetrachlorodibenzo-p-dioxin developmental toxicity in zebrafish - + 30517 Gene Aryl hydrocarbon receptor 2 - + MESH:D064420 Disease - - developmental toxicity + + toxicity - - MESH:D013749 + + MESH:D000072317 Chemical 2,3,7,8-tetrachlorodibenzo-p-dioxin @@ -6145,7 +6174,7 @@ surname:Porteus;given-names:MH 39234 Comparison of zinc finger nucleases for use in gene targeting in mammalian cells - + 9606 Species @@ -6188,24 +6217,18 @@ surname:Nacci;given-names:D 39358 Genetic variation at aryl hydrocarbon receptor (AHR) loci in populations of Atlantic killifish (Fundulus heteroclitus) inhabiting polluted and reference habitats - + 8078 Species Atlantic killifish - + 8078 Species Fundulus heteroclitus - - MESH:D006838 - Chemical - - aryl hydrocarbon - surname:Uchino;given-names:K @@ -6360,7 +6383,7 @@ surname:Uchino;given-names:K 40074 Efficient TALEN construction for Bombyx mori gene targeting - + 7091 Species @@ -6438,7 +6461,7 @@ surname:Wang;given-names:Y 40388 High efficiency system for construction and evaluation of customized TALENs for silkworm genome editing - + 7091 Species @@ -6458,7 +6481,7 @@ 33 40492 Direct gene disruption by TALENs in medaka embryos - + 8090 Species @@ -6503,11 +6526,6 @@ surname:Kilgore;given-names:KH 40651 Use of a novel induced spawning technique for the first reported captive spawning of Tetraodon nigroviridis - - Disease - - Tetraodon nigroviridis - 64 @@ -6521,13 +6539,13 @@ 2085 40759 The evolutionary radiation of diverse osmotolerant physiologies in killifish (Fundulus sp.) - + 8078 Species killifish - + 8078 Species @@ -6549,13 +6567,13 @@ 1305 40851 Salinity- and population-dependent genome regulatory response during osmotic acclimation in the killifish (Fundulus heteroclitus) gill - + 8078 Species killifish - + 8078 Species @@ -6607,13 +6625,13 @@ 250 41156 Heritable targeted inactivation of the rainbow trout (Oncorhynchus mykiss) master sex-determining gene using zinc-finger nucleases - + 8022 Species rainbow trout - + 8022 Species @@ -6637,7 +6655,7 @@ surname:Yamamoto;given-names:T 41287 Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases - + 7719 Species @@ -6671,12 +6689,17 @@ surname:Faraji;given-names:FM 41362 Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases - + 8364 Species Xenopus tropicalis + + Disease + + soma + surname:Naruse;given-names:K @@ -6694,7 +6717,7 @@ surname:Hong;given-names:Y 41493 Parameters and efficiency of direct gene disruption by zinc finger nucleases in medaka embryos - + 8090 Species @@ -6724,42 +6747,42 @@ fig_caption 42247 ZFN-mediated mutagenesis of killifish AHR2a. A. ZFN protein expression in killifish embryos. Uninjected (-) or ZFN-injected (+) embryos were sampled at 3, 6, 9, and 24 hours after micro-injection. Homogenates were resolved on SDS-PAGE and probed with a Flag-tag antibody. Lysate from COS-7 cells transfected with the ZFN expression plasmid was run as a positive control. B. Surveyor nuclease detection of mutations in the ZFN target region of AHR2a. Each lane represents a pool of 5 embryos from which a 335 bp genomic DNA fragment was amplified. U: uninjected control, ZFN: injected embryos. Lanes 1 and 2 are two representative samples that were positive in the mutation detection assay. Approximate sizes of the Surveyor nuclease-digested fragments containing the deletions are shown (240 and 95 bp). The full-length PCR product from sample 2 was cloned and sequenced. C. AHR2a exon 3 sequence surrounding the ZFN target site (in red). Four types of deletion mutants were observed among the sequenced clones (deletions of 2, 4, 5, and 28 nucleotides). - + 8078 Species killifish - + 8078 Species killifish - + 29278 Species expression - + 29278 Species expression - - MESH:C032259 - Chemical - - SDS - - + CVCL:0224 CellLine COS-7 + + MESH:D012967 + Chemical + + SDS + FIG @@ -6768,7 +6791,7 @@ fig_caption 43304 CRISPR-Cas9-mediated mutagenesis of killifish AHR2a targeting exon 3. A. Surveyor nuclease detection of mutations in the CRISPR-Cas9 target region of AHR2a exon 3. Each lane represents a pool of 5 embryos from which a 255 bp genomic DNA fragment was amplified. U: uninjected, lanes 1 and 2: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (160 and 95 bp). The full-length PCR products from samples 1 and 2 were pooled, cloned, and sequenced. B. AHR2a exon 3 sequence surrounding the CRISPR-Cas9 target site (in red). Ten different deletion mutants were observed among the sequenced clones. - + 8078 Species @@ -6782,31 +6805,31 @@ fig_caption 43970 CRISPR-Cas9-mediated mutagenesis of killifish AHR2b targeting exon 2 and exon 3. A. Surveyor nuclease detection of mutations in the AHR2b CRISPR-Cas9 target region. Each lane represents a pool of 5 embryos from which a 309 bp genomic DNA fragment was amplified. U: uninjected control, lanes 1-4: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (175 and 135 bp). The full-length PCR products from samples 1-4 were pooled, cloned, and sequenced. B. AHR2b exon 2 sequence surrounding the CRISPR-Cas9 target site (in red). Five types of deletion mutants and 3 types of insertions were observed among the sequenced clones. C. Surveyor nuclease detection of mutations in the CRISPR-Cas9 target region of AHR2b exon 3. Each lane represents a pool of 5 embryos from which a 198 bp genomic DNA fragment was amplified. U: uninjected, lanes 1 and 2: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (110 and 90 bp). The full-length PCR products from samples 1 and 2 were pooled, cloned, and sequenced. D. AHR2b exon 3 sequence surrounding the CRISPR-Cas9 target site (in red). Twenty different types of deletion mutants and seven types of insertion mutants were observed among the sequenced clones. - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 8078 Species @@ -6820,37 +6843,37 @@ fig_caption 45313 AHR2b CRISPR-Cas9 off-target analysis. A. Nucleotide sequence alignment of the AHR2b CRISPR-Cas9 target sequence with the corresponding regions of closely related genes. AHR1a, AHR2a, AHR1b, and AHRR have 4, 4, 6, and 3 mismatches to AHR2b in this region, respectively. B. Surveyor nuclease detection of mutations in AHR1a, AHR2a, AHR1b, and AHRR in the AHR2b CRISPR-Cas9 target region. Digested fragments are indicated by a *. Each lane represents a pool of 5 embryos from which genomic DNA fragments were amplified. U: uninjected control, 1-4: CRISPR-Cas9 injected embryos. AHR1a, AHR2a, and AHRR PCR products from uninjected and injected embryos were sequenced. - + 105937229 Gene AHR2 - + 105937229 Gene AHR2 - + 105920361 Gene AHRR - + 105920361 Gene AHRR - + 105937229 Gene AHR2 - + 105920361 Gene @@ -6872,150 +6895,174 @@ T1.xml 46057 Gene-editingapproach Species Reference ZFN Monarch butterfly (Danaus plexippus)Two-spotted cricket (Gryllus bimaculatus)Sea urchin (Hemicentrotus pulcherrimus)Ascidian (Ciona intestinalis)Western clawed frog (Xenopus tropicalis)Yellow catfish (Pelteobagrus fulvidraco)Medaka (Oryzias melastigma)Rainbow trout (Oncorhynchus mykiss) TALEN Nematode (Pristionchus pacificus)Marine polycheate (Platyneries dumerilii)Culicine mosquito (Aedes aegypti)Silk worm (Bombyx mori)Sea urchin (Hemicentrotus pulcherrimus)Ascidian (Ciona intestinalis)Western clawed frog (Xenopus tropicalis)Iberian ribbed newts (Pleurodeles waltl)Tilapia (Oreochromis niloticus)Medaka (Oryzias melastigma)Yellow catfish (Pelteobagrus fulvidraco) CRISPR-Cas9 Western clawed frog (Xenopus tropicalis)Axolotl (Ambystoma mexicanum)Atlantic Salmon (Salmo salar)Tilapia (Oreochromis mossambicus) (Ji-Feng et al., 2014) - + 13037 Species Monarch butterfly - + 13037 Species Danaus plexippus - + 6999 Species Two-spotted cricket - + + 6999 + Species + + Gryllus bimaculatus + + + 7656 + Species + + Sea urchin + + + 7650 + Species + + Hemicentrotus pulcherrimus + + 7719 Species Ciona intestinalis - + 8364 Species Western clawed frog - + 8364 Species Xenopus tropicalis - + 1234273 Species Yellow catfish - + 1234273 Species Pelteobagrus fulvidraco - + 8090 Species Medaka - - 30732 + + 54126 Species - - Oryzias melastigma + + Pristionchus pacificus - - 8022 + + 7159 Species - - Rainbow trout + + Aedes aegypti - - 8022 + + 7091 Species - - Oncorhynchus mykiss + + Bombyx mori - - 54126 + + 7656 Species - - Pristionchus pacificus + + Sea urchin - + + 7650 + Species + + Hemicentrotus pulcherrimus + + 7719 Species Ciona intestinalis - + 8364 Species Western clawed frog - + 8364 Species Xenopus tropicalis - + 8319 Species Iberian ribbed newts - + 8319 Species Pleurodeles waltl - + 1234273 Species Pelteobagrus fulvidraco - + 8364 Species Western clawed frog - + + 8364 + Species + + Xenopus tropicalis + + + 8296 + Species + + Axolotl + + 8127 Species Oreochromis mossambicus - - MESH:D009041 - Disease - - Aedes aegypti)Silk worm (Bombyx mori)Sea urchin (Hemicentrotus pulcherrimus)Ascidian - - - MESH:D009041 - Disease - - Gryllus bimaculatus)Sea urchin (Hemicentrotus pulcherrimus)Ascidian - - + Disease - - Xenopus tropicalis)Axolotl (Ambystoma mexicanum)Atlantic Salmon (Salmo salar)Tilapia + + Oryzias melastigma)Rainbow trout (Oncorhynchus mykiss - + MESH:D015004 Disease @@ -7029,6 +7076,12 @@ table_caption 47232 AHR2a and AHR2b target regions and oligonucleotides used in ZFN and CRISPR-Cas9-based mutagenesis. The ZFN target site is shown in capital letters. The sequence flanking the target sites are recognized by the ZFN proteins. + + MESH:D009841 + Chemical + + oligonucleotides + TABLE @@ -7037,48 +7090,36 @@ T2.xml 47455 Name Description Sequence AHR2a exon 2 ZFN target site ctgcgcctcagcgtgGGATAcctgagggtcaagag AHR2a exon 3 ZFN target site gactcattcagcttcTCTGAaggagagctgctgct AHR2a exon 2 CRISPR-Cas9 target site GGAGCGAATCTCATCAGGGA AHR2a exon 3 CRISPR-Cas9 target site GGGTGGCTCCTGGAGTCAAC AHR2b exon2 CRISPR-Cas9 target site GGACCGTCTGAACGGCGAGC AHR2b exon3 CRISPR-Cas9 target site GGGCGAAGGTCTGAGCGGTC 2-ex3Fwd AHR2a ZFN pair3 screening TGTTTCACCTCCTGCACAGCTTCC 1XRR1 AHR2a ZFN pair3 screening TACTAGCTAAACGTGACCTGTCG 2a-ex3CrF AHR2a CRISPR-Cas exon3 screening CTGTTCTATCAGGTGGAGAGTCC 2a-ex3CrR AHR2a CRISPR-Cas exon3 screening GTTGTTGGATGAGTATGCAGGTGG AHR2b-CrF AHR2b CRISPR-Cas9 exon2 screening AATCTTTGATCCGGTCTGATCTGG AHR2b-CrR AHR2b CRISPR-Cas9 exon2 screening AGACACAACCAAGCAGCAACACG 2bex3-Fwd AHR2b CRISPR-Cas9 exon3 screening ATCGCCCAGCCTTAGCTTCCAGTCG 2bex3-Rev AHR2b CRISPR-Cas9 exon3 screening GCGTTTTTGCTTTGTCTGCTGTCGG Fh2A-ex3-CR1 AHR2a exon3 gRNA oligo1 TAGGGTGGCTCCTGGAGTCAAC Fh2A-ex3-CR2 AHR2a exon3 gRNA oligo2 AAACGTTGACTCCAGGAGCCAC FhAHR2b-CR1 AHR2b exon2 gRNA oligo1 TAGGACCGTCTGAACGGCGAGC FhAHR2b-CR2 AHR2b exon2 gRNA oligo2 AAACGCTCGCCGTTCAGACGGT Fh2B-ex3-CR1 AHR2b exon3 gRNA oligo1 TAGGGCGAAGGTCTGAGCGGTC Fh2B-ex3-CR2 AHR2b exon3 gRNA oligo2 AAACGACCGCTCAGACCTTCGC 2b-1aF AHR1a off-target screening GCTTAATCCTGCTTTCGTCTCTGC Fh1a-ex2rev AHR1a off-target screening AAGAAGTGTTTGGCTCGTAGGTAGC 2b-2aF AHR2a off-target screening TTTTACCCCTCACGGAAGAAGATGG Fh2a-ex2rev AHR2a off-target screening GTAGCTCTTGACCCTCAGGTATCC 2b-1bF AHR1b off-target screening TTTCCTCCTCTCCAGAGCCAAACC Fh1b-ex2rev AHR1b off-target screening AAGAAGCTCTTGGTGCGCAGGTAGC RR-ex2fwd AHRR off-target screening CTCATCCCTTCTGCCGTCTCGTCC RR-ex2rev AHRR off-target screening AAGAAGCTTTTGACGCGGAGGTAGG - + 105920361 Gene AHRR - + 105920361 Gene AHRR - + 2498238 Species CR2 - + 2498238 Species CR2 - + 2498238 Species CR2 - - MESH:D002857 - Chemical - - CrF - - - MESH:D002857 - Chemical - - CrF - REF @@ -7091,7 +7132,7 @@ paragraph 51075 AHR2 genes were mutated without detectable off-target effects in paralogs. - + 105937229 Gene @@ -7160,11 +7201,6 @@ abstract 181 A validated and reliable instrument was developed to knowledge, attitudes and behaviours with respect to evidence-based practice (EBB-KABQ) in medical trainees but requires further adaptation and validation to be applied across different health professionals. - - Chemical - - EBB-KABQ - ABSTRACT @@ -7177,7 +7213,7 @@ abstract 449 A modified 33-item evidence-based practice scale (EBP-KABQ) was developed to evaluate EBP perceptions and behaviors in clinicians. An international sample of 673 clinicians interested in treatment of pain (mean age = 45 years, 48% occupational therapists/physical therapists, 25% had more than 5 years of clinical training) completed an online English version of the questionnaire and demographics. Scaling properties (internal consistency, floor/ceiling effects) and construct validity (association with EBP activities, comparator constructs) were examined. A confirmatory factor analysis was used to assess the 4-domain structure EBP knowledge, attitudes, behavior, outcomes/decisions). - + MESH:D010146 Disease @@ -7231,13 +7267,13 @@ paragraph 1751 Evidence-based practice (EBP) is defined as the integration of the best research evidence with patients' interests and clinical circumstances in decision making. As EBP is associated with improved clinical decision-making and patient care, health professional organizations have advocated for increased training in EBP for all health care professionals at all levels of education. Understanding how EBP is understood and implemented across different health professionals can identify educational needs and outcomes, and predict where new research evidence is more likely to be implemented. As such, a validated and reliable instrument is required to evaluate an individual's perceptions of EBP. - + 9606 Species patients - + 9606 Species @@ -7249,7 +7285,7 @@ paragraph 2450 A systematic review which studied 104 instruments on EBP suggested that evaluation of EBP could be divided into the following definable components: EBP knowledge, attitudes toward EBP, application/use of EBP and practitioners' EBP behaviors in the clinical setting. Knowledge about EBP means that clinicians have knowledge of fundamental EBP concepts and terminology and concepts related to quality or levels of evidence. It also includes the ability to search the literature and critically appraise the evidence for its validity, impact and applicability. Attitude toward EBP includes the intuitive appeal of EBP, the likelihood of adopting EBP given professional requirements to do so, openness to new practices, and the perceived divergence between research-based/academically developed interventions versus current practice. Application and use of EBP refers to whether health professionals are able to apply their EBP knowledge to the specific clinical scenarios. This includes: capability to generate clinical question(s) regarding disease prevention, diagnosis and management as well as implementation of evidence with integrity of clinical circumstances. EBP behaviors refer to practitioners' performance of the instrumental activities associated with EBP such as searching and obtaining higher quality evidence in their own practice. - + MESH:D005597 Disease @@ -7267,6 +7303,12 @@ paragraph 4375 Among available scales, one that has taken a multi-dimensional approach and shown early promise is the The knowledge, attitude and behavior questionnaire (KAB) originally developed by Johnson and colleagues. The KAB scale was designed to evaluate EBP teaching and learning in the undergraduate medical education setting. With permission from the developers, two study authors (JMD and ML) developed a modified KAB scale (EBP-KABQ), to be applicable to health professionals other than physicians using expert review and pilot testing. This process resulted in removal of items that were perceived by users as redundant or unclear. + + MESH:C537366 + Disease + + ML + Disease @@ -7278,12 +7320,6 @@ paragraph 5005 The goal of this study was to validate the modified scale (EBP-KABQ) for use in a multidisciplinary group of clinicians by determining: (1) Scaling properties- internal consistency, floor/ceiling effects, and (2) Construct validity- based on predetermined hypotheses on the relationship of subcomponents of EBP, and (3) Structural validity: the integrity of a 4-domain structure based on confirmatory factor analysis. - - MESH:D020914 - Disease - - Structural validity - METHODS @@ -7296,7 +7332,7 @@ paragraph 5431 The EBP-KABQ incorporates 33 items in four domains of EBP: knowledge (8 items, 6 ordinal items), attitudes (14 items, 14 ordinal items), behaviour (8 items, 5 ordinal items) and outcomes/decisions (3 items, 3 ordinal items) (KABQ). The knowledge items retain a 7-point Likert scale with lower scores indicating a lower level of EBP knowledge. The Attitudes towards EBP items retain a 7-point Likert scale. High scores indicate positive attitude after several items were reversely scored. For EBP behaviour, lower scores indicate a lower frequency of using EBP in current practice. A 6-point Likert scale is used for responses to the items in the outcomes/decisions domain. Lower scores indicate unfavorable patient outcomes and poor clinical evidence-based decision making. Detail of the EBP-KABQ scale and a summary of the changes to original scale are presented in Additional files 1 and 2. - + 9606 Species @@ -7314,25 +7350,25 @@ paragraph 6364 All participants were recruited from a clinical trial assessing use of pain research evidence about pain. Eligible practitioners were (1) physicians, nurses, occupational therapists (OTs), physical therapists (PTs), or psychologists who were currently working in clinical practice at least one day/week; (2) fluent in English; (3) able to access a computer at home or at work that provided unrestricted access to the World Wide Web; (4) possessed an active email account;(5) consent to participate in this research studyA total of 870 clinicians met the inclusion criteria and were invited to participate. From August 2011 to February 2013, 673 clinicians (physicians, nurses, OTs/PTs, psychologists etc.) completed an online EBP-KABQ scale prior to receiving new pain information. Demographic and practice characteristics were also obtained. The study received Ethics Approval from the McMaster University Research Ethics Board. - + 9606 Species participants - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - + MESH:D010146 Disease @@ -7380,12 +7416,6 @@ title_2 9592 Structural validity - - MESH:D020914 - Disease - - Structural validity - METHODS @@ -7398,14 +7428,14 @@ paragraph 10699 All analyses except CFA were conducted by SAS (version 9.3, SAS Institute Inc, Cary, NC, USA). We used IBM SPSS v20 Amos statistical software for CFA. - + 6302 Gene 4359 SAS - + 6302 Gene 4359 @@ -7438,7 +7468,7 @@ table_caption 10980 Characteristics of 673 participants of EBP-KABQ study - + 9606 Species @@ -7452,13 +7482,7 @@ Tab1.xml 11036 Characteristics N (%) Age 20-35 178 (26.4) 36-45 158 (23.4) 46-55 221 (32.8) 56+ 116 (17.2) Clinical designation MD 131 (19.5) OT/PT 326 (48.4) RN 127 (18.8) RPsych or CPsych 52 (7.7) Others 37 (5.5) Highest education level Diploma/BA 234 (34.8) MA/MSC 222 (33.0) MD 122 (18.1) Ph.D. 95 (14.1) Received advanced clinical certifications 364 (54.1) Years of clinical training Less than 2 years 190 (28.2) 2-5 years 295 (43.8) Above 5 years 188 (27.9) Location of practice Urban 463 (68.8) Rural 101 (15.0) Both 109 (16.2) - - MESH:D009584 - Chemical - - N - - + Disease Diploma @@ -7469,13 +7493,13 @@ paragraph 11588 In total, 673 health professionals completed EBP-KABQ questionnaire. The description of demographic characteristics is presented in Table 1. Half of participants were age 45 or younger. Nearly half of clinicians were OTs or PTs, while 1/4 were nurses and 1/5 were physicians. One quarter of the sample had more than 5 years of clinical training; and they had a mean time in clinical practice of almost 18 years. Most participants practiced in an urban setting, while 15% were in a rural practice area. - + 9606 Species participants - + 9606 Species @@ -7539,7 +7563,7 @@ table 14777 Q 1: How often do you now look up evidence immediately before, or during patient treatment visit per week? - + 9606 Species @@ -7573,7 +7597,13 @@ Tab3.xml 15095 Theoretical constructs A priori hypotheses Results 1. EBP knowledge is more easily affected than other other aspects of EBP Mean item score in "knowledge" > other domains Knowledge: 5.91 Behaviour: 2.24 Outcome/Decision : 4.18 Attitude: 4.96 2. "Outcome/Decision" is correlated to other 3 domains Correlation coefficients between "outcome" and "knowledge"/"application"/"attitude" > other correlation coefficients. routcome-knowledge=0.54*, routcome-behaviour=0.40*, routcome-attitude=0.57*; rattitude-knowledge=0.41*, rknowledge-application=0.33*, rapplication-attitude=0.26*; 3. MEBP subscale scores are correlated with corresponding EBP activities Correlation coefficients between "application" and 3 external questions evaluating EBP application > other correlation coefficients. rapplication-Q1=0.32*, rknowledge-Q1=0.19*, routcome-Q1=0.28*; rattitude-Q1=0.19*; rapplication-Q2=0.41*, rknowledge-Q2=0.24*, routcome-Q2=0.30*; rattitude-Q2=0.19*; rapplication-Q3=0.35*, rknowledge-Q3=0.24*, routcome-Q3=0.26*; rattitude-Q3=0.16*; 4. Demographic variables would be associated with total MEBP scale score Age, highest education level, possession of advanced clinical training are significant factors are associated with in multivariate modeling Adjusted beta coefficients of following variable: Age: beta = -0.32 Higher education level (ref: diploma/BA): beta =4.63* Years of clinical training (ref: less than 2 years): beta =2.36* Advanced clinical training (ref: No): beta =4.37* Practice setting (ref: urban): beta =1.87* - + + - + Chemical + + routcome + + Disease diploma @@ -7600,7 +7630,7 @@ Tab4.xml 16822 Unadjusted Adjusted Characteristics B SD P value B SD P value Age (years) -0.32 0.59 0.60 -- -- -- Education (ref: diploma/BA) 5.08 0.73 <0.01* 4.63 0.73 <0.01* Clinical designation (ref: MD) 0.06 0.06 0.27 -- -- -- Years of clinical training (ref: less than 2 years) 3.57 0.73 <0.01* 2.36 0.81 <0.01* Advanced clinical training (ref: No) 5.68 1.23 <0.01* 4.37 1.21 <0.01* Practice setting (ref: urban) 1.24 0.82 0.13 1.87 0.79 0.02* Constant -- -- -- 106.2 2.43 <0.01* - + Disease diploma @@ -7611,21 +7641,21 @@ paragraph 17331 Details of the construct validity testing and a priori hypotheses were provided in Table 3. As we expected, mean item score in "knowledge" was 5.91, significantly higher than the rest of the domains (p < 0.05). Our constructed hypotheses were supported in that the correlation coefficients between "outcomes/decision" and "knowledge", "behaviour" and "attitude" were 0.54, 0.40 and 0.57 respectively, which were higher correlations than observed between other subscales. Construct validity was also supported in that there was a significant relationship between the frequency of searching reported by clinicians and the "behaviour" score, with correlation coefficient ranges from 0.32 to 0.41 (hypothesis 3). Regression analyses supported our a priori hypothesis that health professionals who had higher levels of education (beta = 4.63, P < 0.01), longer years in clinical training (beta = 2.36, P < 0.01) and possession of advanced clinical training (beta = 4.37, P < 0.01) were more likely to use EBP (Table 4). Although younger age was related to EBP practice in the direction anticipated, it did not reach statistical significance (beta = -0.32, P = 0.06). - + 10242 Gene 4257 beta = 2 - + 10381 Gene 68503 beta = 4 - + 10381 Gene 68503 @@ -7638,12 +7668,6 @@ title_2 18549 Structural validity - - MESH:D020914 - Disease - - Structural validity - FIG @@ -7652,24 +7676,36 @@ fig_caption 18569 Standardized parameter estimates for the refined EBP-KABQ factor structure model. Rectangles represent the scale items and ellipses represent the proposed factor constructs. Values on the single-headed arrows leading from the factors are standardized factor loadings. Values on the curved double-headed arrows between rectangles are correlations between error terms. Values on the curved double-headed arrows between ellipses are correlations between latent variables. - + 301928 Species ellipses - + 301928 Species ellipses + + MESH:D000069451 + Disease + + error terms + RESULTS paragraph 19039 The Initial second-order model demonstrated poor model fit (x2 = 1838.24, df = 269, P < 0.001, CFI = 0.73, TLI = 0.70, RMSEA = 0.093). Modification indices suggested overall model fit would be improved by adding the correlation of six pairs of error terms (item 4 & 5 within "knowledge", 12 & 13 in "application", 21 & 24, 23 & 31, 27 & 30, and 31 & 32 in "attitude"). After the modification was executed, statistical fit of the model was improved to as follows: x2 = 1205.20, df = 312, P < 0.001, CFI = 0.86, TLI = 0.84, RMSEA = 0.065. Although the overall fit improved, model fit indices especially CFI and TLI were still inadequate. We observed factor loading (beta = 0.05) of the item 3 ("Clinical trials and observational methods are equally valid in establishing treatment effectiveness") was significantly lower than the other five items on the dimension of knowledge. After removing this item from the scale, goodness-of-fit statistics improved to x2 = 1056.65, df = 287, P < 0.001, CFI = 0.89, TLI = 0.86, RMSEA = 0.06 (Figure 1) which was very close to our a priori threshold (CFI/TLI >= 0.90, RMSEA < 0.08). + + MESH:D000069451 + Disease + + error terms + DISCUSS @@ -7682,6 +7718,12 @@ paragraph 20271 This study provided support for the use of a modified EBQ-KABQ questionnaire to understand different aspects of EBP knowledge, attitudes, behavior and outcomes/decisions in a variety of healthcare professionals with respect to EBP. We confirmed that the 26 ordinal items in the modified EBP-KABQ exhibit a four-domain construct consistent with the proposed four aspects of EBP. Our scale was modified based on our need to change wording to make the scale more broadly applicable to different disciplines since the original version targeted medical students. We also made changes based our experiences in pilot testing the measure since an expert committee and pilot users found some items to be redundant or difficult to understand. Our work builds on that of the developers who targeted medical trainees by providing a more broadly applicable and validated version. The newly proposed subscale construct of "outcomes/decisions" contains the items previously termed "future use" in the original scale. Outcomes/decisions more accurately reflect the item content and the targeting of the EBP-KABQ. Whereas, as the original instrument was focused on trainees who might be responding about future use, experienced clinicians will be reporting how they use EBP in current clinical decision-making and whether they attribute better outcomes to their evidence-based decisions. This domain is considered an important aspect of self-reported EBP since its focuses on the impact on practice and outcomes. We found the "outcomes/decisions" domain was moderately correlated with the other three domains, suggesting it played a role in perception of EBP. The shorter measure has improved measurement characteristics, retains conceptual domains and may be save administration time. + + MESH:D008569 + Disease + + wording + DISCUSS @@ -7802,6 +7844,12 @@ paragraph 29346 CFA + + - + Chemical + + CFA + ABBR @@ -7892,6 +7940,12 @@ footnote 29643 JCM, ML and RBH conceived the study. QS and JCM designed the study. QS created the analytic model with contributions from JCM and BC. QS undertook the statistical analysis. QS contributed to the writing of the first draft of the manuscript. All of the authors contributed to and have approved the final manuscript. + + MESH:C537366 + Disease + + ML + REF @@ -7914,7 +7968,7 @@ 38 29969 Clinical expertise in the era of evidence-based medicine and patient choice - + 9606 Species @@ -7987,11 +8041,6 @@ surname:Landsverk;given-names:J 30315 Psychometric properties and US national norms of the evidence-based practice attitude scale (EBPAS) - - Chemical - - EBPAS - surname:Tin;given-names:KY @@ -8027,7 +8076,7 @@ ref 30569 "Push" versus "Pull" for mobilizing pain evidence into practice across different health professions: a protocol for a randomized trial - + MESH:D010146 Disease @@ -8289,7 +8338,7 @@ surname:Vinson;given-names:DC 31917 Obstacles to answering doctors' questions about patient care with evidence: qualitative study - + 9606 Species @@ -8326,11 +8375,6 @@ REF 32121 Critical appraisal skills training for health care professionals: a randomized controlled trial [ISRCTN46272378] - - Chemical - - ISRCTN46272378 - surname:Khan;given-names:K @@ -9198,7 +9242,19 @@ paragraph 10233 It is notable that ambient temperature cannot be used as a surrogate for MKT when assessing whether, or for how long, lorazepam can be stored in an EMS unit. MKT is a dynamic variable that accounts for the potential stress caused by changing temperatures, which is one reason that the pharmacology literature supports the use of MKT, rather than simple temperature means, for evaluating heat stability of drugs. These data support the notion that EMS agencies should take multiple variables into consideration, including temperature exposure and length of field deployment, when determining medication storage and restocking policies. - + + - + Chemical + + MKT + + + - + Chemical + + MKT + + MESH:D008140 Chemical @@ -9210,31 +9266,31 @@ paragraph 10868 Based on our findings here and previously, EMS systems choosing to deploy lorazepam should employ lorazepam storage methods to limit high heat exposures and maintain controlled room temperature (MKT < 25 C) environments. Otherwise, it may be prudent to limit field deployment time of lorazepam to 30 days to minimize degradation risks. In some systems, preferential use of midazolam or diazepam may be warranted. - + MESH:D008140 Chemical lorazepam - + MESH:D008874 Chemical midazolam - + MESH:D008140 Chemical lorazepam - + MESH:D003975 Chemical diazepam - + MESH:D008140 Chemical @@ -9252,13 +9308,13 @@ paragraph 11314 This study has limitations. First, baseline (day 0) samples were not obtained because our previous work demonstrated consistent baseline concentrations for midazolam and lorazepam. Furthermore, the differences in the pair of samples taken from each EMS unit for testing were insignificant. This is consistent with the quality controls and USP specifications expected in these commercial pharmaceuticals. - + MESH:D008874 Chemical midazolam - + MESH:D008140 Chemical @@ -9276,13 +9332,13 @@ paragraph 12028 Finally, this study was not designed to evaluate the impact of different vehicle deployment or medication storage practices on drug degradation. The impact of medication refrigerators and other techniques to control heat exposure and MKT may be more important for lorazepam than the other benzodiazepines and is a topic suitable for further study. - + MESH:D001569 Chemical benzodiazepines - + MESH:D008140 Chemical @@ -9306,19 +9362,19 @@ paragraph 12490 Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. In contrast, lorazepam degraded significantly over this time and appeared especially sensitive to higher mean kinetic temperatures. - + MESH:D008140 Chemical lorazepam - + MESH:D008874 Chemical Midazolam - + MESH:D003975 Chemical @@ -9384,42 +9440,18 @@ paragraph 13594 Harney, MPH, MSW, Donna Harsh, MS, Joy Pinkerton, BSN, RN, MS, Allison Kade, BA, Nicholas Siewert, BA, Ashley Pinawin, BS, Catherin Ring, Phebe Brenne - - MESH:C480682 - Chemical - - BSN - - - MESH:C041626 - Chemical - - MPH - METHODS paragraph 13745 National EMS Coordinator - - MESH:D001259 - Disease - - EMS Coordinator - METHODS paragraph 13770 Kay Vonderschmidt, MPA, MS-EM, NREMT-P - - MESH:C051945 - Chemical - - MPA - METHODS @@ -9432,24 +9464,6 @@ paragraph 13844 Valerie Durkalski, PhD, Yuko Palesch, PhD, Catherine Dillon, Keith Pauls, Qi Wu, Wenle Zhao, PhD - - MESH:C411141 - Chemical - - PhD - - - MESH:C411141 - Chemical - - PhD - - - MESH:C411141 - Chemical - - PhD - METHODS @@ -9462,30 +9476,12 @@ paragraph 13971 Robin Conwit, MD, Scott Janis, PhD, David Jett, PhD, Brandy Fureman, PhD - + 9188 Species Robin - - MESH:C411141 - Chemical - - PhD - - - MESH:C411141 - Chemical - - PhD - - - MESH:C411141 - Chemical - - PhD - METHODS @@ -9504,46 +9500,23 @@ paragraph 14119 Hub Principal Investigator: Robert D. Welch, MD, MS Primary Study Coordinators: Lynnmarie Mango, MPH, Valerie H. Mika, MS EMS Director(s)/Coordinator: Jenny Atas, MD Other Site Investigators: Robert Dunne, MD, Douglas Wheaton, MD, Phillip Levy, MD, MPH, Marc-Anthony Velilla, MD, Robert Sherwin, MD, Brian O'Neil, MD, Angela Groves, MD, Marc Rosenthal, DO, PhD Participating EMS Service: Detroit EMS - + 29780 Species Mango - + 301693 Species Atas - + + MESH:D004881 Disease - - Detroit EMS - - - MESH:C041626 - Chemical - - MPH - - - MESH:C041626 - Chemical - - MPH - - - MESH:C411141 - Chemical - - PhD - - - MESH:D008455 - Chemical - - O'Neil + + Marc-Anthony Velilla @@ -9557,29 +9530,11 @@ paragraph 14550 Hub Principal Investigator: Arthur Pancioli, MD Primary Study Coordinators: Irene Ewing, RN, BSN, Peggy Waymeyer, RN EMS Director(s)/Coordinator: M. Kay Vonderschmidt, MPA, MS-EM, NREMT-P, Jason McMullan, MD Other Site Investigators: Hamilton Schwartz, MD, Brian Stettler, MD, William Knight, MD, Opeolu Adeoye, MD, Rhonda Cadena, MD, Jordan Bonomo, MD, Erin Grise, MD, Laura Heitsch, MD, George Shaw, MD, Nick Gagai, CCRP, Pamela Schmit, RN BSN, Sara Stark, Med, Traci Doellman, RN Participating EMS Services: Cincinnati Fire Department, BlueAsh Fire Department, Forest Park Fire Department, Green Township Fire Department, Florence Fire Department, Independence Fire Department - + CellLine Laura Heitsch - - MESH:C480682 - Chemical - - BSN - - - MESH:C480682 - Chemical - - BSN - - - MESH:C051945 - Chemical - - MPA - METHODS @@ -9592,34 +9547,11 @@ paragraph 15275 Hub Principal Investigator: J. Claude Hemphill, III, MD, MAS Primary Study Coordinators: Michele Meeker, RN, BSN, Kelley Rosborough, BA EMS Director(s)/Coordinator: Jeany Duncan EMT-P Other Site Investigators: Karl Sporer, MD, FACEP, FACP, Alan Gelb, MD; Wade Smith, MD, PhD, Prasanthi Ramanujam, MD, Kazuma Nakagawa, MD, Asma Moheet, MD, Hooman Kamel, MD, Bharath Naravetla, MD, Mary Mercer, MD, Christine Wong, MD Participating EMS Services: San Francisco Fire Department, EMS Division - - MESH:D005359 - Disease - - MAS - - - MESH:C411141 - Chemical - - PhD - - - MESH:D009436 - Disease - - MD Participating EMS - - + + MESH:D020526 Disease - - EMS Division - - - MESH:C480682 - Chemical - - BSN + + Claude Hemphill @@ -9633,27 +9565,11 @@ paragraph 15800 Hub Principal Investigator: Elizabeth Jones, MD Trial Principal Investigator: Truman J. Milling, MD Primary Study Coordinators: Misty Ottman, RN, BSN, Ben King, Laura LaChance EMS Directors/Coordinators: Jeff Brockman, RN, Pete Didonato, EMT-P Other Site Investigator: Paul Hinchey, MD Participating EMS Service: Austin-Travis County EMS - - Chemical - - Didonato - - - MESH:C480682 - Chemical - - BSN - - - MESH:D009436 - Disease - - MD Participating EMS - - - Disease - - Laura LaChance EMS Directors + + CVCL:0030 + CellLine + + Laura La @@ -9667,24 +9583,6 @@ paragraph 16160 Hub Principal Investigator: David W. Wright, MD Trial Principal Investigators: Matthew D. Bitner, MD, Gerald W. Beltran, DO Primary Study Coordinator: Harriet Nevarez, RN, CCRC EMS Director/Coordinator: Rachel Barnhard, Andrea G. McDougal Other Site Investigators: Jeffrey F. Linzer Sr, MD, Lisa H. Merck, MD MPH, Tamara Espinoza, MD Participating EMS Service: Grady EMS - - MESH:D009436 - Disease - - MD Participating EMS - - - MESH:D013324 - Chemical - - Sr - - - MESH:C041626 - Chemical - - MPH - METHODS @@ -9697,29 +9595,6 @@ paragraph 16561 Hub Principal Investigator: Christopher A. Lewandowski, MD Trial Principal Investigator: Taher T. Vohra, MD Primary Study Coordinators: Paula L. Crouse, RN, BSN., MA., Anna E. Baker, RN, BSN EMS Director/Coordinator: Dean R. Creech EMT-P, I/C Other Site Investigator: Andrew N. Russman, DO, Joseph B. Miller, MD, Jumana Nagarwala, MD, Daniel J. Miller, MD, Raymond Fowkes, MD, Anne Marie Lundell, RN, BSN Participating EMS Services: Detroit EMS, West Bloom-field Fire and EMS Services - - MESH:C480682 - Chemical - - BSN - - - MESH:C436350 - Chemical - - BSN EMS - - - MESH:C480682 - Chemical - - BSN - - - Disease - - Detroit EMS - METHODS @@ -9732,18 +9607,12 @@ paragraph 17071 Hub Principal Investigator: James V. Quinn, MD, MS Primary Study Coordinators: Stephanie Casal. RN, CNS, Anke Hebig, Mark Liao EMS Director/Coordinator: Peter D'souza, MD Participating EMS Services: Palo Alto Fire Department, San Jose Fire Department, Redwood City Fire Department, San Mateo Fire Department - + 28980 Species Redwood - - MESH:D009436 - Disease - - MD Participating EMS - METHODS @@ -9756,18 +9625,6 @@ paragraph 17406 Hub Principal Investigator: Kurt R. Denninghoff, MD Trial Principal Investigator: Daniel W. Spaite, MD Primary Study Coordinator: Bruce Barnhart, RN, CEP EMS Director(s)/Coordinator: Willie Haro, CEP Other Site Investigator: Bentley J. Bobrow, MD Participating EMS Service: Glendale Fire Department - - MESH:D009436 - Disease - - MD Participating EMS - - - MESH:C436350 - Chemical - - CEP EMS - METHODS @@ -9786,12 +9643,6 @@ paragraph 17837 EMS Director/Coordinator: Alan D. Payne, CCEMTP Other Site Investigators: Alan R. Towne, MD, Michael C. Kurz, MD, John T. Carmack, MD Participating EMS Service: Richmond Ambulance Authority - - MESH:D009436 - Disease - - MD Participating EMS - METHODS @@ -9804,12 +9655,6 @@ paragraph 18056 Hub Principal Investigator: Michelle Biros, MD Trial Principal Investigator: Brian Mahoney, MD Primary Study Coordinators: Corey Sargent, Kathleen Miller, BSN, CCRC Other Site Investigators: David Hildebrandt, Chris Kummer, Doug Gesme Participating EMS Services: Hennepin County EMS - - MESH:C480682 - Chemical - - BSN - METHODS @@ -9822,23 +9667,10 @@ paragraph 18373 Hub Principal Investigator: Tom P. Aufderheide, MD Primary Study Coordinator: Joseph T. Brandt Jr., BS, EMT-P EMS Director/Coordinator: M. Riccardo Colella, DO Other Site Investigators: Ron Pirrallo, MD, MHSA, Walter Bialkowski, MS, Benjamin Hermanson, BS, Christopher Sandoval, BS, EMT-P, Kevin Morrow, MFA, Kelly McCormick, BS, MBA, Katherine Burpee, BA, Geri Price, BS, Dawn Kawa, BA Participating EMS Services: Milwaukee County EMS, Milwaukee Fire Department, Franklin Fire Department, Greenfield Fire Department, North Shore Fire Department, Oak Creek Fire Department, South Milwaukee Fire Department, Wauwatosa Fire Department, West Allis Fire Department - - MESH:D017827 + Disease - - Joseph T - - - MESH:C111608 - Chemical - - MBA - - - MESH:C032354 - Chemical - - MFA + + Riccardo Colella @@ -9852,18 +9684,6 @@ paragraph 19062 Hub Principal Investigator: Roger L. Humphries, MD Primary Study Coordinator: Linda Dechtenberg, RN, BSN, CCRC EMS Director/Coordinator: Christofer Sweat Other Site Investigator: L. Creed Pettigrew, MD,MPH Participating EMS Service: Lexington-Fayette Urban County Government Division of Fire & Emergency Services - - MESH:C480682 - Chemical - - BSN - - - MESH:C041626 - Chemical - - MPH - METHODS @@ -9876,18 +9696,12 @@ paragraph 19407 Hub Principal Investigator: Jill M. Baren, MD, MBE Trial Principal Investigator: R. Daniel Bledsoe, MD Primary Study Coordinator: Barbie Stahlman, MS, Katherine Lamond, BA, Pamela G. Nathanson, MBE Other Site Investigator: Scott E. Kasner, MD, MSCE, Peter D. Le Roux, MD Participating EMS Services: York Hospital Medic 97, White Rose Ambulance, Grantley Fire Company, Jacobus Lions Ambulance Club, West York Ambulance - + 9689 Species Lions - - MESH:D009436 - Disease - - MD Participating EMS - METHODS @@ -9900,18 +9714,6 @@ paragraph 19865 Hub Principal Investigators: Craig R. Warden, MD, MPH, Robert A. Lowe, MD, MPH Primary Study Coordinator: Rachel N. Stone, CCRP Participating EMS Service: Clackamas Fire District #1 - - MESH:C041626 - Chemical - - MPH - - - MESH:C041626 - Chemical - - MPH - METHODS @@ -9924,18 +9726,6 @@ paragraph 20083 Hub Principal Investigator: Stephan Mayer, MD, FCCM Trial Principal Investigator: Neal Flomenbaum, MD Primary Study Coordinators: M. Cristina Falo, PhD, Lisa-Vanessa Magitbay, RN, Chirag Surti EMS Directors/Coordinators: Heidi Cordi, MD, Daniel Ribaudo Other Site Investigators: Axel Rosengart, MD, PhD, Matthew Vibbert, MD, Santiago Ortega-Gutierrez, MD, H. Alex Choi, MD, Emily Gilmore, MD, Rishi Malhotra, MD, Lawrence Berger Participating EMS Services: New York Presbyterian - - MESH:C411141 - Chemical - - PhD - - - MESH:C411141 - Chemical - - PhD - METHODS @@ -9948,11 +9738,11 @@ paragraph 20583 Hub Principal Investigator: Nina T. Gentile, MD Trial Principal Investigators: Alvin Wang, DO, Christopher Vates, MD, Ben Usatch, MD Primary Study Coordinators: Brent B. Freeman, Stacey L. Cleary Participating EMS Services: Volunteer Medical Services Corps of Lower Merion and Narberth (Narberth Ambulance), Life Lion EMS - - MESH:D057768 - Disease - - Life Lion EMS + + 9689 + Species + + Lion @@ -9966,24 +9756,12 @@ paragraph 20932 Hub Principal Investigator: Barney Stern, MD Trial Principal Investigators: Tricia Ting, MD, Gregory Krauss, MD Primary Study Coordinators: Virginia Ganley, RN, Susan Rice, RN, Jennifer Ronald EMS Director/Coordinator: Michelle Stevens, RN Other Site Investigators: Brian Browne, MD, Robert Rosenthal, MD, Peter Hill, MD Participating EMS Services: Maryland Institute for Emergency Medical Services Systems (MIEMSS), Baltimore City EMS - + 4530 Species Rice - - MESH:D009436 - Disease - - MD Participating EMS - - - MESH:C015161 - Chemical - - Robert Rosenthal - REF @@ -10013,19 +9791,19 @@ surname:Allen;given-names:F 21379 A comparison of lorazepam, diazepam, and placebo for the treatment of out-of-hospital status epilepticus. - + MESH:D013226 Disease status epilepticus - + MESH:D003975 Chemical diazepam - + MESH:D008140 Chemical @@ -10050,7 +9828,7 @@ surname:Pancioli;given-names:A 21485 Intramuscular versus intravenous therapy for prehospital status epilepticus. - + MESH:D013226 Disease @@ -10072,25 +9850,25 @@ 82 21562 Midazolam versus diazepam for the treatment of status epilepticus in children and young adults: a meta-analysis. - + 9606 Species children - + MESH:D013226 Disease status epilepticus - + MESH:D003975 Chemical diazepam - + MESH:D008874 Chemical @@ -10143,7 +9921,7 @@ surname:Cap;given-names:AP 21878 Stability of tranexamic acid after 12-week storage at temperatures from -20 degrees C to 50 degrees C. - + MESH:D014148 Chemical @@ -10186,13 +9964,13 @@ surname:Corry;given-names:MD 22135 Prehospi tal stability of diazepam and lorazepam. - + MESH:D003975 Chemical diazepam - + MESH:D008140 Chemical @@ -10218,13 +9996,13 @@ surname:Siewart;given-names:N 22185 The 60-day temperature-dependent degradation of midazolam and lorazepam in the prehospital environment. - + MESH:D008140 Chemical lorazepam - + MESH:D008874 Chemical @@ -10254,7 +10032,7 @@ fig_caption 22370 Relative concentrations of benzodiazepines at 120 days as a function of cumulative mean kinetic temperatures (MKT). - + MESH:D001569 Chemical @@ -10268,7 +10046,7 @@ fig_caption 22486 Comparison by site of relative concentration of lorazepam and cumulative mean kinetic temperature (MKT) at 30, 60, 90, and 120 days. - + MESH:D008140 Chemical @@ -10312,7 +10090,7 @@ table_caption 22768 Average mean kinetic temperature (MKT) and relative concentration of benzodiazepines compared to label at each measured timepoint. - + MESH:D001569 Chemical @@ -10326,19 +10104,19 @@ T1.xml 22899 Concentration, mean % (95% CI) 30-day 60-day 90-day 120-day Diazepam 97.0 (96.3-97.6) 97.1 (96.6-97.6) 97.4 (96.6-98.3) 97.0 (95.7-98.2) Lorazepam 101.0 (99.0-102.9) 95.6* (91.6-99.5) 90.3** (85.2-95.4) 86.5** (80.7-92.3) Midazolam 101.0 (99.8-101.4) 100.6 (99.8-101.4) 99.0 (98.0-99.9) 99.0 (98.1-100.2) MKT 28.1 (24.9-31.2) 30.4 (26.0-34.7) 31.0 (26.8-35.1) 31.6 (27.1-36.1) - + MESH:D008874 Chemical Midazolam - + MESH:D008140 Chemical Lorazepam - + MESH:D003975 Chemical @@ -10746,257 +10524,251 @@ paragraph 3944 The miR-206/133b cluster is not expressed in the heart, but its expression is confined to developing skeletal muscle. In adult muscles miR-206/133b are preferentially found in slow myofibers. Expression of miR-206/133b is controlled by a network of myogenic regulatory genes, including MyoD, which binds to the miR-206/133b locus in C2C12 cells. The miR-206/133b locus not only encodes the miRNAs miR-206 and miR-133b, but also the long non-coding RNA linc-MD1, which is expressed during muscle development similar to miR-206 and miR-133b and assumed to act as a competing endogenous RNA or miRNA decoy. Genomic deletion of miR-206 did not cause an obvious clinical phenotype in mice, although it was determined that miR-206 is required for re-innervation of muscle tissue in mice with mutant Sod1. It was proposed that miR-1/206 repress Hdac4, which in turn might regulate genes involved in controlling muscle-derived signals that enhance synapse formation under pathological conditions. More recently, miR-206 was claimed to act as a modifier of muscular dystrophy. Deletion of miR-206 in the mdx mouse model of Duchenne muscular dystrophy led to increased lethality in compound mutants and accumulation of degenerated muscle fibers. Surprisingly, no defects in muscle innervation were observed after the loss of miR-206, although muscle fibers are continuously lost and replaced in mdx mice. The dramatic enhancement of muscle dystrophy in miR-206/mdx compound mutants was attributed to impaired differentiation of myofibers due to the up-regulation of Pax7, which contains identical binding sites for miR-206 and miR-1 in its 3' untranslated region (UTR). The presence of miR-206 target sites in Pax7 did also motivate other researchers to investigate a regulation of Pax7 by miR-206, revealing that overexpression or knock-down of miR-206 can affect Pax7 levels and thereby muscle differentiation in vitro. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 17927 Gene 7857 MyoD - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 105661730 Gene linc-MD1 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 20655 Gene 392 Sod1 - + 208727 Gene 55946 Hdac4 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 10090 Species mice - + 10090 Species mice - + 10090 Species mouse - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - + CVCL:0188 CellLine C2C12 - + MESH:D009136 Disease muscle dystrophy - - MESH:D006732 - Disease - - muscle innervation - - + MESH:D009136 Disease muscular dystrophy - + MESH:D020388 Disease @@ -11008,92 +10780,92 @@ paragraph 5858 Here, we analyzed mice with a targeted deletion of the miR-206/133b cluster. miR-206/133b are processed from a common precursor and thus might be regarded as a functional unit similar to the miR-1/133a clusters. Concomitant loss of miR-206 and miR-133b neither leads to an obvious clinical phenotype or causes detectable molecular changes in skeletal muscles nor impairs muscle regeneration in the MDX mouse model of muscular dystrophy. Surprisingly, lack of miR-206/133b and the miR-133 decoy, contained in the third exon of linc-MD1, did not have obvious effects on satellite cell proliferation and differentiation. Our findings differ from a previous analysis of miR-206 mutants, which might suggest that other products processed from the primary miR-206/133b transcript balance effects of miR-206 in vivo. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 105661730 Gene linc-MD1 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 MDX - + 10090 Species mice - + 10090 Species mouse - + 735281 Gene miR - + 735281 Gene miR - + MESH:D009136 Disease @@ -11123,7 +10895,7 @@ title_2 6920 Deletion of miR-206/133b - + 387202 Gene @@ -11135,81 +10907,80 @@ paragraph 6945 The miR-206/133b coding region was deleted from the mouse genome using a vector containing genomic sequences flanking the miRNA miR-206/133b gene and a PGK-neomycin selection cassette. The short arm (2.3 kb) used for recombination encompassed the miR-206 5' flanking region from TTAGGCATATAAAGTTTGCACGACC to GATATAAAGAAGCATGTGGCCTGGG). The long arm of the targeting vector consisted of a 5.9 kb NcoI fragment directly downstream of miR-133b. A DTA selection cassette was used for negative selection against nonhomologous insertion of the vector into the genome. Homologous recombination in mouse embryonic stem (ES) cells was ascertained by Southern blot analysis using a SphI digest and a 453 bp 5' probe amplified by the oligonucleotides (TAAGTCCTGATGCTTCTCAATACCC; GTTGATAAAGAAACTGTGTGTTACG), resulting in an 8.4-kb WT and an 11.8-kb knock-out signal. Although the inserted beta-galactosidase coding (lacZ) cassette contained an IRES sequence, no beta-galactosidase was expressed from the engineered locus. Genotyping of mdx mice followed the protocol described by Shin et al.. Primary sequences of mature miRNAs were derived from mirBase20 (MIMAT0000239, MIMAT0000123, MIMAT0000769 and MIMAT0000145). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 12091 Gene 47922 beta-galactosidase - + 12091 Gene 47922 beta-galactosidase - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mice - + 13405 Gene 20856 mdx - + 387202 Gene miR-206 - + + Disease + + MIMAT0000239, MIMAT0000123, MIMAT0000769 + + MESH:D009355 Chemical neomycin - - MESH:C522116 - Chemical - - DTA - METHODS @@ -11222,72 +10993,90 @@ paragraph 8194 Total RNA was isolated from M. soleus of PBS perfused animals using the TRIZOL method (Invitrogen, Thermo Fisher Scientific, Darmstadt, Germany). The RNase protection assay was performed using the mirVana Probe construction Kit (Ambion, Thermo Fisher Scientific, Darmstadt, Germany; Cat#1550) using oligos (miR-206: TGGAATGTAAGGAAGTGTGTGG, miR-145: GTCCAGTTTTCCCAGGAATCCCTTTTTTTT) together with the mirVana miRNA Detection Kit (Ambion#1552) following the manufacturer's instructions. A polyT at the control oligo was added to allow size resolution of the control oligo. The sequence CCTGTCTC was added to the 3' end of the oligos to allow binding of the Ambion T7-Promoter Primer. Amersham alpha-32P UTP (GE Healthcare, Freiburg, Germany; PB20383, 800 Ci/mmol) was used for labeling. Probes were purified using a 15% Invitrogen TBE-Urea Gel (EC68855BOX), the gel was exposed to a Kodak X-ray film, labeled probes were excised und eluted 30 min in elution buffer. 5 mug of total RNA isolated from WT, heterozygous or KO-muscle or yeast-RNA (Roche, Mannheim, Germany), respectively was hybridized at 42 C overnight with approximately 104 cpm of probe in hybridization buffer in a total volume of 20 mul. Samples were analyzed using 15% TBE-Urea gels after RNase A/T1 digestion. Signals were detected using Kodak X-Ray film. Northern blot analysis was performed as previously described. Antisense oligonucleotides specific for miR-206 (CCACACACTTCCTTACATTCCA), miR-1 (ATACATACTTCTTTACATTCCA) and U6 (ATATGGAACGCTTCACGAATT) were used. - + 387202 Gene miR-206 - + 387163 Gene miR-145 - + 387202 Gene miR-206 - + 4932 Species yeast - + 735281 Gene miR - - MESH:C531972 - Chemical - - TBE-Urea Gel - - c.1A>T - DNAMutation - - A/T1 + MESH:D009841 + Chemical + + oligonucleotides - MESH:C531972 + - Chemical - - TBE-Urea + + TBE - MESH:D011071 + MESH:D014508 Chemical - - polyT + + Urea - MESH:D010634 + MESH:D014508 Chemical - - PB20383 + + Urea - MESH:D014544 + - Chemical alpha-32P UTP + + - + Chemical + + TBE + + + MESH:D007854 + Chemical + + PBS + + + c.1A>T + DNAMutation + + A/T1 + + + MESH:C411644 + Chemical + + TRIZOL + METHODS @@ -11300,61 +11089,49 @@ paragraph 9817 RNA quality was analyzed with the Agilent Bioanalyzer and the RNA 6000 Nano Kit (Agilent Technologies, Santa Clara, CA). For RNA expression analysis, the Affymetrix GeneChip MouseGenome 430 2.0 Array was used, employing the one-cycle target labeling protocol. Data were analyzed using the Affymetrix expression console and the RMA algorithm. Microarray data have been submitted to Arrayexpress (Acc#E-MTAB-2439). TaqMan MicroRNA Expression Assays and the Applied Biosystems StepOnePlus system were used to quantify miR-206 and miR-1 expression. The RT reaction was done using the miR-206 and the U6 specific primer with the Taqman MicroRNA Reverse Transcription Kit (#4366596). qPCR assays were performed with three independent samples each with Taqman Universal PCR Master Mix (#4324018), FAM labeled miR-206 or miR-1 assay (#000510; #002222) and VIC labeled U6 assay (#001973) for normalization. Relative expression was calculated using the Ct method. RNA isolated from a pool of M. soleus of five male C57Bl6 mice was sequenced using IonTorrent sequencing and analyzed as described. Data were submitted to the NCBI's Gene Expression Omnibus (Acc#GSE63342). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13615 Gene 1477 VIC - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - - MESH:D019578 - Disease - - Biosystems StepOnePlus system - - - MESH:C073349 - Chemical - - MTAB-2439 - METHODS @@ -11367,13 +11144,19 @@ paragraph 11019 For whole mount in-situ hybridization, a myogenin cDNA corresponding to 307 to 1393 bp of ENSMUST00000027730 was used to synthesize a DIG-labeled antisense probe by T7-RNA-polymerase. E10.5 embryos were isolated and genotyped after mating of heterozygous parents. - + 17928 Gene 1854 myogenin + + - + Chemical + + DIG + METHODS @@ -11386,98 +11169,128 @@ paragraph 11320 For histological analysis, muscle tissue was perfused in situ with 4% PFA, dissected, washed in PBS, dehydrated and embedded in paraffin, followed by preparation of 10 mum sections, deparaffinization and hydration in distilled water. Von Kossa staining was performed by incubating slides with 1% silver nitrate solution and exposed to light for 30 min. Slides were washed two times for 3 min in H2O and incubated in 5% sodium thiosulfate solution for 5 min. Thereafter, slides were washed two times in H2O, followed by counterstaining for 7 min in 0.1% EosinG (Merck, Darmstadt, Germany) with 0.05% acetic acid. Slides were dehydrated and mounted using Entellan. For Sirius red staining, slides were stained in Weigerts Iron-Hematoxylin solution (Sigma, Munich, Germany; Cat#HT1079) for 8 min, dipped twice in distilled water and stained one hour in 0.1% Direct Red 80 (Sigma#365548)/saturated picric acid. Subsequently, slides were washed using 0.5% acetic acid, dipped in distilled water, dehydrated and mounted using Entellan. - + MESH:D014867 Chemical H2O - - MESH:C005858 + + MESH:D014867 Chemical - - picric acid + + water - - MESH:C101815 + + MESH:C009798 Chemical - - PFA + + Direct Red 80 - - MESH:D004801 + + MESH:D007854 Chemical - - EosinG + + PBS - - MESH:D014867 + + MESH:D017245 Chemical - - H2O + + PFA - + MESH:D012835 Chemical silver nitrate - - MESH:D006416 - Chemical - - Iron-Hematoxylin - - + CVCL:5008 CellLine HT1079 - + + MESH:D014867 + Chemical + + water + + MESH:C017717 Chemical sodium thiosulfate - + MESH:D019342 Chemical - + acetic acid - + + MESH:C005858 + Chemical + + picric acid + + + MESH:D014867 + Chemical + + water + + + - + Chemical + + Sirius red + + + MESH:D010232 + Chemical + + paraffin + + + MESH:D014867 + Chemical + + H2O + + MESH:D019342 Chemical - + acetic acid + + - + Chemical + + Iron-Hematoxylin + METHODS paragraph 12356 For determination and quantification of fiber types in the muscle tissue, tissue was perfused in situ with 4% PFA, isolated and incubated in 15% and 30% sucrose/PBS for 2 hours and overnight, respectively. Tissue was frozen on dry ice and cryotome-sectioned. Sections were mounted on Superfrost slides. Tissue was dried and subsequently treated with 4% PFA/0.1% sodium desoxycholate/0.02% NP-40 for 5 min, washed 3 times with PBS for 5 min, blocked in 2% FCS, 0.5% NP-40/PBS for 1 h and then incubated with monoclonal anti-myosin (skeletal, slow; Sigma, M8421) in 2% FCS/0.5% NP-40/PBS overnight at 4 C. Slides were washed three times with PBS for 5 min and then incubated with biotinylated secondary antibody (Vector Labs, Burlingame, CA; BA-1400) for 2 h at RT and further processed according to the Vectastain Elite ABC Kit. - - MESH:C101815 + + MESH:D007854 Chemical - - PFA + + PBS - + MESH:D013395 Chemical sucrose - - MESH:D012964 - Chemical - - sodium desoxycholate - - - MESH:C101815 + + MESH:D017245 Chemical PFA @@ -11494,81 +11307,117 @@ paragraph 13228 Satellite cells were isolated from the hind leg muscles of approximately 20 to 25-week-old wildtype and miR-206/133b knock-out animals by using the skeletal muscle dissociation kit (Miltenyi Biotech, Bergisch Gladbach, Germany, 130-098-305) and enriched by the satellite cell isolation Kit, mouse (Miltenyi Biotech, 130-104-268) according to the manufacturer's instructions. Isolated cells were plated on gelatin-coated muclear 96-well plates (Sigma#M0562). Satellite cells were grown in proliferation medium (40% DMEM, 40% Ham F-10; 20% FCS; Pen/Strep; 2.5 ng/ml human FGF-2, Miltenyi Biotech#130-093-840) for 3 days, followed by switch to differentiation medium (DMEM, 2% horse serum, and Pen/Strep). Cells were incubated for 5 min in fixative (4% PFA/PBS, 0.1% sodium desoxycholate, 0.2% NP-40), washed 3 times in PBS, blocked in carrier (PBS, 5% BSA, 0.5% NP-40) for 1 h and then incubated with MF 20 supernatant in carrier (1:100, DSHB, Iowa City, Iowa). Secondary antibody was goat anti-mouse IgG1 Alexa 488 (1:2000, Life technologies). - + 27344 Gene 8315 Pen - + 2247 Gene 1521 FGF-2 - + 27344 Gene 8315 Pen - + 16017 Gene IgG1 - + 387202 Gene miR-206 - + 10090 Species mouse - + 9606 Species human - + 9796 Species horse - + 9925 Species goat - + 10090 Species mouse - - MESH:D012964 + + MESH:D007854 + Chemical + + PBS + + + - + Chemical + + DMEM + + + MESH:D003840 Chemical sodium desoxycholate - - MESH:C101815 + + MESH:D007854 + Chemical + + PBS + + + MESH:D007854 + Chemical + + PBS + + + - + Chemical + + Alexa 488 + + + MESH:D017245 Chemical PFA + + - + Chemical + + DMEM + METHODS @@ -11581,100 +11430,94 @@ paragraph 14301 Soleus muscle was dissected from adult 13C6Lys-labeled (SILAC) WT and from nonlabeled WT and miR-206/133b mutant littermates. Protein extracts were processed and mass spectrometry was performed as described previously. Briefly, 30 mug protein extracts of soleus muscle of labeled and the respective unlabeled muscle were combined, size-fractionated into 10 fractions using SDS-PAGE, and the proteins were digested in-gel using the endoproteinase LysC. After digestion peptides were eluted from gel-fractions using an acetonitrile gradient. Peptides were desalted prior to LC-MS/MS analysis and loaded onto a 15 cm in-house packed column (75 mum ID, C18 Beads 3 mum diameter, Dr. Maisch, Germany) and eluted by an linear increase of buffer B within a binary buffer system: A) 0.5% acetic acid and B) 80% ACN, 0.5% acetic acid. The flow-rate was set to 200 nL/min and gradient time to 150 min. The Agilent 1200 nanoflow HPLC system was coupled to an LQT-Orbitrap XL (Thermo Scientific) via an electrospray ionization source (Thermo Fisher Scientific). The mass spectrometer was operated in the data-dependent mode to automatically measure full MS scans and MS/MS spectra. In detail, survey scans were measured after accumulation of 1E6 ions at a resolution of 60,000 and subsequently isolation and fragmentation (CID, 35% norm. collision energy, target value: 5,000) of the top five intense peaks was performed in the linear ion trap. Peptides were identified by correlation of acquired MS/MS spectra against the mouse IPI (ftp://ftp.ebi.ac.uk/pub/databases/IPI) using MaxQuant software. A FDR cutoff of 1% was set on the protein and peptide level. Two mis-cleavages were allowed. Amidomethylation at cysteine residues was set as fixed modification, while oxidation on methione residues and acetylation at the N-term of proteins were defined as variable modifications. Quantification of proteins was performed by MaxQuant software. Only peptides with a minimal length of six amino acids and at least two ratio counts for SILAC pairs were used for quantification. Using SILAC as an internal standard, ratios miR-206/133b KO/13C6Lys-WT and 13C6Lys-WT/WT were obtained and were used for calculation of the direct KO/WT ratios for relative protein abundance. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mouse - + + - + Chemical + + 13C6Lys + + Disease CID - + MESH:D019342 Chemical acetic acid - - MESH:C026591 - Chemical - - 13C6Lys - - - MESH:C026591 - Chemical - - LysC - - - MESH:C026591 - Chemical - - 13C6Lys - - + MESH:D003545 Chemical cysteine - + MESH:D019342 Chemical acetic acid - - MESH:C026591 + + MESH:C084683 Chemical - - 13C6Lys + + ACN - - MESH:C032159 + + - Chemical - - acetonitrile + + 13C6Lys - - MESH:C032259 + + MESH:D012967 Chemical SDS - - MESH:C078466 + + MESH:C032159 Chemical - - IPI + + acetonitrile - - MESH:C529911 + + - Chemical methione - - MESH:C078466 + + - Chemical - - IPI + + 13C6Lys + + + MESH:D004194 + Disease + + ionization @@ -11688,52 +11531,41 @@ paragraph 16573 For western-blot analysis of embryonic tissue, the trunk of E10.5 embryos posterior of the otic placode was used. A total of 10 mug of protein extract were loaded onto a 9% SDS-PAGE and blotted onto nitrocellulose membranes. The following antibodies were used: anti-Pax7 (supernatant of DSHB#P3U1), anti-pan-actin (1:1000, CST, Frankfurt, Germany; Cat#4968), anti-GAPDH (1:1000, CST; Cat#2118), secondary antibodies were anti-mouse-HRP (1:1000, eBioscience, Affymetrix, Frankfurt Germany, Cat#18-8816-31) and anti-rabbit-HRP (1:5000, Pierce, Thermo Fisher Scientific, Darmstadt, Germany, Cat#1858415). - + 18509 Gene 55665 Pax7 - + 12854 Gene 997 CST - + 12854 Gene 997 CST - + 10090 Species mouse - + 9986 Species rabbit - - MESH:C465308 - Chemical - - Cat#18-8816-31 - - - Disease - - eBioscience - - - MESH:C032259 + + MESH:D012967 Chemical SDS @@ -11762,7 +11594,7 @@ title_2 17638 Deletion of the miR-206/133b cluster does not alter skeletal muscle morphology - + 387202 Gene @@ -11776,103 +11608,103 @@ fig_caption 17717 Generation of miR-206/133b knock-out mice. The miRNAs miR-206, miR-1 and 133a/b are encoded in three clusters on mouse chromosome 1, 2 and 18. The primary sequence (A) of mature miR-1/133a encoded on chromosome 2 and 18 is identical, miR-206 differs in four bases from miR-1, and miR-133b differs in one base from miR-133a (red). The seed sequence, which is instrumental for target binding of miRNAs, is identical between the corresponding miRNAs (blue). Schematic representation of the miR-206/133b genomic region (B) with deleted parts indicated (red). Southern blot analysis (C) of genomic DNA isolated from wildtype (WT), heterozygous, and homozygous mutant animals proves deletion of the miR-206/133b locus. An SphI digest of genomic DNA and a probe hybridizing 5' prime of the targeting vector were used to monitor targeting of the miR-206/133b locus by homologous recombination. RNase protection assay (D) and northern blot (E) analysis demonstrates the loss of miR-206 in the soleus muscle of knock-out animals. Expression of miR-1 is not changed in soleus muscle of miR-206/133b knock-out mice. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mice - + 10090 Species mouse - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene @@ -11884,52 +11716,52 @@ table 18824 Deletion of the miR-206/133b cluster does not lead to abnormalities in the Mendelian distribution of mutant animals (a) at the time of weaning. Additional mdx mutation of the dystrophin gene does not lead to further lethality in double mutant animals (b). Distribution of male offspring after mating of male miR-206/133b+/- mdxy/+ or miR-206/133b+/- mdx+/+ to female miR-206/133b+/- mdx+/- is presented. - + 387202 Gene miR-206 - + 13405 Gene 20856 dystrophin - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 @@ -11952,37 +11784,37 @@ Tab1.xml 19304 Genotype N % observed Expected miR-206/133b+/+ mdxy/+ 29 13.0% 12.5% miR-206/133b+/+ mdxy/- 25 11.2% 12.5% miR-206/133b+/- mdxy/+ 57 25.6% 25.0% miR-206/133b+/- mdxy/- 41 18.4% 25.0% miR-206/133b-/- mdxy/+ (a) 35 15.7% 12.5% miR-206/133b-/- mdxy/- (b) 36 16.1% 12.5% Total 223 100.0% 100.0% - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene @@ -11996,19 +11828,19 @@ fig_caption 19630 Deletion of miR-206/133b does not impair myogenesis during somite development. miR-206/133b knock-out animals are born with the expected Mendelian distribution (see text), whole mount in situ hybridization indicates normal myogenin expression in wildtype (WT; A, C) and knock-out embryos (KO; B, D) at embryonic day 10.5. Sections (C, D) are at the level of the forelimbs. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 17928 Gene 1854 @@ -12021,155 +11853,155 @@ paragraph 20004 To analyze the function of the miR-206/133b miRNA cluster we deleted the coding region of the miR-206/133b gene on mouse chromosome 1 by homologous recombination (Figure 1A, B). The targeting strategy also removed the last exon of the ncRNA linc-MD1 including the putative miR-133a/b binding site in linc-MD1, which is identical to the miR-133b* sequence. Neither the putative miR-135 binding sites in the second exon of linc-MD1, nor the described distal or proximal regulatory elements, identified by myogenin or myoD ChIP-seq approaches in C2C12 myocytes, were affected by the targeting strategy. Targeted deletion of the miR-206/133b coding region was validated by southern blot analysis (Figure 1C). Viable knock-out mice were obtained by breeding of heterozygous animals. Deletion of the miR-206/133b cluster did not impair embryonic or early postnatal survival of homozygous animals (Table 1; mdxY/+ mice). To exclude a function of miR-206/133b in early development that might be compensated during later stages we analyzed the spatial distribution of myogenin transcripts at the embryonic stage E10.5. No differences in somite morphology and myogenin expression were detected (Figure 2). Development of body weights and survival rate of miR-206/133b knock-out mice was indistinguishable from WT animals. RNase protection analysis as well as northern blot analysis using RNA isolated from hind limb muscle confirmed the absence of miR-206 in homozygous mutant animals (Figure 1D, E). Due to the presence of miR-133a in skeletal muscles, which differs by only one nucleotide from miR-133b, it was not possible to demonstrate the absence of mature miR-133b in homozygous mutant animals. - + 387202 Gene miR-206 - + 387202;723817 Gene miR-206/133b - + 105661730 Gene linc-MD1 - + 723817 Gene miR-133b - + 105661730 Gene linc-MD1 - + 17928 Gene 1854 myogenin - + 17927 Gene 7857 myoD - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 17928 Gene 1854 myogenin - + 17928 Gene 1854 myogenin - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 723817 Gene miR-133b - + 10090 Species mouse - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 105661730 Gene linc-MD1 - + CVCL:0188 CellLine @@ -12183,141 +12015,141 @@ fig_caption 21701 Expression of miR-206 is confined to skeletal muscle. Expression of miR-206 (A) is detected by quantitative reverse transcription PCR (qRT-PCR) in tissues isolated from wildtype (WT) (open bars), miR-206/133b knock-out and mdxy/- (gray bars) mice. Expression of miR-206 is detected in skeletal muscle of the WT mice. miR-206 is not detected in muscles isolated from knock-out mice, confirming the loss of miR-206 in the knock-out tissue and specificity of the Taqman assay. miR-206 was at the qRT-PCR detection limit in heart and brain of WT mice. In WT mice, miR-206 was strongly expressed in the soleus muscle, which has a high content of type 1/oxidative skeletal muscle fibers. A lower amount of miR-206 was detected in the diaphragm, as well as in other skeletal muscles. In mdx mice, the expression of miR-206 was not increased in soleus muscle. Increased miR-206 expression was detected in the diaphragm and other skeletal muscles. Expression of miR-1 (B) was also analyzed in the respective muscles and appeared unchanged in the soleus muscle of miR-206/133b knock-out mice (black bar). Mdx mutation did not cause significant changes in the expression of miR-1. Samples were isolated from >=3 different animals each, and qRT-PCR was performed in triplicate for each of the samples. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 100316783 Gene miR-1 (B) - + 387202 Gene miR-206 - + 13405 Gene 20856 Mdx - + 13405 Gene 20856 mdx - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 735281 Gene @@ -12331,25 +12163,25 @@ fig_caption 22993 No alterations in fiber type identity and fibers size in miR-206/133b knock-out mice. Sections of the soleus muscle isolated from adult wildtype (WT) (A) and mutant mice (B) were stained for markers of slow myofibers. No changes in fiber size of slow and fast muscle fibers were detected (C; n >=8 per group, 30 fibers per animals). The ratio of slow/fast muscle fibers was unchanged in the soleus muscle of knock-out compared to WT mice (D). The scale bar in A corresponds to 100 mum in A and B. - + 387202 Gene miR-206 - + 10090 Species mice - + 10090 Species mice - + 10090 Species @@ -12361,98 +12193,92 @@ paragraph 23493 The miR-206/133b cluster is predominantly expressed in muscles with a high content of oxidative type I myofibers such as the soleus muscle and less abundant in muscles with only few oxidative fibers (Figure 3A). Analysis of published small RNA sequencing results reveals that miR-206 is much less abundant in skeletal muscle than the related miR-1 (GSM539875; 20124 miR-206 versus 745064 miR-1 tags per million miRNA tags). Small RNA sequencing of mouse soleus muscle revealed that even in the muscle with highest expression of miR-206 miR-1 is by far the most abundant miRNA (GSE63342; 30812 miR-206 versus 281944 miR-1 tags per million miRNA tags). The expression of miR-1 was not significantly altered after loss of miR-206 (Figure 3B, M. soleus samples). Histological examination of M. soleus did not reveal obvious morphological alterations. We detected no changes in fiber size, fiber type distribution (Figure 4) or localization of myofiber nuclei. Similar results were obtained for other skeletal muscles (data not shown). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mouse - + 387202 Gene miR-206 - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - - MESH:C581775 - Chemical - - GSM539875 - RESULTS title_2 24527 miR-206/133b mutant skeletal muscles do not show significant molecular abnormalities - + 387202 Gene miR-206 - - MESH:D030342 + + MESH:C567116 Disease molecular abnormalities @@ -12465,25 +12291,25 @@ fig_caption 24612 No transcriptional changes in the soleus muscle of miR-206/133b knock-out (KO) mice. The scatter blot of the median of the log2 signals of wildtype (WT) (n = 9) versus KO (n = 8) indicates only few genes with expression changes more than twofold (green lines). The probe set 1446563_at detects RNA originating from the genome between miR-206 and miR-133b most likely the pri-miRNA of these miRNAs. The weak signal for this probe set in WT samples is reduced to the detection limit in the KO samples. Only a few of the detected changes in gene expression are significant after statistical analysis using Student's t-test. None of these changes could be verified by quantitative reverse transcription PCR (qRT-PCR) or analysis of protein expression. After false discovery rate correction by the Benjamini-Hochberg algorithm, only one gene directly downstream of the manipulated genomic locus significantly changed expression. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 10090 Species @@ -12495,56 +12321,56 @@ paragraph 25547 To investigate whether the loss of expression of miR-206/133b leads to changes of gene expression levels we performed Affymetrix GeneChip analysis using RNA isolated from M. soleus and mouse genome 430 2.0 arrays (n = 9 WT, n = 8 KO). Only few and subtle changes were detected by this type of analysis (Figure 5). The strongly and significantly downregulated probe set 1446563_at detects RNA transcribed from the miR-206/133b locus with the probes binding between the both miRNAs but not within the known linc-MD1 exons. This finding additionally confirms the successful knock-out strategy and suggests that this probe set detects the common miR-206/133b precursor RNA. The only gene that was significantly (2.5-fold) upregulated after false discovery rate correction was IL-17a, which is located downstream of the miR-206/133b locus, suggesting a cis-regulatory mechanism caused by manipulation of the neighboring miRNA locus. No other gene was significantly deregulated in soleus muscle. In particular, we did not observe up-regulation of predicted miR-206 or miR-133b target genes. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 105661730 Gene linc-MD1 - + 16171 Gene 1651 IL-17a - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 10090 Species @@ -12558,156 +12384,156 @@ fig_caption 26641 miR-206/133b does not modulate abundance of PAX7 in E10.5 embryos. SILAC-based screening of changes in protein abundance based on two independent pairs of miR-206/133b knock-out / wildtype (WT) samples revealed few changes that were greater than 1.5 fold up or down in miR-206/133b knock-out versus WT M. soleus (A). Not one of the proteins expected to be potentially upregulated was a predicted target of miR-206 or miR-133b (microRNA.org). In accordance with transcriptome data, no change in protein abundance could be confirmed in western blot analysis. Loss of miR-206/133b does not change the abundance of PAX7 protein in the trunk of E10.5 embryos (B, C). Analysis of the 3' untranslated region (UTR) of Pax7 using miRanda indicates identical binding sites for miR-206 and miR-1. Differences in the primary sequence of miR-206 and miR-1 do not lead to substantial changes in the predicted miRNA-Pax7 UTR interaction (D). - + 387202;723817 Gene miR-206/133b - + 18509 Gene 55665 PAX7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 18509 Gene 55665 PAX7 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 735281 Gene miR - + 735281 Gene miR - - MESH:C537402 - Disease - - miRanda - RESULTS paragraph 27571 Since microRNAs regulate expression on the transcript or protein level, we performed an unbiased SILAC based proteomics screen to detect potential miRNA targets that are regulated at the translational level. Proteins were isolated from M. soleus of WT and miR-206/133b mutant mice and combined with proteins obtained from soleus muscles of 13C6Lys-labled mice, which serve as a reference for relative quantification. In total, we identified 1,500 proteins and 805 were quantified in two independent experiments. SILAC ratios of WT/13C6-Lys-WT and in knock-out/13C6-Lys-WT were used to calculate the direct KO/WT ratios for each of the experiments. In accordance with our transcriptome analysis, only few changes in protein concentrations between knock-out and WT soleus muscle were observed (Figure 6A). Although we quantified 94 potential miR-206 targets that displayed a good score according to the miRanda based microRNA.org target prediction database (August 2010 release), none of the potential target protein was upregulated more than 1.5 fold in the knock-out tissue. Likewise, none of the 57 proteins found in our SILAC data set that were predicted as miR-133b targets according to the microRNA.org database were upregulated. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 10090 Species mice - + 10090 Species mice - - MESH:C026591 + + - Chemical - - 13C6-Lys + + 13C6 - - MESH:C537402 - Disease - - miRanda + + MESH:D008239 + Chemical + + Lys - - MESH:C026591 + + MESH:D008239 Chemical - - 13C6-Lys + + Lys - - MESH:C026591 + + - + Chemical + + 13C6 + + + - Chemical 13C6Lys @@ -12718,131 +12544,125 @@ paragraph 28806 Although no alterations in skeletal muscle development of miR-206/133b mutants were apparent, we investigated the potential regulatory interaction between miR-206 and Pax7 in somites of E10.5 embryos. No changes in Pax7 protein expression were detected in mutant compared to WT embryos indicating that miR-206 is not required to suppress expression of Pax7 in somites (Figure 6B, C), probably due to the presence of miR-1, which shows a similar expression pattern as miR-206 during skeletal muscle development but is more widely expressed compared to miR-206 in adult muscles. In fact, the seed sequences of miR-1 and miR-206 are identical, which predicts a comparable repression of Pax7 by either miR-206 or miR-1 (Figure 6D) according to target algorithms like miRanda. Taken together, neither transcriptome nor proteome analysis revealed significant changes in gene expression between WT and KO mice, which is in line with the lack of a muscle specific phenotype in miR-206/133b knock-out mice. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mice - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - - MESH:C537402 - Disease - - miRanda - RESULTS title_2 29806 Loss of the miR-206/133 cluster does not affect skeletal muscle regeneration in Duchenne muscular dystrophy model mice - + 387202 Gene miR-206 - + MESH:D020388 Disease @@ -12856,71 +12676,71 @@ fig_caption 29925 No difference in body weight and survival of miR-206/133b and mdx mutant mice. (A) The body weight of male animals is not different at 6 weeks of age (n >=11/group); however, at later ages (B), the mdx mutation leads to an increase in body weight that is not further modified by mutation of the miRNA cluster (wildtype (WT): miR-206/133b+/+, knock-out (KO): miR-206/133b-/-; MDX: mdxY/-; n >=5/group). (C) Survival of miR-206/133b mutants is not impaired. The mdx mutation does not affect survival of miR-206/133b knock-out mice. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 mdx - + 13405 Gene 20856 MDX - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 10090 Species mice - + 10090 Species @@ -12934,81 +12754,63 @@ fig_caption 30463 Loss of miR-206/133b does not change serum creatine kinase activity. Creatine kinase activity in the serum in miR-206/133b mice knock-out mice is not altered. The mdx mutation leads to strong increase of creatine kinase activity. No differences were observed in mdx versus miR-206/133b KO / MDX mice (n >=6/group). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 MDX - + 387202 Gene miR-206 - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - - MESH:D003401 - Chemical - - Creatine - - - MESH:D003401 - Chemical - - creatine - - - MESH:D003401 - Chemical - - creatine - FIG @@ -13017,500 +12819,518 @@ fig_caption 30780 miR-206/133b mutation does not modulate muscle loss or regeneration in Mdx mice. No histological changes are observed in diaphragm or M. quadriceps after loss of miR-206/133b. Mutation of the mouse dystrophin gene (MDX) leads to increased muscle mass, calcifications (von Kossa staining) and increased fibrosis (red signal in Sirius red staining). Additional loss of miR-206/133b does not lead to changes of the mdx phenotype. Sections obtained from 12 weeks old animals are shown, similar results are obtained using tissue from 6-week- and 9-month-old mice. The scale bar corresponds to 200 mum in the upper three rows and to 50 mum in the lower two rows. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 dystrophin - + 387202 Gene miR-206 - + 13405 Gene 20856 MDX - + 13405 Gene 20856 mdx - + 13405 Gene 20856 Mdx - + 10090 Species mice - + 10090 Species mouse - + 10090 Species mice - + MESH:D005355 Disease fibrosis - + MESH:D009133 Disease muscle loss + + MESH:D014842 + Disease + + von + RESULTS paragraph 31441 Previously, miR-206 has been implicated in muscle fiber stability and regeneration of skeletal muscles, although the miR-206 seems dispensable for specification, proliferation and differentiation of skeletal muscle. To analyze the potential function of miR-206/133b miRNAs and their targets in maintenance and regeneration of muscle fibers we generated miR-206/133b-/-//mdxY/- compound mutant mice. Mdx mice carry a point mutation in the dystrophin gene on the X-chromosome leading to constant damage and loss of muscle fibers, which is compensated by increased regeneration. miR-206/133b-/-/mdx mutant mice were born at the expected Mendelian ratio (Table 1) and developed normally, which is in contrast to the previously described miR-206-/-/mdx mutant mice. Body weight development (Figure 7A, B) and survival (Figure 7C) of compound mutant animals was indistinguishable from mdx mice. Regeneration of skeletal muscles in mdx mice leads to increased expression of the miR-206/133b cluster in muscles with a low content of type I muscle fibers that express only low levels of miR-206/133b under normal conditions (Figure 3A,). In contrast we observed a moderate, not significant reduction of miR-1 in some mdx muscles (Figure 3B), possibly related to the emergence of other non-muscle cell types in the dystrophic muscles. Hence, it seems possible that such muscle might be affected by inactivation of miR-206/133b in conditions of constant regeneration. Loss of dystrophin expression results in the release of muscle creatine kinase (MCK) from muscle fibers into the blood, which serves as an indicator of muscle injury or muscle degenerative processes. To access the degree of muscle damage in miR-206/133b-/-/mdx mutant mice we determined MCK activity in serum of adult mice. Only very low MCK activity was detected in WT and miR-206/133b mice, which did not differ significantly between WT (297 +- 85 U/l, n = 6) and mutants (152 +- 62 U/l, n = 11). Importantly, we observed much higher MCK activities in mdx mice (8590 +- 1507 U/l, n = 6), which were not increased any further in miR-206/133b/mdx mutants (8024 +- 2464 U/l, n = 6; Figure 8) indicating that compound mutants do not suffer from increased muscle fiber damage compared to mdx mice. Next, we performed a comprehensive histological analysis of different muscles of the hind limb, as well as of the diaphragm from WT, miR-206/133b-/-, mdxY/- and miR-206/133b-/-//mdxY/- mice. Trichrome- and Sirius Red-, and von Kossa staining were used to determine myofiber organization, fibrosis, and intramuscular calcifications often present in mdx mice (Figure 9). No differences were found between WT and miR-206/133b skeletal muscle based on Trichrome- (not shown), von Kossa or Sirius red staining including absence of fibrosis and calcifications. Mutation of dystrophin caused the typical signs of muscle dystrophy, namely massive thickening of the diaphragm, enhanced fibrosis and patches of calcified tissue within the diaphragm and the M. quadriceps (Figure 9). No calcified fibers were observed in the soleus muscle. Deletion of miR-206/133b in mdx mice did not cause a more severe phenotype in the soleus, quadriceps or diaphragm muscles compared to single mutant mdx mice. Together these data indicate that loss of miR-206/133b cluster does not increase muscle damage in mdx mice or compromise skeletal muscle regeneration in constantly renewing muscle tissue. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 dystrophin - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 dystrophin - + 12715 Gene 20432 muscle creatine kinase - + 12715 Gene 20432 MCK - + 12715 Gene 20432 MCK - + 12715 Gene 20432 MCK - + 387202 Gene miR-206 - + 12715 Gene 20432 MCK - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 dystrophin - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 mdx - + 13405 Gene 20856 Mdx - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 735281 Gene miR - - MESH:D009135 - Disease - - damage and loss of muscle fibers - - + MESH:D009136 Disease dystrophic muscles - + MESH:D009136 Disease muscle dystrophy - + MESH:D005355 Disease fibrosis - - MESH:D009135 + + MESH:D014842 Disease - + + von + + + MESH:D009135 + Disease + muscle injury - + + - + Chemical + + Sirius red + + MESH:D005355 Disease fibrosis - + MESH:D005355 Disease fibrosis - + MESH:D009135 Disease muscle damage - + MESH:D009135 Disease muscle damage + + MESH:D014842 + Disease + + von Kossa + FIG @@ -13519,94 +13339,100 @@ fig_caption 34932 No differences in differentiation of muscle progenitor cells in vitro . Proliferating muscle stem cells isolated from hind limb muscles of wildtype (WT) and miR-206/133b knock-out mice (KO) were induced to differentiate and stained after 3 or 5 days in differentiation medium (DM) for expression of myosin (green: MF20 staining, blue: nuclear staining; the scale bar corresponds to 50 mum). No differences in cell morphology or differences in the expression of myosin were detected between WT and mutant cells (A). Formation of fused myotubes from differentiating myoblasts was not impaired by the loss of the miR-206/133b cluster (B). Pax7 expression in differentiating myogenic progenitor cells was detected by western blot. No differences in Pax7 expression were detected after loss of miR-206/133b (C). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 10090 Species mice + + MESH:D009223 + Disease + + DM + RESULTS paragraph 35742 To further analyze a possible function of the miR-206/133b cluster in the differentiation of muscle stem cells, we isolated muscle satellite cells from WT and miR-206/133b mutant mice. Loss of the miR-206/133b cluster did not impair the ability of muscle stem cells to differentiate to myotubes in vitro (Figure 10A). We did not observe reduced differentiation as accessed by the number of MF20 positive cells nor changes in the fusion of myocytes as accessed by the number of myotubes with more than one nucleus (Figure 10B). Moreover, we found that the lack of miR-206/133b did not increase the concentration of the potential miR-206 and miR-1 target Pax7 in differentiating satellite cells on the 5th day after induction of differentiation (Figure 10C). - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 10090 Species mice - + 735281 Gene @@ -13624,122 +13450,122 @@ paragraph 36513 Physiological properties of skeletal muscle cells including specification, differentiation, function and regeneration are controlled by an extensive network of regulatory molecules. Many key genes have been identified that are essential for specification and differentiation of muscle precursor cells during development. Commitment of muscle cells is controlled by a core program of myogenic regulatory factors of the MyoD family, which also control transcription of the miR-1/206/133 miRNA clusters resulting in a prominent expression of these miRNAs in muscle cells. Yet, important differences in the expression pattern of miR-1/206/133 miRNA clusters exist: the miR-1/133a clusters are expressed in heart and all skeletal muscles, while expression of miR-206/133b becomes restricted to a subset of oxidative muscle fibers by hitherto unknown molecular mechanisms. Since the two miR-1/133a clusters display virtually identical expression patterns, partially overlapping or redundant functions of these miRNAs seem likely. In contrast, the miR-206/133b cluster shows a more restricted expression, which might argue for a distinct function of this cluster albeit muscle cells expressing miR-206/133b also transcribe miR-1/133a. In addition, the primary sequence of miR-206 differs from miR-1 while miR-133b is very similar to miR-133a. However, the seed sequences of miR-206 and miR-1 important for target recognition are identical, indicating only limited differences in the target specificity of miR-206 and miR-1. The miR-206/133b locus also directs expression of the long non-coding RNA linc-MD1, which was postulated to act as a competing endogenous RNA or miRNA decoy adding a further potential function to the miR-206/133b locus. - + 17927 Gene 7857 MyoD - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 105661730 Gene linc-MD1 - + 387202 Gene miR-206 - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene @@ -13751,115 +13577,115 @@ paragraph 38250 We have generated a miR-206/133b knock-out allele by deletion of the genomic region spanning from miR-206 to miR-133b and resulting in removal of the third exon of the linc-MD1 that might act as a miR-133 sponge. Surprisingly, homozygous deletion of the miR-206/133b locus did not cause an apparent phenotype in adult mice. No changes in the morphology and distribution of muscle fiber types were apparent and the transcriptional and proteomics signatures of miR-206/133b mutant muscles were virtually unchanged, ruling out that miR-206 and/or 133b alone play an essential role in muscle physiology. These observations are consistent with to observations made in miR-206 knock-out mice, which also show no obvious phenotype under baseline conditions. Apparently, the concomitant loss of miR-206, miR-133b and the potential miR-133 sponge in linc-MD1 is compatible with normal muscle development and functions, which might be explained by expression of the miR-1/133a clusters in type I myofibers compensating for deletion of miR-206/133b. Theoretically, the inactivation of miR-133b might also be counteracted by the loss of the miR-133 sponge function, although the physiological relevance of endogenous competing RNAs has been questioned making this explanation less likely. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 105661730 Gene linc-MD1 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 105661730 Gene linc-MD1 - + 387202 Gene miR-206 - + 723817 Gene miR-133b - + 10090 Species mice - + 10090 Species mice - + 387202 Gene miR-206 - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene @@ -13871,259 +13697,253 @@ paragraph 39527 We found a more widespread and abundant expression of the miR-206/133b cluster in regenerating muscles, which confirms previous findings. It seems possible that the expression of miR-206 in regenerating muscle reflects a specific function of the miR-206/133b in the regenerative response. However, it is important to point out that miR-206/133b is broadly expressed during development in immature muscle cells. Hence, it is difficult to exclude that the increased expression of miR-206/133b during skeletal muscle regeneration simply mirrors the immature state of newly formed myofibers that have not yet terminated expression of the miR-206/133b cluster in the majority of mature type II fibers. The lack of a more severe phenotype in miR-206/133b-/-//mdxY/- compound mutants compared to mdxY/- mice clearly argues against an important role of the miR-206/133b cluster for skeletal muscle regeneration. Moreover, we did not see an increase in lethality or an increase in serum creatine kinase levels in compound mutant mice, which also excludes an important role of the miR-206/133b cluster in restricting myofiber damage. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mice - + 10090 Species mice - - MESH:D003401 - Chemical - - creatine - DISCUSS paragraph 40655 miR-206 has been claimed to suppress Pax7 expression thereby regulating muscle stem cell renewal and differentiation. The normal expression of Pax7 in isolated miR-206/133b mutant satellite cells and the normal differentiation of mutant satellite cells to myotubes in vitro do not argue for an important role of miR-206 in Pax7 expression. These results are in contrast to previous publications mostly relying on the inhibition of miR-206 by anti-miRs, overexpression of miR-206, or by correlation of expression profiles. It seems possible that previous anti-miR approaches were biased by off-target effects or affected expression of miR-1, which can also suppress Pax7. Generation of skeletal muscle specific miR-1/miR133a//miR-206/133b triple mutants will probably solve this controversy in the future. We do not have a convincing explanation for the attenuation of Pax7 down-regulation in differentiating miR-206-deficient satellite cells reported by Liu et al.. The same authors also described that miR-206/mdx mutant mice suffer from increased muscle degeneration, impaired regeneration and premature death, which was all not seen in our miR-206/133b mutants. In principle, it is feasible that the concomitant loss of mir-206 and miR-133b in our model accounts for these differences, which would strengthen the notion that the miR-206/133b cluster acts as a functional unit targeting similar biological processes sometimes in parallel and sometimes in opposing directions. Alternatively, subtle differences in the genetic background and/or strain-depended differences in the expression of miR-1/133a compensating for the loss of miR-206/133b might differentially affect the course of muscular dystrophy in miR-206/133b-/-//mdxy/- and miR-206-/-//mdxy/- compound mutants. - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 18509 Gene 55665 Pax7 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene mir-206 - + 723817 Gene miR-133b - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 735281 Gene miR - + 13405 Gene 20856 mdx - + 10090 Species mice - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene miR - - MESH:D003643 + + MESH:D009422 Disease - - death + + impaired regeneration and premature death - + MESH:D009135 Disease muscle degeneration - + MESH:D009136 Disease @@ -14141,43 +13961,43 @@ paragraph 42450 Taken together, our results question a major role of the miR-206/133b cluster in development, function and regeneration of skeletal muscle in mouse and argue for overlapping functions of the miR-206/133b and miR-1/133a gene clusters during myogenesis. Our findings are in line with the hypothesis that individual miRNAs often fine-tune biological processes and play modulatory roles. Further genetic studies targeting the miR-1/133a together with the miR-206/133b gene clusters specifically in skeletal muscle will probably reveal the prevalent physiological function of miR-1/206/133 in mammals. - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 387202 Gene miR-206 - + 10090 Species mouse - + 735281 Gene miR - + 735281 Gene miR - + 735281 Gene @@ -14201,7 +14021,7 @@ paragraph 43065 bovine serum albumin - + 9913 Species @@ -14225,6 +14045,12 @@ paragraph 43121 DM + + MESH:D009223 + Disease + + DM + ABBR @@ -14237,25 +14063,26 @@ paragraph 43147 DSHB + + - + Chemical + + DSHB + ABBR paragraph 43152 developmental studies hybridoma bank - - Disease - - hybridoma bank - ABBR paragraph 43189 DTA - - MESH:C522116 + + MESH:C042899 Chemical DTA @@ -14290,7 +14117,7 @@ paragraph 43244 fetal calf serum - + 9913 Species @@ -14320,7 +14147,7 @@ paragraph 43290 horseradish peroxidase - + 3704 Species @@ -14362,7 +14189,7 @@ paragraph 43365 beta-galactosidase coding - + 12091 Gene 47922 @@ -14381,12 +14208,6 @@ paragraph 43395 creatine kinase, muscle - - MESH:D003401 - Chemical - - creatine - ABBR @@ -14399,19 +14220,13 @@ paragraph 43423 mouse model of Duchenne Muscular Dystrophy, X-linked - + 10090 Species mouse - - MESH:D040181 - Disease - - X-linked - - + MESH:D020388 Disease @@ -14435,17 +14250,23 @@ paragraph 43486 PBS + + MESH:D007854 + Chemical + + PBS + ABBR paragraph 43490 phosphate buffered saline - - MESH:D010710 + + - Chemical - - phosphate + + phosphate buffered saline @@ -14453,8 +14274,8 @@ paragraph 43516 PFA - - MESH:C101815 + + MESH:D017245 Chemical PFA @@ -14465,7 +14286,7 @@ paragraph 43520 paraformaldehyde - + MESH:C003043 Chemical @@ -14489,8 +14310,8 @@ paragraph 43606 SDS-PAGE - - MESH:C032259 + + MESH:D012967 Chemical SDS @@ -14501,11 +14322,17 @@ paragraph 43615 sodium dodecyl sulfate polyacrylamide gel electrophoresis - + + MESH:C016679 + Chemical + + polyacrylamide + + MESH:D012967 Chemical - - sodium dodecyl sulfate polyacrylamide gel + + sodium dodecyl sulfate @@ -14579,13 +14406,7 @@ footnote 43905 TBo, SW, and HN conducted the experiments. TBo and TBr designed and interpreted the experiments and wrote the manuscript. All authors read and approved the final manuscript. - - MESH:C009802 - Chemical - - HN - - + CVCL:R777 CellLine @@ -14628,14 +14449,14 @@ surname:Braun;given-names:T 44145 miR-1/133a clusters cooperatively specify the cardiomyogenic lineage by adjustment of myocardin levels during embryonic heart development - + 214384 Gene 17043 myocardin - + 735281 Gene @@ -14661,7 +14482,7 @@ surname:Ang;given-names:YS 44283 microRNA-1 regulates sarcomere formation and suppresses smooth muscle gene expression in the mammalian heart - + 9606 Species @@ -14712,20 +14533,20 @@ surname:Hulver;given-names:MW 44502 Mice lacking microRNA 133a develop dynamin 2-dependent centronuclear myopathy - + 13430 Gene 90883 dynamin 2 - + 10090 Species Mice - + MESH:D009135 Disease @@ -14769,19 +14590,19 @@ surname:Braun;given-names:T 44651 Specific requirements of MRFs for the expression of muscle specific microRNAs, miR-1, miR-206 and miR-133 - + 387202 Gene miR-206 - + 735281 Gene miR - + 735281 Gene @@ -14829,13 +14650,13 @@ surname:Gingeras;given-names:T 44757 An encyclopedia of mouse DNA elements (Mouse ENCODE) - + 10090 Species mouse - + 10090 Species @@ -14926,7 +14747,7 @@ surname:Shelton;given-names:JM 45097 microRNA-206 promotes skeletal muscle regeneration and delays progression of Duchenne muscular dystrophy in mice - + MESH:D020388 Disease @@ -14947,13 +14768,13 @@ 453 45210 The value of mammalian models for duchenne muscular dystrophy in developing therapeutic strategies - + 9606 Species mammalian - + MESH:D020388 Disease @@ -14978,20 +14799,20 @@ surname:Darlison;given-names:MG 45309 The molecular basis of muscular dystrophy in the mdx mouse: a point mutation - + 13405 Gene 20856 mdx - + 10090 Species mouse - + MESH:D009136 Disease @@ -15013,7 +14834,7 @@ 214 45386 miR-206 and -486 induce myoblast differentiation by downregulating Pax7 - + 387202;723876 Gene @@ -15056,14 +14877,14 @@ 286 45581 Genotyping mdx, mdx3cv, and mdx4cv mice by primer competition polymerase chain reaction - + 13405 Gene 20856 mdx - + 10090 Species @@ -15104,13 +14925,13 @@ surname:Kruger;given-names:M 45735 Acquisition of the contractile phenotype by murine arterial smooth muscle cells depends on the Mir143/145 gene cluster - + 387161;387163 Gene Mir143/145 - + 10090 Species @@ -15157,14 +14978,14 @@ surname:Luber;given-names:CA 45916 SILAC mouse for quantitative proteomics uncovers kindlin-3 as an essential factor for red blood cell function - + 108101 Gene 12877 kindlin-3 - + 10090 Species @@ -15265,7 +15086,7 @@ 840 46373 Mammalian microRNAs predominantly act to decrease target mRNA levels - + 9606 Species @@ -15304,11 +15125,6 @@ surname:Sander;given-names:C 46519 MicroRNA targets in Drosophila - - Disease - - Drosophila - surname:Marks;given-names:DS @@ -15344,15 +15160,14 @@ 1192 46600 X chromosome-linked muscular dystrophy (mdx) in the mouse - + 13405 Gene 20856 mdx - - MESH:D009136 + Disease X chromosome-linked muscular dystrophy @@ -15373,7 +15188,7 @@ 767 46658 Biochemical markers of muscular damage - + MESH:D009135 Disease @@ -15412,28 +15227,28 @@ surname:Tapscott;given-names:SJ 46777 MyoD inhibits Fstl1 and Utrn expression by inducing transcription of miR-206 - + 17927 Gene 7857 MyoD - + 14314 Gene 5144 Fstl1 - + 22288 Gene 21398 Utrn - + 735281 Gene @@ -15476,7 +15291,7 @@ surname:Takeda;given-names:S 46946 MicroRNA-206 is highly expressed in newly formed muscle fibers: implications regarding potential for muscle regeneration and maturation in muscular dystrophy - + MESH:D009136 Disease @@ -15501,13 +15316,13 @@ surname:Adachi;given-names:N 47104 Acceleration of muscle regeneration by local injection of muscle-specific microRNAs in rat skeletal muscle injury model - + 10116 Species rat - + MESH:D009135 Disease @@ -15536,7 +15351,7 @@ surname:Maccatrozzo;given-names:L 47224 Myogenic potential of canine craniofacial satellite cells - + 9615 Species @@ -15569,19 +15384,13 @@ surname:Lewis;given-names:S 47282 Impaired regeneration in LGMD2A supported by increased PAX7-positive satellite cell content and muscle-specific microrna dysregulation - + 18509 Gene 55665 PAX7 - - MESH:D021081 - Disease - - microrna dysregulation - @@ -15698,108 +15507,103 @@ paragraph 1906 Sheep are valuable resources for the animal fibre industry. Identifying genes which regulate wool growth offers the opportunity to improve the wool production efficiency, product quality and diversity in breeding programs. It can also offer the opportunity to develop transgenic lines and to develop therapeutic agents that can be used to tailor for desirable fibre attributes by altering gene expression. The genetic polymorphisms and their mechanisms of wool and cashmere growth and regulation have been thoroughly studied. In mammals, several gene families, such as WNTs, tumor necrosis factors (TNFs), fibroblast growth factors (FGFs) and transforming growth factor(TGFs), have been implicated in hair follicle initiation, morphogenesis and cycling. Recently, the molecular characteristics of primary wool follicle initiation in Merino sheep have been reported recently. - + 9940 Species Sheep - + 9940 Species sheep - + MESH:D009336 Disease tumor necrosis - - Chemical - - Merino - INTRO paragraph 2781 Transcriptomic research such as microarray analysis, has been successfully applied to investigate the characteristics of hair follicle stem cells in mice. Microarray studies have been reported for different traits in sheep and goats, such as resistance to parasites, mammary development and milk quality, wool follicle development, natural fleece rot resistance and pigmentation traits of skin and wool. A subset of skin-expressed microRNAs with possible roles in goat and sheep hair growth has also been reported. Several studies have demonstrated the usefulness of cDNA microarray for expression profiling of wool follicle growth cycles in whole skin. More recently the RNA-seq method was also used to determine the genes differentially expressed among various tissues (including whole skin) of sheep. - + 10090 Species mice - + 9940 Species sheep - + 9925 Species goats - + 9925 Species goat - - 9940 - Species - - sheep - - + 9940 Species sheep + + MESH:D012757 + Disease + + sheep hair growth + INTRO paragraph 3585 The Aohan fine wool sheep, bred in Inner Mongolia, is an outstanding breed, providing both wool and meat. The major characteristics of this breed are high quality wool,high disease resistance, and high adaptability. According to figures, Aohan fine wool sheep can provide up to 9 kg of quality wool per year (fiber length up to 10.5 cm, fibre diameter less than 22 mum). Therefore, Aohan fine wool sheep are considered as a valuable genetic resource for fine wool production. Wang et al. demonstrated that seasonal factors significantly influenced the wool growth of Aohan fine wool sheep. The peak of the growth rate occurs in summer and the low in winter. The expression profiling of immune genes and type I inner root sheath (IRS) keratin genes in the whole skin of Aohan fine wool sheep has previously been reported by our laboratory team. - + 9940 Species sheep - + 9940 Species sheep - + 1214577 Species to 9 - + 9940 Species sheep - + 9940 Species sheep - + 9940 Species @@ -15811,13 +15615,13 @@ paragraph 4432 However, to our knowledge, as of yet no microarray or proteomic study at a genome-wide level has been reported on protein-coding genes which are possibly responsible for regulating hair growth of adult sheep so far. The aims of the present work are to investigate and compare the gene expression level of the body side skin and groin skin using microarray and proteomic technology, and to identify the possible genes and proteins responsible for the wool growth regulation of Aohan fine wool sheep. - + 9940 Species sheep - + 9940 Species @@ -15841,42 +15645,36 @@ paragraph 4970 A total of 1494 probes were differentially expressed comparing the body side to groin skins (S/G) in Aohan fine wool sheep, including 602 up-regulated and 892 down-regulated probes, as shown in Additional file 1: Table S1. Most probes (1110) were not assigned to unique transcripts, due to the lack of information. The number of distinct genes/transcripts (annotated) was 331, of which 112 were up-regulated and 219 were down-regulated. In S/G, 7 genes (CYP1A1, LOC100137068, LOC443300, LOC101106865, Connexin 43, SCD and LOC101122398) were down-regulated by more than 10-fold. - + 100170113 Gene CYP1A1 - + 443455 Gene Connexin 43 - + 443185 Gene SCD - + 101122398 Gene LOC101122398 - + 9940 Species sheep - - MESH:C492399 - Chemical - - LOC443300 - RESULTS @@ -15897,126 +15695,114 @@ fig_caption 5868 Q-PCR validation of the microarray data. P values (T-test) of the Q-PCR data are 0.036 (FGF10), 0.019 (LOC443300), 0.028 (FGF18), 0.018 (Connexin 43), 0.044 (SCD) and 0.019 (ZO1), respectively. S/G represent body side skin group/groin skin group, respectively. - + 443074 Gene FGF10 - + 780460 Gene FGF18 - + 443455 Gene Connexin 43 - + 443185 Gene SCD - + 443200 Gene ZO1 - - MESH:C492399 - Chemical - - LOC443300 - RESULTS paragraph 6130 In order to verify the microarray results, we selected 10 genes, namely FGF10, LOC443300, FGF18, Connexin43, SCD, ZO1, MMP2, ITGB1, PAG11 and CRYAB, to comparatively analyze their expression patterns by qPCR. As shown in Figure 1, the qPCR results for the selected seven genes were consistent with the microarray results, except for FGF10, ZO1 and CRYAB, thus reflecting the reliability of our microarray data. - + 443074 Gene FGF10 - + 780460 Gene FGF18 - + 443455 Gene Connexin43 - + 443185 Gene SCD - + 443200 Gene ZO1 - + 443115 Gene MMP2 - + 443141 Gene ITGB1 - + 100034671 Gene PAG11 - + 497274 Gene CRYAB - + 443074 Gene FGF10 - + 443200 Gene ZO1 - + 497274 Gene CRYAB - - MESH:C492399 - Chemical - - LOC443300 - RESULTS @@ -16031,13 +15817,13 @@ fig_caption 6647 Hierarchical cluster analysis of data between different skin areas of the Aohan fine wool sheep in anagen phase. Each column represents one sheep, and each horizontal line refers to a gene. Color legend is on the top of the figure. Red indicates genes with a greater expression relative to the geometrical means, green indicates genes with a lower expression relative to the geometrical means. S1, S2 and S3 represent 3 repeats of body side skin group, and G1, G2, G3 represent 3 repeats of groin skin group. - + 9940 Species sheep - + 9940 Species @@ -16083,49 +15869,49 @@ paragraph 9092 We speculate that wool growth regulation shared similar gene networks with hair. So particular differentially expressed genes, which also appeared in the conclusion of factors with known hair growth regulatory roles, were included in the putative gene lists of wool growth regulation (Figure 4). In the networks, 3 genes (FGF7, IGFBP3 and PRL) were contained in the category of anagen promotion and maintenance, 3 genes (IL1A, IL6 and TAC1) were contained in the category of catagen-telogen promotion and maintenance, while 2 genes (CDKN1B and FGF18) were contained in the category of function unknown. - + 443095 Gene FGF7 - + 100286737 Gene IGFBP3 - + 443317 Gene PRL - + 443404 Gene IL1A - + 443406 Gene IL6 - + 100034669 Gene TAC1 - + 100127219 Gene CDKN1B - + 780460 Gene @@ -16145,14 +15931,8 @@ fig_caption 9769 Representative image of 2-DE silver stained polyacrylamide gel. - - MESH:D012834 - Chemical - - silver - - - MESH:C040388 + + MESH:C016679 Chemical polyacrylamide @@ -16205,13 +15985,13 @@ paragraph 12382 The technical approach (microarrays) is another limitation of the study. First, the number of the probes (15208) designed by Agilent is limited compared to the RNA-seq method. Most of the probes were inadequately annotated. The full utility of the microarray data depends on the complete annotation of the whole sheep genome. Second, the specificity of the probes is relatively lower than RNA-sequencing, although using 60-mers long probes in our study. False positive and false negative phenomena exsit unavoidably in the microarray data. - + 9940 Species sheep - + 1335626 Species @@ -16229,43 +16009,43 @@ paragraph 13003 In the present study, we investigated the molecular events possibly related to sheep hair growth control using microarray and proteomic technologies. Transcriptomic analysis identified hundreds of differentially expressed genes displaying over 2-fold difference between the two sampled skin regions of Aohan fine wool sheep at December time point, and the number of less highly expressed transcripts (892) in body side skin was greater than that of the more highly expressed ones (602). The qPCR results validated the reliability of our microarray data. - - 9940 - Species - - sheep - - + 9940 Species sheep + + MESH:D012757 + Disease + + sheep hair + DISCUSS paragraph 13557 IL-1A and IL-1B inhibit hair growth in vitro as reported previously, but IL-1A has shown downregulation in body side skin in our study. One possibility is in vivo IL-1A controls wool growth in a time dependent or a negative feedback manner. - + 443404 Gene IL-1A - + 443539 Gene IL-1B - + 443404 Gene IL-1A - + 443404 Gene @@ -16277,32 +16057,32 @@ paragraph 13798 The skin of skin-specific SCD1 knock-out (SKO) mice exhibited variable orthokeratotic hyperkeratosis and parakeratotic hyperkeratosis, occasional mal-aligned hair follicles, and instances of protrusion of hair shafts out of the hair follicle and into the surrounding connective tissue, with an infiltration of inflammatory cells surrounding the exposed hair shaft and keratin material. In our study, SCD was less highly expressed at 11.91 fold change in body side skin. This maybe indicate that the SCD could play an important role in groin skin. - + 20249 Gene 74538 SCD1 - + 443185 Gene SCD - + 443185 Gene SCD - + 10090 Species mice - + MESH:D017488 Disease @@ -16314,81 +16094,75 @@ paragraph 14345 MMP2 activity is associated with the disappearance of collagen VII during the invasion of epithelial cords of hair follicles and sweat glands in human skin. MMP2 plays a role in hair growth-associated extracellular matrix remodeling and cell migration, and may be a downstream effector through which thymosin ss4 exerts its effect on hair growth. However, MMP2 showed significant lower expression in body side skin. This may suggest sheep has a different hair growth regulatory mechanism than human. - + 4313 Gene 3329 MMP2 - + 443115 Gene MMP2 - + 443115 Gene MMP2 - + 9606 Species human - + 9940 Species sheep - + 9606 Species human - - MESH:D013947 - Chemical - - thymosin - DISCUSS paragraph 14845 There is a log linear relationship between the relative level of beta 1 integrins (ITGB1) on the cell surface and proliferative capacity in keratinocytes. ITGB1-mediated signalling is also important in human hair growth control. Skin and hair follicle integrity is crucially dependent on ITGB1 expression in keratinocytes. In our study, however,ITGB1 expression in the body side skin was less highly expressed. The mechanism how it happens deserves further investigation. - + 443141 Gene ITGB1 - + 3688 Gene 22999 ITGB1 - + 443141 Gene ITGB1 - + 443141 Gene ITGB1 - + 9606 Species @@ -16400,13 +16174,13 @@ paragraph 15317 Rowe et al. confirmed that predominantly CYP1A1 was located to the sebaceous gland surrounding the hair shaft. It is not known why CYP1A1 expression was less highly expressed more than 10 folds in the body side skin from our data. This interesting phenomenon is worth pursuing in future studies. - + 100170113 Gene CYP1A1 - + 100170113 Gene @@ -16418,74 +16192,80 @@ paragraph 15613 As development proceeds, GluD1 expression becomes restricted to the hippocampus, cochlear and vestibular hair cells, and spiral ganglion cells. Hair follicles are also derived from ectoderm. So why GLUD1 was less highly expressed in body side skin really deserves further investigation. - + 443239 Gene GluD1 - + 443239 Gene GLUD1 + + MESH:D015837 + Disease + + vestibular hair cells + DISCUSS paragraph 15900 Connexin 43 (Cx43) is a gap junction protein expressed in the follicular dermal papilla (DP). A recent mouse model research suggested an important role for Cx43 in hair regeneration, growth, and cuticle formation. However, Connexin 43 was less highly expressed in body side skin in the microarray analysis of this study, and it was also confirmed by qPCR. - + 443455 Gene Connexin 43 - + 443455 Gene Cx43 - + 14611 Gene 9670 Cx43 - + 443455 Gene Connexin 43 - + 10090 Species mouse + + MESH:D010211 + Disease + + follicular dermal papilla + DISCUSS paragraph 16256 Several papers indicated that prostaglandin induced hair growth, but in our results, one prostaglandin synthase (PGFS) were less highly expressed in body side skin. This interesting contradiction remains to be further elucidated. - + 443108 Gene PGFS - - MESH:D011453 - Chemical - - prostaglandin - - + MESH:D011453 Chemical @@ -16497,47 +16277,53 @@ paragraph 16486 We have indeed tried our best to find out differentially expressed genes associated with LCE and MOGAT as reported in a Science paper. However, we could find only a few genes (DGAT2, LPL, ZO1, GJB2 and ITGB1) that have the possibility to play roles in epidermal development complex (EDC) or skin lipid metabolism. One reason for limited DE genes is that what we compared were different regions of sheep skin tissue (which might share similar principles in EDC regulation and lipid synthesis), not distinct tissues such as rumen and skin. That might also be caused by inadequate annotation of the probes or less gene coverage redundancy of the Agilent microarray. - + 101110535 Gene DGAT2 - + 443408 Gene LPL - + 443200 Gene ZO1 - + 443345 Gene GJB2 - + 443141 Gene ITGB1 - + 9940 Species sheep - - MESH:D052439 - Disease - - lipid metabolism + + MESH:D008055 + Chemical + + lipid + + + MESH:D008055 + Chemical + + lipid @@ -16557,13 +16343,13 @@ paragraph 17319 Cryab protein was detected in 2DE in groin skin but not in body side skin. But the Cryab transcripts was more highly expressed in body side skin by the microarray study, though the differential expression was not validated by qPCR. This gene, downregulated in the transcriptional level during the depilation-induced hair cycle, was considered to play a role in regulation of apoptosis. - + 497274 Gene Cryab - + 497274 Gene @@ -16575,19 +16361,19 @@ paragraph 17705 FGF18 protein was less highly expressed in body side skin compared to the control. But the FGF18 transcripts was more highly expressed by the microarray study. Consistent to our microarray results, FGF18 is highly expressed in hair follicles and capable of inducing anagen from telogen stage hair follicles. - + 780460 Gene FGF18 - + 780460 Gene FGF18 - + 780460 Gene @@ -16599,13 +16385,13 @@ paragraph 18013 KRT14 and KRT2.11 proteins were both less highly expressed in body side skin compared to the control. But these proteins were expected to play roles in wool follicles. - + 100303604 Gene KRT14 - + 443415 Gene @@ -16617,34 +16403,34 @@ paragraph 18181 PADI3 played roles in the assembly of a globular S100A3 homotetramer,a putative Ca2+ modulator maturing human hair cuticle. But the PADI3 protein was less highly expressed in body side skin compared to the control in our study. This interesting contradiction remains to be further elucidated. - + 51702 Gene 7882 PADI3 - + 443369 Gene PADI3 - + 6274 Gene 2223 S100A3 - + 9606 Species human - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ @@ -16655,13 +16441,13 @@ paragraph 18474 FGFR3 RNA was detected in precuticle cells in the periphery of the hair bulb. But FGFR3 protein was less highly expressed in body side skin compared to the control in our study. - + 554324 Gene FGFR3 - + 554324 Gene @@ -16697,54 +16483,36 @@ paragraph 19536 For the Agilent microarray, the probe number of each gene is indefinite, from 1 to 5. There exists alternative splicing after gene transcription, so the inconsistency between different probes for the same gene is sometimes reasonable. Certainly the confidence is higher if all probes for the same gene were differentially expressed. For example, there are 4 probes for FGF7 and LOC443300(YWHAE) separately, of which 2 were both less highly expressed in S/G. So we speculated that the reliabilities of differential expression of FGF7 and LOC443300 were close to each other. While MMP2 differential expression was only detected in one of three probes, then its confidence might be lower than the former two genes (FGF7 and LOC443300). Nevertheless, because of the inadequate annotation of most probes, we found it difficult to calculate probe number of all the differentially expressed genes. - + 443095 Gene FGF7 - + 443300 Gene YWHAE - + 443095 Gene FGF7 - + 443115 Gene MMP2 - + 443095 Gene FGF7 - - MESH:C492399 - Chemical - - LOC443300 - - - MESH:C492399 - Chemical - - LOC443300 - - - MESH:C492399 - Chemical - - LOC443300 - DISCUSS @@ -16757,49 +16525,49 @@ paragraph 20441 Figure 5 listed some of the factors which possibly play roles in wool growth regulation. We speculate that lower temperature and shorter day photoperiod were key causes of the downregulation of wool growth rate. So we could speculate that those 8 genes (IL1A,FGF7,TAC1,IGFBP3,CDKN1B,IL6,FGF18 and PRL) played roles in wool growth regulation during winter. - + 443404 Gene IL1A - + 443095 Gene FGF7 - + 100034669 Gene TAC1 - + 100286737 Gene IGFBP3 - + 100127219 Gene CDKN1B - + 443406 Gene IL6 - + 780460 Gene FGF18 - + 443317 Gene @@ -16817,7 +16585,7 @@ paragraph 20810 In summary, the data presented in this study suggested that the body side skin displays a differentially expressed pattern in comparison with the groin skin at December time point. The majority of these genes possibly related to the wool growth control, and they could be assigned into the categories including regulation of receptor binding, multicellular organismal process, protein binding and macromolecular complex. Several potential gene families might participate in hair growth regulation, including fibroblast growth factors, transforming growth factor-beta, insulin-like growth factor, and so on. Proteomic analysis also identified hundreds of differentially expressed proteins. This systematic analysis could lead to a better understanding of the wool growth control mechanism in Aohan fine wool sheep. - + 9940 Species @@ -16847,19 +16615,19 @@ paragraph 21889 Sampling methods were described previously. One ram and two ewes of 16-month-old Aohan fine wool sheep were used in the microarray study. These animals were half sibs (sharing the same father). In December 2010, two areas of full-thickness skin were sampled from the same animal under local anaesthesia: body side skin (wool bearing) and groin skin (non-wool bearing) for microarray and proteomic experiments. The area of each sample was about 1 cm2. All samples were immediately put into collection tubes and stored in liquid nitrogen for RNA and protein extraction. A total of 15, 208 probes were spotted on this Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA). - + 9940 Species sheep - + 9940 Species Sheep - + MESH:D009584 Chemical @@ -16877,13 +16645,25 @@ paragraph 22614 TRIzol (Invitrogen) was used for total RNA extractions according to the manufacturer's protocol. RNA quantity and quality were measured by NanoDrop ND-1000. Its OD260/OD280 ratio was confirmed to be higher than 1.8. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. The RNA samples were sent to Kangchen Biotechnology Limited Company (Shanghai, China) for hybridization to the Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA). Each RNA sample was hybridized to one microarray slide. 1 mug of total RNA from each sample was amplified and transcribed into fluorescent cRNA with using the manufacturer's Agilent's Quick Amp Labeling protocol (version 5.7, Agilent Technologies). The labeled cRNAs were hybridized onto the Whole Genome Oligo Array (4x44K, Agilent Technologies). - + 9940 Species Sheep - + + MESH:D012685 + Chemical + + agarose + + + MESH:D000249 + Chemical + + Amp + + MESH:C411644 Chemical @@ -16901,13 +16681,13 @@ paragraph 23466 After hybridization and washing, the microarray slides were scanned with the Agilent Scanner G2505B. The resulting text files extracted from Agilent Feature Extraction Software (version 10.5.1.1) were imported into the Agilent GeneSpring GX software (version 11.0) for further analysis. Quantile normalization, probe annotation and subsequent data processing were performed using the GeneSpring GX v11.0 software package (Agilent Technologies). After Quantile normalization of the raw data, genes that at least 1 out of 6 samples have flags in Present ("All Targets Value") were chosen for differentially expressed genes screening. Gene expression levels were quantified relative to the expression of GAPDH. Differentially expressed genes were identified through fold-change screening. The fold-change of 2.0 and a false discovery rate of approximately 5% were set as the threshold. All data have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE62552 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc). Clustering analysis of all differentially expressed genes was performed using Cluster 3.0 to analyze the similarity in the expression patterns among different skin sites. The functional annotation of differentially expressed genes was performed by the DAVID (The Database for Annotation, Visualization and Integrated Discovery) gene annotation tool (http://david.abcc.ncifcrf.gov/). The KEGG pathway analysis was done manually (http://www.genome.jp/kegg/). - + 443005 Gene GAPDH - + 34205 Species @@ -16937,7 +16717,7 @@ table 25488 bRNA levels of GAPDH was assayed for normalization during quantitative PCR. - + 443005 Gene @@ -16959,79 +16739,67 @@ Tab1.xml 25601 Gene Primer sequence (5'-3') Tm ( C) a Target size (bp) GAPDHb Forward: ATGCCTCCTGCACCACCA 60 76 Reverse: AGTCCCTCCACGATGCCAA FGF10 Forward: GATCCGAGAAAGGAGCGAGG 60 554 Reverse: TCCAGGATACTGTACGGGCA LOC443300 Forward: ACCAACACATCCCATTCGCT 60 140 Reverse: CACTCAGCGTGTCCAGTTCT FGF18 Forward: AAGTCCGGATCAAGGGCAAG 60 98 Reverse: CACACACTCTTTGCTGGTGC Connexin43 Forward: GTCGTGTCGTTGGTGTCTCT 60 291 Reverse: CACTCAGCGTGTCCAGTTCT SCD Forward: AAGAGTGGCTGAGTTTCTGGTC 60 277 Reverse: GAAAGGAAGGTGATAGGGACAA Zo1 Forward: AGATAGCCCTGCAGCCAAAG 60 117 Reverse: GGGAGGTCAAGCAGGAAGAG MMP2 Forward: AACGCCATCCCTGATAACCT 60 126 Reverse: GCTTCCGAACTTCACGCTC ITGB1 Forward: AGCACGGATGAGGTGAACAG 60 407 Reverse: CCAAGGCAGGTCTGACACAT PAG11 Forward: AGCGTCGCCTACGAATCTG 60 120 Reverse: CTCAAACCCATATTCCGTCACA CRYAB Forward: CACCCAGCTGGATTGACACT 60 147 Reverse: CCTCATGTTTGCCATGCACC - + 443074 Gene FGF10 - + 780460 Gene FGF18 - + 443455 Gene Connexin43 - + 443115 Gene MMP2 - + 443141 Gene ITGB1 - + 100034671 Gene PAG11 - + 497274 Gene CRYAB - + 443200 Gene Zo1 - + 443185 Gene SCD - - MESH:C492399 - Chemical - - LOC443300 - - - MESH:D013932 - Chemical - - (5'-3') Tm - METHODS paragraph 26479 The total RNA samples prepared for microarray analysis were also used for qPCR analysis. Reverse transcriptions were performed using RevertAid First Strand cDNA Synthesis Kit (MBI Fermentas, Vilnius, Lithuania) according to the manufacturer's protocols. The primers were designed with the Oligo 6.0 program. The primer sequences, melting temperatures and product sizes are shown in Table 1. Gene expression levels were quantified relative to the expression of GAPDH. - + 443005 Gene @@ -17049,12 +16817,36 @@ paragraph 26975 Lysis buffer preparation: 42% Urea, 15.2% Thiourea, 4% CHAPS, 1% DTT. Sampled tissues were homogenziated in lysis buffer (containing 1% cocktail and 2% IPG-buffer, added right before use) at the ratio of 1:7 (weight/volume). The tissues were cut into small pieces by ophthalmic scissors, and left at 4 C for 1 h, vortexed it every 15 min. Then, the tissue homogenate was centrifuge at 40,000 g for 30 min. Supernatants were collected and stored at -80 C. Protein concerntrations were determined by Bradford method. - + + MESH:C028213 + Chemical + + CHAPS + + + MESH:D004229 + Chemical + + DTT + + MESH:D013890 Chemical Thiourea + + MESH:D014508 + Chemical + + Urea + + + - + Chemical + + IPG + METHODS @@ -17067,11 +16859,11 @@ paragraph 27530 0.5% IPG-buffer was added into each 150 mug protein sample (in a final volume of 400-600 muL), and was loaded in the One-dimensional electrophoresis instrument. The progamme is as follows: Step-n-hold (S1, 30 V for 6 h; S2, 60 V for 6 h); Gradient (S3, 500 V for 1 h;S4,1000 V for 1 h; S5, 3000 V for 3 h; S6, 8000 V for 3 h); Step-n-hold (S7, 8000 V for 20 h). - - MESH:C012008 + + - Chemical - - S6 + + IPG @@ -17079,8 +16871,8 @@ title_2 27908 2-dimensional (2-D) SDS-PAGE preparation - - MESH:C032259 + + MESH:D012967 Chemical SDS @@ -17091,50 +16883,74 @@ paragraph 27949 Tris-HCl (PH = 8.8), Monomer storage (30% Acrylamid and 0.8% NN'-methy lenebisacry lamid), 10 x electrophoresis buffer (3.03% Tris-Base, 14.4% Glycine, 1% SDS), balanced solution (36.05% Urea, 5% Tris-HCl, 2% SDS, 34.5% Glycerine). - - MESH:C032259 - Chemical - - SDS - - + MESH:D005990 Chemical Glycerine - - MESH:C014843 + + MESH:D005998 + Chemical + + Glycine + + + MESH:D014325 + Chemical + + Tris-Base + + + - Chemical Tris-HCl + + MESH:D012967 + Chemical + + SDS + - MESH:D005998 + - Chemical - - Glycine + + NN'-methy lenebisacry lamid - MESH:C032259 + MESH:D012967 Chemical SDS - MESH:C014843 + MESH:D020106 + Chemical + + Acrylamid + + + - Chemical Tris-HCl + + MESH:D014508 + Chemical + + Urea + METHODS title_2 28196 The second dimensional SDS-PAGE - - MESH:C032259 + + MESH:D012967 Chemical SDS @@ -17157,13 +16973,13 @@ paragraph 28351 Fixative preparation: 40% Ethanol and 10% Acetic acid. - + MESH:D019342 Chemical Acetic acid - + MESH:D000431 Chemical @@ -17193,12 +17009,6 @@ title_2 28628 Staining and visualization - - MESH:D014786 - Disease - - visualization - METHODS @@ -17206,7 +17016,7 @@ 28655 Sensitizing solution preparation: 30% Ethanol, 0.314% Na2S2O3, 6.8%NaAc. - MESH:D012964 + - Chemical Na2S2O3 @@ -17218,7 +17028,7 @@ Ethanol - MESH:D012964 + - Chemical NaAc @@ -17236,7 +17046,7 @@ 28779 Washing: the gels were washed for three times using ddH2O. Ten minutes each time. - MESH:C062226 + - Chemical ddH2O @@ -17247,29 +17057,23 @@ paragraph 28861 Ilver staining: Silver staining solution was prepared (1.25 AgNO3 and 200 muL Formaldehyde in 500 mL ddH2O). The gels were stained by the solution for 20 min. - - MESH:C062226 + + MESH:D012835 Chemical - - ddH2O + + AgNO3 - + MESH:D005557 Chemical Formaldehyde - - MESH:D012834 - Chemical - - Silver - - - MESH:D012834 + + - Chemical - - AgNO3 + + ddH2O @@ -17277,8 +17081,8 @@ paragraph 29023 Washing: the gels were washed for 2 min using ddH2O. - - MESH:C062226 + + - Chemical ddH2O @@ -17289,19 +17093,19 @@ paragraph 29077 Visualizing solution preparation: 12.5 g Na2CO3 and 100 muL Formaldehyde in 500 mL ddH2O. - - MESH:D012964 + + MESH:C005686 Chemical Na2CO3 - - MESH:C062226 + + - Chemical ddH2O - + MESH:D005557 Chemical @@ -17313,13 +17117,13 @@ paragraph 29170 Termination solution preparation: 2 g Glycine in 50 ml ddH2O. - - MESH:C062226 + + - Chemical ddH2O - + MESH:D005998 Chemical @@ -17343,18 +17147,6 @@ paragraph 29356 Gels were then scanned and analyzed using ImageMaster TM 2D platinum software (Version 5.0, GE Healthcare, San Francisco, CA, USA). The expression level was determined by the relative volume of each spot in the gel and expressed as %Vol (%Vol = [spot volume/Sigmavolumes of all spots resolved in the gel]). The means and standard deviations of both sample groups were calculated. Statistical significance with Student's t-tests using ImageMaster TM 2D platinum software. P values <0.05 were considered statistically significant. - - MESH:D010984 - Chemical - - platinum - - - MESH:D010984 - Chemical - - platinum - METHODS @@ -17367,35 +17159,19 @@ paragraph 29966 Protein spots with significant differences between the two groups were excised, dehydrated in acetonitrile, and dried at room temperature. Gel pieces were denatured, alkylated, trypsin digested and analyzed by an Ultraflex II MALDI-TOF-TOF mass spectrometer (Bruker Daltonics GmbH, Bremen, Germany) under the control of FlexControl TM 2.4 software (Bruker Daltonics GmbH). Acquired peptide mass fingerprint (PMF) were processed using the software FlexAnalysis 3.0 (Bruker Daltonics, Bremen, Germany). The peak detection algorithm was: SNAP (Sort Neaten Assign and Place); S/N threshold: 1.5; Quality Factor Threshold: 50. The tryptic auto-digestion ion picks (trypsin [108-115] 842.5094 Da, trypsin [58-77] 2211.104 Da) were used as internal standards. The resulting peptide mass lists were used to search the Matrixscience database (http://www.matrixscience.com). The following search parameter criteria were used: mass tolerance 100 ppm, miss cleavage 1, modification comprises carbamidomethyl and methionine oxidation. Matched peptides number between experimental PMF and theoretical PMF 5. All MS data have been deposited in PeptideAtlas and are accessible through Dataset Identifier PASS00597 (http://www.peptideatlas.org/PASS/PASS00597). - - Chemical - - PASS00597 - - - MESH:D009584 - Chemical - - N - - - MESH:C018860 + + - Chemical carbamidomethyl - + MESH:C032159 Chemical acetonitrile - - Chemical - - PASS00597 - - + MESH:D008715 Chemical @@ -17419,16 +17195,6 @@ paragraph 31463 Proteomic data have been deposited in PeptideAtlas and are accessible through Dataset Identifier PASS00597 (http://www.peptideatlas.org/PASS/PASS00597). - - Chemical - - PASS00597 - - - Chemical - - PASS00597 - SUPPL @@ -17465,13 +17231,13 @@ footnote 31793 JZ and HL carried out data collection and analysis and manuscript preparation. KL carried out data analysis. JY participated in data analysis. MC, participated in data collection. WD participated in data discussion and manuscript preparation. RB participated in manuscript preparation. JL and GH participated in discussion of the study. NL participated in the design of the study, coordination, data analysis and manuscript preparation. All authors read and approved the final manuscript. - + CVCL:2492 CellLine HL - + MESH:D006527 Disease @@ -17516,11 +17282,6 @@ surname:Lenane;given-names:I 32363 A genome scan for QTL affecting fleece and wool traits in Merino sheep - - Chemical - - Merino - surname:Cano;given-names:EM @@ -17540,11 +17301,6 @@ surname:Taddeo;given-names:HR 32434 Wool quantitative trait loci in Merino sheep - - Chemical - - Merino - surname:Bidinost;given-names:F @@ -17567,12 +17323,6 @@ surname:Abad;given-names:M 32479 QTL affecting fleece traits in Angora goats - - MESH:D015511 - Disease - - Angora goats - surname:McKenzie;given-names:GW @@ -17664,7 +17414,7 @@ 1949 32795 Lhx2 maintains stem cell character in hair follicles - + 101108898 Gene @@ -17684,7 +17434,7 @@ 22 32848 Gene expression profiling gets to the root of human hair follicle stem cells - + 9606 Species @@ -17712,7 +17462,7 @@ surname:Pise-Masison;given-names:CA 32925 Characterization and isolation of stem cell-enriched human hair follicle bulge cells - + 9606 Species @@ -17739,13 +17489,13 @@ surname:Greer;given-names:GJ 33010 Gene expression profiling of naive sheep genetically resistant and susceptible to gastrointestinal nematodes - + 9940 Species sheep - + MESH:D009349 Disease @@ -17768,17 +17518,12 @@ 220 33119 Microarray analysis reveals difference in gene expression profiles of hair and wool sheep infected with Haemonchus contortus - + MESH:D012757 Disease sheep infected - - Disease - - Haemonchus contortus - surname:Bueno;given-names:S @@ -17798,7 +17543,7 @@ surname:Valentini;given-names:A 33244 A tool for sheep product quality: custom microarrays from public databases - + 9940 Species @@ -17826,7 +17571,7 @@ surname:Aubert;given-names:J 33319 Terminal differentiation of goat mammary tissue during pregnancy requires the expression of genes involved in immune functions - + 9925 Species @@ -17849,18 +17594,12 @@ surname:Leroux;given-names:C 33446 Mammary transcriptome analysis of food-deprived lactating goats highlights genes involved in milk secretion and programmed cell death - + 9925 Species goats - - MESH:D003643 - Disease - - death - surname:Cam;given-names:GR @@ -17878,7 +17617,7 @@ surname:Reverter;given-names:A 33580 Gene expression profiling of ovine skin and wool follicle development using a combined ovine-bovine skin cDNA microarray - + 9913 Species @@ -17906,23 +17645,18 @@ surname:McWilliam;given-names:SM 33701 A genomics-informed, SNP association study reveals FBLN1 and FABP4 as contributing to resistance to fleece rot in Australian Merino sheep - + 100037704 Gene FBLN1 - + 100137067 Gene FABP4 - - Chemical - - Merino - surname:Robello;given-names:C @@ -17941,12 +17675,6 @@ surname:Urioste;given-names:JI 33839 Gene expression analysis identifies new candidate genes associated with the development of black skin spots in Corriedale sheep - - MESH:D012757 - Disease - - Corriedale sheep - surname:Yashizawa;given-names:M @@ -17964,24 +17692,24 @@ 396 33967 A subset of skin-expressed microRNAs with possible roles in goat and sheep hair growth based on expression profiling of mammalian microRNAs - + 9925 Species goat - - 9940 - Species - - sheep - - + 9606 Species mammalian + + MESH:D012757 + Disease + + sheep hair growth + surname:Nixon;given-names:AJ @@ -18022,18 +17750,6 @@ ref 34234 Liu N, Li H, Liu K, Yu J, Cheng M, De W, Liu J, Shi S, He Y, Zhao J: Differential expression of genes and proteins associated with wool follicle cycling.Mol Biol Rep 2014, published online in advance of the print edition. - - MESH:D008094 - Chemical - - Li H - - - MESH:D009584 - Chemical - - N - surname:Talbot;given-names:R @@ -18078,17 +17794,17 @@ surname:Maddox;given-names:JF 34456 The sheep genome illuminates biology of the rumen and lipid metabolism - + 9940 Species sheep - - MESH:D052439 - Disease - - lipid metabolism + + MESH:D008055 + Chemical + + lipid @@ -18200,25 +17916,31 @@ surname:Sullivan;given-names:R 34906 Skin-specific deletion of stearoyl-CoA desaturase-1 alters skin lipid composition and protects mice from high fat diet-induced obesity - + 20249 Gene 74538 stearoyl-CoA desaturase-1 - + 10090 Species mice - + MESH:D009765 Disease obesity + + MESH:D008055 + Chemical + + lipid + 114 @@ -18235,14 +17957,14 @@ 375 35041 Basement membrane zone remodeling during appendageal development in human fetal skin: the absence of type VII collagen is associated with gelatinase-A (MMP2) activity - + 4313 Gene 3329 MMP2 - + 9606 Species @@ -18268,12 +17990,18 @@ surname:Villa;given-names:AM 35208 Thymosin beta4 increases hair growth by activation of hair follicle stem cells - + 101106384 Gene Thymosin beta4 + + MESH:D006130 + Disease + + increases hair growth + 73 @@ -18289,7 +18017,7 @@ 724 35287 Separation of human epidermal stem cells from transit amplifying cells on the basis of differences in integrin function and expression - + 9606 Species @@ -18316,14 +18044,14 @@ surname:Humphries;given-names:MJ 35422 Functional role of beta 1 integrin-mediated signalling in the human hair follicle - + 3688 Gene 22999 beta 1 integrin - + 9606 Species @@ -18354,7 +18082,7 @@ surname:Jorcano;given-names:JL 35504 Skin and hair follicle integrity is crucially dependent on beta 1 integrin expression on keratinocytes - + 3688 Gene 22999 @@ -18380,7 +18108,7 @@ surname:Brown;given-names:K 35607 Illuminating role of CYP1A1 in skin function - + 100170113 Gene @@ -18446,7 +18174,7 @@ surname:Bensinger;given-names:A 35728 Genome-wide atlas of gene expression in the adult mouse brain - + 10090 Species @@ -18480,14 +18208,14 @@ surname:Lehman;given-names:K 35790 BGEM: an in situ hybridization database of gene expression in the embryonic and adult mouse nervous system - + 10090 Species mouse - - MESH:D018236 + + MESH:D009373 Disease embryonic @@ -18510,6 +18238,12 @@ surname:Jahoda;given-names:CA 35897 Modelling the hair follicle dermal papilla using spheroid cell cultures + + MESH:D010211 + Disease + + Modelling the hair follicle dermal papilla + surname:Laird;given-names:DW @@ -18527,32 +18261,38 @@ 2204 35969 The G60S connexin43 mutant regulates hair growth and hair fiber morphology in a mouse model of human oculodentodigital dysplasia - + 14609 Gene 136 connexin43 - + 10090 Species mouse - + 9606 Species human - + p.G60S ProteinMutation G60S - + + MESH:D006201 + Disease + + hair growth and hair fiber morphology + + MESH:C563160 Disease @@ -18575,13 +18315,13 @@ 769 36098 Prostaglandin metabolism in human hair follicle - + 9606 Species human - + MESH:D011453 Chemical @@ -18603,7 +18343,7 @@ S202 36146 Prostaglandin-induced hair growth - + MESH:D011453 Chemical @@ -18623,7 +18363,7 @@ 1977 36180 Time-course expression profiles of hair cycle-associated genes in male mini rats after depilation of telogen-phase hairs - + 10116 Species @@ -18644,7 +18384,7 @@ 177 36301 Characterization of a hair (wool) keratin intermediate filament gene domain - + 100141298 Gene @@ -18671,7 +18411,7 @@ surname:Rogers;given-names:MA 36377 Expression patterns of keratin intermediate filament and keratin associated protein genes in wool follicles - + 100141298 Gene @@ -18694,24 +18434,24 @@ 343 36485 A case of dermatopathia pigmentosa reticularis with wiry scalp hair and digital fibromatosis resulting from a recurrent KRT14 mutation - + 100303604 Gene KRT14 - + + MESH:C563765 + Disease + + wiry scalp hair and digital fibromatosis + + MESH:C535374 Disease dermatopathia pigmentosa reticularis - - MESH:D005350 - Disease - - digital fibromatosis - surname:Holland;given-names:K @@ -18739,24 +18479,24 @@ surname:Huson;given-names:S 36620 Naegeli-Franceschetti-Jadassohn syndrome and dermatopathia pigmentosa reticularis: two allelic ectodermal dysplasias caused by dominant mutations in KRT14 - + MESH:C538331 Disease Naegeli-Franceschetti-Jadassohn syndrome - + + MESH:D004476 + Disease + + allelic ectodermal dysplasias + + MESH:C535374 Disease dermatopathia pigmentosa reticularis - - MESH:D004476 - Disease - - ectodermal dysplasias - surname:Lynch;given-names:PJ @@ -18772,6 +18512,12 @@ ref 36775 Dowling-Degos disease involving the vulva and back: case report and review of the literature + + MESH:C562924 + Disease + + Dowling-Degos disease + surname:Kleinert;given-names:P @@ -18791,21 +18537,21 @@ surname:Mochida;given-names:U 36868 Specific citrullination causes assembly of a globular S100A3 homotetramer: a putative Ca2+ modulator matures human hair cuticle - + 6274 Gene 2223 S100A3 - + 9606 Species human - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ @@ -18835,12 +18581,6 @@ surname:Stieler;given-names:KM 36996 Profiling of alopecia areata autoantigens based on protein microarray technology - - MESH:D000506 - Disease - - alopecia areata - 205 @@ -18856,7 +18596,7 @@ 386 37077 Fibroblast growth factor signalling in the hair growth cycle: expression of the fibroblast growth factor receptor and ligand genes in the murine hair follicle - + 10090 Species @@ -18893,7 +18633,7 @@ surname:Chen;given-names:Y 37236 Molecular characteristics of non-small cell lung cancer - + MESH:D002289 Disease @@ -18940,12 +18680,6 @@ surname:Gao;given-names:W 37385 DAVID: database for annotation, visualization, and integrated discovery - - MESH:D014786 - Disease - - visualization - 20 @@ -19024,25 +18758,31 @@ abstract 87 Proctitis after radiation therapy for prostate cancer remains an ongoing clinical challenge and critical quality of life issue. SBRT could minimize rectal toxicity by reducing the volume of rectum receiving high radiation doses and offers the potential radiobiologic benefits of hypofractionation. This study sought to evaluate the incidence and severity of proctitis following SBRT for prostate cancer. - + + MESH:D011349 + Disease + + proctitis + + MESH:D064420 Disease toxicity - + MESH:D011471 Disease prostate cancer - + MESH:D011471 Disease prostate cancer - + MESH:D011349 Disease @@ -19060,42 +18800,48 @@ abstract 499 Between February 2008 and July 2011, 269 men with clinically localized prostate cancer were treated definitively with SBRT monotherapy at Georgetown University Hospital. All patients were treated to 35-36.25Gy in 5 fractions delivered with the CyberKnife Radiosurgical System (Accuray). Rectal bleeding was recorded and scored using the CTCAE v.4. Telangiectasias were graded using the Vienna Rectoscopy Score (VRS). Proctitis was assessed via the Bowel domain of the Expanded Prostate Index Composite (EPIC)-26 at baseline and at 1, 3, 6, 9, 12, 18 and 24 months post-SBRT. - + 9606 Species men - + 9606 Species patients - + MESH:D011471 Disease - - prostate cancer + + localized prostate cancer - + MESH:D011349 Disease Proctitis - - MESH:D012002 + + MESH:D006470 Disease Rectal bleeding - + MESH:D013684 Disease Telangiectasias + + - + Chemical + + SBRT + ABSTRACT @@ -19108,36 +18854,60 @@ abstract 1083 The median age was 69 years with a median prostate volume of 39 cc. The median follow-up was 3.9 years with a minimum follow-up of two years. The 2-year actuarial incidence of late rectal bleeding >= grade 2 was 1.5%. Endoscopy revealed VRS Grade 2 rectal telangiectasias in 11% of patients. All proctitis symptoms increased at one month post-SBRT but returned to near-baseline with longer follow-up. The most bothersome symptoms were bowel urgency and frequency. At one month post-SBRT, 11.2% and 8.5% of patients reported a moderate to big problem with bowel urgency and frequency, respectively. The EPIC bowel summary scores declined transiently at 1 month and experienced a second, more protracted decline between 6 months and 18 months before returning to near-baseline at two years post-SBRT. Prior to treatment, 4.1% of men felt their bowel function was a moderate to big problem which increased to 11.5% one month post-SBRT but returned to near-baseline at two years post-SBRT. - + 9606 Species patients - + 9606 Species patients - + 9606 Species men - - MESH:D012002 + + MESH:D011349 + Disease + + proctitis + + + MESH:D006470 Disease late rectal bleeding - + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel urgency + + MESH:D013684 Disease telangiectasias + + MESH:D015212 + Disease + + bowel function + ABSTRACT @@ -19150,12 +18920,18 @@ abstract 2082 In this single institution cohort, the rate and severity of proctitis observed following SBRT is low. QOL decreased on follow-up; however, our results compare favorably to those reported for patients treated with alternative radiation modalities. Future prospective randomized studies are needed to confirm these observations. - + 9606 Species patients + + MESH:D011349 + Disease + + proctitis + INTRO @@ -19168,167 +18944,221 @@ paragraph 2420 Post-treatment bowel function is a primary determinant of quality of life following radiation therapy for prostate cancer. Late proctitis occurs at a frequency of 5-20% with conventionally fractionated treatment. The incidence of proctitis is dependent on the assessment method and whether it is patient or physician reported. Patients with radiation-induced proctitis report increased bowel frequency/urgency, incontinence, bleeding and pain. These symptoms occur months to years after treatment with the large majority of patients reporting symptoms within two years following radiation therapy. Patient characteristics such as age, comorbidities, hemorrhoids, inflammatory bowel disease and/or anticoagulation may increase an individual's risk of radiation-induced proctitis. Unfortunately, treatment options for radiation proctitis are limited and of unclear clinical benefit. - + 9606 Species patient - + 9606 Species Patients - + 9606 Species patients - + 9606 Species Patient - - MESH:D010146 - Disease - - pain - - - MESH:D014549 - Disease - - incontinence - - - MESH:D011349 - Disease - - radiation proctitis - - + MESH:D015212 Disease inflammatory bowel disease - + MESH:D006484 Disease hemorrhoids - + MESH:D011471 Disease prostate cancer - + MESH:D006470 Disease bleeding + + MESH:D011349 + Disease + + proctitis + + + MESH:D015212 + Disease + + bowel function + + + MESH:D011349 + Disease + + proctitis + + + MESH:D014549 + Disease + + incontinence + + + MESH:D010146 + Disease + + pain + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis + INTRO paragraph 3305 Proctitis is the principle dose-limiting toxicity of prostate radiotherapy. Acute proctitis has been attributed to radiation-induced injury to the epithelial lining of the rectal mucosa, leading to mucosal sloughing, acute inflammatory infiltrates, and vascular permeability leading to edema. These early changes are associated with bowel frequency, bowel urgency and rectal pain. Chronic proctitis results from epithelial atrophy and obliterative endarteritis, resulting in tissue ischemia, rectal fibrosis, stricture formation, and neovascularization; this presents clinically with bowel frequency, urgency and rectal bleeding. Historically, endoscopic findings in patients with proctitis include telangiectasia, congested mucosa, and ulcers. Telangiectasias are observed in 20-80% of patients receiving conventionally fractionated radiation therapy. - + 9606 Species patients - + 9606 Species patients - - MESH:D011349 - Disease - - Chronic proctitis - - - MESH:D014456 - Disease - - ulcers - - - MESH:D004692 + + MESH:D010146 Disease - - obliterative endarteritis + + pain - - MESH:D011349 + + MESH:D058186 Disease - - Proctitis + + injury - + MESH:D013684 Disease telangiectasia - + + MESH:D011349 + Disease + + proctitis + + MESH:D005355 Disease fibrosis - + + MESH:D015212 + Disease + + bowel urgency + + MESH:D064420 Disease toxicity - - MESH:C563475 - Disease - - rectal pain - - - MESH:D012002 - Disease - - rectal bleeding - - + MESH:D007511 Disease ischemia - - MESH:D013684 + + MESH:D006470 Disease - - Telangiectasias + + bleeding - + + MESH:D002277 + Disease + + epithelial atrophy + + MESH:D004487 Disease edema - - MESH:D001284 + + MESH:D011349 Disease - - atrophy + + proctitis + + + MESH:D011349 + Disease + + Chronic proctitis + + + MESH:D014456 + Disease + + ulcers + + + MESH:D011349 + Disease + + Proctitis + + + MESH:D004692 + Disease + + endarteritis + + + MESH:D013684 + Disease + + Telangiectasias @@ -19336,19 +19166,25 @@ paragraph 4158 The risk of proctitis appears to be dependent upon both the total radiation dose and the volume of the rectum in the high dose area. Technical factors such as treatment of the proximal seminal vesicles and expansion of planning target volume (PTV) to compensate for intra-fraction prostate motion may contribute to the severity of rectal toxicities. To minimize bowel toxicity, it is currently recommended that the volume of the rectum receiving > 75 Gray (Gy) be limited to < 10% with conventionally fractionated radiation therapy. Studies have shown that advanced radiation technologies such as intensity modulated radiation therapy (IMRT) may decrease the dose to the rectum and potentially decrease proctitis. - + MESH:D012002 Disease rectal toxicities - - MESH:D011472 + + MESH:D011349 Disease - - intra-fraction prostate motion + + proctitis - + + MESH:D011349 + Disease + + proctitis + + MESH:D015212 Disease @@ -19360,25 +19196,31 @@ paragraph 4880 Recent data suggest that large radiation fraction sizes are radiobiologically favorable over lower fraction sizes in prostate cancer. The alpha/beta for prostate cancer may be as low as 1.5 Gy. If the alpha/beta for prostate cancer is less than 3-5 Gy, which is generally the value accepted for late rectal complications, the linear-quadratic model predicts that delivering large radiation fraction sizes will result in improved local control with a similar rate of bowel complications. Therefore, using large fraction sizes in SBRT provides an attractive modality to leverage the potential radiobiologic benefits of hypofractionation with the minimal invasiveness of an external-beam treatment modality. Furthermore, by reducing the number of treatment visits, hypofractionated therapy can also be logistically favorable for patients and reduce financial burden, ultimately decreasing barriers to care. - + 9606 Species patients - + + MESH:D015212 + Disease + + bowel complications + + MESH:D011471 Disease prostate cancer - + MESH:D011471 Disease prostate cancer - + MESH:D011471 Disease @@ -19390,24 +19232,24 @@ paragraph 5784 The safety and efficacy of SBRT in the treatment of clinically localized prostate cancer has been established in a number of studies. The use of large fraction sizes in SBRT offers the potential radiobiological benefits of hypofractionation and potentially may minimize radiation proctitis by reducing the volume of the rectum receiving high radiation doses. The CyberKnife robotic radiosurgical system uses image guidance to track implanted fiducials to account for intrafraction prostatic motion. This reduces the uncertainty of the location of the prostate and allows treatment to be delivered with a smaller PTV expansion, which may reduce the doses delivered to the anterior rectal wall. The goal of this study is to report the bowel outcomes following SBRT for clinically localized prostate cancer. - + + MESH:D011349 + Disease + + proctitis + + MESH:D011471 Disease localized prostate cancer - + MESH:D011471 Disease localized prostate cancer - - MESH:D011349 - Disease - - radiation proctitis - METHODS @@ -19420,7 +19262,7 @@ title_2 6597 Patient selection - + 9606 Species @@ -19432,13 +19274,13 @@ paragraph 6615 Patients eligible for study inclusion had histologically-confirmed prostate cancer treated per our institutional protocol. Clinical stage was defined according to the 6th edition of the American Joint Committee on Cancer criteria. Risk groups were defined using the D'Amico criteria. Internal Review Board (IRB)-approval was obtained for retrospective review of data that was prospectively collected in our institutional database. - + 9606 Species Patients - + MESH:D011471 Disease @@ -19456,43 +19298,49 @@ paragraph 7085 SBRT was delivered using the CyberKnife robotic radiosurgical system (Accuray Inc., Sunnyvale, CA). The fiducial placement and computed tomography (CT)/magnetic resonance (MR) simulation procedures have been previously described in Lei et al.. The clinical target volume (CTV) was defined as the prostatic capsule and proximal seminal vesicles (to the point where the seminal vesicles separate). To cover areas of potential ECE, the expansion from the CTV to the PTV was 5 mm in all directions except 3 mm posteriorly into the rectum. There was no difference in the target volume delineation for different risk groups. Fiducial-based tracking was used to account for interfraction and intrafraction prostate motion. Treatment planning was performed using Multiplan (Accuray Inc., Sunnyvale, CA). Patients were treated with 35 or 36.25 Gy of radiotherapy delivered in 5 fractions of 7-7.25 Gy each to the PTV, administered over a timespan of 1-2 weeks. This corresponds to a tumor equivalent dose in 2-Gy fractions (EQD2) of approximately 85-90 Gy assuming an alpha/beta ratio of 1.5. The rectum was contoured as a solid structure from the anus (at the level of the ischial tuberosities) to the rectosigmoid flexure and evaluated with dose-volume histogram analysis during treatment planning using Multiplan (Accuray Inc., Sunnyvale, CA) inverse treatment planning. Rectal volume receiving 36 Gy was limited to <= 1 cc. Assuming an alpha/beta of 3 Gy for late bowel complications, this is biologically equivalent to approximately 74 Gy administered in 2 Gy fractions. The rectal dose-volume histogram (DVH) goals were <50% rectal volume receiving 50% of the prescribed dose, <20% receiving 80% of the dose, <10% receiving 90% of the dose, and < 5% receiving 100% of the dose. Typical dose distributions have been previously described. Patients were placed on a low-residual diet and given enemas prior to simulation and treatment delivery to maximize the potential distance between the prostate and the rectal wall and minimize intrafraction prostate motion. - + 1889 Gene 1068 ECE - + 9606 Species Patients - + 9606 Species Patients - + + MESH:D015212 + Disease + + late bowel complications + + + - + Chemical + + PTV + + MESH:D009369 Disease tumor - + MESH:D014402 Disease ischial tuberosities - - MESH:C564570 - Disease - - MR - METHODS @@ -19505,71 +19353,77 @@ paragraph 9189 Rectal bleeding was prospectively documented at follow-up visits using the National Cancer Institute (NCI) Common Toxicity Criteria (CTC) version 4.0. Acute rectal bleeding was defined as experiencing toxicity within 6 months of radiation therapy. Late rectal bleeding was defined as occurring at least 6 months after delivery of radiation therapy. Grade 1 represents minimal bleeding not requiring medications for symptom control. Grade 2 indicates rectal bleeding requiring new medication (i.e. steroid suppository) or minor laser coagulation. Telangiectasia were graded using the Vienna Rectoscopy Score (VRS): Grade 1 (a single telangiectasia), Grade 2 (multiple non-confluent telangiectasia) and Grade 3 (multiple confluent telangiectasia). - + MESH:D064420 Disease toxicity - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D013684 Disease telangiectasia - + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + MESH:D013684 Disease telangiectasia - + MESH:D064420 Disease - - Toxicity + + Toxicity Criteria - + MESH:D006470 Disease bleeding - - MESH:D012002 - Disease - - Rectal bleeding - - + MESH:D013684 Disease Telangiectasia - - MESH:D013684 - Disease - - telangiectasia + + MESH:D013256 + Chemical + + steroid - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + Rectal bleeding - - MESH:D012002 + + MESH:D013684 Disease - - rectal bleeding + + telangiectasia @@ -19577,7 +19431,7 @@ paragraph 9937 Proctitis was assessed via the bowel domain of the Expanded Prostate Index Composite (EPIC)-26 at baseline and at 1, 3, 6, 9, 12, 18 and 24 months. The timing of the quality of life (QOL) assessments was relative to the last day of SBRT treatment. The EPIC-26 bowel domain includes five questions related to individual symptoms (questions 6a-e: urgency, frequency, pain, bloody stool, incontinence) and one question (question 7) related to overall bother (degree of interference or annoyance caused by bowel symptoms. - + MESH:D011349 Disease @@ -19589,18 +19443,30 @@ paragraph 10456 Wilcoxon signed-rank test and chi-square analysis were used to assess differences in ongoing quality of life scores in comparison to baseline. For each EPIC question, the responses were grouped into three clinically relevant categories (no problem, very small-small problem and moderate to big problem). To statistically compare changes between time points, the levels of responses were assigned a score and the significance of the mean changes in the scores was assessed by Wilcoxon rank test. EPIC scores for the bowel domain and its individual questions range from 0 - 100 with lower values representing worsening bowel symptoms. The minimally important difference (MID) in EPIC score was defined as a change of one-half standard deviation (SD) from the baseline. To limit the effect of attrition bias, statistical analysis was limited to time points in which >= 80% of the patient data were available. - + 9606 Species patient - + Disease MID - + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D019217 + Disease + + attrition bias + + MESH:D029461 Disease @@ -19620,7 +19486,7 @@ table_caption 11375 Baseline patient characteristics and treatment - + 9606 Species @@ -19634,7 +19500,7 @@ Tab1.xml 11424 % Patients (N = 269) Age (y/o) Median 69 (44-90) <60 8.20% 60-69 42.40% 70-79 41.30% >=80 8.20% Race White 55.80% Black 37.20% Other 7.10% Prostate volume (cc) Median 39.04 (11.56-138.69) Charlson comorbidity index (CCI) 0 66.9% 1 22.7% 2 6.7% >=3 3.7% Body mass index BMI <25 21.9% 25 <= BMI < 30 47.2% 30 <= BMI < 35 21.9% BMI >=35 8.9% Use of anticoagulants Yes 35.3% No 64.7% Risk group Low 36.80% Intermediate 53.20% High 10.00% Hormonal therapy Yes 16.40% No 83.60% Dose 36.25 Gy 83.30% 35 Gy 16.40% Other 0.40% - + 9606 Species @@ -19646,37 +19512,37 @@ paragraph 12029 From February 2008 to July 2011, 269 prostate cancer patients were treated per our institutional SBRT monotherapy protocol (Table 1). The median follow-up was 3.9 years. The median age was 69 years and 44.2% were of non-Caucasian ancestry. Comorbidities were common with 35% were taking anticoagulants (including aspirin) prior to treatment. The median prostate volume was 39 cc. By D'Amico classification, 99 patients were low-, 143 intermediate-, and 27 high-risk. 16.4% also received androgen deprivation therapy (ADT) at the discretion of the treating Urologist. 83.3% of the patients were treated with 36.25 Gy in five 7.25 Gy fractions. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D000728 + + - Chemical - - androgen + + ADT - + MESH:D011471 Disease prostate cancer - + MESH:D001241 Chemical @@ -19688,79 +19554,73 @@ paragraph 12680 A total of 61 (22.7%) patients reported rectal bleeding after SBRT, with 28 (10.4%) patients reporting acute bleeding and 38 patients (14.1%) reporting late bleeding. Overall, Grade 2 acute and late rectal bleeding were observed in 0 (0%) and 4 (1.5%) of patients, respectively. There was no Grade 3 or higher rectal bleeds. The majority of acute rectal bleeding occurred within 1-month post-SBRT treatment (79%). The bleeding completely resolved in the majority of the patients by the subsequent follow-up visit. Likewise, the majority of late rectal bleeding was observed at one specific follow-up appointment and did not persist on subsequent follow-ups. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D012002 - Disease - - rectal bleeding - - + MESH:D006470 Disease - + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D006470 Disease - + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - late rectal bleeding + + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeds + + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D006470 Disease @@ -19772,73 +19632,73 @@ paragraph 13338 Seventy-three patients had one or more rectal endoscopies during the follow-up period. Endoscopy was performed as a part of normal colorectal cancer screening or if patient had clinically significant rectal bleeding. The median interval from completion of SBRT to endoscopy was 24 months (3 months to 53 months). On endoscopy, rectal telangiectasias were found in 8 (11.0%) patients. All 8 patients were observed to have multiple non-confluent telangiectasias (VRS Grade 2). No patient had confluent or circumferential telangiectasia. Minor laser coagulation was performed in three patients. No rectal ulcerations, strictures, or fistulas were observed. - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + MESH:D013684 Disease telangiectasia - + MESH:D013684 Disease telangiectasias - + MESH:D015179 Disease colorectal cancer - + MESH:D005402 Disease fistulas - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D013684 Disease @@ -19850,29 +19710,29 @@ table 13995 Patient-reported responses to EPIC-26 questions 6A (Urgency to have a bowel movement), 6B (Frequency of bowel movements), 6C (Losing control of your stools), 6D (Bloody stools) and 6E (Abdominal, pelvic or rectal pain). - + 9606 Species Patient - - MESH:C563475 + + MESH:D009069 Disease - - rectal pain + + bowel movements - - MESH:D002244 - Chemical - - 6C + + MESH:D009069 + Disease + + bowel movement - - MESH:D001895 - Chemical - - 6B + + MESH:D010146 + Disease + + pain @@ -19882,7 +19742,7 @@ table_caption 14215 Bowel symptoms following SBRT for prostate cancer - + MESH:D011471 Disease @@ -19896,7 +19756,7 @@ Tab2.xml 14267 Start 1 3 6 9 12 18 24 Bowel urgency No problem 77.2% 49.8% 62.0% 60.2% 64.5% 62.3% 64.7% 66.8% Very small-small problem 21.0% 39.0% 33.7% 34.1% 29.8% 31.6% 29.5% 31.1% Moderate-big problem 1.9% 11.2% 4.3% 5.6% 5.6% 6.1% 5.8% 2.1% p value <0.0001 0.0002 <0.0001 0.0002 0.0001 0.0004 0.01568 Bowel frequency No problem 82.0% 57.0% 71.9% 71.5% 70.6% 72.5% 76.3% 77.4% Very small-small problem 15.7% 34.5% 25.0% 25.3% 23.4% 23.0% 18.8% 19.7% Moderate-big problem 2.2% 8.5% 3.1% 3.2% 6.0% 4.5% 4.9% 2.9% p value <0.0001 0.0079 0.0038 0.0031 0.0214 0.0169 0.2508 Incontinence No problem 94.8% 79.5% 85.2% 86.7% 88.3% 85.2% 85.7% 89.5% Very small-small problem 4.1% 17.0% 14.5% 11.2% 9.7% 12.3% 11.6% 8.8% Moderate-big problem 1.1% 3.5% 0.4% 2.0% 2.0% 2.5% 2.7% 1.7% p value <0.0001 0.0033 0.0021 0.03 0.0001 0.0001 0.0147 Bloody stools No problem 98.1% 90.8% 96.9% 95.6% 96.0% 96.3% 94.2% 94.2% Very small-small problem 1.9% 8.1% 2.3% 3.6% 2.4% 2.5% 5.4% 5.8% Moderate-big problem 0.0% 1.2% 0.8% 0.8% 1.6% 1.2% 0.4% 0.0% p value 0.0002 0.1677 0.0934 0.0574 0.0906 0.0066 0.0258 Pain (abdominal, pelvic or rectal) No problem 86.1% 76.8% 89.8% 86.7% 86.7% 88.5% 86.6% 90.3% Very small-small problem 12.4% 17.8% 7.8% 10.4% 9.7% 8.2% 12.1% 7.1% Moderate-big problem 1.5% 5.4% 2.3% 2.8% 3.6% 3.3% 1.3% 2.5% p value 0.0002 0.5516 0.6606 0.232 0.5713 0.8565 0.7498 N= 267 259 255 249 248 244 224 238 - + MESH:D010146 Disease @@ -19910,7 +19770,7 @@ table_caption 15733 Changes in EPIC bowel summary and overall bowel bother scores following SBRT for prostate cancer - + MESH:D011471 Disease @@ -19932,49 +19792,67 @@ fig_caption 16256 Individual EPIC-26 bowel symptoms (Questions 6a-e). Average individual symptom EPIC bother scores at baseline and following SBRT for prostate cancer: (a) urgency to have a bowel movement-Question 6a of the EPIC-26; (b) increase frequency of bowel movements- Question 6b of the EPIC-26; (c) losing control of stools- Question 6c of the EPIC-26; (d) bloody stools- Question 6d of the EPIC-26; (e) abdominal/pelvic/rectal pain- Question 6e of the EPIC-26. Thresholds for clinically significant changes in scores (1/2 standard deviation above and below the baseline) are marked with dashed lines. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. - - MESH:C563475 + + MESH:D009069 Disease - - rectal pain + + bowel movements - + + MESH:D010146 + Disease + + pain + + MESH:D011471 Disease prostate cancer + + MESH:D009069 + Disease + + bowel movement + + + MESH:D051271 + Disease + + bowel symptoms + RESULTS paragraph 16949 At baseline, 1.9% of our cohort reported some level of annoyance due to bloody stools, however no patient felt it was a moderate to big problem (Table 2). The mean changes in EPIC bloody stool bother scores from baseline to 2 years of follow-up are shown in Table 3. The mean EPIC bloody stool bother score was 99.5 at baseline (Table 3). Bloody stool bother increased following treatment with the mean score decreasing to 96.6 at 1 month post-treatment (mean change, -2.9) (p = 0.0002) (Table 3, Figure 1d). However, only 1.2% of patients felt that that bloody stools were a moderate to big problem at 1 month following treatment (Table 2). Although bloody stool bother declined quickly, a second late increase in bloody stool bother was observed with the mean bloody stool bother score decreasing to 97.4 at 18 months (mean change from baseline, -2.1) (p = 0.0066) (Table 3, Figure 1d). Both declines met the threshold for clinically significant change (MID = 1.7). However, only 1.2% of patients felt that that bloody stools were a moderate to big problem at 12 months following treatment (Table 2). By two years following SBRT, bloody stool bother returned to near-baseline with bloody stool bother score of 98.1 (mean change from baseline, -1.4). By two years post-SBRT, no patient felt that blood stools was a moderate to big problem (Table 2). - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + Disease MID @@ -19985,24 +19863,48 @@ paragraph 18327 Baseline proctitis symptoms were uncommon in our patients (Table 2). All proctitis symptoms increased at one month post-SBRT. The most bothersome symptoms were bowel urgency and frequency. At one month post SBRT, 11.2% and 8.5% of patients reported moderate to big problems with bowel urgency and frequency, respectively (Table 2). 9.7% of patients required anti-diarrheals within the first month post-SBRT. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis symptoms + + + MESH:D015212 + Disease + + bowel urgency + FIG @@ -20011,7 +19913,7 @@ fig_caption 18737 Average EPIC bowel summary scores at baseline and following SBRT for prostate cancer. Thresholds for clinically significant changes in scores (1/2 standard deviation above and below the baseline) are marked with dashed lines. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. - + MESH:D011471 Disease @@ -20023,11 +19925,17 @@ paragraph 19063 Likewise, the EPIC bowel summary score declined transiently at 1 month (mean change, -9.8) (Table 3). In addition, a second more protracted decline occurred between 6 and 18 months (mean change from baseline at 12 months, -3.5). Bowel declines at 1 month and 12 months were statistically significant (p < 0.0001); however, only the 1 month change met the threshold for clinically significant change (MID = 4.6) (Figure 2). The EPIC bowel summary score returned to near-baseline at two years post-SBRT (mean change from baseline, -1.6). Individual bowel symptoms changed with time in a similar manner (Figure 1). - + Disease MID + + MESH:D051271 + Disease + + bowel symptoms + TABLE @@ -20036,13 +19944,13 @@ table_caption 19692 Overall bowel bother following SBRT for prostate cancer (Patient-reported responses to Question 7 of the EPIC-26) - + 9606 Species Patient - + MESH:D011471 Disease @@ -20070,41 +19978,71 @@ paragraph 20484 Overall bowel bother showed a similar pattern as the bowel summary score. At baseline, 24.0% of patients reported some level of annoyance due to bowel symptoms with 4.1% of patients feeling that bowel function was a moderate to big problem (Table 4). The mean EPIC bowel bother score was 90.7 at baseline (Table 3). Overall bowel bother worsened post-treatment and the mean score decreased to 75 at 1 month (mean change, -14.8) (Table 3, Figure 3). However, only 11.5% of patients felt that their bowel function was a moderate to big problem at 1 month following SBRT (Table 4). Bowel bother scores worsened over a second, more protracted time period (Figure 3). Once again, only the first decline met the threshold for clinically significant change (MID = 9.8). Only 7% of patients felt that bowel symptoms were a moderate to big problem at 12 month following treatment (Table 4). By two years following SBRT, overall bowel bother returned to near baseline with a bowel bother score of 89.8 (mean change from baseline, -0.9) (Table 3) and 3.3% of patients feeling bowel symptoms were a moderate to big problem (Table 4). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + Disease MID + + MESH:D015212 + Disease + + bowel function + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D051271 + Disease + + feeling bowel symptoms + + + MESH:D015212 + Disease + + bowel function + DISCUSS @@ -20117,41 +20055,65 @@ paragraph 21633 Proctitis following prostate cancer radiotherapy is a critical quality of life issue and the principle dose-limiting toxicity. Currently, there is limited data on proctitis following SBRT for prostate cancer. A better understanding of bowel symptoms following SBRT would enable clinicians to provide more realistic expectations to patients as they weigh their treatment options. Our prior paper reported the prevalence, severity and overall incidence of bowel frequency/urgency, rectal pain and rectal bleeding following SBRT. However, it is reliant on physician reporting which may under report the incidence of bowel symptoms and provides no information on the associated bother. In this study, we utilized the EPIC-26 bowel domain to evaluate bowel symptoms but also assess related bother. - + 9606 Species patients - - MESH:D012002 + + MESH:D051271 Disease - - rectal bleeding + + bowel symptoms - + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D010146 + Disease + + pain + + MESH:D064420 Disease toxicity - + MESH:D011471 Disease + + prostate cancer + + + MESH:D011349 + Disease Proctitis following prostate cancer - - MESH:D011471 + + MESH:D011349 Disease - - prostate cancer + + proctitis - - MESH:C563475 + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D006470 Disease - - rectal pain + + bleeding @@ -20159,71 +20121,65 @@ paragraph 22426 Stereotactic body radiation therapy (SBRT) offers to minimize radiation-associated rectal toxicity by reducing the volume of rectum receiving high radiation doses. The overall incidence of post-SBRT rectal bleeding in this series was 23%. The low rate of late Grade >= 2 rectal bleeding (1.5%) seen in this study is consistent with the results from a prior review by our institution and results from other institutions which all report a rate of late Grade >= 2 rectal bleeding of < 5%. In fact, many of the bleeds were acute and transient suggestive of acute anal irritation or hemorrhoid exacerbation due to bowel frequency. Furthermore, the incidence of telangiectasia in patients who completed post-SBRT rectal endoscopy for the assessment of rectal bleeding or cancer screening was 11% which is significantly lower than the rates reported in prospective studies that looked at endoscopic outcomes after conventionally fractionated radiation therapy. - + 9606 Species patients - - MESH:D001005 - Disease - - acute anal irritation - - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D006484 Disease hemorrhoid - + MESH:D013684 Disease telangiectasia - + MESH:D009369 Disease cancer - + MESH:D006470 Disease - - bleeds + + bleeding - - MESH:D012002 + + MESH:D001523 Disease - - rectal toxicity + + irritation - - MESH:D012002 + + MESH:D064420 Disease - - SBRT rectal bleeding + + toxicity - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding @@ -20231,53 +20187,53 @@ paragraph 23395 These findings are particularly significant given that rectal bleeding is one of the principle dose-limiting toxicities, and is thus a potential barrier to administering radiation at appropriately therapeutic levels. In the work by Sanda et al, the proportion of patients reporting rectal bleeding as a moderate-big problem on the EPIC 26 survey progressively increased over time (reaching 5% at 12 and 24 months), ultimately manifesting as a late complication post-treatment. However, we observed that with SBRT rectal bleeding occurred at lower rates and presented as a relatively earlier complication: the incidence of patients reporting rectal bleeding as a moderate-big problem in this study reached a peak at 9 months at 1.6%, and by 24 months the incidence dropped to null. Additionally, in our series the rate of Grade 2-3 late rectal bleeding was 1.5%. This compares favorably to the 5%-15% seen with contemporary high-dose IMRT. We speculate that this reassuring profile with SBRT may be a result of the enhanced accuracy with fiducial tracking and narrowed target volumes, thus enhancing our efforts to spare normal tissue from inadvertent irradiation. - + 9606 Species patients - + 9606 Species patients - + MESH:D064420 Disease toxicities - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - + MESH:D006470 Disease - - bleeding + + late rectal bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - SBRT rectal bleeding + + bleeding - - MESH:D012002 + + MESH:D006470 Disease - - rectal bleeding + + bleeding @@ -20287,7 +20243,7 @@ fig_caption 24562 Radar plots showing the distribution of individual symptom bother following SBRT for prostate cancer. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. Points further out from the center indicate higher levels of bother with a given symptom. - + MESH:D011471 Disease @@ -20299,49 +20255,79 @@ paragraph 24855 The pattern seen in bowel QOL after SBRT in our study is similar to the pattern seen after conventionally fractionated radiotherapy, proton therapy or brachytherapy. The bowel QOL score is at its lowest 1 or 2 months after treatment, but improves slowly thereafter to near baseline by 1-2 years after treatment. In our series, most moderate to big problems were seen at 1 month post-SBRT with approximately 10% of patients reporting moderate to big problems with bowel urgency and/or frequency (Figure 4). This compares favorably with 15-19% incidence of moderate to severe bowel urgency and frequency seen two months after conventionally fractionated external beam radiation therapy or brachytherapy. Unlike conventional modalities, this increase in bowel bother was transient and returned to near baseline by 3 months post-SBRT. A second transient increase in bowel bother occurred at 12 months with approximately 7% of patients reporting moderate to big problems. Unlike the pattern seen with conventional modalities, post-SBRT bowel bother returned to near baseline by two years. The mean bowel summary score change from baseline to 24 months in this study was -1.6. This change compares favorably to that seen at 24 months with conventionally fractionated IMRT and proton therapy -7.4 and -3.7, respectively. - + 9606 Species patients - + 9606 Species patients + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel QOL + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel QOL + DISCUSS paragraph 26176 The present study has several identifiable limitations. The patient population was derived from a single high-volume institution cohort that can limit the translation of our work to the general population. Nevertheless, our work utilized a fairly large patient population with excellent follow-up rates, and was heterogeneous with respect to ethnicity and risk stratification, and considered key patient characteristics such as comorbidity, body mass index, and use of anticoagulants and hormonal therapy. It is conceivable, however, that institutions without the same experience may have a learning curve before they can achieve similar results. Also noteworthy is that our analysis was conducted without a concurrent comparator arm and thus must be carefully weighed against previously published work. Reassuringly, given that the observed toxicity rates in the acute phase are within range of previous studies, the superior toxicity profile at two years follow-up with SBRT compared to other modalities offers intriguing insight to guide subsequent trials for a more comprehensive assessments of SBRT-related proctitis in the future. - + 9606 Species patient - + 9606 Species patient - + 9606 Species patient - + MESH:D064420 Disease toxicity - + + MESH:D011349 + Disease + + proctitis + + MESH:D064420 Disease @@ -20359,12 +20345,18 @@ paragraph 27325 In this single institution cohort, the rate and severity of proctitis observed following SBRT is low. QOL tended to improve with longer follow-up and was near baseline at two years post-SBRT. Our results compare favorably to those reported for patients treated with alternative radiation modalities. Future prospective randomized studies are needed to confirm these observations. - + 9606 Species patients + + MESH:D011349 + Disease + + proctitis + CONCL @@ -20389,18 +20381,18 @@ paragraph 27872 ADT + + - + Chemical + + ADT + ABBR paragraph 27876 Androgen deprivation therapy - - MESH:D000728 - Chemical - - Androgen - ABBR @@ -20425,7 +20417,7 @@ paragraph 27932 Common toxicity criteria - + MESH:D064420 Disease @@ -20449,6 +20441,12 @@ paragraph 27984 DVH + + - + Chemical + + DVH + ABBR @@ -20485,6 +20483,12 @@ paragraph 28088 GTV + + - + Chemical + + GTV + ABBR @@ -20497,6 +20501,12 @@ paragraph 28112 Gy + + - + Chemical + + Gy + ABBR @@ -20533,6 +20543,12 @@ paragraph 28189 PTV + + - + Chemical + + PTV + ABBR @@ -20557,7 +20573,7 @@ paragraph 28236 MID - + Disease MID @@ -20574,12 +20590,6 @@ paragraph 28271 MR - - MESH:C564570 - Disease - - MR - ABBR @@ -20598,7 +20608,7 @@ paragraph 28297 National cancer institute - + MESH:D009369 Disease @@ -20610,7 +20620,7 @@ paragraph 28323 SD - + MESH:D029461 Disease @@ -20652,6 +20662,12 @@ footnote 28414 Daniel Y Joh and Leonard N Chen contributed equally to this work. + + MESH:C536108 + Disease + + Leonard N + ABBR @@ -20676,25 +20692,19 @@ footnote 28798 DJ and LC are lead authors, who participated in data collection, data analysis, manuscript drafting, table/figure creation, and manuscript revision. GP participated in data analysis and manuscript drafting. AB, SS, RM, and JK aided in the quality of life data collection and maintaining the patient database. TY aided in clinical data collection. SL is the dosimetrist who developed the majority of patients' treatment plans, and contributed to the dosimetric data analysis and interpretation. BC and AJ participated in the design and coordination of the study. SS is a senior author who collected the dosimetric data, participated in its analysis, and helped draft the manuscript. JC, AD, and JL are senior authors who aided in drafting the manuscript. SC was the principal investigator who initially developed the concept of the study and the design, aided in data collection, drafted and revised the manuscript. All authors read and approved the final manuscript. - + 9606 Species patient - + 9606 Species patients - - MESH:C441720 - Chemical - - JL - - + MESH:D000544 Disease @@ -20724,8 +20734,8 @@ surname:Kawachi;given-names:I 29778 The association of treatment-related symptoms with quality-of-life outcomes for localized prostate carcinoma patients - - MESH:D011471 + + MESH:D011472 Disease localized prostate carcinoma @@ -20751,7 +20761,7 @@ surname:Muller;given-names:T 29896 Late rectal symptoms and quality of life after conformal radiation therapy for prostate cancer - + MESH:D011471 Disease @@ -20790,7 +20800,7 @@ surname:Northouse;given-names:L 29991 Quality of life and satisfaction with outcome among prostate-cancer survivors - + MESH:D011471 Disease @@ -20823,13 +20833,13 @@ surname:Sandler;given-names:HM 30069 Patient-reported outcomes after 3-dimensional conformal, intensity-modulated, or proton beam radiotherapy for localized prostate cancer - + 9606 Species Patient - + MESH:D011471 Disease @@ -20862,13 +20872,13 @@ surname:Henderson;given-names:RH 30205 Comparative effectiveness study of patient-reported outcomes after proton therapy or intensity-modulated radiotherapy for prostate cancer - + 9606 Species patient - + MESH:D011471 Disease @@ -20893,14 +20903,14 @@ surname:Butler;given-names:EB 30343 Radiation proctopathy in the treatment of prostate cancer - + MESH:D011471 Disease prostate cancer - - MESH:D011832 + + MESH:D004194 Disease Radiation proctopathy @@ -20923,7 +20933,7 @@ surname:Fransson;given-names:P 30401 Late radiation effects to the rectum and anus after treatment for prostate cancer; validity of the LENT/SOMA score - + MESH:D011471 Disease @@ -20946,13 +20956,13 @@ 2302 30516 Differing perceptions of quality of life in patients with prostate cancer and their doctors - + 9606 Species patients - + MESH:D011471 Disease @@ -20975,13 +20985,13 @@ 1300 30608 Time course and incidence of late complications in patients treated with radiation therapy for FIGO stage IB carcinoma of the uterine cervix - + 9606 Species patients - + MESH:D002277 Disease @@ -21014,23 +21024,17 @@ surname:Jackson;given-names:W 30749 Age and comorbid illness are associated with late rectal toxicity following dose-escalated radiation therapy for prostate cancer - + MESH:D011471 Disease prostate cancer - - MESH:D064420 - Disease - - toxicity - - - MESH:D002908 + + MESH:D012002 Disease - - illness + + late rectal toxicity @@ -21052,30 +21056,30 @@ surname:Goldberg;given-names:S 30878 Acute and late toxicity of patients with inflammatory bowel disease undergoing irradiation for abdominal and pelvic neoplasms - + 9606 Species patients - + + MESH:D009369 + Disease + + neoplasms + + MESH:D064420 Disease toxicity - + MESH:D015212 Disease inflammatory bowel disease - - MESH:D010386 - Disease - - pelvic neoplasms - 76 @@ -21092,19 +21096,19 @@ 760 31004 External beam radiotherapy for prostate cancer patients on anticoagulation therapy: how significant is the bleeding toxicity? - + 9606 Species patients - - MESH:D064420 + + MESH:D006470 Disease bleeding toxicity - + MESH:D011471 Disease @@ -21125,11 +21129,11 @@ 729 31130 Treatment of radiation proctitis - + MESH:D011349 Disease - - radiation proctitis + + proctitis @@ -21152,7 +21156,7 @@ surname:Huang;given-names:EH 31163 Long-term results of the M. D. Anderson randomized dose-escalation trial for prostate cancer - + MESH:D011471 Disease @@ -21188,13 +21192,13 @@ surname:Bottomley;given-names:D 31256 Escalated-dose versus standard-dose conformal radiotherapy in prostate cancer: first results from the MRC RT01 randomised controlled trial - + CVCL:D923 CellLine MRC RT01 - + MESH:D011471 Disease @@ -21221,7 +21225,7 @@ surname:Slot;given-names:A 31395 Update of Dutch multicenter dose-escalation trial of radiotherapy for localized prostate cancer - + MESH:D011471 Disease @@ -21254,19 +21258,19 @@ surname:Wachter;given-names:S 31491 Proctitis after external-beam radiotherapy for prostate cancer classified by Vienna Rectoscopy Score and correlated with EORTC/RTOG score for late rectal toxicity: results of a prospective multicenter study of 166 patients - + MESH:D011349 Disease Proctitis - - MESH:D012002 + + MESH:D064420 Disease - - rectal toxicity + + toxicity - + MESH:D011471 Disease @@ -21290,7 +21294,7 @@ surname:Deasy;given-names:JO 31714 Radiation dose-volume effects in radiation-induced rectal injury - + MESH:D012002 Disease @@ -21349,7 +21353,7 @@ surname:Winter;given-names:K 31932 Do intermediate radiation doses contribute to late rectal toxicity? An analysis of data from radiation therapy oncology group protocol 94-06 - + MESH:D064420 Disease @@ -21381,13 +21385,13 @@ surname:Sandler;given-names:HM 32073 Multi-institutional prospective evaluation of bowel quality of life after prostate external beam radiation therapy identifies patient and treatment factors associated with patient-reported outcomes: the PROSTQA experience - + 9606 Species patient - + 9606 Species @@ -21413,18 +21417,18 @@ surname:Shippy;given-names:AM 32295 Incidence of late rectal and urinary toxicities after three-dimensional conformal radiotherapy and intensity-modulated radiotherapy for localized prostate cancer - - MESH:D064420 - Disease - - toxicities - - + MESH:D011471 Disease localized prostate cancer + + MESH:D001745 + Disease + + urinary toxicities + surname:Bosch;given-names:WR @@ -21448,13 +21452,13 @@ surname:Winter;given-names:K 32457 Preliminary toxicity analysis of 3-dimensional conformal radiation therapy versus intensity modulated radiation therapy on the high-dose arm of the Radiation Therapy Oncology Group 0126 prostate cancer trial - + MESH:D064420 Disease toxicity - + MESH:D011471 Disease @@ -21474,7 +21478,7 @@ 276 32665 The radiobiology of prostate cancer including new aspects of fractionated radiotherapy - + MESH:D011471 Disease @@ -21495,8 +21499,8 @@ 1101 32752 Fractionation and protraction for radiotherapy of prostate carcinoma - - MESH:D011471 + + MESH:D011472 Disease prostate carcinoma @@ -21519,11 +21523,11 @@ surname:Petereit;given-names:D 32821 Hypofractionation for prostate cancer - + MESH:D011471 Disease - - Hypofractionation for prostate cancer + + prostate cancer @@ -21559,7 +21563,7 @@ surname:Ju;given-names:AW 32859 Stereotactic body radiation therapy (SBRT) for clinically localized prostate cancer: the Georgetown University experience - + MESH:D011471 Disease @@ -21581,7 +21585,7 @@ ref 32981 Stereotactic body radiotherapy for localized prostate cancer: disease control and quality of life at 6 years - + MESH:D011471 Disease @@ -21607,7 +21611,7 @@ surname:Kupelian;given-names:P 33091 Health Related Quality of Life after Stereotactic Body Radiotherapy for Localized Prostate Cancer: Results from a Multi-Institutional Consortium of Prospective Trials - + MESH:D011471 Disease @@ -21636,7 +21640,7 @@ surname:Bolzicco;given-names:G 33258 Stereotactic Body Radiotherapy for Localized Prostate Cancer: Pooled Analysis of Multi-Insitutional Prospective Trials - + MESH:D011471 Disease @@ -21659,7 +21663,7 @@ 882 33377 Long-term outcomes from a prospective trial of stereotactic body radiotherapy for low-risk prostate cancer - + MESH:D011471 Disease @@ -21686,7 +21690,7 @@ surname:Tapella;given-names:P 33484 Hypofractionated stereotactic body radiotherapy in low-risk prostate adenocarcinoma: Preliminary results of a multi-institutional phase 1 feasibility trial - + MESH:D011471 Disease @@ -21716,7 +21720,7 @@ surname:Oermann;given-names:EK 33640 Hypofractionated stereotactic body radiation therapy as monotherapy for intermediate-risk prostate cancer. Radiation Oncology - + MESH:D011471 Disease @@ -21792,7 +21796,7 @@ surname:Blank;given-names:K 33957 Biochemical outcome after radical prostatectomy, external beam radiation therapy, or interstitial radiation therapy for clinically localized prostate cancer - + MESH:D011471 Disease @@ -21821,7 +21825,7 @@ surname:Uhm;given-names:S 34114 Rectal endoscopy findings following stereotactic body radiation therapy for clinically localized prostate cancer - + MESH:D011471 Disease @@ -21846,18 +21850,18 @@ surname:Wambersie;given-names:A 34227 Endoscopic scoring of late rectal mucosal damage after conformal radiotherapy for prostatic carcinoma - - MESH:D012002 - Disease - - late rectal mucosal damage - - - MESH:D011471 + + MESH:D011472 Disease prostatic carcinoma + + MESH:D009059 + Disease + + mucosal damage + surname:Sandler;given-names:HM @@ -21876,19 +21880,19 @@ surname:Sanda;given-names:MG 34329 Development and validation of the expanded prostate cancer index composite (EPIC) for comprehensive assessment of health-related quality of life in men with prostate cancer - + 9606 Species men - + MESH:D011471 Disease prostate cancer - + MESH:D011471 Disease @@ -21910,7 +21914,7 @@ 603 34502 Should function and bother be measured and reported separately for prostate cancer quality-of-life domains? - + MESH:D011471 Disease @@ -21950,7 +21954,7 @@ surname:Krupski;given-names:TL 34728 Correlates of bother following treatment for clinically localized prostate cancer - + MESH:D011471 Disease @@ -21976,7 +21980,7 @@ surname:Lee;given-names:SP 34810 Quality of life after surgery, external beam irradiation, or brachytherapy for early-stage prostate cancer - + MESH:D011471 Disease @@ -21999,7 +22003,7 @@ surname:Witten;given-names:M 34917 Stereotactic body radiotherapy for organ-confined prostate cancer - + MESH:D011471 Disease @@ -22034,7 +22038,7 @@ surname:Macchia;given-names:G 34983 Early proctoscopy is a surrogate endpoint of late rectal toxicity in prostate cancer treated with radiotherapy - + MESH:D011471 Disease @@ -22061,7 +22065,7 @@ surname:van der Vight;given-names:LP 35094 Reduced late rectal mucosal changes after prostate three-dimensional conformal radiotherapy with endorectal balloon as observed in repeated endoscopy - + MESH:D054549 Disease @@ -22086,19 +22090,19 @@ surname:Cox;given-names:B 35244 Long-term survival and toxicity in patients treated with high-dose intensity modulated radiation therapy for localized prostate cancer - + 9606 Species patients - + MESH:D064420 Disease toxicity - + MESH:D011471 Disease @@ -22160,47 +22164,47 @@ rhesus monkeys - MESH:D000661 - Chemical - - D-amphetamine - - MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:C100294 Chemical phendimetrazine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine + + MESH:D003913 + Chemical + + D-amphetamine + ABSTRACT @@ -22225,18 +22229,24 @@ abstract 977 GML models provided an excellent fit of the cocaine choice dose-effect functions in individual monkeys. Reductions in cocaine choice by both pharmacological and environmental manipulations were principally produced by systematic decreases in sensitivity to reinforcer price and non-systematic changes in sensitivity to reinforcer magnitude. - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine + + - + Chemical + + GML + ABSTRACT @@ -22249,36 +22259,36 @@ abstract 1330 The modeling approach used provides a theoretical link between the experimental analysis of choice and pharmacological treatments being evaluated as candidate 'agonist-based' medications for cocaine addiction. The analysis suggests that monoamine releaser treatment efficacy to decrease cocaine choice was mediated by selectively increasing the relative price of cocaine. Overall, the net behavioral effect of these pharmacological treatments was to increase substitutability of food pellets, a nondrug reinforcer, for cocaine. - + MESH:D003042 Chemical cocaine - - MESH:D003042 + + - Chemical - - cocaine + + monoamine - + MESH:D003042 Chemical - + cocaine - + + MESH:D019970 + Disease + + cocaine addiction + + MESH:D003042 Chemical cocaine - - MESH:D015306 - Chemical - - monoamine - INTRO @@ -22297,7 +22307,7 @@ paragraph 2624 Previous studies have examined the determinants of drug reinforcement in choice procedures in which subjects choose between drug (e.g., cocaine) and an alternative nondrug reinforcer (e.g., food; for review,). This literature body supports the general conclusion that choice between drug and food is sensitive to the magnitude, price, frequency, and delay dimensions of reinforcers. Although this research suggests that drug vs. food choice is sensitive to the relative value of drug reinforcement, relatively few studies have integrated these results with quantitative theories of operant choice. - + MESH:D003042 Chemical @@ -22309,55 +22319,73 @@ paragraph 3222 The generalized matching law (GML) is a quantitative framework, which predicts that behavior will be allocated among different reinforcers in proportion to their relative value. Relative reinforcer value has been quantified most effectively by the concatenated GML, which predicts that value is determined by a multiplicative combination of each reinforcer dimension listed above. A version of the GML suitable for drug vs. food choice may be written + + - + Chemical + + GML + INTRO paragraph 3673 where B represents behavior allocated to cocaine (Bc) or food (Bf), M represents the magnitude dimension of each reinforcer, and P represents the price dimension (fixed-ratio schedule) of each reinforcer. For cocaine, Mc equals the unit dose (mg/kg/injection) and for food Mf equals the value of food scaled in dose units of cocaine (Mf Mc). On the basis of preliminary model simulations, the proportionality constant was set to 0.032 in the present study; consequently, the magnitude of a 1-g food pellet was set equal to a 0.032-mg/kg-cocaine injection. Furthermore, we have previously demonstrated comparable reinforcing effects of a 0.032-mg/kg cocaine injection and a 1-g food pellet in both progressive-ratio and choice procedures. The present approach of fixing the scaling parameter at a single value is equivalent to previous applications of the GML using a bias parameter to account for choice between qualitatively different reinforcers (e.g.,). The free parameters sm (sensitivity to magnitude) and sp (sensitivity to price) capture the extent of changes in drug vs. food choice with changes in the relative magnitude and price of cocaine vs. food for an individual subject. For example, in the case of strict matching, sm = sp = 1, the proportion of responses allocated to the drug alternative is equal to the multiplicative combination of magnitude and price proportions. Fig. 1 shows cocaine choice dose-effect functions simulated by Eq. (1) with relative reinforcer magnitude and price equal to the values used in the present study. The main focus of these simulations is to demonstrate that the slope and horizontal position of the cocaine choice dose-effect functions are determined independently by sensitivity to reinforcer magnitude and reinforcer price, respectively. - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - - MESH:D003042 + + - Chemical - - cocaine + + GML - + + MESH:D008748 + Chemical + + Mc + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical - + cocaine - + MESH:D003042 Chemical - + cocaine - + + MESH:C000604007 + Chemical + + sp + + MESH:D003042 Chemical @@ -22369,59 +22397,53 @@ paragraph 5469 The purpose of the present investigation was to determine monoamine releaser treatment effects in a cocaine vs. food choice procedure on sensitivity parameters of the GML. We reanalyzed data from demonstrating reductions in cocaine vs. food choice produced by continuous 14-day D-amphetamine, phendimetrazine, and phenmetrazine treatment. A version of the concatenated GML incorporating the relative magnitude and price of cocaine and food reinforcers was fit to the cocaine choice dose-effect curves of individual monkeys. A multi-model comparison approach was used to evaluate competing hypotheses regarding dose- and time-course effects on sensitivity to reinforcement. For comparison, we also modeled 7-day effects of extinction on cocaine- or food-maintained responding during the choice procedure. - + MESH:D003042 Chemical cocaine - + + MESH:D003913 + Chemical + + D-amphetamine + + MESH:C100294 Chemical phendimetrazine - - MESH:D003042 - Chemical - - cocaine - - - MESH:D015306 + + - Chemical monoamine - + MESH:D003042 Chemical cocaine - + MESH:D010633 Chemical phenmetrazine - - MESH:D000661 - Chemical - - D-amphetamine - - - MESH:D003042 + + - Chemical - - cocaine + + GML - - MESH:D003042 + + - Chemical - - cocaine + + GML @@ -22441,13 +22463,13 @@ paragraph 6314 The experimental methods have been previously described in detail in). Briefly, studies were conducted in four-six adult male rhesus monkeys (Macaca mulatta) surgically implanted with a double-lumen catheter inserted in a major vein. The catheter was connected to a fluid swivel attached to the top of each subject's cage and was protected by a custom jacket and stainless steel tether. Animal research and maintenance were conducted according to the 8th edition of the Guide for the Care and Use of Laboratory Animals as adopted and promulgated by the National Institutes of Health (National Academies Press, 2011). Animal facilities were licensed by the United States Department of Agriculture and accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. The Institutional Animal Care and Use Committee approved the research protocol. Monkeys had visual, auditory, and olfactory contact with other monkeys throughout the study. Operant procedures and foraging toys were provided for environmental manipulation and enrichment. Videos were played daily in animal housing rooms to provide additional environmental enrichment. - + 9544 Species rhesus monkeys - + 9544 Species @@ -22459,37 +22481,31 @@ paragraph 7476 Experimental sessions were conducted in each monkey's home cage. Monkeys responded in daily 2 h choice sessions (0900-1100 h) that consisted of a five-component concurrent schedule of food pellet and intravenous cocaine availability as described in detail previously. During each component, responses on the left key were reinforced with food (1-g banana-flavored pellets; Test Diets, Richmond, IN) according to a fixed-ratio (FR) 100 schedule, and responses on the right key were reinforced with intravenous cocaine (0-0.1 mg/kg/injection) according to an FR 10 schedule. A response on one key reset the ratio requirement on the alternative key. Each reinforcer delivery was followed by a 3-s timeout during which all stimulus lights were extinguished, and responding had no programmed consequences. During each component, the food key was transilluminated red. The stimulus lights for the cocaine key were flashed on and off in 3 s cycles, and longer flashes were associated with higher unit cocaine doses. Across components of the choice procedure, a different unit cocaine dose was available (0, 0.0032, 0.01, 0.032, and 0.1 mg/kg/injection during components 1-5, respectively) by manipulating the injection volume (0, 0.01, 0.03, 0.1, and 0.3 ml/injection, respectively) delivered via the 'cocaine' pump. Each component was in effect until 10 total reinforcers were earned or 20 min elapsed, whichever occurred first. - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - - MESH:D003042 - Chemical - - cocaine - - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical @@ -22501,42 +22517,42 @@ paragraph 8911 Once cocaine choice was stable, tests sessions were initiated. Subsequently, a 14-day treatment period was initiated during which a test solution was administered via the 'treatment' pump for 23 h/day. The treatment solutions and doses examined were D-amphetamine (0.032-0.1 mg/kg/h), (+)-phendimetrazine (0.32-1.0 mg/kg/h) or (+)-phenmetrazine (0.1-0.32 mg/kg/h). At the conclusion of each 14-day test period, a saline control treatment period was reinstated for at least 5 days and until cocaine choice returned to pretest levels. For the extinction studies, separate 7-day treatment periods were initiated where either cocaine was removed from the syringe or food pellets were removed from the dispenser. All other component-correlated stimuli were retained. - - MESH:D003042 + + MESH:D012965 Chemical - - cocaine + + saline - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D010633 Chemical (+)-phenmetrazine - - MESH:D000661 - Chemical - - D-amphetamine - - + MESH:C100294 Chemical (+)-phendimetrazine + + MESH:D003913 + Chemical + + D-amphetamine + METHODS @@ -22549,65 +22565,95 @@ paragraph 9729 The dependent measure of interest was the proportion of cocaine choices (number of completed ratios on the cocaine-associated key/total completed ratios) for days 5-7 (7 days) and days 12-14 (14 days) of the continuous 14-day treatments or days 5-7 for the 7-day extinction condition. These data were then plotted as a function of unit cocaine dose. In the modeling presented below, the value of a single food pellet was held constant at 0.032 mg/kg/injection cocaine as in Fig. 1. Changes in response allocation produced by removal of cocaine or food availability were modeled using baseline reinforcer magnitudes. This approach allowed reinforcer sensitivity parameters to account for changes in response allocation and facilitated comparison with pharmacological manipulations. Parameters of Eq. (1) were estimated for individual monkeys by minimizing the residual sum of squares (RSS) using the Solver add-in of Microsoft Excel for Mac 2011. Model fitting was conducted separately for each manipulation. Data from a monkey were not included if (1) the monkey did not complete each dose of a treatment drug and (2) a choice proportion could not be calculated for two or more cocaine doses. The number of data points fit for individual monkeys are included in Table 1. Within a monkey and a treatment drug, a total RSS was computed by summing the RSS across each dose and treatment week. Therefore, if a monkey completed 14-day treatments for each test drug dose, the total RSS was computed from five cocaine choice dose-effect functions. - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - - MESH:D005517 - Disease - - food availability - - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + + - + Chemical + + RSS + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + + - + Chemical + + RSS + + MESH:D003042 Chemical cocaine + + - + Chemical + + RSS + METHODS paragraph 11276 Different versions of Eq. (1) were fit to the data for days 5-7 (7 days) and days 12-14 (14 days) of each continuous 14-day pharmacological treatment to test different hypotheses regarding treatment effects on sensitivity to reinforcement. Thus, different combinations of the free parameters (k) were either allowed to vary or were constrained to determine whether dose had no effect on sensitivity to reinforcement (the Null model, H0), affected sensitivity to both reinforcer dimensions (Full model), or selectively affected sensitivity to a single reinforcer dimension. In the Null model, sensitivity to reinforcer magnitude and price (si) in Eq. (1) were constrained (e.g., k = 2; s(BL) = s(Dose 1) = s(Dose)). The Full model placed no restrictions on estimated sensitivity to reinforcement parameters (k = 10). The Magnitude model allowed sensitivity to reinforcer magnitude (sm(BL) / sm(Dose 1) / sm(Dose 2)) to vary across doses while sensitivity to reinforcer price (sp(BL) sp(Dose 1) sp(Dose 2)) was constrained (k = 6). The Price model constrained sensitivity to reinforcer magnitude (sm(BL) = sm(Dose 1) = sm(Dose 2)) while sensitivity to reinforcer price (sp(BL) / sp(Dose 1) / sp(Dose 2)) was allowed to vary across doses (k = 6). - - MESH:C027727 + + MESH:C000604007 Chemical - - H0 + + sp + + + MESH:C000604007 + Chemical + + sp + + + MESH:C000604007 + Chemical + + sp + + + MESH:C000604007 + Chemical + + sp @@ -22615,13 +22661,7 @@ paragraph 12548 The minimum corrected Akaike information criterion (AICc) score indicates the best model after correcting for the number of free parameters (see Supplement for further explanation). Multi-model comparison was used to characterize dose-effects of pharmacological treatments on sensitivity to reinforcement. The corrected Akaike information criterion (AICc) was used to compare different models and DeltaAICc [i.e., DeltaAICc (H0) - DeltaAICc (H1) > 4.0] was used to select the best model for each monkey. The model-comparison approach is analogous to a significance test; therefore, the parameters of the selected model were treated as a significant effect of a specific experimental manipulation for an individual monkey. Because an alternative model was the selected model in 18/19 cases, we focused on the magnitude of systematic parameter changes below. Cocaine vs. food choice and sensitivity to reinforcement dose-effect curves were constructed by averaging the individual, best-fitting parameter estimates. - - MESH:C027727 - Chemical - - H0 - - + MESH:D003042 Chemical @@ -22645,46 +22685,52 @@ paragraph 14193 Fig. 2 shows continuous 14-day D-amphetamine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of D-amphetamine dose. The GML model accurately described the cocaine choice dose-effect functions across each D-amphetamine dose and each treatment week (Table 1). The best-fit model for D-amphetamine effects differed somewhat between monkeys with the Full and Price models being selected. In 5/6 monkeys that were sensitive to pharmacological treatment, however, 0.1 mg/kg/h D-amphetamine produced an approximate 10-fold decrease in sensitivity to reinforcer price. Changes were less systematic across monkeys producing a 1.5-fold increase in sensitivity to reinforcer magnitude. - - MESH:D003042 + + - Chemical - - cocaine + + GML - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - + + MESH:D003913 + Chemical + + D-amphetamine + + MESH:D003042 Chemical - + cocaine - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - - MESH:D000661 + + MESH:D003042 Chemical - - D-amphetamine + + cocaine - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine @@ -22693,47 +22739,53 @@ paragraph 14959 Fig. 3 shows continuous 14-day (+)-phendimetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of phendimetrazine dose. The GML model provided an accurate description of the cocaine choice dose-effect functions across each (+)-phendimetrazine dose and each treatment week (Table 1). The best-fit model was consistent for four of the five monkeys and indicated that (+)-phendimetrazine produced a selective decrease in sensitivity to reinforcer price. The best-fit model for the other monkey suggested (+)-phendimetrazine increased sensitivity to reinforcer magnitude during 0.32/mg/kg/h treatment and decreased sensitivity to reinforcer magnitude during 1.0/mg/kg/h treatment. The multi-model comparison suggested that sensitivity to reinforcer price decreased dose-dependently and these decreases were more pronounced after 14 days of treatment. - + MESH:C100294 Chemical (+)-phendimetrazine - + MESH:C100294 Chemical - + (+)-phendimetrazine - + + MESH:C100294 + Chemical + + phendimetrazine + + MESH:D003042 Chemical cocaine - + MESH:C100294 Chemical - + (+)-phendimetrazine - - MESH:C100294 + + - Chemical - - (+)-phendimetrazine + + GML - + MESH:D003042 Chemical cocaine - + MESH:C100294 Chemical - - phendimetrazine + + (+)-phendimetrazine @@ -22741,37 +22793,43 @@ paragraph 15901 Fig. 4 shows continuous 14-day (+)-phenmetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of phenmetrazine dose The GML model provided an accurate description of the cocaine choice dose-effect functions across each (+)-phenmetrazine dose and each treatment week (Table 1). The best-fit model for three monkeys was the Full model and for the other three monkeys was the price sensitivity model. Decreases in sensitivity to reinforcer magnitude and price were apparent at 0.32 mg/kg/h (+)-phenmetrazine following 14 treatment days. - + MESH:D010633 Chemical (+)-phenmetrazine - + + - + Chemical + + GML + + MESH:D010633 Chemical phenmetrazine - + MESH:D003042 Chemical cocaine - + MESH:D010633 Chemical (+)-phenmetrazine - + MESH:D003042 Chemical cocaine - + MESH:D010633 Chemical @@ -22783,31 +22841,37 @@ paragraph 16529 Fig. 5 shows the effects of extinguishing either cocaine- or food-maintained responding for 7 days on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1). Substituting 0.001 for food or cocaine in the GML model during removal of each reinforcer provided an accurate description of the cocaine choice dose-effect functions (Table 1). The best-fit model was consistent between monkeys (note that the AICc values for full and price models are not significantly different for monkey M1414). Cocaine removal produced a greater than 8-fold decrease in sensitivity to reinforcer price, whereas food removal produced an approximately 2-fold increase in sensitivity to reinforcer price. - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical Cocaine - + + - + Chemical + + GML + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical @@ -22825,19 +22889,37 @@ paragraph 17265 The aim of the present investigation was to determine monoamine releaser treatment effects on sensitivity to reinforcement parameters of the GML. A multi-model comparison approach was used to evaluate competing hypotheses regarding dose- and time-course effects on sensitivity to relative reinforcer magnitude and price. There were two main findings. First, the GML models provided excellent fits to the cocaine choice dose-effect functions. In addition, in 18/19 fits to individual monkey data models assuming an effect of a manipulation on sensitivity to reinforcement were superior to the null hypothesis model justifying the use of additional model parameters. Second, pharmacological and environmental manipulations altered cocaine vs. food choice in 18/19 monkeys and did so by decreasing sensitivity to reinforcer price in 16/18 monkeys. - - MESH:D015306 + + MESH:D012640 + Disease + + fits + + + - + Chemical + + GML + + + - Chemical monoamine - - MESH:D003042 + + - Chemical - - cocaine + + GML - + + MESH:D012640 + Disease + + fits + + MESH:D003042 Chemical @@ -22855,7 +22937,7 @@ paragraph 18174 Previous studies have employed the model comparison approach to determine the specificity of pharmacological and neurobiological manipulations on reinforcement-related processes. The present study applied a well-known model of choice that incorporates various reinforcer dimensions to construct a relative value of the available reinforcers. provided rhesus monkeys a choice between alternatives that differed in both reinforcer magnitude and price, and GML model comparison allowed us to determine which reinforcer dimension(s) were affected by pharmacological and environmental manipulations. Taken together, these studies suggest the model comparison provides a unique approach for testing competing hypotheses regarding the behavioral mechanisms of pharmacological and environmental manipulations. - + 9544 Species @@ -22867,13 +22949,19 @@ paragraph 18976 The results of the present study are consistent with prior reports demonstrating GML framework applicability to drug choice procedures with rhesus monkeys (e.g.,). The estimates of sensitivity to reinforcer magnitude obtained in the present study, however, differ markedly from modal values of sensitivity to reinforcer rate established in earlier reviews (e.g.,). Sensitivity to reinforcer magnitude estimates displayed substantial overmatching, often 5-10-fold greater than in previous reports (cf.,). There are at least two possible reasons for this difference. First, this study employed qualitatively different reinforcers, which may have enhanced discrimination of the choice alternatives. A previous study by, however, reported undermatching (s < 1.0) to reinforcer rate in a cocaine vs. food choice procedure employing concurrent variable interval schedules (see also,). Second, the present procedure employed ratio schedules of reinforcement and an additional reset contingency following changeover responses. Concurrent ratio schedules generally produce exclusive preference for the choice alternative associated with the lower ratio requirement (; see also). In studies where choices are made between qualitatively different reinforcers, preference is determined by the relative magnitude and price of each reinforcer. The present findings are consistent with concurrent ratio schedules producing behavior allocation that maximizes relative reinforcer value. - + 9544 Species rhesus monkeys - + + - + Chemical + + GML + + MESH:D003042 Chemical @@ -22885,31 +22973,43 @@ paragraph 20448 Two potential limitations from the present GML analysis should be considered. One limitation is the scaling factor for the food reinforcer was held constant across all monkeys and, therefore, did not permit an assessment of individual differences. Although we chose the scaling factor based on previous studies, exploratory modeling with scaling factors of 0.032 +- 0.5 log units produced similar relative changes in parameter estimates, but provided a poorer fit to the data. A second limitation concerns the approach to modeling the effects of extinction. To model data from extinction and facilitate comparison to the pharmacological manipulations, we used non-zero values to predict the observed nonexclusive preference. Moreover, a single reinforcer magnitude (e.g., 0.001 mg/kg/injection for saline) could not predict the cocaine dose dependency still evident in the cocaine choice dose-effect function after 7 days of cocaine and food removal. - + MESH:D003042 Chemical cocaine - + + - + Chemical + + GML + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine + + MESH:D012965 + Chemical + + saline + DISCUSS paragraph 21399 Therefore, while this ad hoc approach does not provide insight into the mechanisms by which the component-correlated stimuli maintained dose-dependent cocaine choice, model comparison suggests that the effects of extinction on price sensitivity were similar to the effects of pharmacological manipulations. - + MESH:D003042 Chemical @@ -22927,31 +23027,37 @@ paragraph 21781 Pharmacological manipulations that decreased cocaine vs. food preference did so mostly by decreasing sensitivity to reinforcer price. Changes in sensitivity to reinforcer magnitude were less consistent across monkeys compared to price changes. When changes in magnitude sensitivity did occur in individual monkeys, magnitude sensitivity decreased suggesting that total reinforcer consumption decreased (see below).) proposed that the sensitivity parameter in the GML might serve as an index of economic substitutability between qualitatively different reinforcers (see also). The cocaine vs. food choice procedure employed in the present study systematically varied unit cocaine price (UP, fixed ratio/dose) by increasing the available cocaine dose and provided a measure of the own-price elasticity of cocaine demand and the cross-price elasticity of food demand. Therefore, changes in sensitivity to reinforcer price observed in the present study may be related to parameter values in behavioral economic models of choice. - + MESH:D003042 Chemical cocaine - + + - + Chemical + + GML + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical @@ -22963,77 +23069,83 @@ paragraph 22806 To examine the relationship between GML and economic approaches, choice data from each 7-day period of the continuous 14-day D-amphetamine (0.032-0.1 mg/kg/h) treatment are presented in Fig. 6 as demand curves; consumption is plotted as a function of the unit price of cocaine. The continuous curves are the predictions of the GML model and the interpolated unit cocaine price (UP50) at which cocaine and food consumption are predicted to be equal is given in each panel. Consistent with previous demand analyses of cocaine vs. food choice procedures, cocaine consumption was a decreasing function of UP of cocaine. At the end of the 14-day treatment, 0.1-mg/kg/h D-amphetamine produced a ten-fold decrease in UP50 and the GML model predicted an increase in cocaine elasticity and a corresponding decrease in food elasticity. Therefore, according to the substitutability interpretation of GML sensitivity to reinforcement, cocaine and food were imperfect economic substitutes under baseline conditions, and the effects of pharmacological and environmental manipulations increased the elasticity of cocaine preference. - - MESH:D014397 - Disease - - food consumption - - + MESH:D003042 Chemical cocaine - - MESH:D000661 - Chemical - - D-amphetamine - - - MESH:D000661 + + MESH:D003913 Chemical D-amphetamine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - + + - + Chemical + + GML + + MESH:D003042 Chemical cocaine - - MESH:D003042 + + - Chemical - - cocaine + + GML - + MESH:D003042 Chemical - + cocaine - + MESH:D003042 Chemical cocaine - + + MESH:D003913 + Chemical + + D-amphetamine + + MESH:D003042 Chemical cocaine - - MESH:D003042 + + - Chemical - - cocaine + + GML + + + - + Chemical + + GML @@ -23041,19 +23153,25 @@ paragraph 23926 There was concordance among the effects of pharmacological treatment and extinction of cocaine-maintained behavior on sensitivity to reinforcer price. Consistent with previous studies, elasticity of demand for saline was greater than demand for drug. Model comparison suggested that sensitivity to reinforcer magnitude was not altered over the 7 days of each extinction manipulation. This result is consistent with the dose-dependent increase in cocaine choice maintained by the component-correlated stimuli throughout extinction. Taken together, these findings suggest that pharmacological treatments and extinction may affect behavior allocation in the presence of cocaine-associated stimuli through a common behavioral mechanism. - + MESH:D003042 Chemical cocaine - + + MESH:D012965 + Chemical + + saline + + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical @@ -23065,31 +23183,13 @@ paragraph 24659 Finally, the extension of the GML presented here is closely related to Shizgal and colleagues' 'reinforcement mountain' model of brain stimulation reinforcement in intracranial self-stimulation (ICSS) procedures. Their approach employs a single-operant matching model along with generalized matching reinforcement sensitivity parameters (see also). An intriguing similarity between the present and Shizgal approaches is that matching law analyses suggest that pharmacological manipulations affecting dopaminergic neurotransmission selectively alter sensitivity to reinforcement. For example, reported that the cocaine-induced increases in low baseline rates of responding maintained by low frequencies of brain stimulation were consistent with a decreased sensitivity to the price of brain stimulation. Cocaine-induced facilitation of ICSS also correlates with self-administration procedures and has been interpreted as an abuse-related effect. In turn, the blunted expression of abuse-related cocaine effects on ICSS produced by chronic D-amphetamine treatment may also be attributable to altered reinforcer price sensitivity, consistent with the effects reported here on cocaine self-administration. - - MESH:D003042 - Chemical - - cocaine - - - MESH:D000661 + + MESH:D003913 Chemical D-amphetamine - - MESH:D003042 - Chemical - - cocaine - - - MESH:D003042 - Chemical - - cocaine - - + MESH:D003042 Chemical @@ -23107,49 +23207,49 @@ paragraph 25980 Cocaine addiction continues to present a significant public health problem, and the development of effective medications remains a priority of preclinical drug addiction research. The goal of 'agonist-based' medication research is to develop pharmacotherapies that decrease drug use by identifying compounds that share mechanisms of action with cocaine. In a recent review, Haney and Spealman concluded that preclinical self-administration procedures are the most reliable predictor of clinical medication outcomes (; see also). This conclusion suggests the success of preclinical self-administration procedures to accurately predict medication efficacy in humans resides in the neuropharmacological and behavioral mechanisms of reinforcement. Model comparison results of the present study suggest that chronic D-amphetamine and phendimetrazine treatment may function as economic substitutes that selectively increase cocaine demand elasticity by increasing the effective price. Moreover, when a substitutable non-drug reinforcer is concurrently available during chronic D-amphetamine and phendimetrazine treatment, preference for an alternative therapeutic reinforcer will reciprocally increase. - + 9606 Species humans - + MESH:C100294 Chemical phendimetrazine - - MESH:D000661 - Chemical - - D-amphetamine - - + MESH:D003042 Chemical Cocaine - + + MESH:D003913 + Chemical + + D-amphetamine + + MESH:C100294 Chemical phendimetrazine - + + MESH:D003913 + Chemical + + D-amphetamine + + MESH:D003042 Chemical cocaine - - MESH:D000661 - Chemical - - D-amphetamine - - + MESH:D003042 Chemical @@ -23236,7 +23336,7 @@ 326 27973 The generalized matching law as a predictor of choice between cocaine and food in rhesus monkeys - + MESH:D003042 Chemical @@ -23289,8 +23389,8 @@ surname:Chavez;given-names:CL 28304 Modeling operant behavior in the Parkinsonian rat - - MESH:D010300 + + MESH:D010302 Disease Parkinsonian @@ -23310,13 +23410,13 @@ 2470 28354 The effect of chronic amphetamine treatment on cocaine-induced facilitation of intracranial self-stimulation in rats - + MESH:D000661 Chemical amphetamine - + MESH:D003042 Chemical @@ -23337,31 +23437,31 @@ 836 28471 Effects of monoamine releasers with varying selectivity for releasing dopamine/norepinephrine versus serotonin on choice between cocaine and food in rhesus monkeys - + MESH:D009638 Chemical norepinephrine - - MESH:D015306 + + - Chemical monoamine - + MESH:D003042 Chemical cocaine - + MESH:D012701 Chemical serotonin - + MESH:D004298 Chemical @@ -23393,7 +23493,7 @@ 404 28730 Interaction between behavioral and pharmacological treatment strategies to decrease cocaine choice in rhesus monkeys - + MESH:D003042 Chemical @@ -23414,13 +23514,13 @@ 213 28847 Effects of 14-day treatment with the schedule III anorectic phendimetrazine on choice between cocaine and food in rhesus monkeys - + MESH:C100294 Chemical phendimetrazine - + MESH:D003042 Chemical @@ -23493,7 +23593,7 @@ surname:Zacny;given-names:JP 29138 The role of human drug self-administration procedures in the development of medications - + 9606 Species @@ -23556,8 +23656,8 @@ 195 29486 Assessing unit-price related remifentanil choice in rhesus monkeys - - MESH:C071741 + + MESH:D000077208 Chemical remifentanil @@ -23605,7 +23705,7 @@ ref 29655 At what stage of neural processing does cocaine act to boost pursuit of rewards? - + MESH:D003042 Chemical @@ -23638,19 +23738,19 @@ 130 29790 Inelastic preference for ethanol in rats: an analysis of ethanol's reinforcing effects - + 10116 Species rats - + MESH:D000431 Chemical ethanol - + MESH:D000431 Chemical @@ -23746,8 +23846,8 @@ 330 30155 Delay discounting of food and remifentanil in rhesus monkeys - - MESH:C071741 + + MESH:D000077208 Chemical remifentanil @@ -23780,7 +23880,13 @@ 424 30341 Preclinical evaluation of pharmacotherapies for treatment of cocaine and opioid abuse using drug self-administration procedures - + + MESH:D009293 + Disease + + opioid abuse + + MESH:D003042 Chemical @@ -23853,7 +23959,7 @@ 199 30738 Behavioral economics and drug choice: effects of unit price on cocaine self-administration by monkeys - + MESH:D003042 Chemical @@ -23887,7 +23993,7 @@ 300 30958 Effects of increasing response requirement on choice between cocaine and food in rhesus monkeys - + MESH:D003042 Chemical @@ -23907,13 +24013,13 @@ 332 31054 Effects of chronic D-amphetamine treatment on cocaine-and food-maintained responding under a progressive-ratio schedule in rhesus monkeys - - MESH:D000661 + + MESH:D003913 Chemical D-amphetamine - + MESH:D003042 Chemical @@ -23946,17 +24052,17 @@ ref 31259 Mu/kappa opioid interactions in rhesus monkeys: implications for analgesia and abuse liability - + 9544 Species rhesus monkeys - + MESH:D000699 Disease - - analgesia and abuse liability + + analgesia @@ -23976,7 +24082,7 @@ surname:Cheung;given-names:TH 31354 A new criterion for acquisition of nicotine self-administration in rats - + MESH:D009538 Chemical @@ -24037,17 +24143,6 @@ 1364 31544 A new view of the effect of dopamine receptor antagonism on operant performance for rewarding brain stimulation in the rat - - MESH:D004298 - Chemical - - dopamine - - - Chemical - - antagonism - 127 @@ -24061,7 +24156,7 @@ 203 31667 Intravenous self-administration of cocaine under concurrent VI schedules of reinforcement - + MESH:D003042 Chemical @@ -24081,7 +24176,7 @@ 106 31757 Effects of delay to reinforcement on the choice between cocaine and food in rhesus monkeys - + MESH:D003042 Chemical @@ -24107,19 +24202,13 @@ fig_caption 32019 Simulated cocaine vs. food choice dose-effect functions. (A) The effects of changing sensitivity to magnitude with sensitivity to price held constant at 1.0. (B) The effects of changing sensitivity to price with sensitivity to magnitude held constant at 5.0. Ordinates: proportion of cocaine choices. Abscissae: unit cocaine dose (mg/kg/injection). - + MESH:D003042 Chemical cocaine - - MESH:D003042 - Chemical - - cocaine - - + MESH:D003042 Chemical @@ -24133,37 +24222,31 @@ fig_caption 32368 Continuous 14-day D-amphetamine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of D-amphetamine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: D-amphetamine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). - + MESH:D003042 Chemical cocaine - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - - MESH:D003042 - Chemical - - cocaine - - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - - MESH:D000661 + + MESH:D003913 Chemical - + D-amphetamine - + MESH:D003042 Chemical @@ -24177,37 +24260,31 @@ fig_caption 33047 Continuous 14-day (+)-phendimetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of (+)-phendimetrazine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phendimetrazine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). - + MESH:C100294 Chemical (+)-phendimetrazine - - MESH:D003042 - Chemical - - cocaine - - + MESH:C100294 Chemical (+)-phendimetrazine - + MESH:D003042 Chemical cocaine - + MESH:C100294 Chemical (+)-phendimetrazine - + MESH:D003042 Chemical @@ -24221,37 +24298,31 @@ fig_caption 33744 Continuous 14-day (+)-phenmetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of (+)-phenmetrazine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phenmetrazine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). - - MESH:D003042 - Chemical - - cocaine - - + MESH:D010633 Chemical (+)-phenmetrazine - + MESH:D010633 Chemical (+)-phenmetrazine - + MESH:D010633 Chemical (+)-phenmetrazine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical @@ -24265,31 +24336,25 @@ fig_caption 34435 Effect of 7-day cocaine or food removal on (A) cocaine vs. food choice and (B) and (C) best-fitting parameter estimates of Eq. (1) plotted as a function of extinction manipulation. (A) Ordinate: obtained and predicted (solid curves) proportion cocaine choice. Abscissa: unit dose cocaine in mg/kg/injection (log scale). (B) and (C) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phenmetrazine dose in mg/kg/h. Model predictions (A) and parameter estimates (B) and (C) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (B) and (C). - + MESH:D003042 Chemical cocaine - - MESH:D003042 - Chemical - - cocaine - - + MESH:D003042 Chemical cocaine - + MESH:D010633 Chemical (+)-phenmetrazine - + MESH:D003042 Chemical @@ -24303,36 +24368,30 @@ fig_caption 35087 Continuous 14-day D-amphetamine treatment effects on demand for cocaine and food. Model predictions were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Ordinates: normalized consumption (reinforcers earned/total reinforcers). Abscissae: unit price of cocaine (UP = FR/dose). The unit price at which cocaine and food consumption are predicted to be equal is indicated in the insets as UP50. - + MESH:D003042 Chemical cocaine - - MESH:D014397 - Disease - - food consumption + + MESH:D003913 + Chemical + + D-amphetamine - + MESH:D003042 Chemical cocaine - + MESH:D003042 Chemical cocaine - - MESH:D000661 - Chemical - - D-amphetamine - TABLE @@ -24367,47 +24426,17 @@ T1.xml 36447 Mean M1452 M1473 M1504 M1488 M1478 M1480 M1412 M1414 D-Amphetamine AICc (Null, H0) -428.2 -62.4 -99.4 -239.3 -104.5 -57.0 -63.0 Delta (Full, sm, sp) 602.0 22.4 107.1 -12.9 122.8 262.2 129.0 Delta (Magnitude, sm) -4.4 18.0 -24.0 -4.4 13.3 3.1 -3.2 Delta (Price sp) 473.5 15.1 78.2 -1.7 47.1 291.7 167.0 n 15 25 19 25 25 24 R2 0.999 0.983 0.996 0.999 0.991 0.999 0.999 (+)-Phendimetrazine AICc (Null, H0) -276.7 -67.6 -50.8 -136.8 -70.3 Delta (Full, sm, sp) 271.1 57.8 118.1 172.7 46.9 Delta (Magnitude, sm) 63.5 0.2 131.0 -1.1 -3.3 Delta (Price sp) 298.2 93.2 89.3 184.5 58.8 n 20 23 25 25 R2 0.991 0.999 0.999 0.999 0.977 (+)-Phenmetrazine AICc (Null, H0) -425.8 -106.8 -74.4 -73.8 -104.6 -83.5 Delta (Full, sm, sp) 163.7 30.7 170.5 3.0 128.1 145.5 Delta (Magnitude, sm) 58.2 28.2 4.2 4.7 -4.5 144.4 Delta (Price sp) 203.8 30.4 136.0 14.8 139.9 93.1 n 25 25 25 25 25 R2 0.997 0.982 0.999 0.999 0.999 0.997 Extinction AICc (Null, H0) -234.5 -30.4 -29.1 -45.0 -33.9 Delta (Full, sm, sp) 235.6 150.1 83.2 226.4 21.3 Delta (Magnitude, sm) 13.2 -1.6 -8.4 34.2 3.1 Delta (Price sp) 239.1 157.3 90.5 238.0 18.4 n 15 15 15 15 R2 0.996 0.999 0.999 0.999 0.900 - - MESH:C027727 - Chemical - - H0 - - - MESH:C027727 - Chemical - - H0 - - - MESH:D000661 - Chemical - - D-Amphetamine AICc - - - MESH:C027727 - Chemical - - H0 - - + MESH:D010633 Chemical - - Phenmetrazine AICc - - - MESH:C027727 - Chemical - - H0 + + (+)-Phenmetrazine - - MESH:C100294 + + MESH:C000604007 Chemical - - Phendimetrazine AICc + + sp @@ -24440,144 +24469,156 @@ Cholon DM, Quinney NL, Fulcher ML, Esther CR, Das J, Dokholyan NV, Randell SH, Boucher RC, Gentzsch M 0 Potentiator Ivacaftor Abrogates Pharmacological Correction of DeltaF508 CFTR in Cystic Fibrosis - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease Cystic Fibrosis + + MESH:C545203 + Chemical + + Ivacaftor + ABSTRACT abstract 96 Cystic Fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR). Newly developed "correctors" such as lumacaftor (VX-809) that improve CFTR maturation and trafficking and "potentiators" such as ivacaftor (VX-770) that enhance channel activity may provide important advances in CF therapy. Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation (G551D) that affects only channel activity, a single compound is not sufficient to treat patients with the more common CFTR mutation, DeltaF508. Thus, patients with DeltaF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit. However, whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro, the impact of chronic therapy has not been established. In studies of human primary airway epithelial cells, we found that both acute and chronic treatment with VX-770 improved CFTR function in cells with the G551D mutation, consistent with clinical studies. In contrast, chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated CFTR correction in DeltaF508 homozygous cultures. This result reflected the destabilization of corrected DeltaF508 CFTR by VX-770, dramatically increasing its turnover rate. Chronic VX-770 treatment also reduced mature wild-type CFTR levels and function. These findings demonstrate that chronic treatment with CFTR potentiators and correctors may have unexpected effects that cannot be predicted from short-term studies. Combining of these drugs to maximize rescue of DeltaF508 CFTR may require changes in dosing and/or development of new potentiator compounds that do not interfere with CFTR stability. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species human - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation G551D - + MESH:D003550 Disease Cystic Fibrosis - - p.508delF - DNAMutation - - DeltaF508 + + MESH:C545203 + Chemical + + ivacaftor - + + MESH:C569105 + Chemical + + VX-809 + + + MESH:C569105 + Chemical + + lumacaftor + + p.G551D;RS#:75527207 ProteinMutation @@ -24595,110 +24636,110 @@ paragraph 1885 The most common autosomal recessive genetic disease of the Caucasian population in the United States and Europe, cystic fibrosis (CF), is characterized by abnormal epithelial ion transport. Mutations in the CF transmembrane conductance regulator (CFTR) result in loss of CFTR-mediated Cl- and HCO3- transport by secretory and absorptive epithelial cells in multiple organs, including lungs, pancreas, liver, and intestine. In the lung, disturbances of airway surface liquid homeostasis produce thick and viscous mucus that leads to mucus stasis, airway obstruction, persistent infection, inflammation, and a progressive decline in lung function. These features are the hallmarks of CF lung disease and result in limited life expectancy. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + + MESH:D001639 + Chemical + + HCO3 + + MESH:D007239 Disease infection - + + MESH:D003550 + Disease + + CF lung disease + + MESH:D003550 Disease cystic fibrosis - + MESH:D000402 Disease airway obstruction - - MESH:D003550 - Disease - - hallmarks of CF lung disease - - + MESH:D010190 Disease pancreas - + MESH:D007249 Disease inflammation - + MESH:D030342 Disease autosomal recessive genetic disease - - MESH:C504136 - Chemical - - HCO3 - INTRO paragraph 2626 In 1989, the identification of the CFTR gene on chromosome 7 and its most common mutation, DeltaF508 CFTR, raised hope for a cure that would address the underlying cause of CF. Intense high-throughput screening approaches over the last decade have yielded compounds that modulate mutant CFTR function. Small-molecule compounds that rescue mutant CFTR can be assigned to 2 groups: 1) "corrector" compounds that promote maturation and delivery of CFTR proteins to the apical surface and 2) "potentiator" compounds that activate apical CFTR by increasing the open time of the channel. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + c.508DELTA>CFTR DNAMutation @@ -24710,208 +24751,184 @@ paragraph 3213 The FDA recently approved the CFTR potentiator compound VX-770 (ivacaftor; trade name Kalydeco) as the first drug that directly restores CFTR activity in CF patients who carry a G551D mutation. G551D CFTR reaches the plasma membrane of epithelial cells, but the protein exhibits a gating defect that abolishes ATP-dependent channel opening and causes severe CF. In patients carrying a G551D mutation, VX-770 has proven to be effective in clinical trials, in which treated patients exhibited marked improvements in sweat chloride values and pulmonary function. The development of a CFTR-targeted drug that benefits CF patients marked a breakthrough in the treatment of CF. Unfortunately, because less than 5% of the CF population have the G551D mutation, this specific therapy helps only a limited number of patients. 90% of CF patients carry the DeltaF508 mutation, which produces a protein that does not mature normally and does not traffic to the plasma membrane. VX-770 treatment did not benefit CF subjects with the DeltaF508 mutation, likely because this compound only acts on protein that has trafficked to the plasma membrane. Based on these findings, an attractive therapeutic strategy for the DeltaF508 CF patient population is to promote transfer of the ER-retained DeltaF508 CFTR protein to the plasma membrane using small-molecule corrector compounds. Studies have estimated that the extent of correction in DeltaF508 airway epithelial cells must approximate 10-25% of wild-type (WT) CFTR function to provide therapeutic benefit. In vitro treatment of CF airway epithelial cultures homozygous for the DeltaF508 mutation with the most promising corrector compound, VX-809 (lumacaftor), resulted in CFTR function of ~14% relative to non-CF ("wild-type") human airway epithelial cells. However, administration of VX-809 did not provide a significant therapeutic benefit for DeltaF508 CF patients in recent clinical trials, most likely because DeltaF508 CFTR correction in vivo was less than 10% of wild-type levels, the lower limit of detection, and thus no mature DeltaF508 CFTR protein was observed. Therefore, a logical next step was to combine corrector and potentiator therapies to rescue DeltaF508 and increase protein function. One of the most promising current clinical trials designed to optimize DeltaF508 CFTR function involved the administration of the corrector VX-809 with the potentiator VX-770. Increases in VX-809-rescued DeltaF508 CFTR function have been demonstrated after acute administration of VX-770 in primary human airway epithelial cells from CF patients and human organoids derived from CF (DeltaF508/DeltaF508) intestinal tissue. Surprisingly, chronic co-administration of VX-809 and VX-770 in Phase 2 and 3 studies produced only small improvements in lung function in CF patients homozygous for the DeltaF508 CFTR mutation. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species human - + 9606 Species patients - + 9606 Species human - + 9606 Species patients - + 9606 Species human - + 9606 Species patients - - p.508delF - DNAMutation - - DeltaF508 - - - p.G551D;RS#:75527207 - ProteinMutation - - G551D - - - p.508delF - DNAMutation - - DeltaF508 + + MESH:C569105 + Chemical + + VX-809 - + p.G551D;RS#:75527207 ProteinMutation G551D - + MESH:D002712 Chemical chloride - - p.508delF - DNAMutation - - DeltaF508 - - - MESH:D000255 - Chemical - - ATP - - p.G551D;RS#:75527207 - ProteinMutation - - G551D + MESH:C545203 + Chemical + + VX-770 c.508DELTA>CFTR @@ -24920,18 +24937,42 @@ DeltaF508 CFTR - p.508delF - DNAMutation - - DeltaF508 - - MESH:C545203 Chemical - - Kalydeco + + ivacaftor + + + MESH:D003550 + Disease + + benefits CF + MESH:C569105 + Chemical + + VX-809 + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C569105 + Chemical + + lumacaftor + + p.G551D;RS#:75527207 ProteinMutation @@ -24943,76 +24984,82 @@ paragraph 6075 The aim of this study was to elucidate the molecular mechanism(s) underlying the limited improvement in DeltaF508 CFTR function when a corrector, VX-809 and a potentiator, VX-770 were co-administered to CF patients. We therefore investigated whether there were unexpected effects of chronically exposing CF cultures in vitro to VX-809 and VX-770, as would be achieved by oral dosing in clinical trials. A combination of CFTR bioelectric and biochemical approaches were utilized to investigate this interaction. Human bronchial epithelial (HBE) cells were used for these studies and exposed for 48 hrs to clinically relevant concentrations of both compounds. In addition, because of the success of VX-770 in CF patients with the G551D mutation, it has recently been suggested that treatment with VX-770 may be a pharmacological approach to enhance CFTR function in patients with chronic obstructive pulmonary disease (COPD). Accordingly, similar experimental approaches were utilized to explore the effects of VX-770 on WT CFTR, which matures normally and traffics to the plasma membrane. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species Human - + 9606 Species patients - + 9606 Species patients - + 1080 Gene 55465 CFTR - + MESH:D029424 Disease COPD - + p.G551D;RS#:75527207 ProteinMutation G551D - + MESH:D029424 Disease chronic obstructive pulmonary disease + + MESH:C545203 + Chemical + + VX-770 + RESULTS @@ -25025,14 +25072,14 @@ title_2 7171 Acute and chronic VX-770 treatments rescue G551D CFTR function - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation @@ -25044,113 +25091,107 @@ paragraph 7234 It has been recently demonstrated that acute VX-770 administration increased CFTR function in cell lines expressing G551D CFTR and augmented Cl- secretion in primary human bronchial epithelial (HBE) cells derived from CF patients with the G551D mutation on one allele and the DeltaF508 mutation on the other allele. To compare the effects of chronic versus acute VX-770 drug administration in CF airway epithelia with a G551D mutation, we used well-differentiated primary CF HBE cultures (G551D/DeltaF508) as a model. Cultures were treated chronically for 48 hrs with VX-770 or vehicle in the basolateral medium, and then transepithelial short-circuit currents (ISC) were measured in Ussing chambers (Fig. 1). Cultures were exposed to amiloride to inhibit the epithelial Na+ channel (ENaC) and subsequently forskolin to stimulate Cl- secretion by CFTR. As previously reported, acute administration of VX-770 (aVX770) raised Cl- secretion after forskolin administration (Fig. 1A). Chronic VX-770 (cVX770) administration raised forskolin responsiveness but eliminated subsequent responses to acute VX-770 administration (Fig. 1A,B,C, Table S1). Cultures chronically treated with VX-770 exhibited total CFTR-mediated responses (Fig. 1D) and inhibition with CFTRinh-172 (Fig. 1E) equal to cultures treated with forskolin and acute VX-770. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species human - + 9606 Species patients - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical forskolin - + MESH:D005576 Chemical forskolin - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation G551D - + MESH:D005576 Chemical forskolin - + + MESH:C482900 + Chemical + + CFTRinh-172 + + MESH:D000584 Chemical amiloride - + MESH:D005576 Chemical forskolin - + p.G551D;RS#:75527207 ProteinMutation G551D - + MESH:D003550 Disease CF airway epithelia - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation @@ -25162,40 +25203,40 @@ paragraph 8569 Chronically VX-770-treated G551D/DeltaF508 HBE cultures also exhibited a decrease in amiloride-sensitive currents (Fig. S1A, Table S2), suggesting decreased ENaC function. This finding is consistent with restoration of CFTR-mediated ENaC inhibitory activity because cleavage of ENaC was diminished in chronically VX-770-treated CF cultures (Fig. S1B). The average UTP responsiveness, an index of Ca2+ activated Cl- channel (CaCC) activity, was reduced with chronic as compared to acute VX-770 administration (Fig. S1C, Table S2). - + 1080 Gene 55465 CFTR - + 55107 Gene 75079 Ca2+ activated Cl- channel - + 55107 Gene 75079 CaCC - + MESH:D014544 Chemical UTP - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + MESH:D000584 Chemical @@ -25207,14 +25248,14 @@ paragraph 9099 In sum, these results demonstrate that both acute and chronic treatment with VX-770 improved CFTR function in HBE cells with the G551D mutation, consistent with clinical studies. - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation @@ -25232,115 +25273,133 @@ paragraph 9348 CF patients harboring the DeltaF508 CFTR mutation, which produces protein maturation and trafficking defects, have little/no CFTR at the cell surface. Consequently, treatment with a corrector compound, such as VX-809, is required for VX-770 to potentiate surface-localized DeltaF508 CFTR. To mimic clinical administration of VX-809 and VX-770 as a combination therapy for the DeltaF508 CF patients, primary CF HBE cultures (DeltaF508/DeltaF508) were treated with both pharmacological agents for 48 hrs and then Ussing chamber experiments were performed to measure DeltaF508 CFTR function (Fig. 2, Table S3). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patients - + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + c.508DELTA>CFTR DNAMutation DeltaF508 CFTR - + CVCL:0287 CellLine CF HBE + + MESH:C569105 + Chemical + + VX-809 + RESULTS paragraph 9956 Anion efflux across the apical epithelial membrane of airway epithelia in response to cAMP is mediated by CFTR and was not substantial in vehicle-treated DeltaF508/DeltaF508 CF HBE cultures Fig. 2A,B; Vehicle). However, VX-809 administration produced DeltaF508 CFTR correction as evidenced by stimulation of Cl- secretion (Fig. 2). Specifically, correction by VX-809 produced responses to forskolin (Fig. 2A, B; VX809) that were enhanced by acute administration of VX-770 (Fig. 2A,C). These data indicate that acutely applied VX-770 further activated ("potentiated") VX-809-rescued DeltaF508 CFTR. However, the CFTR-mediated ISC increase after addition of VX-770 to corrector-rescued DeltaF508 CFTR was transient. The ISC decrease over time may be indicative of a rapidly decreasing quantity of functional protein at the apical membrane. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + CVCL:0287 CellLine CF HBE - - MESH:D000242 + + MESH:D005576 + Chemical + + forskolin + + + MESH:C569105 + Chemical + + VX-809 + + + - Chemical cAMP - - MESH:D005576 + + MESH:C569105 Chemical - - forskolin + + VX-809 @@ -25348,27 +25407,33 @@ paragraph 10794 In contrast, rescue of DeltaF508 CFTR function was dramatically decreased in cultures that had been chronically treated with VX-809 and VX-770 compared to VX-809 alone (Fig. 2A: VX809 vs. VX809+VX770). This loss of "corrected" function was reflected in reduced Cl- secretion responses to forskolin (Fig. 2B) and reduced inhibition of stimulated CFTR Cl- secretion with CFTRinh-172 (Fig. 2D). Thus, these data contrast with the significant (P = 0.0177) acute VX-770 responses in VX-809-treated cultures (Fig. 2C). Again, we noticed that the response to UTP-stimulated ISC decreased upon chronic VX-770 treatment (Figs. 2A and S1D). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + MESH:D014544 Chemical UTP - + + MESH:C482900 + Chemical + + CFTRinh-172 + + MESH:D005576 Chemical @@ -25380,32 +25445,32 @@ paragraph 11430 We also tested the impact of chronic VX-770 treatment on DeltaF508 correction in CF HBE cultures (DeltaF508/DeltaF508) by corrector compound VX-661. Similar to VX-809-treated CF cells, VX-661-corrected CF HBE cells showed a drastic reduction in forskolin-mediated CFTR function when VX-770 was chronically added (Fig. S2, Table S4). - + 1080 Gene 55465 CFTR - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical forskolin - - p.508delF - DNAMutation - - DeltaF508 + + MESH:C569105 + Chemical + + VX-809 - + CVCL:0287 CellLine @@ -25423,113 +25488,119 @@ paragraph 11868 To explore the mechanism(s) mediating the VX-770-induced reduction of VX-809-corrected DeltaF508 CFTR function, we used Western blotting techniques to analyze protein maturation and turnover. In normal HBE cells, we detected a mature, complex glycosylated form, band C (Fig. 3A, NL: *), with a substantially greater molecular weight than the immature band B (Fig. 3A, NL: ). In contrast, only band B could be detected in vehicle-treated DeltaF508/DeltaF508 CF HBE cells (Fig. 3A, CF). As previously reported, treatment with VX-809 alone resulted in formation of a modest amount of mature band C in CF HBE cultures, which was not present in vehicle- or VX-770-treated CF cells (Fig. 3A, CF). However, when CF cells were treated chronically with both VX-809 and VX-770, the amount of mature DeltaF508 CFTR was diminished, and instead the DeltaF508 CFTR protein appeared almost exclusively as immature band B. These data suggest that chronic VX-770 treatment impeded correction of DeltaF508 CFTR by VX-809. We investigated the impact of chronic VX-770 treatment on protein stability by measuring the turnover rate of corrected DeltaF508 CFTR. DeltaF508 CFTR was stably expressed in baby hamster kidney (BHK-21) cells and corrected with VX-809 in the presence and absence of VX-770. Rescue with VX-809 was performed at 27 C for 24 hrs because VX-770 prevented VX-809 mediated correction of DeltaF508 CFTR at 37 C (Fig. 3A). After rescue, cells were shifted to 37 C and protein biosynthesis was inhibited by addition of cycloheximide. The amount of remaining mature DeltaF508 CFTR was then measured after 3 and 6 hrs. The turnover rate of rescued DeltaF508 CFTR band C increased and the half-life accordingly decreased by ~2.5 fold in the presence of VX-770 (Fig. 3B,C, Table S5), whereas the decrease in band B levels was not affected by the presence of VX-770 (Fig. 3B). These data clearly show that VX-770 decreased the stability, and thus increased the turnover rate, of VX-809-rescued DeltaF508 CFTR. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 10034 Species hamster - + CVCL:1914 CellLine BHK-21 - + MESH:D003513 Chemical cycloheximide - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE + + MESH:C569105 + Chemical + + VX-809 + RESULTS title_2 13870 VX-770 affects correction of DeltaF508 CFTR in a dose-dependent relationship - + 1080 Gene 55465 @@ -25542,72 +25613,72 @@ paragraph 13947 Lower concentrations of VX-770 were chronically administered to DeltaF508/DeltaF508 CF HBE cells to study the dose effect on VX-809-rescued DeltaF508 CFTR (Fig. 4). To obtain an average measure of CFTR-mediated ISC for the period spanning forskolin stimulation to CFTR inhibition, the area under the curve (AUC) was calculated for this interval. Dividing AUC by time yielded average CFTR DeltaISC between activation and inhibition. When 1 microM VX-770 was administered chronically with VX-809, the forskolin responses of CF HBE cells were intermediate between cells treated with VX-809 alone and cells treated with 5 microM VX-770 and VX-809 (Fig. 4A,B, Table S6). There was a significant reduction in AUC in corrector-treated cells with 1 microM versus 5 microM VX-770 (P =0.0049). Although there was not a significant difference in AUC of VX-809-treated cells with 50 nM VX-770 versus VX-809 alone (Fig. 4B), this low dose of VX-770 caused a rapid decline of the slope after forskolin treatment (Fig 4C, Table S7). Chronic treatment with either 50 nM or 1 microM VX-770 eliminated responses to acute VX-770 (Fig. 4A,C). Western blots to detect mature band C protein in CF HBE cells confirmed that VX-809 rescue was inhibited by VX-770 in a dose-dependent manner. As the concentration of VX-770 increased, the amount of VX-809-corrected DeltaF508 CFTR decreased (Fig. 4D, E, Table S8). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical forskolin - + MESH:D005576 Chemical forskolin - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical @@ -25625,58 +25696,70 @@ paragraph 15406 To investigate the effects of chronic VX-770 treatment on normal (NL) CFTR, we measured anion secretion of NL primary HBE cultures treated with VX-770 for 48 hrs (Fig. 5). Strikingly, administration of 5 microM VX-770 reduced CFTR-mediated Cl- secretion, as reflected by decreased forskolin responses (Fig. 5A,B,C, Table S9) and decreased inhibition of CFTR-mediated current by CFTRinh-172 (Fig. 5A,D). These functional responses were paralleled by a decrease in CFTR band C (Fig. 5E). In cells chronically treated with VX-770, we also observed a consistent reduction in amiloride-sensitive current (Fig. S3A, Table S2) and a substantial inhibition of UTP-sensitive current (Fig. S3B, Table S2) similar to that observed in CF HBE cells (Fig. S1A,C,D, Table S2). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + + MESH:C482900 + Chemical + + CFTRinh-172 + + CVCL:0287 CellLine CF HBE - + MESH:D000584 Chemical amiloride - + MESH:D005576 Chemical forskolin - + MESH:D014544 Chemical UTP + + MESH:C545203 + Chemical + + VX-770 + RESULTS @@ -25689,78 +25772,78 @@ paragraph 16253 As a test for the specificity of chronic VX-770 effects, we analyzed whether fundamental epithelial parameters were altered in HBE cultures chronically treated with VX-770 (Fig. 6). The morphology of highly differentiated ciliated HBE cultures was identical in cells treated with vehicle (DMSO) or VX-770 (Fig. 6A). Transepithelial resistance (Rt) of primary HBE cultures was also not affected by chronic VX-770 exposure (Fig. 6B, Table S10). The inhibition by VX-770 of both Na+ absorption (amiloride-sensitive ISC) and Cl- secretion and currents (CFTR- and CaCC-mediated ISC) raised the possibility that driving forces for ion transport, in part generated by Na+/K+ ATPase activity, were perturbed by VX-770. Nystatin, a polyene antibiotic that enables monovalent cations to permeate biological membranes and raise Na+/K+ ATPase activity, did not produce significantly different ISC responses when applied to vehicle- or VX-770-treated HBE cultures (Fig. 6C, Table S10), suggesting intact Na+/K+ ATPase activity. Measurements of intracellular concentrations of VX-809 and VX-770 by mass spectrometry confirmed the presence of these compounds in treated HBE cultures and indicated that cellular VX-809 concentrations were not affected by the presence of VX-770 (Fig. S4, Table S11). - + 1080 Gene 55465 CFTR - + 55107 Gene 75079 CaCC - + 6267 Gene S11 - + 6204 Gene 788 S10 - + 6204 Gene 788 S10 - + MESH:D004121 Chemical DMSO - + + MESH:D011090 + Chemical + + polyene + + MESH:D000584 Chemical amiloride - + MESH:D009761 Chemical Nystatin - - MESH:D011090 - Chemical - - polyene - RESULTS title_2 17539 Destabilization by VX-770 is beneficial for G551D, but not for wild-type, or DeltaF508 CFTR function - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation @@ -25772,72 +25855,72 @@ paragraph 17640 A high C:B band ratio indicates normal CFTR protein maturation. Biochemical analysis by Western blotting showed that chronic VX-770 treatment dramatically reduced the amount of mature CFTR in both NL cultures expressing wild-type CFTR (Figs. 5E and 7A) and VX-809-rescued DeltaF508 CFTR in CF HBE cells (Figs. 3A and 4E). Indeed, the C:B band ratio decreased with chronic VX-770 exposure by more than 50% in NL cultures (Fig. 7B, NL, Table S12). In contrast, the levels of mature (band C) G551D CFTR detected in G551D/DeltaF508 CF HBE cultures were not significantly diminished by chronic exposure to 5 microM VX-770, suggesting that G551D was resistant to the destabilizing effects of VX-770 (Fig 7A, G551D/DeltaF508). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 6268 Gene S12 - + CVCL:0287 CellLine CF HBE - + p.G551D;RS#:75527207 ProteinMutation G551D - + CVCL:0287 CellLine CF HBE - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + p.G551D;RS#:75527207 ProteinMutation @@ -25849,100 +25932,112 @@ paragraph 18360 To explore the relationship between VX-770, VX-809, and CFTR protein stability, we performed calculations of thermodynamic stability of CFTR protein utilizing a structural homology model. This model revealed that CFTR amino acid F508 is located at the nucleotide binding domain 1- cytoplasmic loop 4 (NBD1-CL4) interface (Fig. 7C), and therefore participates in important interdomain interactions. Thus, in DeltaF508 CFTR, the deletion of amino acid F508 not only reduces the stability of the NBD1 domain, but importantly, may destabilize multidomain assembly of CFTR. In contrast, in this structural model of CFTR, amino acid G551 is positioned between the 2 NBDs (Figs. 7C, S5), and the G551D mutation is thought to contort NBD dimer formation and abolish ATP-dependent channel opening by disrupting the signature sequence in NBD1. To evaluate whether the G551D mutation also affects the overall stability of CFTR by inducing conformational restructuring of the protein, we computationally estimated the DeltaDeltaG for G551D CFTR. We found that G551D had a stabilizing effect on the CFTR protein (DeltaDeltaG = -8.1 kcal/mol). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 100862696 Gene CL4 - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + + - + Chemical + + amino acid F508 + + MESH:D000255 Chemical ATP - + + - + Chemical + + amino acid F508 + + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation @@ -25960,60 +26055,54 @@ paragraph 19501 Potentiator compounds act on mutant CFTR channels that are on the surface of epithelial cells. VX-770 has been approved as a pharmacological agent to treat CF patients with at least one copy of the G551D mutation. However, the most common mutant protein in CF patients, DeltaF508 CFTR, is not found at the cell surface. DeltaF508 CFTR has a folding defect and is retained in the ER but can be partially rescued by corrector compounds that promote delivery of a small proportion of mutant DeltaF508 proteins to the cell surface. Corrector-rescued DeltaF508 CFTR is reported to have a shorter half-life at the cell surface and exhibits increased thermal inactivation as well as a gating defect when compared to WT CFTR. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patients - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation @@ -26025,113 +26114,113 @@ paragraph 20219 The efficacy of orally administered VX-770 was established in clinical trials in G551D CF patients by multiple outcome measurements. The clinical benefit of potentiation of G551D function was predicted from the effectiveness of acute administration of VX-770 in Ussing chambers, which measured rates of Cl- secretion across primary G551D/DeltaF508 CFTR airway epithelial cultures. Our studies confirmed that acute VX-770 administration restored CFTR Cl- secretion activity in HBE cells from patients carrying the G551D mutation. Further, our data demonstrated that G551D/DeltaF508 cultures chronically treated with VX-770 also exhibited increased Cl- secretion via G551D CFTR, but stimulation with forskolin, which raises intracellular cAMP, was required to activate Cl- secretion. This result could suggest a benefit from administering cAMP-raising beta2-adrenergic receptor agonists as a routine part of the treatment for G551D CF patients receiving VX-770. Overall, improvement in G551D CFTR activity with acute VX-770 in vitro was also observed with chronic in vitro VX-770 administration. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D005576 Chemical forskolin - + + - + Chemical + + cAMP + + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation G551D - - MESH:D000242 + + - Chemical - + cAMP - + p.G551D;RS#:75527207 ProteinMutation G551D - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation G551D - - MESH:D000242 - Chemical - - cAMP - - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + p.G551D;RS#:75527207 ProteinMutation @@ -26143,127 +26232,139 @@ paragraph 21313 VX-809 appears to restore approximately 15% of normal function in DeltaF508/DeltaF508 CF HBE cells. However, 10-25% of CFTR function is estimated to be required to overcome CF symptoms. Therefore, a combination of VX-809 with a potentiator compound to further enhance DeltaF508 function may be necessary. Although acute treatment with VX-770 has been reported to enhance VX-809-rescued DeltaF508 activity, our data revealed that chronic application of VX-770 in combination with VX-809 or VX-661 did not. Chronic co-administration of VX-770 with either corrector to DeltaF508/DeltaF508 CF HBE cultures produced Cl- secretory responses that were smaller than responses to corrector alone. Our data suggest that the reduced capacity for Cl- secretion after chronic VX-770/VX-809 exposure reflected an increased turnover rate of corrected DeltaF508 CFTR. The VX-770-induced reduction of DeltaF508 correction observed in primary CF HBE cells was dose-dependent as measured by functional and biochemical approaches. We did not detect alterations in physiological properties of HBE cells that would suggest that toxic effects contributed to CFTR dysfunction after chronic VX-770/VX-809 treatment. Thus, our studies suggest that data describing the effectiveness of acute addition of VX-770 to VX-809-treated DeltaF508 CF HBE cells do not predict the outcome for chronic VX-770/VX-809 administration. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + CVCL:0287 CellLine CF HBE - + + MESH:C569105 + Chemical + + VX-809 + + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - - p.508delF - DNAMutation - - DeltaF508 - - + CVCL:0287 CellLine CF HBE + + MESH:C569105 + Chemical + + VX-809 + + + MESH:D006331 + Disease + + dysfunction + DISCUSS paragraph 22707 DeltaF508 CFTR has been shown to exhibit an increased thermodynamic instability of NBD1 and improper assembly of NBD1 into a complex with intracellular loop 4 (ICL4) of the second membrane-spanning domain (MSD2). The recently published data on CFTR domain fragments strongly suggest that VX-809 targets MSD1 of CFTR to suppress folding defects of DeltaF508 CFTR by enhancing interactions among NBD1, MSD1 and MSD2. Thus, VX-809 is predicted to enhance function of DeltaF508 by increasing its stability (Fig. 7D). Importantly, chronic exposure to VX-770 appeared to reverse the stabilization effect of VX-809 in DeltaF508 CFTR. Chronic VX-770 treatment resulted in a severe reduction in rescued DeltaF508 CFTR protein due to destabilization of rescued protein as reflected by a 2.5x increase in turnover rate. In contrast, G551D CFTR was more resistant to destabilization and loss of mature CFTR protein with chronic VX-770 exposure than WT or DeltaF508 CFTR. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation @@ -26275,91 +26376,91 @@ paragraph 23666 Our CFTR computational structural model allows us to speculate how VX-770 may interact with WT or mutant CFTR proteins to alter protein stability. CFTR requires conformational flexibility to function properly. The flexibility and stability of the CFTR protein is finely tuned and precisely balanced, which is a requirement for its ability to function properly as a regulated ion channel. The inherent increase in stability of the G551D protein may render it too rigid and inflexible (stable) for proper channel opening under basal cAMP-stimulated conditions. However, a decrease in stability mediated by VX-770 may render the G551D molecule more flexible and allow it to function as a cAMP-regulated Cl- channel (Fig. 7D). In contrast, destabilization of WT (ideal stability) or VX-809-rescued DeltaF508 CFTR (low stability) by VX-770 bound to the CFTR molecule resulted in decreased WT CFTR function and absent DeltaF508 CFTR function. These considerations reveal that chemical correction of low stability CFTR mutants is complicated and necessitates precision. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation G551D - - MESH:D000242 + + - Chemical - + cAMP - + p.G551D;RS#:75527207 ProteinMutation G551D - - MESH:D000242 + + - Chemical - + cAMP @@ -26368,92 +26469,92 @@ paragraph 24729 Some airway diseases such as chronic obstructive pulmonary disease (COPD) have been associated with lessened CFTR function and consequently VX-770 has been suggested as a potential therapy. In a study that modeled the effects of VX-770 on COPD patients, reduction of CFTR function was achieved by exposing primary HBE cultures to cigarette smoke extract, and these cultures were subsequently exposed to acute administration of VX-770, which led to augmented CFTR-mediated currents. The finding that acute VX-770 treatment enhanced WT CFTR function contrasts with our finding that chronic treatment with VX-770 reduced WT CFTR function. In addition, forskolin-stimulated ISC in NL HBE cultures chronically treated with VX-770 was not stable over time, as indicated by the downward sloping trace. These functional data, coupled with our observations that the amount of mature WT CFTR was reduced by the continuous presence of VX-770, do not favor VX-770 as a therapy to enhance CFTR function in COPD. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + MESH:D029424 Disease COPD - + MESH:D029424 Disease COPD - + CVCL:0287 CellLine NL HBE - + + MESH:C545203 + Chemical + + VX-770 + + MESH:D029424 Disease COPD - - MESH:D005576 - Chemical - - forskolin - - + MESH:D029424 Disease @@ -26465,60 +26566,48 @@ paragraph 25728 Although potentiation of more rare CFTR mutants with partial defects in CFTR processing was recently detected upon acute VX-770 treatment in Fisher rat thyroid (FRT) cells overexpressing these variants, our data raise the concern that potentiation with chronic VX-770 treatment may not be observed in airway epithelia expressing these rare mutations. To optimize combination therapy for both DeltaF508 and rare CFTR processing mutations, minimizing interference of potentiator with corrector activity is required. One approach may be to finely tune the dosing regimens for potentiator compounds. Studies by Van Goor et al. indicate that the EC50 for acute application of VX-770 differs remarkably in G551D/DeltaF508 (EC50: 236 +- 200 nM) and DeltaF508/DeltaF508 (EC50: 22 +- 10 nM) HBE cultures. These data together with our findings, which demonstrate that inhibition of correction by VX-770 is dose dependent, suggest that drug concentrations are very critical and attempts to optimize potentiator activity on channel function while minimally affecting turnover rate of mutant CFTR should be considered. As a second approach, improved potentiator compounds that do not interfere with DeltaF508 CFTR correction and turnover are needed. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 10116 Species rat - - p.508delF - DNAMutation - - DeltaF508 - - - p.508delF - DNAMutation - - DeltaF508 - - + c.551G>D,D;RS#:75527207 ProteinMutation @@ -26530,56 +26619,56 @@ paragraph 26965 After chronic VX-770 treatment, we also observed diminished Cl- secretory responses to additions of the P2Y2 receptor agonist, UTP. UTP-stimulated Cl- secretion is elevated in CF airway epithelia and may compensate for the lack of CFTR function in vivo. Reduction of UTP-stimulated CaCC activity may constitute a disadvantage in CF airways, particularly if insufficient amounts of CFTR have been rescued. Thus, monitoring CaCC activity may be useful in future clinical corrector/potentiator studies. - + 1080 Gene 55465 CFTR - + 55107 Gene 75079 CaCC - + 1080 Gene 55465 CFTR - + 55107 Gene 75079 CaCC - + MESH:D014544 Chemical UTP - + MESH:D003550 Disease CF airway epithelia - + MESH:D014544 Chemical - + UTP - + MESH:D014544 Chemical - + UTP @@ -26588,33 +26677,33 @@ paragraph 27469 We observed a decrease in amiloride responses in the presence of chronic VX-770. While the effects in CF cells can be explained by the restoration of CFTR inhibiting activity, the inhibition of ENaC-mediated Na+ absorption in NL HBE cells raises the possibility that VX-770 may have off-target effects. Thus, it may be useful to measure ENaC function in future VX-770 clinical trials. Although a decline in ENaC function may be beneficial for CF patients, diminution of Cl- channels other than CFTR may be a disadvantage for maintaining adequate hydration of CF airways. The recent observation that VX-770 has antimicrobial properties in vitro suggests that it may also display off-target effects in vivo. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + CVCL:0287 CellLine NL HBE - + MESH:D000584 Chemical @@ -26626,14 +26715,14 @@ paragraph 28177 A limitation of our studies is that our experiments were performed in primary HBE cultures and not in vivo. However, primary HBE cultures are a well-established, near-physiologic system that is the most relevant model for studying CFTR function in airway epithelia, the tissue most affected by CF. Drug concentrations, turnover, and formation of metabolites may also differ in vitro and in vivo, which are crucial parameters to consider when extrapolating our data to the clinic. We therefore selected in vitro doses that mimicked clinically measured drug concentrations. For example, 5-day VX-770 treatments with 150 mg or 450 mg (administered as one dose/day) in patients resulted in VX-770 concentrations in blood plasma of 1.4 microg/ml and 5.5 microg/ml, respectively, which are equivalent to ~3.5 microM and ~14 microM. Current clinical trials test VX-770 doses in this range (250 mg taken twice per day). Thus, the concentrations tested in our studies in vitro appear relevant to the clinical experience. - + 1080 Gene 55465 CFTR - + 9606 Species @@ -26645,52 +26734,58 @@ paragraph 29189 We measured intracellular concentrations of VX-809 and VX-770, which revealed that these compounds (particularly VX-770) accumulated in cells and reached much higher concentrations than in the surrounding media. To determine whether the presence of VX-770 might have a negative impact on the intracellular concentration of VX-809, we obtained measurements of VX-809 concentrations in cell lysates with increasing concentrations of VX-770 and observed that the intracellular concentration of VX-809 was not affected by the presence of VX-770. Thus, comparisons of drug concentrations from in vitro and in vivo tissues may be useful in the future. + + MESH:C545203 + Chemical + + VX-770 + + + MESH:C569105 + Chemical + + VX-809 + DISCUSS paragraph 29835 Because there are no corrector compounds available that provide sufficient rescue of DeltaF508 in CF airways in vivo to alleviate symptoms of CF, potentiation of the small amount of corrected DeltaF508 CFTR is required. However, combination approaches to restore DeltaF508 CFTR function in CF to date have not considered drug-drug interactions of clinically relevant co-administered modulator compounds. Based on our study and the confirmatory data of Veit et al., knowledge of the interactions and interference between corrector and potentiator compounds is essential for successful therapy of the most prevalent mutation in CF patients, most of whom carry at least one allele of the DeltaF508 mutation. Furthermore, understanding the impact that potentiator compounds, such as VX-770, have on the stability of apical WT CFTR may also be important for other airway diseases that would benefit from augmentation of CFTR function. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - - p.508delF - DNAMutation - - DeltaF508 - METHODS @@ -26709,83 +26804,77 @@ paragraph 30800 Simultaneous treatment with small-molecular compounds, VX-809 or VX-661, together with VX-770 (Selleck Chemicals), is currently being examined as therapy for CF patients with the mutation, DeltaF508. Although in vitro studies examining acute treatment with this drug combination have been conducted, we sought to determine the impact of chronic treatment (48 hrs) with these compounds in primary human bronchial epithelial (HBE) cells. Primary HBE cells from normal (NL) or CF patients were obtained from bronchi of human lung tissue, as previously described. To evaluate CFTR function HBE cultures chronically treated with compounds were mounted in Ussing chambers to measure short-circuit currents (ISC). We examined differentiated primary CF HBE cells from at least 3 individuals for each genotype. To visualize the amount of mature and immature CFTR protein from HBE cultures, CFTR from whole-cell lysates was immunoprecipitated as previously described and Western blots were performed. Previously created baby hamster kidney (BHK-21) stably expressing DeltaF508 CFTR, were used in cycloheximide chase studies to examine the rate of protein turnover. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species human - + 9606 Species patients - + 9606 Species human - + 10034 Species hamster - + CVCL:0287 CellLine CF HBE - + CVCL:1914 CellLine BHK-21 - - p.508delF - DNAMutation - - DeltaF508 - - + MESH:D003513 Chemical @@ -26803,20 +26892,20 @@ paragraph 31968 Primary HBE cells were obtained from bronchi of human lung tissue under a protocol approved by the University of North Carolina Medical School Institutional Review Board. Primary NL and CF HBE cells were seeded at passage 2 on collagen-coated Millicell CM inserts (Millipore) and maintained at an air-liquid interface (ALI) at 37 C in 5% CO2 for 3-4 weeks, which allowed the cells to become fully differentiated. - + 9606 Species human - + CVCL:0287 CellLine CF HBE - - MESH:D002245 + + - Chemical CO2 @@ -26827,25 +26916,25 @@ paragraph 32384 BHK-21 cells were obtained from the American Type Culture Collection and grown at 37 C in 5% CO2. BHK-21 cells stably expressing Extope-DeltaF508 CFTR were created previously and maintained as described. - + 101831516 Gene CFTR - + + - + Chemical + + CO2 + + CVCL:1914 CellLine BHK-21 - - MESH:D002245 - Chemical - - CO2 - - + CVCL:1914 CellLine @@ -26863,58 +26952,64 @@ paragraph 32629 Whole-cell lysates were prepared as described previously. CFTR was immunoprecipitated as described previously and isolated using Protein A/G PLUS agarose (Santa Cruz Biotechnology). Samples were separated on 4-20% gradient SDS-PAGE gels (Bio-Rad) and then transferred to nitrocellulose. Blots were probed with mouse monoclonal anti-CFTR antibodies and then with IR Dye 680-goat anti-mouse IgG (Molecular Probes). Anti-actin (Cell Signaling) or anti-tubulin (LI-COR) was used as a loading control. Protein bands were visualized using an Odyssey Infrared Imaging System (LI-COR). - + 1080 Gene 55465 CFTR - + 6236 Gene 3071 Rad - + 1080 Gene 55465 CFTR - + 10090 Species mouse - + 9925 Species goat - + 10090 Species mouse - - MESH:C032259 + + MESH:D012967 Chemical SDS + + MESH:D012685 + Chemical + + agarose + METHODS title_2 33209 Cycloheximide chase to study turnover of rescued DeltaF508 CFTR - + MESH:D003513 Chemical @@ -26926,25 +27021,31 @@ paragraph 33273 BHK-21 cells expressing Extope-DeltaF508 CFTR were pretreated with compounds (VX-809, 5 microM; VX-770, 5 microM) for 24 hrs at 27 C before treatment with cycloheximide (200 microg/ml; Sigma) in the presence of compounds during chase times at 37 C. Whole-cell lysates were prepared and subjected to Western blotting. - + 1080 Gene 55465 CFTR - + CVCL:1914 CellLine BHK-21 - + MESH:D003513 Chemical cycloheximide + + MESH:C569105 + Chemical + + VX-809 + METHODS @@ -26957,17 +27058,35 @@ paragraph 33615 Primary HBE cultures grown at ALI on Millicell inserts were washed in PBS and fixed in 10% neutral buffered formalin prior to being embedded in paraffin and hematoxylin-eosin stained at the UNC CF Histology Core. Slides were viewed on a Leica DMIRB Inverted Microscope with a 40x 1.0 numerical aperture oil objective. - + + MESH:D011535 + Disease + + PBS + + + MESH:D004801 + Chemical + + eosin + + + MESH:D010232 + Chemical + + paraffin + + MESH:D005557 Chemical formalin - - MESH:D004801 + + MESH:D006416 Chemical - - hematoxylin-eosin + + hematoxylin @@ -26981,117 +27100,153 @@ paragraph 33970 In Ussing chambers (Physiological Instruments) HBE cultures were equilibrated to 37 C in a bilateral bath of Krebs-bicarbonate-Ringer buffer (KBR; in mM: 140 Na+, 120 Cl-, 5.2 K+, 1.2 Ca2+, 1.2 Mg2+, 2.4 HPO42-, 0.4 H2PO4-, 25 HCO3-, and 5 glucose, pH 7.4) and circulated with 95% O2-5% CO2. Short-circuit currents (ISC) were measured as previously described. All NL HBE cultures were measured in bilateral KBR. Unless noted otherwise, CF HBE cultures were measured with high potassium, low chloride (HKLC) buffer applied apically and KBR applied basolaterally, creating a Cl- gradient (5 mM/120 mM). Amiloride (100 microM) was added to block ENaC, followed by forskolin (10 microM), VX-770 (5 microM) and, if applicable, genistein (10 microM) to stimulate CFTR. CFTR was then inhibited with CFTRinh-172 (10 microM) and response to UTP (100 microM) was examined. In some experiments, nystatin (40 microM, apical) was added at the end of the measurements. Data were analyzed using Acquire and Analysis (version 2.3) software (Physiologic Instruments). ISC traces were imported to and processed in Origin 9.0.0. (OriginLab Corporation). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + + - + Chemical + + CO2 + + MESH:D000584 Chemical Amiloride - - MESH:C093415 + + - Chemical - - H2PO4 + + Mg2+ - + CVCL:0287 CellLine CF HBE - - MESH:D013481 + + - Chemical - - O2 + + Krebs - + MESH:D011188 Chemical potassium - - MESH:D002245 - Chemical - - CO2 - - - MESH:D002118 + + MESH:D001639 Chemical - - Ca2+ + + HCO3 - + MESH:D014544 Chemical UTP - + MESH:D019833 Chemical genistein - + + - + Chemical + + HPO42- + + MESH:D005947 Chemical glucose - - MESH:D008274 - Chemical - - Mg2 - - + MESH:D005576 Chemical forskolin - + + MESH:C545203 + Chemical + + VX-770 + + MESH:D001639 Chemical bicarbonate - + + - + Chemical + + H2PO4 + + + - + Chemical + + HKLC + + + MESH:D000069285 + Chemical + + Ca2+ + + + - + Chemical + + O2 + + CVCL:0287 CellLine NL HBE - - MESH:C504136 + + MESH:C482900 Chemical - - HCO3 + + CFTRinh-172 - + + MESH:C039004 + Chemical + + KBR + + MESH:D009761 Chemical nystatin - + MESH:D002712 Chemical @@ -27109,13 +27264,13 @@ paragraph 35136 Mass spectrometric (MS) methods were developed to detect VX-770 and VX-809 using strategies similar to those previously described. VX-770 was detected by monitoring transition of parent to daughter ion of m/z 393.3 171.1 in positive mode MS, with VX-809 detected by monitoring the transition m/z 453.3 197.1. Each compound generated a single peak using previously described liquid-chromatography-tandem MS (LC-MS/MS) methods, with run times of 11.1 and 10.8 minutes, respectively. To quantify drug concentrations in epithelial cells, cell lysates were extracted x 2 with equal volume of MTBE (Sigma), which was then lyophilized to dryness under vacuum centrifugation. Lyophilized samples were resuspended in a volume of 20% methanol in water equal to the original lysate volume, extracted, and 5 microl analyzed by LC-MS/MS as above. To control for matrix effects and variable recovery during extraction, untreated lysates were spiked with known concentrations of VX-770 and VX-809 and extracted in parallel. Concentrations in cell lysates were assessed by examining signal relative to the spiked samples. - - MESH:C043243 + + MESH:D014867 Chemical - - MTBE + + water - + MESH:D000432 Chemical @@ -27133,28 +27288,28 @@ paragraph 36280 We computationally estimated the DeltaDeltaG of mutation for G551D mutant CFTR using the Eris suite as described previously. Eris algorithms re-pack the side chains and evaluate the new free energy according to a physical force field upon the substitution of the relevant residue. - + 1080 Gene 55465 CFTR - + 340061 Gene 18868 Eris - + 340061 Gene 18868 Eris - + p.G551D;RS#:75527207 ProteinMutation @@ -27172,7 +27327,7 @@ paragraph 36582 Results are presented as means of average response per primary HBE cell donor and error bars are the standard error of the mean (SEM). Statistical analysis was performed by an unpaired two-tailed Student's t test in GraphPad Prism version 6.02 (GraphPad Software). P values of < 0.05 were considered to indicate statistical significance. - + 9606 Species @@ -27202,19 +27357,19 @@ footnote 37012 Fig. S1. Chronic VX-770 treatment alters responses to amiloride and UTP in CF HBE cells. - + MESH:D014544 Chemical UTP - + MESH:D000584 Chemical amiloride - + CVCL:0287 CellLine @@ -27232,19 +27387,19 @@ footnote 37186 Fig. S3. Chronic VX-770 treatment alters responses to amiloride and UTP in NL HBE cells. - + MESH:D014544 Chemical UTP - + MESH:D000584 Chemical amiloride - + CVCL:0287 CellLine @@ -27262,7 +27417,7 @@ footnote 37353 Fig. S5. G551D mutation in the NBD1 stabilizes CFTR protein. - + 1080 Gene 55465 @@ -27281,7 +27436,7 @@ footnote 37436 Table S1. VX-770 treatment restores G551D function. - + p.G551D;RS#:75527207 ProteinMutation @@ -27293,13 +27448,13 @@ footnote 37488 Table S2. Chronic VX-770 treatment alters responses to amiloride and UTP. - + MESH:D014544 Chemical UTP - + MESH:D000584 Chemical @@ -27323,7 +27478,7 @@ footnote 37734 Table S5. VX-770 diminishes biochemical correction by increasing turnover of corrected DeltaF508 CFTR. - + 1080 Gene 55465 @@ -27342,13 +27497,13 @@ footnote 37915 Table S7. VX-770 at low dose (50 nM) affects ISC of VX-809-corrected CF HBE cells (DeltaF508/DeltaF508). - + CVCL:0287 CellLine CF HBE - + DeltaF508/DeltaF508 ProteinMutation @@ -27360,13 +27515,13 @@ footnote 38020 Table S8. VX-770 affects C:B band ratio in CF HBE cultures (DeltaF508/DeltaF508). - + CVCL:0287 CellLine CF HBE - + DeltaF508/DeltaF508 ProteinMutation @@ -27378,7 +27533,7 @@ footnote 38102 Table S9. Chronic VX-770 treatment decreases the function of wild-type CFTR. - + 1080 Gene 55465 @@ -27391,14 +27546,14 @@ footnote 38179 Table S10. Transepithelial resistance and nystatin responses were not altered in HBE cultures chronically treated with VX-770. - + 6204 Gene 788 S10 - + MESH:D009761 Chemical @@ -27410,7 +27565,7 @@ footnote 38306 Table S11. VX-770 and VX-809 concentrations were measured in HBE cell lysates. - + 6267 Gene @@ -27422,14 +27577,14 @@ footnote 38385 Table S12. VX-770 reduces stability of CFTR. - + 1080 Gene 55465 CFTR - + 6268 Gene @@ -27447,28 +27602,28 @@ footnote 38618 Author contributions: DMC, NLQ, and MG performed the experiments and analyzed the data. MLF selected and cultured the primary CF and NL HBE cells. MGG, SHR and RCB discussed the overall design of experiments. MGG and DMC wrote the manuscript. CRE measured drug concentrations. JD and NVD determined DeltaDeltaG of the G551D protein. SHR and RCB provided reagents, technical advice and critical evaluation of the manuscript. All authors read and commented on the manuscript. - - MESH:C004944 + + MESH:C023025 Chemical - + DMC - + CVCL:0287 CellLine NL HBE - + p.G551D;RS#:75527207 ProteinMutation G551D - - MESH:C004944 + + MESH:C023025 Chemical - + DMC @@ -27483,7 +27638,7 @@ footnote 39109 Competing interests: RCB serves as the chairman of the board of Parion Sciences, a company which works on therapies designed to hydrate airway surfaces and also studies CFTR correctors; Parion Sciences does not have a potentiator program. The other authors declare that they have no competing interests. - + 1080 Gene 55465 @@ -27514,7 +27669,7 @@ 158 39451 New concepts of the pathogenesis of cystic fibrosis lung disease - + MESH:D003550 Disease @@ -27533,13 +27688,13 @@ 403 39516 Early pulmonary disease in cystic fibrosis - + MESH:D003550 Disease cystic fibrosis - + MESH:D008171 Disease @@ -27558,11 +27713,11 @@ 482 39559 Cystic fibrosis since 1938 - - MESH:D003550 + + MESH:D005355 Disease - - Cystic fibrosis + + fibrosis @@ -27584,7 +27739,7 @@ surname:Cox;given-names:TK 39586 Identification of the cystic fibrosis gene: genetic analysis - + MESH:D003550 Disease @@ -27612,7 +27767,7 @@ surname:Rozmahel;given-names:R 39647 Identification of the cystic fibrosis gene: cloning and characterization of complementary DNA - + MESH:D003550 Disease @@ -27640,7 +27795,7 @@ surname:Melmer;given-names:G 39741 Identification of the cystic fibrosis gene: chromosome walking and jumping - + MESH:D003550 Disease @@ -27679,7 +27834,7 @@ surname:Cao;given-names:D 39816 Rescue of CF airway epithelial cell function in vitro by a CFTR potentiator, VX-770 - + 1080 Gene 55465 @@ -27712,13 +27867,19 @@ surname:Stack;given-names:JH 39900 Correction of the F508del-CFTR protein processing defect in vitro by the investigational drug VX-809 - + 1080 Gene 55465 CFTR + + - + Chemical + + F508del + surname:Gonzalez;given-names:J @@ -27750,20 +27911,20 @@ surname:Hadida;given-names:S 40001 Rescue of DeltaF508-CFTR trafficking and gating in human cystic fibrosis airway primary cultures by small molecules - + 1080 Gene 55465 CFTR - + 9606 Species human - + MESH:D003550 Disease @@ -27788,19 +27949,13 @@ surname:Zegarra-Moran;given-names:O 40117 Small-molecule correctors of defective DeltaF508-CFTR cellular processing identified by high-throughput screening - + 1080 Gene 55465 CFTR - - p.508delF - DNAMutation - - DeltaF508 - surname:Balghi;given-names:H @@ -27826,14 +27981,20 @@ surname:Lesimple;given-names:P 40231 Correction of the Delta phe508 cystic fibrosis transmembrane conductance regulator trafficking defect by the bioavailable compound glafenine - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + + - + Chemical + + phe508 + + MESH:D005897 Chemical @@ -27861,15 +28022,21 @@ surname:Anjos;given-names:SM 40372 Structural analog of sildenafil identified as a novel corrector of the F508del-CFTR trafficking defect - + 1080 Gene 55465 CFTR - - MESH:C101426 + + - + Chemical + + F508del + + + MESH:D000068677 Chemical sildenafil @@ -27893,26 +28060,20 @@ surname:Zegarra-Moran;given-names:O 40475 High-affinity activators of cystic fibrosis transmembrane conductance regulator (CFTR) chloride conductance identified by high-throughput screening - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + 1080 Gene 55465 CFTR - - MESH:D002712 - Chemical - - chloride - surname:Verkman;given-names:AS @@ -27929,31 +28090,25 @@ 392 40623 Novel amino-carbonitrile-pyrazole identified in a small molecule screen activates wild-type and DeltaF508 cystic fibrosis transmembrane conductance regulator in the absence of a cAMP agonist - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - - MESH:D011720 - Chemical - - amino-carbonitrile-pyrazole - - - p.508delF - DNAMutation - - DeltaF508 - - - MESH:D000242 + + - Chemical cAMP + + - + Chemical + + amino-carbonitrile-pyrazole + surname:Wood;given-names:AB @@ -27973,37 +28128,25 @@ surname:Lukacs;given-names:GL 40814 Cyanoquinolines with independent corrector and potentiator activities restore DeltaPhe508-cystic fibrosis transmembrane conductance regulator chloride channel function in cystic fibrosis - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - - MESH:C025253 - Chemical - - DeltaPhe508 - - - MESH:D002712 + + - Chemical - - chloride + + Cyanoquinolines - + MESH:D003550 Disease cystic fibrosis - - MESH:D001190 - Chemical - - Cyanoquinolines - surname:Gopinath;given-names:VS @@ -28028,14 +28171,14 @@ surname:Ma;given-names:T 41001 Nanomolar affinity small molecule correctors of defective Delta F508-CFTR chloride channel gating - + 1080 Gene 55465 CFTR - + p.508delF DNAMutation @@ -28055,6 +28198,12 @@ 350 41099 Ivacaftor + + MESH:C545203 + Chemical + + Ivacaftor + surname:Boyle;given-names:MP @@ -28093,26 +28242,26 @@ surname:Dunitz;given-names:JM 41109 Effect of VX-770 in persons with cystic fibrosis and the G551D-CFTR mutation - + 1080 Gene 55465 CFTR - + 9606 Species persons - + MESH:D003550 Disease cystic fibrosis - + p.G551D;RS#:75527207 ProteinMutation @@ -28149,26 +28298,26 @@ surname:Bell;given-names:SC 41186 A CFTR potentiator in patients with cystic fibrosis and the G551D mutation - + 1080 Gene 55465 CFTR - + 9606 Species patients - + MESH:D003550 Disease cystic fibrosis - + p.G551D;RS#:75527207 ProteinMutation @@ -28187,12 +28336,18 @@ 937 41261 Development, clinical utility, and place of ivacaftor in the treatment of cystic fibrosis - + MESH:D003550 Disease cystic fibrosis + + MESH:C545203 + Chemical + + ivacaftor + surname:Kaniecki;given-names:K @@ -28226,7 +28381,7 @@ surname:Yu;given-names:H 41351 Defining the disease liability of variants in the cystic fibrosis transmembrane conductance regulator gene - + 1080 Gene 55465 @@ -28249,14 +28404,14 @@ 606 41458 Cystic fibrosis: a worldwide analysis of CFTR mutations--correlation with incidence data and application to screening - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -28281,19 +28436,31 @@ surname:Yen;given-names:K 41576 Ivacaftor in subjects with cystic fibrosis who are homozygous for the F508del-CFTR mutation - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease cystic fibrosis + + - + Chemical + + F508del + + + MESH:C545203 + Chemical + + Ivacaftor + 34 @@ -28309,11 +28476,11 @@ 125 41668 Novel pharmacological strategies to treat cystic fibrosis - - MESH:D005355 + + MESH:D003550 Disease - - fibrosis + + cystic fibrosis @@ -28330,14 +28497,14 @@ 248 41726 Insights into the mechanisms underlying CFTR channel activity, the molecular basis for cystic fibrosis and strategies for therapy - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -28357,14 +28524,14 @@ 597 41856 Cystic fibrosis transmembrane conductance regulator protein repair as a therapeutic strategy in cystic fibrosis - + 1080 Gene 55465 Cystic fibrosis transmembrane conductance regulator - + MESH:D003550 Disease @@ -28391,7 +28558,7 @@ surname:Koehler;given-names:DR 41968 Gene transfer of CFTR to airway epithelia: low levels of expression are sufficient to correct Cl- transport and overexpression can generate basolateral CFTR - + 1080 Gene 55465 @@ -28422,24 +28589,24 @@ surname:Yan;given-names:Y 42125 CFTR delivery to 25% of surface epithelial cells restores normal rates of mucus transport to human cystic fibrosis airway epithelium - + 1080 Gene 55465 CFTR - + 9606 Species human - + MESH:D003550 Disease - - cystic fibrosis airway epithelium + + cystic fibrosis @@ -28483,21 +28650,21 @@ surname:Amin;given-names:RS 42258 Results of a phase IIa study of VX-809, an investigational CFTR corrector compound, in subjects with cystic fibrosis homozygous for the F508del-CFTR mutation - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -28517,11 +28684,11 @@ 213 42416 Cystic fibrosis therapeutics: the road ahead - - MESH:D003550 + + MESH:D005355 Disease - - Cystic fibrosis + + fibrosis @@ -28536,7 +28703,7 @@ 402 42461 Managing the underlying cause of cystic fibrosis: a future role for potentiators and correctors - + MESH:D003550 Disease @@ -28569,18 +28736,18 @@ surname:Janssens;given-names:HM 42557 A functional CFTR assay using primary cystic fibrosis intestinal organoids - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease - - cystic fibrosis intestinal organoids + + cystic fibrosis @@ -28621,14 +28788,14 @@ surname:Amaral;given-names:M 42632 Cystic Fibrosis Society Clinical Trial Network Standardisation, CFTR biomarkers: time for promotion to surrogate end-point - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -28660,19 +28827,25 @@ surname:Tang;given-names:LP 42755 A pharmacologic approach to acquired cystic fibrosis transmembrane conductance regulator dysfunction in smoking related lung disease - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + MESH:D008171 Disease lung disease + + MESH:D006331 + Disease + + dysfunction + surname:Garcia-Caballero;given-names:A @@ -28692,7 +28865,7 @@ surname:Suchindran;given-names:H 42888 The cystic fibrosis transmembrane conductance regulator impedes proteolytic stimulation of the epithelial Na+ channel - + 1080 Gene 55465 @@ -28718,25 +28891,19 @@ surname:Cohn;given-names:JA 43006 CFTR as a cAMP-dependent regulator of sodium channels - + 1080 Gene 55465 CFTR - - MESH:D000242 + + - Chemical cAMP - - MESH:D012964 - Chemical - - sodium - surname:Boucher;given-names:RC @@ -28754,7 +28921,7 @@ surname:Ribeiro;given-names:CM 43060 Modelling dysregulated Na+ absorption in airway epithelial cells with mucosal nystatin treatment - + MESH:D009761 Chemical @@ -28779,15 +28946,15 @@ surname:Cui;given-names:L 43157 Phenylalanine-508 mediates a cytoplasmic-membrane domain contact in the CFTR 3D structure crucial to assembly and channel function - + 1080 Gene 55465 CFTR - - MESH:C119108 + + MESH:D010649 Chemical Phenylalanine @@ -28812,14 +28979,14 @@ surname:Aleksandrov;given-names:LA 43288 Multiple membrane-cytoplasmic domain contacts in the cystic fibrosis transmembrane conductance regulator (CFTR) mediate regulation of channel gating - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + 1080 Gene 55465 @@ -28845,7 +29012,7 @@ surname:Jensen;given-names:T 43437 Correctors of DeltaF508 CFTR restore global conformational maturation without thermally stabilizing the mutant protein - + 1080 Gene 55465 @@ -28876,7 +29043,7 @@ surname:Bagdany;given-names:M 43556 Correction of both NBD1 energetics and domain interface is required to restore DeltaF508 CFTR folding and function - + 1080 Gene 55465 @@ -28899,20 +29066,20 @@ 298 43671 G551D and G1349D, two CF-associated mutations in the signature sequences of CFTR, exhibit distinct gating defects - + 1080 Gene 55465 CFTR - + p.G1349D;RS#:193922525 ProteinMutation G1349D - + p.G551D;RS#:75527207 ProteinMutation @@ -28948,7 +29115,7 @@ 467 43853 Eris: an automated estimator of protein stability - + 340061 Gene 18868 @@ -28974,14 +29141,14 @@ surname:Mohamed;given-names:A 43903 The delta F508 mutation decreases the stability of cystic fibrosis transmembrane conductance regulator in the plasma membrane. Determination of functional half-lives on transfected cells - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + p.508delF DNAMutation @@ -29007,7 +29174,7 @@ surname:Du;given-names:K 44090 Peripheral protein quality control removes unfolded CFTR from the plasma membrane - + 1080 Gene 55465 @@ -29030,14 +29197,14 @@ 8950 44172 Conformational and temperature-sensitive stability defects of the delta F508 cystic fibrosis transmembrane conductance regulator in post-endoplasmic reticulum compartments - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - + p.508delF DNAMutation @@ -29063,18 +29230,12 @@ surname:Ozols;given-names:VV 44344 Endocytic trafficking routes of wild type and DeltaF508 cystic fibrosis transmembrane conductance regulator - + MESH:D003550 Disease cystic fibrosis - - p.508delF - DNAMutation - - DeltaF508 - surname:Chen;given-names:L @@ -29095,26 +29256,20 @@ surname:Matalon;given-names:S 44452 Enhanced cell-surface stability of rescued DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) by pharmacological chaperones - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 cystic fibrosis transmembrane conductance regulator - - p.508delF - DNAMutation - - DeltaF508 - surname:Jensen;given-names:T @@ -29161,14 +29316,14 @@ surname:Skalak;given-names:T 44689 Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -29191,18 +29346,12 @@ surname:Kirk;given-names:KL 44821 Thermally unstable gating of the most common cystic fibrosis mutant channel (DeltaF508): "rescue" by suppressor mutations in nucleotide binding domain 1 and by constitutive mutations in the cytosolic loops - + MESH:D003550 Disease cystic fibrosis - - p.508delF - DNAMutation - - DeltaF508 - surname:He;given-names:L @@ -29239,20 +29388,14 @@ 66 45118 Rescue of dysfunctional deltaF508-CFTR chloride channel activity by IBMX - + 1080 Gene 55465 CFTR - - MESH:D015056 - Chemical - - IBMX - - + p.508delF DNAMutation @@ -29280,19 +29423,13 @@ surname:Dott;given-names:K 45191 Altered chloride ion channel kinetics associated with the delta F508 cystic fibrosis mutation - - MESH:D002712 - Chemical - - chloride - - + p.508delF DNAMutation delta F508 - + MESH:D003550 Disease @@ -29313,13 +29450,19 @@ 1282 45285 Ivacaftor in a G551D homozygote with cystic fibrosis - + p.G551D;RS#:75527207 ProteinMutation G551D - + + MESH:C545203 + Chemical + + Ivacaftor + + MESH:D003550 Disease @@ -29339,19 +29482,25 @@ 296 45338 Ivacaftor treatment of cystic fibrosis patients with the G551D mutation: a review of the evidence - + 9606 Species patients - + MESH:D003550 Disease cystic fibrosis - + + MESH:C545203 + Chemical + + Ivacaftor + + p.G551D;RS#:75527207 ProteinMutation @@ -29380,26 +29529,32 @@ surname:Cohen;given-names:M 45436 Optimizing Nasal Potential Difference Analysis for CFTR Modulator Development: Assessment of Ivacaftor in CF Subjects with the G551D-CFTR Mutation - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + p.G551D;RS#:75527207 ProteinMutation G551D + + MESH:C545203 + Chemical + + Ivacaftor + surname:Skach;given-names:WR @@ -29417,14 +29572,14 @@ surname:Dawson;given-names:DC 45583 Thermal Instability of DeltaF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Channel Function: Protection by Single Suppressor Mutations and Inhibiting Channel Activity - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 @@ -29466,7 +29621,7 @@ surname:Sopha;given-names:P 45834 VX-809 corrects folding defects in cystic fibrosis transmembrane conductance regulator protein through action on membrane-spanning domain 1 - + 1080 Gene 55465 @@ -29489,13 +29644,19 @@ 36 45974 Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or function - + 1080 Gene 55465 CFTR + + MESH:C545203 + Chemical + + ivacaftor + surname:Schreiber;given-names:R @@ -29516,24 +29677,24 @@ surname:Seydewitz;given-names:HH 46090 Modulation of Ca2+-activated Cl- secretion by basolateral K+ channels in human normal and cystic fibrosis airway epithelia - + 9606 Species human - - MESH:D002118 - Chemical - - Ca2+ - - + MESH:D003550 Disease cystic fibrosis airway epithelia + + MESH:D000069285 + Chemical + + Ca2+ + surname:Olsen;given-names:JC @@ -29554,21 +29715,21 @@ surname:Gray;given-names:MA 46213 Regulation of murine airway surface liquid volume by CFTR and Ca2+-activated Cl- conductances - + 12638 Gene 55465 CFTR - + 10090 Species murine - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ @@ -29590,7 +29751,7 @@ surname:Lethem;given-names:MI 46309 Expression of nucleotide-regulated Cl(-) currents in CF and normal mouse tracheal epithelial cell lines - + 10090 Species @@ -29611,13 +29772,19 @@ surname:Diekema;given-names:DJ 46415 Antibacterial properties of the CFTR potentiator ivacaftor - + 1080 Gene 55465 CFTR + + MESH:C545203 + Chemical + + ivacaftor + REF @@ -29649,19 +29816,13 @@ surname:Bagdany;given-names:M 46475 Some gating potentiators, including VX-770, diminish DeltaF508-CFTR functional expression - + 1080 Gene 55465 CFTR - - p.508delF - DNAMutation - - DeltaF508 - surname:Yankaskas;given-names:JR @@ -29679,7 +29840,7 @@ surname:Randell;given-names:SH 46565 Well-differentiated human airway epithelial cell cultures - + 9606 Species @@ -29699,7 +29860,7 @@ 121 46623 Human nasal and tracheo-bronchial respiratory epithelial cell culture - + 9606 Species @@ -29727,14 +29888,14 @@ surname:Randell;given-names:SH 46693 SERCA pump inhibitors do not correct biosynthetic arrest of deltaF508 CFTR in cystic fibrosis - + 1080 Gene 55465 CFTR - + MESH:D003550 Disease @@ -29761,13 +29922,13 @@ surname:Gentzsch;given-names:M 46787 Novel human bronchial epithelial cell lines for cystic fibrosis research - + 9606 Species human - + MESH:D003550 Disease @@ -29793,13 +29954,13 @@ surname:Jensen;given-names:TJ 46860 Role of N-linked oligosaccharides in the biosynthetic processing of the cystic fibrosis membrane conductance regulator - - MESH:D009584 + + - Chemical - - N + + N-linked oligosaccharides - + MESH:D003550 Disease @@ -29822,14 +29983,14 @@ surname:Gentzsch;given-names:M 46979 Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures - + 1080 Gene 55465 CFTR - + 9606 Species @@ -29880,7 +30041,7 @@ fig_caption_title 47274 VX-770 treatment restores G551D function - + p.G551D;RS#:75527207 ProteinMutation @@ -29894,53 +30055,53 @@ fig_caption 47315 Electrophysiological properties of G551D/DeltaF508 cultures analyzed in Ussing chambers treated chronically with VX-770 (cVX770, 5 microM for 48 hrs) or with vehicle (0.1% DMSO). (A) Representative recording of ISC measured in Ussing chambers. Quantification of response to treatment with (B) forskolin (significant difference between vehicle and cVX770, *P = 0.0009), (C) acute VX-770 (aVX770) (significant difference between vehicle and cVX770, *P = 0.0054), (D) forskolin + aVX770, (E) CFTRinh-172. Primary CF HBE cultures (G551D/DeltaF508) were derived from 3 different patients, 3-4 replicates were performed per patient for a total of 10 measurements per treatment. - + 9606 Species patients - + 9606 Species patient - + CVCL:0287 CellLine CF HBE - + MESH:D004121 Chemical DMSO - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + MESH:D005576 Chemical forskolin - + MESH:D005576 Chemical forskolin - - p.508delF - DNAMutation - - DeltaF508 + + MESH:C482900 + Chemical + + CFTRinh-172 @@ -29950,12 +30111,6 @@ fig_caption_title 47987 Chronic VX-770 treatment inhibits functional rescue of DeltaF508 - - p.508delF - DNAMutation - - DeltaF508 - FIG @@ -29964,61 +30119,61 @@ fig_caption 48052 (A) Representative ISC traces of CF HBE cells recorded in Ussing chambers. Primary CF HBE cells (DeltaF508/DeltaF508) were treated with vehicle (DMSO) or VX-809 +/-VX-770 for 48 hrs at 5 microM each. (B) DeltaISC response to forskolin observed in VX-809-treated CF HBE cells (*P = 0.0033, VX809 vs. vehicle) was prevented by chronic VX-770 treatment and significantly different from VX-809-treated cells (#P = 0.0147, VX809 vs. VX809+VX770). (C) CF HBE cells treated with VX-809 responded to acute VX-770 exposure (*P = 0.0177, VX809 vs. vehicle). This response was significantly abrogated in VX-809 + VX-770-treated cells (#P = 0.0031, VX809 vs. VX809+VX770). (D) The response to CFTRinh-172 observed in VX-809-treated cells (*P = 0.0209, VX809 vs. vehicle) was significantly decreased in VX809+VX770-treated cells (#P = 0.0006, VX809 vs. VX809+VX770). Primary HBE cultures (DeltaF508/DeltaF508) were derived from 6 different patients, 2-4 replicates were performed per patient for a total of 15 measurements per condition. - + 9606 Species patients - + 9606 Species patient - + MESH:D005576 Chemical forskolin - + CVCL:0287 CellLine CF HBE - + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + CVCL:0287 CellLine CF HBE - + MESH:D004121 Chemical DMSO - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + DeltaF508/DeltaF508 ProteinMutation @@ -30040,27 +30195,27 @@ fig_caption 49169 (A) CFTR Western blot of normal (NL) and CF HBE cultures treated with VX-809 (5 microM) +/-VX-770 (5 microM) for 48 hrs. * indicates the mature, complex glycosylated form of CFTR, band C; indicates the immature band B. (B) Turnover of rescued DeltaF508 in BHK-21 cells. DeltaF508 was rescued at 27 C in the presence of VX-809 +/- VX-770 for 24 hrs. After adding cycloheximide (200 microg/ml, 37 C) cells were lysed at the indicated times and analyzed by Western blotting. (C) Quantification of remaining band C over time, normalized to actin (n = 3). - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + CVCL:1914 CellLine BHK-21 - + MESH:D003513 Chemical @@ -30082,155 +30237,161 @@ fig_caption 49789 (A) ISC traces of CF HBE cells (DeltaF508/DeltaF508) recorded in Ussing chambers. CF HBE cells were treated as indicated (VX-809: 5 microM, VX-770: 1 or 5 microM) for 48 hrs. (B) CFTR function in VX-809-treated cells decreased as chronic VX-770 concentrations increased. Significant reduction of the area under the curve (AUC)/min calculated from the time period between CFTR stimulation by forskolin and CFTR inhibition by CFTRinh-172 (yields average DeltaIsc (microA/cm2)) was observed in CF cells chronically treated for 48 hrs with VX-809 when compared to VX-809 and 1 microM VX-770, (*P = 0.0352). A further reduction was detected when the chronic VX-770 concentration was increased to 5 microM (#P = 0.0049, VX-809 + 1 microM VX-770 vs. VX-809 + 5 microM VX-770). Primary CF HBE cultures were derived from at least 4 different CF patients; 2-5 replicates were performed per patient for a total of at least 14 measurements per condition. (C) In VX-809-corrected CF HBE cultures (DeltaF508/DeltaF508), the presence of chronic VX-770 at 50 nM caused a significant decline of the slope after forskolin treatment (*P = 0.0353, VX-809 vs. VX-809 + 50 nM VX-770). Primary CF HBE cultures were derived from 4 different patients; 3-5 replicates were performed per patient for a total of at least 15 measurements per condition. (D) Quantification of C:B band ratio in CF HBE cultures (DeltaF508/DeltaF508). CFTR C:B band ratio decreased in CF HBE cells as chronic VX-770 concentrations were increased. The C:B band ratio was significantly reduced in CF cells chronically treated for 48 hrs with VX-809 and 1 microM VX-770 compared to VX-809 alone (*P = 0.0181), and a further reduction was detected when the chronic VX-770 concentration was increased from 1 microM to 5 microM (#P = 0.0151, VX-809 + 1 microM VX-770 vs. VX-809 + 5 microM VX-770). Primary CF HBE cultures (DeltaF508/DeltaF508) from 4 different patients were analyzed. (E) Representative Western blot of CF HBE cells (DeltaF508/DeltaF508) showing decrease of VX-809-corrected DeltaF508 as chronic VX-770 concentrations were increased. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical forskolin - + + MESH:C482900 + Chemical + + CFTRinh-172 + + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + MESH:D005576 Chemical forskolin - + DeltaF508/DeltaF508 ProteinMutation DeltaF508/DeltaF508 - + CVCL:0287 CellLine - + CF HBE - + CVCL:0287 CellLine - + CF HBE - + CVCL:0287 CellLine CF HBE - + CVCL:0287 CellLine CF HBE - + DeltaF508/DeltaF508 ProteinMutation @@ -30252,36 +30413,48 @@ fig_caption 51947 (A) Representative ISC traces of NL HBE cells recorded in Ussing chambers. Cultures were treated with vehicle (DMSO) or 5 microM VX-770 for 48 hrs. HBE cells that were chronically treated with VX-770 showed significantly reduced response to (B,C) forskolin (*P = 0.0198 for forskolin peak and *P = 0.0008 for forskolin plateau) and (D) CFTRinh-172 (*P = 0.0014). Primary HBE cultures were derived from 6 different individuals, 2-4 replicates were performed per individual for a total of 17 measurements per condition. (E) Western blot of HBE cultures treated with VX-809 (5 microM) +/-VX-770 (5 microM) for 48 hrs. Mature CFTR was diminished in HBE cells that were chronically treated with VX770. - + 1080 Gene 55465 CFTR - + MESH:D004121 Chemical DMSO - + MESH:D005576 Chemical forskolin - + MESH:D005576 Chemical forskolin - + + MESH:C545203 + Chemical + + VX-770 + + + MESH:C482900 + Chemical + + CFTRinh-172 + + MESH:D005576 Chemical - - forskolin plateau + + forskolin @@ -30299,25 +30472,25 @@ fig_caption 52733 (A) Microscopy after hematoxylin and eosin (H&E) staining of HBE cultures did not reveal a detectable difference between VX-770- or vehicle-treated cells (bar = 10 m). (B) Transepithelial resistance (Rt) of primary HBE cultures was not altered after chronic treatment with VX-770. (C) Nystatin responses were not significantly different in primary HBE cultures that were treated with vehicle or VX-770 (48 hrs, 5 microM). Nystatin was added to the apical side in Ussing chambers. Primary HBE cultures were derived from 5 different individuals, and 2-4 replicates per individual were performed. - + MESH:D006416 Chemical hematoxylin - + MESH:D004801 Chemical eosin - + MESH:D009761 Chemical Nystatin - + MESH:D009761 Chemical @@ -30339,157 +30512,151 @@ fig_caption 53360 (A) The amount of mature CFTR was reduced when NL HBE cells were chronically treated with VX-770 (48 hrs, 5 microM). G551D is more stable than NL CFTR and the amount of mature G551D protein in CF cultures (G551D/DeltaF508) was not significantly reduced by 48 hrs treatment with 5 microM VX-770. (B) Quantification of C:B band ratio with chronic treatment of VX-770 at 5 microM. C:B band ratio was significantly decreased in NL cells chronically treated for 48 hrs with 5 microM VX-770, (*P = 0.008) (n = 3, cultures from 3 different NL individuals). The reduction of C:B band ratio in G551D/DeltaF508 cells chronically treated for 48 hrs with 5 microM VX-770 was not statistically significant (n = 3, cultures from 3 different CF patients (G551D/DeltaF508)). (C) Structural model showing positions of G551D and F508 in the CFTR molecule. (D) Illustration representing the proposed relationship between function and stability of CFTR variants. Wild-type CFTR has an intermediate stability that allows for optimal function. G551D CFTR is a more rigid protein that exhibits increased stability compared to wild-type CFTR but lacks sufficient function, presumably due to decreased flexibility. VX-770 decreases G551D CFTR stability and renders it a more flexible protein, resembling the stability and function of wild-type CFTR. However, VX-770 causes destabilization of wild-type CFTR and VX-809-corrected DeltaF508 CFTR, diminishing their function. VX-809 increases the stability of DeltaF508, bringing it closer to resembling wild-type CFTR, but this increased stability is diminished when VX-770 is present. - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 1080 Gene 55465 CFTR - + 9606 Species patients - + p.G551D;RS#:75527207 ProteinMutation G551D - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation G551D - + + MESH:C545203 + Chemical + + VX-770 + + CVCL:0287 CellLine NL HBE - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + c.551G>D,D;RS#:75527207 ProteinMutation G551D/D - + p.G551D;RS#:75527207 ProteinMutation G551D - + p.G551D;RS#:75527207 ProteinMutation G551D - - p.508delF - DNAMutation - - DeltaF508 - - + p.G551D;RS#:75527207 ProteinMutation @@ -30573,174 +30740,168 @@ abstract 116 The BCL6 gene, which is expressed in certain B- and T-cell human lymphomas, is involved with chromosomal rearrangements and mutations in a number of these neoplasms. Lymphomagenesis is believed to evolve through a multi-step accumulation of genetic alterations in these tumors. We used retroviral insertional mutagenesis in transgenic mice expressing the human BCL6 transgene in order to identify genes that cooperate with BCL6 during lymphomatous transformation. We previously reported PIM1 as the most frequently recurring cooperating gene in this model. We now report three newly identified cooperating genes:GFI1B, EVI5, and MYB:that we identified in the lymphomas of retroviral-injected BCL6 transgenic mice (but not in retroviral-injected non-transgenic controls); mRNA and protein expression of GFI1B and EVI5 were decreased in the murine tumors, whereas MYB mRNA and protein expression were increased or decreased. These findings correlated with protein expression in human lymphomas, both B- and T-cell. Improved therapy of lymphomas may necessitate the development of combinations of drugs that target the alterations specific to each neoplasm. - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 18712 Gene 11214 PIM1 - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 9606 Species human - + 10090 Species transgenic mice - + 9606 Species human - + 10090 Species transgenic mice - + 10090 Species murine - + 9606 Species human - + 10090 Species transgenic - + MESH:D009369 Disease neoplasm - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease tumors - + MESH:D009369 Disease neoplasms - + MESH:D009369 Disease tumors - - MESH:D013967 - Disease - - lymphomatous transformation - - + MESH:D008223 Disease @@ -30758,71 +30919,71 @@ paragraph 1288 It has long been known that chromosomal rearrangements and/or mutations of the BCL6 gene are associated with diffuse large B-cell lymphomas (DLBLs) in humans. BCL6 expression also is known to occur in certain T-cell lymphomas. Because multiple oncogenic hits are believed to accumulate during lymphoma development, we used retroviral insertional mutagenesis in mice containing the human BCL6 transgene in an effort to identify mutated genes that cooperate with BCL6 during lymphoma development. - + 604 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 9606 Species humans - + 10090 Species mice - + 9606 Species human - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphoma - + MESH:D016399 Disease @@ -30834,238 +30995,238 @@ paragraph 1783 We previously reported that retroviral insertional mutagenesis in mice transgenic for human BCL6 leads to enhanced lymphoma development, and we described the gene proviral integration site for Moloney murine leukemia virus 1 (PIM1) as the most frequently recurring gene cooperating with BCL6 to promote lymphoma development in this model. We now report additional recurring retroviral integration sites that we identified in the lymphomas which developed in retroviral-injected BCL6 transgenic mice but not in lymphomas from retroviral-injected non-transgenic controls. The mRNA and proteins encoded by two of the genes involved, growth factor independent 1B (GFI1B) and ecotropic viral integration site 5 (EVI5), each of which we identified in the lymphomas from three transgenic animals (three B-cell and three T-cell, respectively), showed decreased expression in the lymphomas from transgenic (BCL6-positive) mice as compared with the lymphomas from non-transgenic retroviral injected controls (BCL6-negative) as well as in BCL6-positive (as compared with BCL6-negative) human lymphomas. The mRNA and protein encoded by a third involved gene, myeloblastosis oncogene (MYB), which we also identified in the lymphomas from three transgenic animals (two B-cell, one T-cell) but not in controls, was variably increased or decreased. Although these three genes have been associated with lymphoma development, they have not been reported previously to cooperate with BCL6 in lymphomagenesis. - + 604 Gene 7640 BCL6 - + 18712 Gene 11214 PIM1 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 growth factor independent 1B - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 ecotropic viral integration site 5 - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 myeloblastosis oncogene - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 10090 Species mice - + 9606 Species human - + 11801 Species Moloney murine leukemia virus - + 10090 Species transgenic mice - + 10090 Species mice - + 9606 Species human - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease @@ -31083,14 +31244,14 @@ title_2 3295 BCL6 transgenic mice and retroviral insertional mutagenesis - + 12053 Gene 7640 BCL6 - + 10090 Species @@ -31102,95 +31263,100 @@ paragraph 3355 Our transgenic mice (described previously), which express BCL6 constitutively in B- and T-lymphocytes, contain two transgenes: tet-o-BCL6 (human BCL6 cDNA under control of the tetracycline-responsive minimal promoter), and EmuSR-tTA (the tetracycline- transactivating protein under control of the Ig heavy-chain enhancer and the SRalpha promoter). One of our transgenic lines contains four copies of the human BCL6 transgene, the other 20 copies. Under approved institutional protocols, neonatal animals from both transgenic lines and controls (non-transgenic mice of the same background, wild-type or positive for either the EmuSR-tTA or tet-o-BCL6 transgenes, but not both) were injected with 105 infectious units of the retrovirus MOL4070LTR intraperitoneally and followed until the need for euthanasia or natural death. - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 10090 Species transgenic mice - + 9606 Species human - + 9606 Species human - + 10090 Species transgenic mice - + 10090 Species transgenic - + 10090 Species transgenic - - MESH:D013754 + + MESH:D013752 Chemical - + tetracycline - - MESH:D003643 - Disease - - death + + MESH:C010349 + Chemical + + tet - - MESH:D013754 + + MESH:D013752 Chemical - + tetracycline + + Disease + + euthanasia + METHODS @@ -31203,93 +31369,87 @@ paragraph 4269 Processing of tissues, FACS analysis, inverse PCR, cloning, sequencing, database searches, total RNA extraction, and real-time RT-PCR methodologies have been described previously. The classification of murine lymphoid neoplasms was according to the Bethesda proposals. Primers included beta-actin as described or glyceraldehyde-3-phosphate dehydrogenase (Real Time Primers, Elkins Park, PA) and primers common to all transcript variants of the murine GFI1B, EVI5, or MYB genes (GFI1B, forward: 5'-CTCTCCAGGCATGGACACTT-3'; reverse: 5'-GACGTGAGTATGCTGCTCCA-3'; EVI5, forward: 5'-CCCATCAAAGTTG AGTCCAG-3'; reverse: 5'-TTCTTCCCCAGAGAATCCAA-3'; MYB, Real Time Primers, forward: 5'-CTGGACAGAAGAGGAGGACA-3'; reverse: 5'-TTGTT CTTCTGGAAGCTCGT-3'). - + 14081 Gene 37561 FACS - + 14433 Gene 107053 glyceraldehyde-3-phosphate dehydrogenase - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 11461 Gene 110648 beta-actin - + 10090 Species murine - + 10090 Species murine - + MESH:D008223 Disease lymphoid neoplasms - - MESH:C068492 - Chemical - - 5'-CTGGACAGAAGAGGAGGACA - METHODS @@ -31302,152 +31462,152 @@ paragraph 5052 Human lymphoma blocks were retrieved from the surgical pathology archives under an Institutional Review-Board approved protocol. Human and murine tissues were stained with anti-BCL6 as described previously. For additional immunohistochemistry, antigen retrieval was performed in pH 6 solution (Epitomics, Burlingame, CA) in a near boiling waterbath for 40 min. Anti-GFI1B (#HPA007012, for human tissues) and anti-EVI5 (#HPA027339, for human and murine tissues) were affinity-isolated Prestige antibodies produced in rabbit (Sigma-Aldrich, Saint Louis, MO). Anti-GFI1B was diluted 1:20 and anti-EVI5 1:35 for incubation overnight at 4 C. Murine tissues were stained with GFI1B affinity isolated antibody produced in rabbit (Sigma Aldrich, #AV30093), 20 mug/ml, overnight at 4 C. For immunohistochemistry with MYB antibody, human tissues were incubated overnight at 4 C with a 1:20 dilution of a rabbit monoclonal antibody (Epitomics, Burlingame, CA, cat. # 3195-1). Murine tissues were incubated with the same MYB antibody diluted 1:1000 for 1 h at room temperature. For all antibodies, antigen-antibody binding was detected with DAB chromogen, and tissues were counterstained with hematoxylin. Images were taken with a BX41 microscope (Olympus), a DP72 digital camera, and cellSens Standard imaging software (Olympus). - + 12053 Gene 7640 BCL6 - + 8328 Gene 31223 GFI1B - + 7813 Gene 121902 EVI5 - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 14582 Gene 31223 GFI1B - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 9606 Species Human - + 9606 Species Human - + 10090 Species murine - + 9606 Species human - + 9606 Species human - + 10090 Species murine - + 9986 Species rabbit - + 10090 Species Murine - + 9986 Species rabbit - + 9606 Species human - + 9986 Species rabbit - + 10090 Species Murine - - MESH:C047158 + + MESH:D008223 + Disease + + lymphoma + + + MESH:C000469 Chemical - - HPA027339 + + DAB - + MESH:D006416 Chemical hematoxylin - - MESH:D006327 - Disease - - lymphoma blocks - METHODS @@ -31472,6 +31632,12 @@ title_2 6664 Lymphoma development in MOL4070LTR-infected mice + + MESH:D008223 + Disease + + Lymphoma + RESULTS @@ -31547,17 +31713,17 @@ lymphomas - MESH:D005935 - Disease - - T-cell tumors - - MESH:D008223 Disease lymphoma + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemias + MESH:D007938 Disease @@ -31613,10 +31779,10 @@ aggressive tumors - MESH:D054218 + MESH:D016399 Disease - - T-cell lymphoblastic lymphoma/leukemias + + T-cell tumors MESH:D008223 @@ -31868,28 +32034,28 @@ transgenic - MESH:D005935 + MESH:D008223 Disease - - T-cell tumors + + lymphomas MESH:D016399 Disease - - T-cell lymphomas + + T-cell tumors - MESH:D054218 + MESH:D016399 Disease - - T-cell lymphoblastic lymphoma/leukemias + + T-cell tumors - MESH:D005935 + MESH:D016399 Disease - - T-cell tumors + + T-cell lymphoblastic lymphoma/leukemias @@ -31930,35 +32096,35 @@ transgenic - MESH:D054218 + MESH:D016399 Disease T-cell lymphoblastic lymphoma/leukemias - MESH:D054218 - Disease - - T-cell lymphoblastic lymphoma/leukemia - - MESH:D016399 Disease T-cell lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease tumors + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemia + MESH:D008223 Disease @@ -32117,271 +32283,265 @@ paragraph 11351 Tissue sections from the lymphomas of the three transgenic mice containing retroviral inserts near GFI1B (all large B-cell) and from B-cell lymphomas of three randomly selected retroviral-injected nontransgenic controls (no control tumors contained inserts near GFI1B) were studied with anti-GFI1B. Representative sections, shown in Fig. 1b, panel I, indicate that GFI1B staining is decreased in transgenic tumors (nuclear staining is positive for BCL6) as compared with the tumors in non-transgenic retroviral-injected controls (nuclear staining is negative for BCL6). Additionally, sixteen human lymphomas were studied by immunohistochemistry for GFI1B. Nine were B-cell tumors, seven were T-cell neoplasms; eight were BCL6-positive (six B-cell, two T-cell), and eight were BCL6-negative (three B cell, five T-cell). GFI1B expression was decreased in the BCL6-positive lymphomas as compared with the BCL6-negative tumors, which showed strong cytoplasmic expression of GFI1B (Fig. 1b, panel II). Thus, the findings in human lymphomas paralleled those in mice. Additionally, whereas in mice inserts near GFI1B were noted only in BCL6-positive B-cell tumors (which demonstrated decreased expression of GFI1B RNA and protein levels as compared with BCL-negative murine lymphomas), in humans, BCL6-positive T-cell as well as B-cell neoplasms demonstrated decreased expression of GFI1B protein as compared with BCL-negative lymphomas of each cell type (Fig. 1b, panel II). - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 8328 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 604 Gene 7640 BCL6 - + 8328 Gene 31223 GFI1B - + 10090 Species transgenic mice - + 9606 Species human - + 9606 Species human - + 10090 Species mice - + 10090 Species mice - + 10090 Species murine - + 9606 Species humans - - 10090 - Species - - transgenic - - + 10090 Species transgenic - + MESH:D016399 Disease T-cell neoplasms - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumors - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease tumors - - MESH:D009369 - Disease - - tumors - - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease tumors - + MESH:D009369 Disease neoplasms - + + MESH:D009369 + Disease + + transgenic tumors + + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease @@ -32393,264 +32553,260 @@ paragraph 12820 Representative immunohistochemical staining of the murine lymphomas containing retroviral inserts in the EVI5 gene (all precursor T-cell lymphoblastic lymphoma/leukemias) is shown in Fig. 2b, panel I and reveals that EVI5 staining (nuclear) is decreased in the BCL6-positive T-cell neoplasms as compared with the non-transgenic retroviral-injected BCL-negative T-cell controls, none of which contained inserts in or near EVI5. Twenty-seven human lymphomas were studied with anti-BCL6. Eight were negative for BCL6 (3 B-cell, 5 T-cell), 17 were BCL6-positive (13 B, 4 T), and two (T-cell) stained weakly with anti-BCL6. Representative sections (Fig. 2b, panel II) indicate that BCL6-positive lymphomas contain less EVI5 than do BCL-negative neoplasms. As in the case of GFI1B, findings for EVI5 in human lymphomas paralleled the findings in murine lymphomas, and, additionally, whereas in the mouse tumors inserts in the EVI5 gene were noted only in BCL6-positive T-cell tumors (which had decreased expression of EVI5 RNA and protein as compared with BCL-negative T-cell lymphomas), in human lymphomas, decreased levels of EVI5 protein were present in BCL6-positive B- as well as T-cell tumors as compared with BCL-negative controls (Fig. 2b, panel II). Non-neoplastic BCL6-positive germinal center cells also contained less EVI5 than the surrounding BCL-negative tumor cells (Fig. 2b, panel II, 4C). - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + + 12053 + Gene + 7640 + + BCL6 + + 14020 Gene 121902 EVI5 - + 604 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14020 Gene 121902 EVI5 - + 8328 Gene 31223 GFI1B - + 7813 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + + 12053 + Gene + 7640 + + BCL6 + + 14020 Gene 121902 EVI5 - + 7813 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 14020 Gene 121902 EVI5 - + 10090 Species murine - + 9606 Species human - + 9606 Species human - + 10090 Species murine - + 10090 Species mouse - + 9606 Species human - + 10090 Species transgenic - + 12053 Gene 7640 BCL6 - - MESH:D054198 - Disease - - lymphoblastic lymphoma - - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease neoplasms - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - - MESH:D007938 - Disease - - leukemias - - - MESH:C060809 - Chemical - - 4C - - - MESH:C563822 - Disease - - BCL6-positive T-cell neoplasms - - - MESH:D016399 - Disease - - BCL-negative T-cell lymphomas - - + MESH:D009369 Disease tumor - - MESH:D005935 + + MESH:D016399 Disease - - T-cell tumors + + T-cell lymphoblastic lymphoma/leukemias - + MESH:D008223 Disease lymphomas - + + MESH:D009369 + Disease + + neoplasms + + MESH:D008223 Disease lymphomas - + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D016399 + Disease + + T-cell tumors + + MESH:D009369 Disease tumors - - MESH:C513342 - Chemical - - 13 B - - - MESH:C563822 + + MESH:D016399 Disease - - BCL6-positive T-cell tumors + + T-cell lymphomas - + MESH:D008223 Disease @@ -32662,301 +32818,301 @@ paragraph 14220 Representative immunohistochemistry of MYB staining (Fig. 3b, I) of two murine lymphomas from transgenic mice (a B-cell tumor, panel 1 and a T-cell tumor, panel 2), reveals that these BCL6-positive neoplasms also show nuclear staining for MYB, whereas two murine lymphomas from non-transgenic animals (B-cell, panel 3, and T-cell, panel 4) are BCL-negative (column B) as well as MYB-negative (column C). A third transgenic mouse with a large B-cell lymphoma (not shown) showed little to no staining for MYB. Murine lymphomas stained readily with anti-MYB, which was used in a dilution of 1:1000 for 1 h at room temperature. Human lymphomas were more difficult to stain, requiring a 1:20 dilution and overnight incubation, and even then, positive staining was often focal and noted in only six (24%) of the 25 lymphomas studied. Of the 25 human lymphomas (12 B-cell, 13 T-cell) studied, 17 were BCL6-positive (10 B-cell, seven T-cell), five were BCL-negative (one B-cell, four T-cell), and three stained weakly for BCL6 (one B-cell, two T-cell). In four BCL6-positive lymphomas (two B-cell, two T-cell), the MYB-positive cells were also BCL6-positive (Fig. 3b, II, panels 1 and 2); however, in two T-cell lymphomas, one BCL6-positive, but with variably weak BCL6 staining, and one BCL-negative (Fig. 3b, panels 3 and 4, respectively), the MYB-positive cells were BCL-negative. Our observations in human lymphomas paralleled those in murine tumors in the respect that MYB levels could be increased or decreased in BCL6-positive neoplasms, T- as well as B-cell, and in human tumors, MYB positivity could be demonstrated in BCL6-positive as well as BCL-negative neoplastic cells (Fig. 3b, panel II). - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 4602 Gene 31311 MYB - + 604 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 10090 Species murine - + 10090 Species transgenic mice - + 10090 Species murine - + 10090 Species mouse - + 10090 Species Murine - + 9606 Species Human - + 9606 Species human - + 9606 Species human - + 10090 Species murine - + 9606 Species human - + 10090 Species transgenic - + 10090 Species transgenic - + MESH:D009369 Disease tumors - + MESH:D008223 Disease lymphomas - + CVCL:4V27 CellLine 13 T-cell - - MESH:D016399 - Disease - - T-cell lymphomas - - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease neoplasms - + MESH:D009369 Disease tumor - + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + a T-cell tumor + + MESH:D009369 Disease tumors - + MESH:D009369 Disease neoplasms - - MESH:D005935 - Disease - - T-cell tumor - - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease @@ -32968,14 +33124,14 @@ title_2 15916 Recurring sites in BCL6-positive transgenic mice not found in nontransgenic controls - + 12053 Gene 7640 BCL6 - + 10090 Species @@ -32987,39 +33143,39 @@ paragraph 16001 Table 2 lists the genes nearest to recurring sites found in at least two transgenic animals but not in non-transgenic controls. Additional genes that may cooperate with BCL6 occurred in multiple retroviral-injected transgenic mice and also in at least one retroviral-injected non-transgenic (control) animal (e.g., MYC), and thus may have transforming properties on their own. These are not reported here. - + 12053 Gene 7640 BCL6 - + 17869 Gene 31092 MYC - + 10090 Species transgenic mice - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species @@ -33031,67 +33187,67 @@ paragraph 16407 We previously reported that the most frequent retroviral insertions in our BCL6 transgenic mice occurred in or near the PIM1 gene, and we showed overall higher levels of PIM1 RNA and protein (by immunohistochemistry) in lymphomas (B- or T-cell) containing these insertion sites. Here we have studied the genes involved by the next most common insertion sites (in or near GFI1B, EVI5, and MYB, which were observed in lymphomas from three transgenic mice in each case). - + 12053 Gene 7640 BCL6 - + 18712 Gene 11214 PIM1 - + 18712 Gene 11214 PIM1 - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 10090 Species transgenic mice - + 10090 Species transgenic mice - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease @@ -33109,187 +33265,187 @@ paragraph 16886 Although the BCL6 gene is best known for its association with DLBLs in humans (~40% of these neoplasms are associated with chromosomal rearrangements involving BCL6 and ~16% contain mutations disrupting autoregulation of the BCL6 gene), the role of BCL6 in T-cell development and function has gained increased attention. Through the use of retroviral insertional mutagenesis in BCL6 transgenic mice, we have identified three genes that have not been reported previously as cooperating with BCL6 to promote lymphomagenesis. All of these have been recognized as proto-oncogenes. Two (GFI1B and MYB) are transcription factors and the third (EVI5) is considered an essential regulator of cell membrane trafficking. Additionally, immunohistochemistry performed on B- and T-cell human lymphomas (16 in the case of GFI1B, 27 for EVI5, 25 for MYB), both BCL6-positive and negative, confirmed our observations in murine lymphomas at the RNA and protein level: GFI1B and EVI5 are downregulated in BCL6-positive lymphomas as compared with BCL-negative tumors, and MYB can be either up- or down-regulated in BCL6-positive lymphomas as compared with BCL-negative controls. - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 17863 Gene 31311 MYB - + 14020 Gene 121902 EVI5 - + 8328 Gene 31223 GFI1B - + 7813 Gene 121902 EVI5 - + 4602 Gene 31311 MYB - + 604 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 9606 Species humans - + 10090 Species transgenic mice - + 9606 Species human - + 10090 Species murine - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease neoplasms - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D009369 Disease @@ -33301,39 +33457,33 @@ paragraph 18046 The GFI1B gene, located on human chromosome 9q34.13, is not a common target in retroviral insertional mutagenesis. It encodes a transcriptional repressor with an N-terminal SNAG (Snail/Gfi1) domain and six C-terminal zinc fingers that are believed to have an important role in hematopoiesis. Of two transcripts in humans, the most frequent is a 330 amino acid protein which binds DNA and suppresses gene expression through recruitment of histone modifying enzymes at target promoters. It is expressed in myeloid progenitor cells as well as in B- and T-cell subsets, is known to control expression levels of genes critical for B-cell development, and is essential for erythroid and megakaryocytic development. It is not expressed in mature thymocytes. - + 8328 Gene 31223 GFI1B - + 9606 Species human - + 9606 Species humans - + 14581 Gene 3854 Gfi1 - - MESH:D009584 - Chemical - - N - - + MESH:C536227 Disease @@ -33345,140 +33495,140 @@ paragraph 18797 The EVI5 gene, located on human chromosome 1p22, is a common viral integration site in T-cell lymphomas derived from AKXD mice. It encodes a coiled-coil protein that is considered essential for regulation of membrane trafficking and has been believed to be involved in T-cell disease. As we noted in our murine lymphomas, a common viral integration site is in a 3' intron, and integrations are opposite to the transcriptional orientation of the gene. Although real-time RT-PCR revealed decreased RNA expression in all of the lymphomas from the transgenic mice as compared with the nontransgenic controls, the differences were not large; however, it is known that even small perturbations in the expression level of a protein may lead to significant biological effects. Alternatively, as it is known that gene expression can be affected over hundreds of kilobases by retroviral integrations, it is possible that viral insertions in the EVI5 gene could affect another gene, e.g., growth factor independent 1 (GFI1), which is located 18 kb downstream from EVI5 and is known to be a transcription factor that is crucial for normal hematopoietic development. Thus, Schmidt et al., who could not detect a signal with an EVI5 probe on Northern blots prepared from total RNA of lymphomas bearing proviral EVI integrations in MYC/PIM bitransgenic mice, found that the tumors had enhanced GFI1 RNA expression. However, these authors indicate that other targets which are located several hundred kilobases apart also could be activated by EVI5 integrations. - + 7813 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + 14581 Gene 3854 growth factor independent 1 - + 14581 Gene 3854 GFI1 - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + 68151 Gene 11779 EVI - + 14581 Gene 3854 GFI1 - + 14020 Gene 121902 EVI5 - + 17869 Gene 31092 MYC - + 9606 Species human - + 10090 Species mice - + 10090 Species murine - + 10090 Species transgenic mice - + 10090 Species mice - + MESH:D009369 Disease tumors - - MESH:D008223 - Disease - - lymphomas - - + MESH:D016399 Disease T-cell disease - - MESH:D016399 + + MESH:D008223 Disease - - T-cell lymphomas + + lymphomas - + MESH:D008223 Disease - + lymphomas - + MESH:D008223 Disease - + + lymphomas + + + MESH:D008223 + Disease + lymphomas @@ -33487,118 +33637,112 @@ paragraph 20346 MYB, a site of recurrent retroviral insertional mutagenesis in a number of murine hematopoietic malignancies, is a leucine zipper DNA-binding transcription factor with a short half-life that undergoes post-translational modifications, including ubiquitylation, phosphorylation, acetylation, and sumoylation. Human MYB, located on chromosome 6q23.3, contains 15 exons and encodes a family of related proteins. Alternative splicing leads to splice variants that are predicted to encode proteins with differing transcriptional activities and specificity domains. The MYB protein is expressed in all proliferating hematopoietic cells, is involved in the regulation of proliferation and differentiation of bone marrow progenitor cells (also of colon and adult brain), and is required for normal hematopoiesis, T-cell development, pro-B to pre-B transition, and survival of spleen B cells. In humans, expression of MYB is known to be relevant for the lymph node germinal center phenotype, and that expression is sustained by BCL6 repression of microRNA (miR)-155. Although, traditionally, MYB has been touted as a transcriptional activator, several corepressor molecules that interact through various MYB domains imply that MYB could act also as a repressor. In one study, about half of the genes found to be regulated by MYB were repressed. Its target genes have functions in cell cycle progression, cell differentiation, and survival. The gene is known to be translocated in T-cell acute lymphocytic leukemia, and its protein is overexpressed in that disease, in part through gene duplication. - + 17863 Gene 31311 MYB - + 4602 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 4602 Gene 31311 MYB - + 604 Gene 7640 BCL6 - + 406947 Gene microRNA (miR)-155 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 10090 Species murine - + 9606 Species Human - + 9606 Species humans - + 17863 Gene 31311 MYB - - MESH:C038361 - Chemical - - leucine - - + MESH:D015179 Disease colon - + MESH:C536227 Disease hematopoiesis - + MESH:D054218 Disease T-cell acute lymphocytic leukemia - + MESH:D019337 Disease @@ -33610,214 +33754,208 @@ paragraph 21936 In addition to the three genes described above, additional genes of potential interest are listed in Table 2. Almost all of these are known to be involved in hematopoiesis or hematologic malignancies, lending validity to the notion that they may well cooperate with BCL6 in lymphomagenesis. For example, LMO2 is up-regulated in non-Hodgkin lymphomas and has been noted to be translocated in certain human T-cell acute lymphoblastic leukemias; MN1 is deregulated in human acute myeloid leukemia; AHI1 is an oncoprotein that interacts with BCR-ABL and Janus kinase 2 (JAK2) in chronic myelogenous leukemia cells and is expressed in cutaneous T-cell lymphomas with intermediate to poor prognosis; BCOR is part of a BCL6 repression complex that facilitates survival and proliferation of lymphomas; IKZF1 encodes the IKAROS transcription factor, which drives lymphoid development, and IKZF1 deletions have been noted in childhood B-cell precursor acute lymphoblastic leukemia; SYK was identified as a target in acute myeloid leukemia and, when involved in translocations with the interleukin 2-inducible T cell kinase (ITK) gene, it induces a T-cell lymphoproliferative disease in mice mimicking human disease; DNTT encodes a lymphoid regulator; and, finally, GATA1 encodes an erythroid transcription factor. - + 12053 Gene 7640 BCL6 - + 4005 Gene 4072 LMO2 - + 4330 Gene 37620 MN1 - + 54806 Gene 9762 AHI1 - + 25 Gene 3783 BCR-ABL - + 3717 Gene 21033 Janus kinase 2 - + 3717 Gene 21033 JAK2 - + 54880 Gene 9809 BCOR - + 604 Gene 7640 BCL6 - + 10320 Gene 55948 IKZF1 - + 10320 Gene 55948 IKZF1 - + 6850 Gene 2390 SYK - + 3702 Gene 4051 interleukin 2-inducible T cell kinase - + 3702 Gene 4051 ITK - + 1791 Gene 3014 DNTT - + 2623 Gene 1549 GATA1 - + 9606 Species human - + 9606 Species human - + 10090 Species mice - + 9606 Species human - + 22778 Gene 55948 IKAROS - + MESH:D016410 Disease cutaneous T-cell lymphomas - + MESH:D015470 Disease acute myeloid leukemia - + MESH:D054198 Disease lymphoblastic leukemias - + MESH:D007951 Disease myelogenous leukemia - + MESH:D008223 Disease lymphomas - - MESH:D006689 - Disease - - Hodgkin lymphomas - - - MESH:D019337 - Disease - - hematologic malignancies - - + MESH:D015470 Disease acute myeloid leukemia - + MESH:D054198 Disease lymphoblastic leukemia - + MESH:D008232 Disease - - T-cell lymphoproliferative disease + + a T-cell lymphoproliferative disease - - MESH:C536227 + + MESH:D019337 Disease - - hematopoiesis + + hematopoiesis or hematologic malignancies + + + MESH:D008228 + Disease + + non-Hodgkin lymphomas @@ -33825,103 +33963,97 @@ paragraph 23240 BCL6 is believed to exert its function by repressing hundreds of proteins. B- as well as T-cell lymphomas are aggressive hemato-logic neoplasms resulting from the malignant transformation of B- or T-cell progenitors, respectively. These transformation processes are believed to be multi-step events in which a series of heterogeneous genetic alterations cooperate to induce perturbations in normal lymphocyte growth and differentiation. DLBLs and T-cell acute lymphoblastic leukemias, for example, have been reported to harbor multiple genetic abnormalities (in the case of DLBLs, more than 30 clon-ally represented gene alterations per neoplasm) which differ from tumor to tumor, making approaches to therapeutic intervention diffi-cult. Nonetheless, therapies directed against MYB and the proteins encoded by a number of other genes known to be involved in lymphomatous transformation are currently under investigation. Our studies have added GFI1B, EVI5, and MYB to the list of genes cooperating with BCL6 during lymphomagenesis, thus expanding the arsenal of potential therapeutic targets for DLBL. The finding in this study of several down-regulated cooperating genes suggests that efforts to upregulate them, e.g., by target mRNA manipulation, may have some therapeutic benefit. Identification of the alterations in individual lymphomas, with selection of the appropriate combinations of agents targeting the specific lesions, may hold promise for therapy of these neoplasms. - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + MESH:D009369 Disease tumor - - MESH:D013967 + + MESH:D009369 Disease - - lymphomatous transformation + + neoplasms - + MESH:D030342 Disease genetic abnormalities - + MESH:D009369 Disease neoplasm - + MESH:D009369 Disease neoplasms - - MESH:C537032 - Disease - - aggressive hemato-logic neoplasms - - - MESH:D008223 - Disease - - lymphomas - - + MESH:D054218 Disease T-cell acute lymphoblastic leukemias - + MESH:D009369 Disease tumor - + + MESH:D008223 + Disease + + lymphomas + + MESH:D016399 Disease @@ -34011,14 +34143,14 @@ surname:Chaganti;given-names:RS 25547 Mutations of the BCL6 proto-oncogene disrupt its negative autoregulation in diffuse large B-cell lymphoma - + 12053 Gene 7640 BCL6 - + MESH:D008223 Disease @@ -34041,15 +34173,15 @@ surname:Knowles;given-names:DM 25653 BCL6-protein is expressed in precursor T-cell lymphoblastic lymphoma and in prenatal and post-natal thymus - + 12053 Gene 7640 BCL6 - - MESH:D054218 + + MESH:D016399 Disease T-cell lymphoblastic lymphoma @@ -34072,27 +34204,27 @@ surname:Reddy;given-names:PL 25760 PIM1 gene cooperates with human BCL6 gene to promote the development of lymphomas - + 5292 Gene 11214 PIM1 - + 604 Gene 7640 BCL6 - + 9606 Species human - + MESH:D008223 Disease @@ -34112,28 +34244,28 @@ 4414 25842 Growth factor independent 1B (Gfi1b) is an E2A target gene that modulates Gata3 in T-cell lymphomas - + 14582 Gene 31223 Growth factor independent 1B - + 14582 Gene 31223 Gfi1b - + 14462 Gene 1550 Gata3 - + MESH:D016399 Disease @@ -34156,7 +34288,7 @@ surname:Copeland;given-names:NG 25942 Proviral integrations at the Evi5 locus disrupt a novel 90 kDa protein with homology to the Tre2 oncogene and cell-cycle regulatory proteins - + 14020 Gene 121902 @@ -34178,20 +34310,20 @@ surname:Ge;given-names:J 26083 Re-expression of miR-150 induces EBV-positive Burkitt lymphoma differentiation by modulating c-Myb in vitro - + 387168 Gene miR-150 - + 17863 Gene 31311 c-Myb - + MESH:D002051 Disease @@ -34215,20 +34347,20 @@ surname:Baron;given-names:RM 26191 The human BCL6 transgene promotes the development of lymphomas in the mouse - + 604 Gene 7640 BCL6 - + 9606 Species human - + MESH:D008223 Disease @@ -34252,7 +34384,7 @@ surname:Harris;given-names:AW 26267 Bethesda proposals for classification of lymphoid neoplasms in mice - + MESH:D008223 Disease @@ -34287,18 +34419,13 @@ 4971 26393 A Moloney murine leukemia virus-based retrovirus with 4070A long terminal repeat sequences induces a high incidence of myeloid as well as lymphoid neoplasms - + 11801 Species Moloney murine leukemia virus - - Chemical - - 4070A - - + MESH:D008223 Disease @@ -34322,7 +34449,7 @@ surname:Zhang;given-names:QY 26550 BCL6 is required for differentiation of Ig-like transcript 3-Fc-induced CD8+ T suppressor cells - + 12053 Gene 7640 @@ -34347,7 +34474,7 @@ surname:Wang;given-names:A 26646 Bcl6 expression specifies the T follicular helper cell program in vivo - + 12053 Gene 7640 @@ -34387,14 +34514,14 @@ surname:Reimann;given-names:JD 26829 The Evi5 oncogene regulates cyclin accumulation by stabilizing the anaphase-promoting complex inhibitor Emi1 - + 14020 Gene 121902 Evi5 - + 18538 Gene 1945 @@ -34416,7 +34543,7 @@ 3175 26938 Aberrant splicing events that are induced by proviral integration: implications for myb oncogene activation - + 17863 Gene 31311 @@ -34441,7 +34568,7 @@ surname:Carroll;given-names:KJ 27046 Teleost growth factor independence (gfi) genes differentially regulate successive waves of hematopoiesis - + MESH:C536227 Disease @@ -34465,24 +34592,30 @@ surname:Cayuela;given-names:J-M 27151 The C-MYB locus is involved in chromosomal translocation and genomic duplications in human T-cell acute leukemia (T-ALL), the translocation defining a new T-ALL subtype in very young children - + 4602 Gene 31311 C-MYB - + 9606 Species human - + + MESH:D054218 + Disease + + T-cell acute leukemia + + MESH:D054218 Disease - - T-cell acute leukemia (T-ALL) + + T-ALL subtype @@ -34502,7 +34635,7 @@ surname:She;given-names:D 27343 Evi5 promotes cell migration through its Rab-GAP activity - + 14020 Gene 121902 @@ -34527,14 +34660,14 @@ surname:Prekeris;given-names:R 27401 Identification of Rab11 as a small GTPase binding protein for the Evi5 oncogene - + 53869 Gene 37903 Rab11 - + 14020 Gene 121902 @@ -34557,21 +34690,21 @@ 998 27481 Direct transcriptional repression of the genes encoding the zinc-finger proteins Gfi1b and Gfi1 by Gfi1b - + 14582 Gene 31223 Gfi1b - + 14581 Gene 3854 Gfi1 - + 14581 Gene 3854 @@ -34593,21 +34726,21 @@ 1843 27586 Gfi1 and Gfi1b: key regulators of hematopoiesis - + 14581 Gene 3854 Gfi1 - + 14582 Gene 31223 Gfi1b - + MESH:C536227 Disease @@ -34631,7 +34764,7 @@ surname:Amin;given-names:RH 27634 Gfi1b negatively regulates Rag expression directly and via the repression of FoxO1 - + 14582 Gene 31223 @@ -34671,6 +34804,11 @@ 72 27859 Natural variation in cardiac metabolism and gene expression in Fundulus heteroclitus + + Disease + + Fundulus heteroclitus + surname:Thomas;given-names:H @@ -34686,6 +34824,12 @@ 216 27944 Improved conditional expression systems resulting in physiological level of HN4alpha expression confirm HN4alpha induced apoptosis in the pancreatic beta-cell line INS-1 + + MESH:D010195 + Disease + + pancreatic + 87 @@ -34701,24 +34845,24 @@ 173 28114 Long-distance activation of the Myc protooncogene by provirus insertion in Mlvi-1or Mlvi-4 in rat T-cell lymphomas - + 24577 Gene 31092 Myc - + 10116 Species rat - - MESH:D016399 + + MESH:D008223 Disease - - T-cell lymphomas + + lymphomas @@ -34736,27 +34880,27 @@ 2534 28229 MoMuLV proviral integrations identified by Sup-F selection in tumours from infected myc/pim bitransgenic mice correlate with activation of the gfi-1 gene - + 14581 Gene 3854 gfi-1 - + 17869 Gene 31092 myc - + 10090 Species mice - + MESH:D009369 Disease @@ -34776,14 +34920,14 @@ 277 28383 Role and potential for therapeutic targeting of MYB in leukemia - + 17863 Gene 31311 MYB - + MESH:D007938 Disease @@ -34803,7 +34947,7 @@ 3406 28447 The molecular basis of T cell acute lymphoblastic leukemia - + MESH:D054218 Disease @@ -34823,14 +34967,14 @@ 534 28506 MYB function in normal and cancer cells - + 17863 Gene 31311 MYB - + MESH:D009369 Disease @@ -34850,7 +34994,7 @@ 2101 28546 Alternative RNA splicing produces multiple forms of c-Myb with unique transcriptional activities - + 17863 Gene 31311 @@ -34873,14 +35017,14 @@ surname:Teichmann;given-names:S 28643 PRMT4 is a novel coactivator of c-Myb-dependent transcription in haematopoietic cell lines - + 59035 Gene 10990 PRMT4 - + 17863 Gene 31311 @@ -34904,7 +35048,7 @@ surname:Bender;given-names:TP 28734 c-Myb is critical for B cell development and maintenance of follicular B cells - + 17863 Gene 31311 @@ -34929,20 +35073,20 @@ surname:Setty;given-names:M 28813 BCL6 positively regulates AID and germinal center expression via repression of miR-155 - + 12053 Gene 7640 BCL6 - + 735281 Gene miR - + 11628 Gene 7623 @@ -34985,14 +35129,14 @@ surname:Bhatt;given-names:S 29047 Identification of LMO2 transcriptome and interactome in diffuse large B-cell lymphoma - + 16909 Gene 4072 LMO2 - + MESH:D008223 Disease @@ -35016,31 +35160,31 @@ surname:Pieters;given-names:R 29133 Characterization of a pediatric T-cell acute lymphoblastic leukemia patient with simultaneous LYL1 and LMO2 rearrangements - + 4066 Gene 4078 LYL1 - + 4005 Gene 4072 LMO2 - + 9606 Species patient - - MESH:D054218 + + MESH:D054198 Disease - - pediatric T-cell acute lymphoblastic leukemia + + acute lymphoblastic leukemia @@ -35058,19 +35202,13 @@ surname:Terranova;given-names:S 29256 Mapping of MN1 sequences necessary for myeloid transformation - + 433938 Gene 37620 MN1 - - MESH:D002472 - Disease - - myeloid transformation - surname:Sloma;given-names:I @@ -35089,21 +35227,21 @@ surname:Forrest;given-names:DL 29318 Targeting primitive chronic myeloid leukemia cells by effective inhibition of a new AHI-1-BCR-ABL-JAK2 complex - + 16452 Gene 21033 JAK2 - + 25 Gene 3783 BCR-ABL - + MESH:D007951 Disease @@ -35124,25 +35262,25 @@ 966 29429 The role of AHI1 and CDKN1C in cutaneous T-cell lymphoma progression - + 52906 Gene 9762 AHI1 - + 12577 Gene 134519 CDKN1C - + MESH:D016410 Disease - - cutaneous T-cell lymphoma progression + + cutaneous T-cell lymphoma @@ -35162,14 +35300,14 @@ surname:Lev;given-names:P 29498 BCL6 programs lymphoma cells for survival and differentiation through distinct biochemical mechanisms - + 12053 Gene 7640 BCL6 - + MESH:D008223 Disease @@ -35193,13 +35331,19 @@ surname:Binato;given-names:R 29600 Refinement of IKZF1 recombination hotspots in pediatric BCP-ALL patients - + 10320 Gene 55948 IKZF1 + + - + Chemical + + BCP + surname:Silvestri;given-names:D @@ -35215,14 +35359,14 @@ surname:Carrino;given-names:V 29673 What is the relevance of Ikaros gene deletions as prognostic marker in pediatric Philadelphia negative B-cell precursor acute lymphoblastic leukemia? - + 22778 Gene 55948 Ikaros - + MESH:D054198 Disease @@ -35246,14 +35390,14 @@ surname:Clauser;given-names:K 29823 Proteomic and genetic approaches identify Syk as an AML target - + 20963 Gene 2390 Syk - + MESH:D015470 Disease @@ -35277,37 +35421,37 @@ surname:Maurer;given-names:H 29886 The ITK-SYK fusion oncogene induces a T-cell lymphoproliferative disease in mice mimicking human disease - + 16428 Gene 4051 ITK - + 20963 Gene 2390 SYK - + 10090 Species mice - + 9606 Species human - + MESH:D008232 Disease - - T-cell lymphoproliferative disease + + a T-cell lymphoproliferative disease @@ -35327,14 +35471,14 @@ surname:Pasalic;given-names:Z 29991 RUNX1 mutations in cytogenetically normal acute myeloid leukemia are associated with a poor prognosis and up-regulation of lymphoid genes - + 12394 Gene 1331 RUNX1 - + MESH:D007951 Disease @@ -35358,7 +35502,7 @@ surname:Song;given-names:SH 30129 Ldb1-nucleated transcription complexes function as primary mediators of global erythroid gene activation - + 16825 Gene 2891 @@ -35383,7 +35527,7 @@ surname:Rossi;given-names:D 30234 Analysis of the coding genome of diffuse large B-cell lymphoma - + MESH:D008223 Disease @@ -35410,270 +35554,270 @@ fig_caption 30343 GFI1B: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative GFI1B RNA expression from the lymphomas (all large B-cell) of three retroviral-injected BCL6 transgenic mice (black bar) containing inserts 7.9 to 8.7 kb 5' to the GFI1B gene as compared with three randomly selected B-cell lymphomas from retroviral-injected non-transgenic controls that did not contain inserts in or near GFI1B (gray bar). Expression in each transgenic animal was decreased as compared with the controls [range, 1.8 to 5.7-fold (Table 1), overall decrease 2.71-fold; mean +- SEM = 0.041 +- 0.018 in the controls as compared with 0.015 +- 0.005 in the transgenics; P < 0.05]. (b) I, Representative murine lymphomas from (1) a transgenic mouse (nuclei positive for BCL6, column B) and (2) a non-transgenic animal (nuclei negative for BCL6). The cytoplasmic staining, evident in the anti-GFI1B (C) column, is decreased in the transgenic mouse as compared with the non-transgenic control. II, Representative human lymphomas: (1) BCL6-positive (B-cell), (2) BCL6-positive (T-cell), (3) BCL6-negative (B-cell), (4) BCL6-negative (T-cell). As in the murine lymphomas, the GFI1B cytoplasmic staining (column C) is decreased in the BCL6-positive lymphomas as compared with the stronger expression in the BCL6-negative neoplasms. The bar (lowest right panel) indicates 50 mum. - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 14582 Gene 31223 GFI1B - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 10090 Species mice - + 10090 Species murine - + 9606 Species human - + 10090 Species transgenic mice - + 10090 Species murine - + 10090 Species mouse - + 10090 Species mouse - + 9606 Species human - + 10090 Species murine - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + MESH:D009369 Disease neoplasms - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease @@ -35687,303 +35831,303 @@ fig_caption 31830 EVI5: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative EVI5 RNA expression from the lymphomas (all T-cell) of three retroviral-injected BCL6 transgenic mice (black bar) containing inserts within the terminal intron or 3'-untranslated region of the EVI5 gene as compared with three randomly selected T-cell lymphomas from retroviral-injected non-transgenic controls that did not contain inserts in or near EVI5 (gray bar). Expression in each transgenic animal was decreased as compared with the controls [range, 1.3 to 2-fold (Table 1), overall decrease, 1.86-fold; mean +- SEM = 0.007 +- 0.001 in the controls as compared with 0.004 +- 0.0007 in the transgenics; P < 0.05]. (b) I, Representative murine lymphomas from (1) a transgenic mouse (nuclei positive for BCL6, column B) and (2) a non-transgenic mouse (nuclei negative for BCL6). The nuclear staining evident in the anti-BCL6 (C) column is decreased in the transgenic mouse as compared with the non-transgenic control. II, Representative human lymphomas: (1) BCL6-positive B-cell, (2) BCL6-positive (T-cell), (3) BCL-negative (B-cell), (4) BCL-negative (T-cell); the cytoplasm of the tumor cells stains strongly positive with the T-cell marker CD3 (II, 4A). A germinal center in 4A (upper left corner) does not stain with CD3 and is BCL6-positive (column B), whereas the tumor cells around it are BCL-negative. As in the murine lymphomas, the EVI5 nuclear staining (column C) is decreased in BCL6-positive cells (tumors 1 and 2, or germinal center cells, tumor 4) as compared with the stronger expression in the BCL-negative tumor cells [tumors 3 and 4 (outside the germinal center)]. The length of the bar in the lowest right panel depicts 50 mum. - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 604 Gene 7640 BCL6 - + 12501 Gene 586 CD3 - + 12501 Gene 586 CD3 - + 12053 Gene 7640 BCL6 - + 14020 Gene 121902 EVI5 - + 12053 Gene 7640 BCL6 - + 10090 Species mice - + 10090 Species murine - + 9606 Species human - + 10090 Species transgenic mice - + 10090 Species murine - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 9606 Species human - + 10090 Species murine - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + MESH:D008223 Disease lymphomas - - MESH:D008223 - Disease - - lymphomas - - - MESH:D009369 - Disease - - tumor - - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumor - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumor - + + MESH:D009369 + Disease + + tumor + + MESH:D008223 Disease lymphomas - + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + MESH:D009369 Disease - + tumor - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease - + lymphomas - - MESH:D016399 - Disease - - T-cell lymphomas - - + MESH:D008223 Disease - + lymphomas @@ -35994,352 +36138,358 @@ fig_caption 33683 MYB: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative MYB RNA expression in the lymphomas of three retroviral-injected BCL6 transgenic mice (black bars, study mice S1, S2, S3) containing inserts ~35 to ~74 kb 3' to the MYB gene as compared with randomly selected B- or T-cell lymphomas (gray bars) from retroviral-injected control mice that did not contain inserts in or near MYB. S1 is a large B-cell lymphoma from a transgenic mouse whose relative expression is 2-fold higher than the mean of three randomly selected B-cell control tumors (S1 expression = 0.54 vs. control B-cell tumors, mean +- SEM = 0.27 +- 0.07), whereas the relative RNA expression of S2 (0.08), also a large B-cell lymphoma from a transgenic animal, is 3.2-fold decreased as compared with the mean of the three randomly selected control B-cell lymphomas (see above). S3 is a precursor T-cell lymphoblastic lymphoma/ leukemia from a transgenic mouse whose relative RNA expression is 2.2-fold higher than the mean of two randomly selected precursor T-cell lymphoblastic lymphoma/leukemia control tumors; S3 expression = 0.42 vs. control T-cell tumors, mean +- SEM = 0.19 +- 0.07. (b) I, Representative murine lymphomas: tumors (1) and (2) depict the B- and T-cell lymphomas, respectively, from transgenic mice (BCL6-positive nuclear staining, column B) with positive MYB nuclear staining (column C); tumors (3) and (4) show B- and T-cell lymphomas, respectively, from non-transgenic mice (nuclei do not stain with anti-BCL6, column B); staining with anti-MYB is also negative in these animals (column C). (b) II, Representative human lymphomas: tumors (1) and (2) show BCL6-positive (column B) B- and T-cell lymphomas, respectively, which show positive nuclear staining for MYB (column C). Tumor (3) is a T-cell lymphoma that is weakly BCL-6 positive (column B), and tumor (4) is a BCL-negative T-cell neoplasm; in tumors 3 and 4, the cells that are MYB positive (column C) are BCL-negative. The bar (lowest right panel) depicts 50 mum. - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 12053 Gene 7640 BCL6 - + 12053 Gene 7640 BCL6 - + 17863 Gene 31311 MYB - + 604 Gene 7640 BCL6 - + 4602 Gene 31311 MYB - + 12053 Gene 7640 BCL-6 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 10090 Species mice - + 10090 Species murine - + 9606 Species human - + 10090 Species transgenic mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species murine - + 10090 Species transgenic mice - + 10090 Species transgenic mice - + 9606 Species human - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + MESH:D009369 Disease tumor - + MESH:D008223 Disease lymphomas - - MESH:C563311 + + MESH:D009369 Disease - - BCL-negative T-cell neoplasm + + tumors - + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemia + + MESH:D009369 Disease tumors - + MESH:D008223 Disease lymphoma - - MESH:D008223 - Disease - - lymphomas: tumors - - + MESH:D016399 Disease T-cell lymphomas - + MESH:D009369 Disease tumors - + MESH:D007938 Disease leukemia - + MESH:D009369 Disease tumors - + MESH:D008223 Disease lymphoma - + MESH:D009369 Disease tumors - + MESH:D008223 Disease lymphomas - - MESH:D054218 + + MESH:D009369 Disease - - T-cell lymphoblastic lymphoma/leukemia control tumors + + tumors - + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D009369 + Disease + + neoplasm + + MESH:D009369 Disease tumors - + MESH:D016399 Disease T-cell lymphomas - - MESH:D005935 + + MESH:D008223 Disease - - T-cell tumors + + lymphomas - + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma + + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease lymphomas - - MESH:D054198 - Disease - - lymphoblastic lymphoma - - + MESH:D016399 Disease T-cell lymphoma - + MESH:D016399 Disease T-cell lymphomas - + MESH:D016399 Disease T-cell lymphomas - - MESH:D001260 - Disease - - black bars - - + MESH:D008223 Disease @@ -36395,44 +36545,44 @@ table_caption 36069 Location of retroviral inserts in transgenic mice and relative RNA expression levels in lymphomas from BCL6 transgenic mice as compared with lymphomas from retroviral-injected non-transgenic (control) mice. - + 12053 Gene 7640 BCL6 - + 10090 Species transgenic mice - + 10090 Species transgenic mice - + 10090 Species mice - + 10090 Species transgenic - + MESH:D008223 Disease lymphomas - + MESH:D008223 Disease @@ -36446,122 +36596,122 @@ T1.xml 36276 Mouse gene (chromosome) Lymphoma type Location of insert Expression level vs. control GFI1B (Chr 2) B 8.692 kb 5' to GFI1B 5.7-fold decreased GFI1B B 8.686 kb 5' to GFI1B 2.6-fold decreased GFI1B B 7.877 kb 5' to GFI1B 1.8-fold decreased EVI5 (Chr 5) T Terminal intron* 2.0-fold decreased EVI5 T 3' untranslated region** 1.5-fold decreased EVI5 T 3' untranslated region*** 1.3-fold decreased MYB (Chr 10) T 34.794 kb 3' to MYB 2.2-fold increased MYB B 73.684 kb 3' to MYB 2.0-fold increased MYB B 70.687 kb 3' to MYB 3.2-fold decreased - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 GFI1B - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 EVI5 - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 17863 Gene 31311 MYB - + 10090 Species Mouse - + MESH:D008223 Disease - - Lymphoma type + + Lymphoma type Location @@ -36569,7 +36719,7 @@ table_footnote 37634 Sequences have >=98% identity to the murine databases described in the text. - + 10090 Species @@ -36587,28 +36737,28 @@ table_footnote 37744 In one lymphoma, the insert is within SYKb; in the other, SYKb is 61.3 kb 3' to the insert, but there is a gene closer to the insert at the 5' side [40.3 kb away, D/RAS2 (GTP-binding protein DI-RAS2)]. - + 20963 Gene 2390 SYKb - + 20963 Gene 2390 SYKb - + 68203 Gene 56777 GTP-binding protein DI-RAS2 - + MESH:D008223 Disease @@ -36626,31 +36776,25 @@ table_footnote 37982 MPPE (5') is closer to the insert in one lymphoma (17.8 kb vs. 50.2 kb), but IMPA2 (3') is closer to the insert in the other lymphoma (1 kb vs. 33.8 kb). - + 114663 Gene 22799 IMPA2 - + MESH:D008223 Disease lymphoma - + MESH:D008223 Disease lymphoma - - MESH:C508639 - Chemical - - MPPE - TABLE @@ -36671,20 +36815,20 @@ table_caption 38207 Recurring retroviral integration sites in BCL6 transgenic mice not found in non-transgenic controls.* - + 12053 Gene 7640 BCL6 - + 10090 Species transgenic mice - + 10090 Species @@ -36698,192 +36842,175 @@ T2.xml 38309 Murine genes containing inserts or, if insert not within a gene, the nearest characterized gene (unless indicated) Mouse chromosome Number of mice Lymphoma type PIM1 (proviral integration site for Moloney murine leukemia virus 1) 17 7 6T, 1B GFI1B (growth factor independent 1B) 2 3 B EVI5 (ecotropic viral integration site 5) 5 3 T MYB (transcriptional activator MYB isoform 1) 10 3 2B, 1T LMO2 (LIM domain only 2) 2 2 T MN1 (probable tumor suppressor protein MN1) 5 2 T AHI1 (jouberin isoform 1; Abelson helper integration site-1) 10 2 T BCOR (BCL6 corepressor isoform d) 10 2 B IKZF1 (DNA-binding protein Ikaros isoform a and b) 11 2 T SYKb (tyrosine protein kinase SYK) 13 2** T MPPE (metallophosphoesterase 1); IMPA2 (inositol monophosphatase 2) 18 2*** B DNTT (DNA nucleotidylexotransferase) 19 2 T GATA1 (erythroid transcription factor) X 2 B - + 14582 Gene 31223 GFI1B - + 14582 Gene 31223 growth factor independent 1B - + 14020 Gene 121902 EVI5 - + 14020 Gene 121902 ecotropic viral integration site 5 - + 17863 Gene 31311 MYB - + 4005 Gene 4072 LMO2 - + 433938 Gene 37620 MN1 - + 433938 Gene 37620 MN1 - + 52906 Gene 9762 AHI1 - + 54880 Gene 9809 BCOR - + 10320 Gene 55948 IKZF1 - + 20963 Gene 2390 SYKb - + 20963 Gene 2390 SYK - + 114663 Gene 22799 IMPA2 - + 1791 Gene 3014 DNTT - + 2623 Gene 1549 GATA1 - + 10090 Species Murine - + 10090 Species Mouse - + 10090 Species mice - + 11801 Species Moloney murine leukemia virus - + 17863 Gene 31311 MYB - + 22778 Gene 55948 Ikaros - + 12053 Gene 7640 BCL6 - - Chemical - - tyrosine - - + MESH:D008223 Disease Lymphoma type PIM1 - - MESH:C508639 - Chemical - - MPPE - - + MESH:D009369 Disease tumor - - MESH:D007294 - Chemical - - inositol - @@ -36911,6 +37038,12 @@ Tanışman B, Cevizci S, Çelik M, Sevim S 0 Work Stress and Risk Factors For Health Management Trainees in Canakkale, Turkey + + MESH:D004194 + Disease + + Stress + ABSTRACT @@ -36935,14 +37068,14 @@ abstract 231 This cross-sectional study was conducted on Health Management Musters students (N=96) in Canakkale Onsekiz Mart University Health Sciences Institute, May-June 2014. A total of 58 students who voluntarily participated in the study were reached (60.42%). Participants completed a 22-question sociodemographic survey form and a 12-item General Health Questionnaire in a face-to-face interview. Data were analyzed using the SPSS software version 20.0. - + 5414 Gene 6107 Mart - + 9606 Species @@ -36960,66 +37093,72 @@ abstract 688 The average age of participants was 36.4+-6.2 (Min:24-Max:62) years. Thirty five of the participants were female (60.3%), 23 were male (39.7%). The number of people using cigarettes and alcohol were 23 (39.7%) and 9 (15.8%) respectively. In our study group according to GHQ scale 32 people (55.2%) were in the group at risk of depression. Eighty-six percent of participants reported experiencing work stress. The most frequently reported sources of stress were superiors (56.8%), work itself (41.3%), and work colleagues (25.8%). There was no significant difference between those at risk of depression and those not at risk in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family time and vacations (p>0.05). - + 9606 Species participants - + 9606 Species participants - + 9606 Species people - + 9606 Species people - + 9606 Species participants - - MESH:D000431 - Chemical - - alcohol - - - MESH:D003866 + + MESH:D000275 Disease depression - - MESH:D003866 - Disease - - depression - - - MESH:D000431 + + MESH:D000438 Chemical alcohol - + + - + Chemical + + GHQ + + + MESH:D000275 + Disease + + depression + + MESH:D002908 Disease chronic disease + + MESH:D000438 + Chemical + + alcohol + ABSTRACT @@ -37032,14 +37171,14 @@ abstract 1648 Our study results indicated that majority of participants reported experiencing work stress with more than half at high risk of developing depression. The most reported risk factors were superiors, the work itself and colleagues in the present study. Psychosocial risk factors at work environment should be investigated in terms of psychological, sociological and ergonomics in more detail to reduce the risk of health management trainees experiencing work stress and mental health problems. - + 9606 Species participants - - MESH:D003866 + + MESH:D000275 Disease depression @@ -37056,7 +37195,13 @@ paragraph 2156 Stress, a psychosocial factor affecting work health and work productivity, is an important public health problem today. Work stress is an employee health problem that negatively affects the health of workers, disturbs public peace, and may cause harmful results for employee and employer. Work stress affects 22% of employees and threatens work health and safety. Physical and mental health problems damage the health of employees, and if necessary precautions are not taken against it, loss of work productivity may cause an increase in health spending. Work stress comprises 21.5% of health spending, 40% of hiring entry-exit costs, 50% of falls in work productivity, 50% of unplanned absences (absences caused by illness, etc.) and 33% of loss of manpower and employee compensation costs. - + + MESH:D004194 + Disease + + Stress + + MESH:C535569 Disease @@ -37068,65 +37213,47 @@ paragraph 2948 Currently there is a significant increase in the psychological, psychiatric and physical diseases caused by work stress. Previous studies have shown that work stress is the primary cause of work-related health problems and may cause chronic diseases like cardiovascular diseases, musculoskeletal diseases, migraine and depression. Depression developing linked to work is one of the most important causes of long-term chronic diseases. Investigations of the relationship between depression and work environment have found an important increase. The cause of this increase is the continuous presence of depressive disorders in the work environment and it has an excessive negative effect on work productivity, performance, work absences and disability costs. - - MESH:D003866 - Disease - - Depression - - - MESH:D008881 - Disease - - migraine - - - MESH:D009140 - Disease - - musculoskeletal diseases - - MESH:D001523 + MESH:D000275 Disease - - psychiatric + + depressive disorders - MESH:D002908 + MESH:D008881 Disease - - chronic diseases + + migraine and depression - MESH:D003866 + MESH:D000275 Disease depression - MESH:D003866 + MESH:D000275 Disease - - depressive disorders + + Depression - MESH:D002908 + MESH:D009140 Disease - - chronic diseases + + musculoskeletal diseases - MESH:D003866 + MESH:D001523 Disease - - depression + + psychiatric MESH:D002318 Disease - - cardiovascular diseases + + chronic diseases like cardiovascular diseases @@ -37134,32 +37261,44 @@ paragraph 3705 Work stress studies of occupational groups at risk have shown that physical, mental and social health problems that may develop after stress can be prevented. A study in Europe found that the occupational group with greatest risk of work stress was health personnel. To reduce the stress experienced linked to work by health personnel, especially nurses, or to prevent negative effects, it is very important to monitor risks in the work environment, increase work fulfilment and provide social support. Reducing the administration and clinical stress load of employees responsible for health management is advantageous to monitor work environment stressors to protect mental health. Studies of health workers and nurses in with work environment frequently use the 12-item General Health Questionnaire (GHQ-12) form to monitor risk of developing depression and anxiety, mental health situation and stress levels. Stress and depression resulting from working conditions are important mental health problems among health professionals. One of the groups at risk of developing this health problem in their working life is health management trainees. - + MESH:D001008 Disease anxiety - - MESH:D003866 + + MESH:D000275 + Disease + + depression + + + MESH:D004194 Disease - - Stress and depression + + Stress - - MESH:D003866 + + MESH:D000275 Disease depression + + - + Chemical + + GHQ-12 + INTRO paragraph 4851 The aim of this study is to investigate the relationship between stress, risk of developing depression and risk factors of health management trainees. - - MESH:D003866 + + MESH:D000275 Disease depression @@ -37176,7 +37315,7 @@ paragraph 5013 This was a cross-sectional study which was performed in the Health Science Institute of Canakkale Onsekiz Mart University, Canakkale, covering the period May-Jun 2014. - + 5414 Gene 6107 @@ -37195,7 +37334,7 @@ paragraph 5211 The population of this cross-sectional study comprised 96 students enrolled in Canakkale Onsekiz Mart University Health Sciences Institute Health Management Department Non-Thesis Masters program. Sampling was not completed as the aim was to reach the whole population. Our study included 58 students (response rate for our study was 60.42%). - + 5414 Gene 6107 @@ -37214,20 +37353,20 @@ paragraph 5569 Participants completed a 22-question sociodemographic survey form and a 12-item General Health Questionnaire in a face-to-face interview. The sociodemographic survey form included questions about age, gender, educational level, marital status, cigarette and alcohol use, presence of chronic disease, working hours, work stress and causes, regular exercise and nutrition. - + 9606 Species Participants - + MESH:D002908 Disease chronic disease - - MESH:D000431 + + MESH:D000438 Chemical alcohol @@ -37238,18 +37377,30 @@ paragraph 5940 The General Health Questionnaire developed by Goldberg and Hillier (1979) is a short, easy-to-apply, self-evaluating survey for epidemiological research defining mental health problems used for societal scanning and first-stage health services. The Turkish validity and reliability studies were completed by Kilic. The GHQ-12 is widely used to research mental discomfort like depression or anxiety that may be triggered by stress factors. This study used the GHQ point method (0-0-1-1); according to this method a score of 2 points or above is accepted as the "high risk group" for mental health problems. - - MESH:D003866 - Disease - - depression + + - + Chemical + + GHQ - + + - + Chemical + + GHQ-12 + + MESH:D001008 Disease anxiety + + MESH:D000275 + Disease + + depression + METHODS @@ -37262,7 +37413,7 @@ paragraph 6560 The study received written permission from Canakkale Onsekiz Mart University Clinical Research Ethics Committee dated 28.05.2014 and numbered 2014/10-06. - + 5414 Gene 6107 @@ -37281,24 +37432,18 @@ paragraph 6735 Data were analyzed using the SPSS software version 20.0. Descriptive statistics (frequencies, percentages, means, standard deviations) were used to describe groups of numerical data and the basic features of the data. The variables were investigated using visual (histograms, probability plots) and analytical methods (Kolmogorov-Simirnov/Shapiro-Wilk's test) to determine whether or not they are normally distributed. Chi-square test was used to examine the association between categorical variables. A p-value of less than 0.05 was considered statistically significant. In the present study, the dependent variable was "developing risk of mental health problem according to GHQ-12 scale. Independent variables were sociodemographic characteristics (gender, marital status, education, age), work-related factors (Daily work, computer use, duration of sitting at desk), having a chronic disease, healthy lifestyle behaviors (sleep duration, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations). - + MESH:D002908 Disease chronic disease - - MESH:D000431 + + MESH:D000438 Chemical alcohol - - MESH:D002653 - Disease - - healthy lifestyle behaviors - RESULTS @@ -37311,108 +37456,96 @@ paragraph 7822 The average age of participants was 36.4+-6.2 (Min:24-Max:62) years. Thirty five of the participants were female (60.3%), 23 were male (39.7%) with 48 married (82.8%), 10 single (17.2%) and 55 graduates of university (94.8%). When the substance use of participants was examined the number of people using cigarettes and alcohol were 23 (39.7%) and 9 (15.8%) respectively (Table 1). The rate of those considering giving up alcohol was 37.5% with 28.6% feeling guilty for using alcohol. There were 10 people with chronic diseases (17.2%). The most frequently reported chronic diseases were 3 people with hypertension (30%), 3 with asthma (30%), 2 with thyroid function disorders (20%) and 1 with diabetes (10%). When the occupations of participants were examined 22 (37.9%) were nurses, 6 (10.3%) were midwives, 4 (6.9%) were health officers and 26 (44.9%) were health workers from different occupational groups. When lifestyles were examined 16 participated in regular exercise (28.1%), 43 ate regular meals (75.4%) and 27 reported eating fast food (46.6%). Of participants 47 reported spending sufficient time with their families (81.0%). The workplace environmental factors were examined in the working group and the average working day, time at desk, computer use and sleeping durations are presented in Figure 1. - + 9606 Species participants - + 9606 Species participants - + 9606 Species participants - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species participants - + 9606 Species participants - + + MESH:D003920 + Disease + + diabetes + + MESH:D013959 Disease thyroid function disorders - - MESH:D000431 - Chemical - - alcohol - - - MESH:D002908 + + MESH:D006973 Disease - - chronic diseases + + hypertension - - MESH:D000431 + + MESH:D000438 Chemical alcohol - - MESH:D000431 + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 Chemical alcohol - + MESH:D001249 Disease asthma - - MESH:D002908 - Disease - - chronic diseases - - - MESH:D003920 - Disease - - diabetes - - - MESH:D006973 - Disease - - hypertension - TABLE @@ -37435,97 +37568,97 @@ paragraph 9337 There were 39 people in the study group who had weekend holidays (68.4%). The average holiday duration was 1.7+-0.4 days. When the last date of blood pressure reading was questioned 6 people (10.3%) couldn't answer the question, 11 people (18.9%) couldn't remember, 2 people (3.4%) had never measured it, 1 person (1.7%) said 4 years previous, 2 people (3.4%) said 3 years previous, 3 people (5.1%) said 1 year previous, 2 people (3.4%) said 6 months previous, 1 person (1.7%) said 5 months previous, 3 people (5.1%) said 3 months previous, 3 people (5.1%) said 2 months previous, 7 people (12.0%) said 1 month previous, 5 people (8.6%) said 2-3 weeks previous, 5 people (8.6%) said 1 week previous and 7 people (12.0%) said within the last week. The number of the people who had consulted a physician regularly was 24 (41.4%). - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species person - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species person - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species people - + 9606 Species @@ -37537,66 +37670,78 @@ paragraph 10169 In our study group according to GHQ points from the GHQ scale 32 people (55.2%) were in the group at risk of depression. Statistically, there was no significant difference between those at risk of depression and those not at risk according to the GSA scale in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations (p>0.05). When experience work stress in work environment was questioned, 86.0% of participants reported experiencing work stress. Of participants in the study 33 reported superiors (56.8%), 24 said work itself (41.3%), and 15 said work colleagues (25.8%) had caused them to experience work stress. Insufficient definition of duties, human resources, profession and distance from occupation were other reported stressors result in work stress among participants (Figure 2). - + 2778 Gene 55534 GSA - + 9606 Species people - + 9606 Species participants - + 9606 Species participants - + 9606 Species human - + 9606 Species participants - + + MESH:D000275 + Disease + + depression + + + MESH:D000438 + Chemical + + alcohol + + MESH:D002908 Disease chronic disease - - MESH:D000431 + + - Chemical - - alcohol + + GHQ - - MESH:D003866 + + MESH:D000275 Disease depression - - MESH:D003866 - Disease - - depression + + - + Chemical + + GHQ @@ -37618,14 +37763,14 @@ paragraph 11322 Our study aimed to investigate the general mental health and sociodemographic characteristics, work-related factors and factors like work stress in individuals pursuing a masters degree in health management. With this aim students in the Health Management Masters program of Canakkale Onsekiz Mart University, Health Sciences Institute, Health Management Department were included in the study. After the 2-year masters program is completed the health personnel will take the title Health management. The majority of participants were interviewed face-to-face and evaluated for general mental health with the 12-item General Health Questionnaire. Additionally sociodemographic factors and work-related factors that may affect mental health were investigated. - + 5414 Gene 6107 Mart - + 9606 Species @@ -37637,127 +37782,121 @@ paragraph 12080 In our working group there were more women than men and more married than single. When the occupations of participants are examined the majority were nurses, midwives, health officers and other health workers. The cigarette use among participants was high (39.7%) while the use of alcohol was lower (15.8%). When other behavior related to healthy lifestyle was examined 16 took regular exercises, 43 ate regular meals and 27 ate fast-food. It has been reported that work stress, high workload and depression can negatively affect the eating habits of workers and may cause obesity, especially in female workers. Sagara et al. showed a significant relationship between increase in weight and development of mental health complaints (high GHQ-12 point) in male employees. In the literature it is reported that increasing exercise and limiting calorie intake can prevent weight gain and development of psychological heatlh problems. In our study, different to the literature, there was no statistically significant difference between those at risk of developing depression according to GHQ-12 scores and those not at risk in terms of regular physical activity. We believe that as health personnel are societal role models their healthy lifestyle and behavior attracts more attention and it may be beneficial, especially for personnel who will play a role in management of health services such as our working group, to participate in internal education on behavior to protect and enhance health. While chronic disease was present in at least 10 people in our study group, the most frequently reported chronic diseases were hypertension, asthma, thyroid function disorders and diabetes. In the literature it is reported that work stress may lead to chronic diseases such as cardiovascular diseases and musculoskeletal diseases. In our study, we also asked to our participants the last date of blood pressure measurement and the last date of general checkup. The minority of the participants reported that they had measured their blood pressure and also consulted a physician regularly. This finding indicated that health management trainees overlooked their health such as measuring blood pressure. Therefore all health professions in this study must be educated to recognize their own health checks and health-promoting opportunities. - + 9606 Species women - + 9606 Species men - + 9606 Species participants - + 9606 Species participants - + 9606 Species people - + 9606 Species participants - + 9606 Species participants - + MESH:D002318 Disease cardiovascular diseases - + MESH:D009140 Disease musculoskeletal diseases - - MESH:D002908 - Disease - - chronic diseases - - - MESH:D003866 - Disease - - depression - - - MESH:D000431 - Chemical - - alcohol - - + MESH:D001249 Disease asthma - + MESH:D009765 Disease obesity - + + MESH:D000438 + Chemical + + alcohol + + MESH:D006973 Disease hypertension - + MESH:D003920 Disease diabetes - - MESH:D015430 + + MESH:D000275 Disease - - weight gain + + depression - - MESH:D003866 + + MESH:D000275 Disease depression - - MESH:D002908 + + - + Chemical + + GHQ-12 + + + MESH:D015430 Disease - - chronic diseases + + weight gain - + MESH:D002908 Disease chronic disease - + MESH:D013959 Disease @@ -37769,132 +37908,162 @@ paragraph 14411 In a study evaluating the psychological health of health workers in New Zealand, the GHQ-12 was used and while the highest points were for pharmacists, general practitioners and surgeons had similar average points. In our study according to the GHQ socres 32 people (55.2%) were found to have high risk of developing depression and anxiety. Statistically, there was no significant difference between those at risk of depression and those not at risk according to the GSA scale in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations (p>0.05). Tabolli et al. in as study of biomedical researchers at a research institute in Rome identified that according to GHQ-12 scores 15.1% of participants were at risk of depression and anxiety, 77.4% were at risk of developing stress and 66% were dissatisfied with work. The most important determinants of work dissatisfaction are reported to be work opportunities, communication between departments, material rewards and participation in departmental organization. Work dissatisfaction related to economic rewards was a risk factor for developing depression independent of gender, age and duration of work. Another Italian study used the GHQ-12 to investigate the psychological status of health workers and found one third of participants were at risk of developing depression and anxiety. The most basic solution to prevent and reduce mental health problems such as development of depression and anxiety linked to psychological stress in the work environment, is to reduce workload while increasing work fulfillment and social supports. A study of personnel managing a hospital in Rome used the GHQ-12 to investigate psychological health and found 37.8% of participants were at high risk of developing depression and anxiety. The same study found that determinants of work dissatisfaction were low control of work and lack of communication between departments. Job dissatisfaction among management personnel was independent of age, gender and duration of work and was reported to be associated with high levels of risk of developing depression/anxiety, low work control and weak communication. - + 2778 Gene 55534 GSA - + 9606 Species people - + 9606 Species participants - + 9606 Species participants - + 9606 Species participants - - MESH:D003866 + + MESH:D000275 Disease depression - - MESH:D003866 + + MESH:D000275 Disease - + depression - + + - + Chemical + + GHQ-12 + + + MESH:D000275 + Disease + + depression + + MESH:D001008 Disease anxiety - + + MESH:D000275 + Disease + + depression + + MESH:D001008 Disease anxiety - - MESH:D003866 + + MESH:D000275 Disease - + depression - - MESH:D000431 + + MESH:D000438 Chemical alcohol - - MESH:D003866 + + MESH:D000275 Disease - + depression - + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ-12 + + MESH:D001008 Disease anxiety - - MESH:D003866 + + MESH:D000275 Disease - + depression - + + MESH:D004194 + Disease + + anxiety linked to psychological stress + + MESH:D001008 Disease anxiety - - MESH:D003866 - Disease - - depression + + - + Chemical + + GHQ-12 - - MESH:D003866 - Disease - - depression + + - + Chemical + + GHQ-12 - + MESH:D002908 Disease chronic disease - + MESH:D001008 Disease anxiety - - MESH:D003866 - Disease - - depression - - - MESH:D001008 - Disease - - anxiety + + - + Chemical + + GHQ @@ -37902,109 +38071,115 @@ paragraph 16804 In the health sector, work stress presents as an important risk factor for health workers and at the same time is an important problem negatively affecting the quality of health services. In our study group, the majority of the participants reported that they had experienced work-related stress. Previous studies have emphasized the necessity of researching stress factors linked to work and psychosocial working environment in health workers, a high risk group for developing burnout, role conflict and work dissatisfaction. As a result of the stress factors linked to work in health workers (long working hours, stressful work conditions, etc.) they remain at risk of developing many mental health problems such as work stress, burnout, depression, anxiety and post-traumatic stress disorder. According to a study in an Italian hospital, working environment and ergonomy, shift-work and disruption of workflow are among the factors causing work-related stress. A study of 36 hospitals in America examined the stress factors in nurses and found that the top three stressors were workload, organizational pressure and role conflict. The most frequently reported sources of stress were superiors, the work itself and colleagues. Unclear description of duties, human resources, and distance from occupation and workplace were other reported causes of work stress. A study in Hungary emphasized that there is an important relationship between burnout, role conflict and mental health in the psychosocial work environment of health workers. A study of nurses in Belgium showed that workload, burnout, work satisfaction and expectations were factors affecting the relationship between workers in the work environment and between workers and management. - + 9606 Species participants - + 9606 Species human - - MESH:D001008 - Disease - - anxiety and post-traumatic stress disorder - - + MESH:C535569 Disease psychosocial - - MESH:D003866 + + MESH:D001008 Disease - - depression + + anxiety - + MESH:C535569 Disease psychosocial + + MESH:D004194 + Disease + + traumatic stress disorder + + + MESH:D000275 + Disease + + depression + DISCUSS paragraph 18553 In our study, workplace factors were examined and the average working day was 9.3+-3.3 hours, with 4.8+-1.9 hours sitting at a desk and 4.9+-3.1 hours computer use. In the literature it is reported that work environment factors like the duration of the working day, time spent sitting at work and time working on a computer may cause psychosocial factors (work stress, depression, anxiety, etc) and musculoskeletal diseases. In our study while the time for sitting at a desk and using computers were at acceptable levels, the average work day of participants was above the legal limit, more than 8 hours. But long working hours negatively affect the health of workers and has been shown to cause harmful results in sensitive individuals. Additionally increased working hours may trigger risk factors for developing depression such as heavy workload, smoking and sleep disorders. In our study the duration of sleep was 6.9+-0.1 hours. When the active working hours of health professionals are considered, heavy workloads, such as being on call, can reduce the duration of sleep and sleep quality. It is reported that middle managers in hospitals could be rewarded for intense workloads, heavy responsibilities and long working hours. However, heavy workloads may cause role conflict, tiredness, burnout and mistakes, interrupting patient care and negatively affecting family life. In our working group 39 of the participants had a weekend holiday and 47 declared they created sufficient time for family. Needs such as weekend holidays and sufficient time with family, which are relaxing and important from the point of view of social support, can reduce work stress and the risk of developing depression, aiding in protecting mental health. - + 9606 Species participants - + 9606 Species patient - + 9606 Species participants - + MESH:D001008 Disease anxiety - - MESH:D003866 + + MESH:D000275 Disease - + depression - - MESH:D003866 + + MESH:D000275 Disease - + depression - - MESH:D003866 + + MESH:D000275 Disease - + depression - + + MESH:C535569 + Disease + + psychosocial + + MESH:D005221 Disease tiredness - - MESH:C535569 - Disease - - psychosocial factors - - + MESH:D009140 Disease musculoskeletal diseases - + MESH:D012893 Disease @@ -38016,14 +38191,14 @@ paragraph 20293 In a study by Lao, factors affecting work satisfaction among health workers included solving conflict in the workplace, relationships with colleagues and organizational structure. In our working group among the leading causes of work stress were colleagues and superiors. Interpersonal communication and increased social opportunities may solve these problems. Recently new methods of managing hospitals have begun to be developed, and studies have been completed on groups of individuals with high work satisfaction, programs with experienced managers and improving communication levels to increase social support in the workplace and reduce stress factors. A study of managers in Switzerland found that though improving communication between workers and with patients with positive support for widespread communication at all levels, some health organizations may encounter violent consequences linked to not taking sufficient care with communication. According to a study in India, the most important motivation factor not requiring material resources that must be considered by managers and political structures was reported as the opportunity for workers to develop skills in the work environment. - + 259307 Gene 22567 Lao - + 9606 Species @@ -38035,8 +38210,8 @@ paragraph 21496 The result of this study was that those at risk of developing mental health problems like depression or those with other physical or social health problems were directed to psychiatry and other related branches. To protect work health, an important part of societal health, it is important to monitor risk groups. Additionally employers can periodically monitor and measure work environment factors in the workplace to prevent physical and mental chronic health problems that may accompany the development of work stress risk. Organizations and managers can organise social activities to strengthen inter team communication and reduce the stress levels among workers in the workplace. - - MESH:D003866 + + MESH:D000275 Disease depression @@ -38053,29 +38228,29 @@ paragraph 22195 In this study, the majority of participants reported experiencing work stress with more than half at high risk of developing depression according to the GHQ-12 scale. The top three risk factors affecting stress among individuals in the study were superiors, work and colleagues. In our study group while there was no difference found between those at risk of depression and those not at risk in terms of sociodemographic characteristics, work-related factors and health behavior, to reduce the risk of health management trainees experiencing work stress and mental health problems psychosocial risk factors in the work environment should be investigated in more detail. - + 9606 Species participants - - MESH:D003866 + + MESH:D000275 Disease - + depression - - MESH:D003866 + + MESH:C535569 Disease - - depression + + psychosocial - - MESH:D008607 + + MESH:D000275 Disease - - mental health problems psychosocial + + depression @@ -38119,11 +38294,6 @@ surname:Bilgin;given-names:AC 23001 Isstresi olceginin Izmir Konak Belediyesi' nde calisan erkek iscilerde gecerlik ve guvenilirliginin incelenmesi - - Chemical - - guvenilirliginin - REF @@ -38131,7 +38301,7 @@ ref 23129 Work-related stress: new challenges in a changing workplace. the European Risk Observatory (ERO), part of EU-OSHA, unveiled the emerging risks related to workplaces - + 54719 Species @@ -38160,6 +38330,12 @@ ref 23435 Stress in the Workplace. What's eating your company? + + MESH:D004194 + Disease + + Stress + 20 @@ -38175,12 +38351,6 @@ 254 23488 A tipi kisilik oruntusunde bilissel ve duygusal zekanin stresle basa cikma ve stress belirtileri ile iliskisi - - MESH:C535750 - Disease - - cikma ve stress belirtileri ile iliskisi - surname:Bongers;given-names:PM @@ -38197,7 +38367,7 @@ surname:van der Beek;given-names:AJ 23609 Stay@Work: Participatory Ergonomics to prevent low back and neck pain among workers: design of a randomised controlled trial to evaluate the (cost-)effectiveness - + MESH:D017116 Disease @@ -38248,8 +38418,8 @@ 260 24024 Work-related depression - - MESH:D003866 + + MESH:D000275 Disease depression @@ -38282,8 +38452,8 @@ 79 24113 Depression and the workplace: a progress report - - MESH:D003866 + + MESH:D000275 Disease Depression @@ -38303,8 +38473,8 @@ 410 24161 What does research tell us about depression, job performance, and work productivity? - - MESH:D003866 + + MESH:D000275 Disease depression @@ -38350,6 +38520,12 @@ ref 24489 Stress, Cognitive Appraisal and Psychological Health: Testing Instruments for Health Professionals + + MESH:D004194 + Disease + + Stress + surname:Yu;given-names:N @@ -38368,14 +38544,14 @@ surname:Crisp;given-names:J 24588 Enhancing the resilience of nurses and midwives: pilot of a mindfulness-based program for increased health, sense of coherence and decreased depression, anxiety and stress - + MESH:D001008 Disease anxiety - - MESH:D003866 + + MESH:D000275 Disease decreased depression @@ -38396,7 +38572,7 @@ ref 24760 Trauma and mental health of medics in eastern Myanmar's conflict zones: a cross-sectional and mixed methods investigation - + MESH:D014947 Disease @@ -38443,7 +38619,7 @@ 50 25001 Eating behavior related to obesity and job stress in male Japanese workers - + MESH:D009765 Disease @@ -38504,12 +38680,6 @@ surname:Sjogaard;given-names:G 25246 Physical activity, job demand-control, perceived stress-energy, and salivary cortisol in white-collar workers - - MESH:D006854 - Chemical - - cortisol - 11 @@ -38557,18 +38727,18 @@ 250 25517 Job satisfaction, depressive and anxiety disorders among hospital personnel at IDI Sanita, Rome - + + MESH:D000275 + Disease + + depressive + + MESH:D001008 Disease anxiety disorders - - MESH:D003866 - Disease - - depressive - surname:Renzi;given-names:C @@ -38616,7 +38786,7 @@ 318 25743 Burnout, role conflict, job satisfaction and psychosocial health among Hungarian health care staff: a questionnaire survey - + MESH:C535569 Disease @@ -38707,7 +38877,7 @@ 1524 26230 Nurse practice environment, workload, burnout, job outcomes, and quality of care in psychiatric hospitals: a structural equation model approach - + MESH:D001523 Disease @@ -38730,19 +38900,19 @@ surname:Kochen;given-names:MM 26374 Depression and anxiety as major determinants of neck pain: a cross-sectional study in general practice - - MESH:D003866 + + MESH:D000275 Disease Depression - + MESH:D001008 Disease anxiety - + MESH:D019547 Disease @@ -38763,6 +38933,12 @@ ref 26477 Musculoskeletal symptoms of the upper extremities and the neck: A cross-sectional study on prevalence and symptom-predicting factors at visual display terminal (VDT) workstations + + MESH:D009140 + Disease + + Musculoskeletal symptoms of the upper extremities + surname:Toomingas;given-names:A @@ -38781,6 +38957,12 @@ surname:Tornqvist;given-names:EW 26656 Musculoskeletal symptoms in relation to work exposures at call centre companies in Sweden + + MESH:D009140 + Disease + + Musculoskeletal symptoms + surname:Heitmann;given-names:A @@ -38853,8 +39035,8 @@ surname:Haratani;given-names:T 26947 Correlates of depressive symptoms among workers in small- and medium-scale manufacturing enterprises in Japan - - MESH:D003866 + + MESH:D000275 Disease depressive symptoms @@ -38895,7 +39077,7 @@ surname:Outavong;given-names:P 27109 Job satisfaction of health-care workers at health centers in Vientiane Capital and Bolikhamsai Province, Lao PDR - + 259307 Gene 22567 @@ -38989,31 +39171,25 @@ abstract 117 Our objective is to show the number of patients with postoperative pulmonary thromboembolism (PPTE) treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from June 1, 2011 - June 1, 2014 and to indicate the importance of various surgical operations in the development of pulmonary thromboembolism (PTE). - + 9606 Species patients - + MESH:D008171 Disease pulmonary diseases - + MESH:D011655 Disease pulmonary thromboembolism - - MESH:C539179 - Chemical - - PTE - - + MESH:D011655 Disease @@ -39031,59 +39207,59 @@ abstract 500 This is the retrospective study which shows the number of patients with PPTE treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from 01.06.2011.-01.06.2014. It represents the number of these patients, per cent of patients with PPTE of total patients with PTE, age and sex of patients, type of surgery, period expressed in days from surgery to diagnosis of PTE, presence of deep venous thrombosis (DVT) of lower extremities, massiveness of PPTE e.g. level of pulmonary artery with embolus. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE + + MESH:D004617 + Disease + + embolus - + MESH:D020246 Disease deep venous thrombosis - + MESH:D008171 Disease pulmonary diseases - - MESH:C539179 - Chemical - - PTE + + OMIM:612862 + Disease + + DVT @@ -39158,16 +39334,28 @@ pulmonary embolus - MESH:C539179 - Chemical - - PTE + OMIM:612862 + Disease + + DVT - MESH:C539179 + - Chemical - - PTE + + PPTE + + + - + Chemical + + PPTE + + + OMIM:612862 + Disease + + DVT @@ -39181,23 +39369,17 @@ abstract 1843 Different surgical operations are the big risk factor for the development of PTE. There is great significance of anticoagulant prophylaxis before surgery even in patients with no anamnesis of previous DVT or PTE. - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - MESH:C539179 - Chemical - - PTE + OMIM:612862 + Disease + + DVT @@ -39211,25 +39393,31 @@ paragraph 2072 Embolism of the pulmonary artery or one of its branches is the most striking and characteristic appearance of thromboembolic disease. According to textbooks and most of literature in most cases the starting point of the embolus is phlebothrombosis or thrombophlebitis of veins of lower extremities. - + MESH:D011655 Disease Embolism of the pulmonary artery - + + MESH:D004617 + Disease + + embolus + + MESH:D013924 Disease thrombophlebitis - + MESH:D013923 Disease thromboembolic disease - + MESH:D020246 Disease @@ -39241,30 +39429,24 @@ paragraph 2371 It is often difficult to determine the starting point of pulmonary emboli. Pulmonary embolism is very common nowadays, and favoring factors for its occurrence are the chronic diseases and therefore a long lying (bed rest), which leads to the development of phlebothrombosis and a large number of different surgical operations - + 100653365 Gene bed - + MESH:D020246 Disease phlebothrombosis - + MESH:D011655 Disease Pulmonary embolism - - MESH:D011655 - Disease - - pulmonary emboli - INTRO @@ -39283,11 +39465,17 @@ paragraph 2746 Phlebothrombosis and thrombophlebitis of lower extremities veins - + MESH:D020246 Disease - - Phlebothrombosis and thrombophlebitis of lower extremities veins + + Phlebothrombosis + + + MESH:D013924 + Disease + + thrombophlebitis of lower extremities veins @@ -39295,7 +39483,7 @@ paragraph 2815 Phlebothrombosis of pelvic and abdominal veins usually after surgery - + MESH:D020246 Disease @@ -39307,17 +39495,23 @@ paragraph 2888 Thrombophlebitis and phlebothrombosis of veins of lower extremities after trauma and various surgical operations - + + MESH:D013924 + Disease + + Thrombophlebitis + + MESH:D014947 Disease trauma - + MESH:D020246 Disease - - Thrombophlebitis and phlebothrombosis + + phlebothrombosis of veins of lower extremities @@ -39325,7 +39519,7 @@ paragraph 3005 Intracardiac thrombosis of the right cardiac ventricle - + MESH:D013927 Disease @@ -39337,7 +39531,7 @@ paragraph 3064 Thrombosis of the pulmonary artery or some of its branches in situ - + MESH:D013927 Disease @@ -39349,25 +39543,31 @@ paragraph 3135 There are other, less common types of emboli: e.g. tissue cells, fat, oil, gas embolism and the like - + MESH:D004618 Disease gas embolism + + MESH:D009821 + Chemical + + oil + INTRO title_2 3241 1.2. Risk factors for deep vein thrombosis and pulmonary thromboembolism - + MESH:D020246 Disease deep vein thrombosis - + MESH:D011655 Disease @@ -39379,7 +39579,7 @@ paragraph 3314 Factors dependent on the patient: - + 9606 Species @@ -39397,25 +39597,19 @@ paragraph 3370 Obesity - - MESH:D009765 - Disease - - Obesity - INTRO paragraph 3382 Immobilization / bed rest/- lying in bed for longer than 4 days - + 100653365 Gene bed - + 100653365 Gene @@ -39427,13 +39621,13 @@ paragraph 3450 Deep vein thrombosis or pulmonary embolism in history - + MESH:D011655 Disease pulmonary embolism - + MESH:D020246 Disease @@ -39445,16 +39639,21 @@ paragraph 3508 Thrombophilia, deficiency of antithrombin(AT) III factor, protein C, protein S; lupus anticoagulant, resistance to activated protein C, hemocistinemia - + + Disease + + AT + + Disease hemocistinemia - - MESH:D020152 + + MESH:D019851 Disease - - Thrombophilia, deficiency of antithrombin(AT) + + Thrombophilia, deficiency @@ -39468,7 +39667,7 @@ paragraph 3716 Trauma or surgery: especially the pelvis, hip, leg - + MESH:D014947 Disease @@ -39486,7 +39685,7 @@ paragraph 3854 Heart failure - + MESH:D006333 Disease @@ -39498,7 +39697,7 @@ paragraph 3872 Recent myocardial infarction - + MESH:D009203 Disease @@ -39510,49 +39709,25 @@ paragraph 3905 Hormonal therapy (estrogen, progesterone) - - MESH:D004967 - Chemical - - estrogen - - - MESH:D011374 - Chemical - - progesterone - INTRO paragraph 3951 Using the contraceptive pills - - MESH:D003270 - Chemical - - contraceptive - INTRO paragraph 3985 Paraplegia - - MESH:D010264 - Disease - - Paraplegia - INTRO paragraph 4000 Severe infection - + MESH:D007239 Disease @@ -39564,19 +39739,13 @@ paragraph 4021 Intestinal inflammations - - MESH:D007249 - Disease - - inflammations - INTRO paragraph 4050 Polycythemia - + MESH:D011086 Disease @@ -39588,7 +39757,7 @@ paragraph 4067 Paraproteinemia - + MESH:D010265 Disease @@ -39600,23 +39769,17 @@ paragraph 4087 Behcet's disease - - MESH:D010300 - Disease - - s disease - INTRO paragraph 4108 Paroxysmal nocturnal hemoglobinuria - + MESH:D006457 Disease - - nocturnal hemoglobinuria + + Paroxysmal nocturnal hemoglobinuria @@ -39624,13 +39787,13 @@ paragraph 4149 Although etiology and common risk factors for pulmonary thromboembolism are well known there is little data about frequency of postoperative pulmonary thromboembolism after different surgical operations. - + MESH:D011655 Disease pulmonary thromboembolism - + MESH:D011655 Disease @@ -39648,31 +39811,25 @@ paragraph 4366 Our objective is to show the number of patients with postoperative pulmonary thromboembolism (PPTE) treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from June 1, 2011 - June 1, 2014 and to indicate the importance of various surgical operations in the development of pulmonary thromboembolism (PTE). - + 9606 Species patients - + MESH:D011655 Disease pulmonary thromboembolism - + MESH:D008171 Disease pulmonary diseases - - MESH:C539179 - Chemical - - PTE - - + MESH:D011655 Disease @@ -39690,67 +39847,85 @@ paragraph 4751 This is the retrospective study which shows the number of patients with postoperative pulmonary thromboembolism treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period: 01.06.2011.-01.06.2014. It represents the number of these patients, per cent of patients with postoperative pulmonary thromboembolism of total patients with pulmonary thromboembolism, age and sex of patients, type of surgery, period expressed in days from surgery to clinical presence of pulmonary thromboembolism, presence of deep venous thrombosis (DVT) of lower extremities, massiveness of PPTE e.g. level of pulmonary artery with embolus: segmental, lobar, main branches of pulmonary artery; unilateral, bilateral. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D020246 Disease deep venous thrombosis - - MESH:D011655 - Disease - - pulmonary thromboembolism + + - + Chemical + + PPTE - + MESH:D008171 Disease pulmonary diseases - + MESH:D011655 Disease pulmonary thromboembolism - + + MESH:D004617 + Disease + + embolus + + + OMIM:612862 + Disease + + DVT + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + MESH:D011655 Disease postoperative pulmonary thromboembolism - + MESH:D011655 Disease @@ -39768,53 +39943,59 @@ paragraph 5503 In three-year period 232 patients with PTE were treated in Intensive care unit of Clinic "Podhrastovi". 60 of them or 25.86% were patients with postoperative PTE. Among them there were 24 males or 40% middle-aged 58.5 years, and 36 females or 60% middle-aged 56.3 years. - + 9606 Species patients - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE - RESULTS paragraph 5778 Only one man had DVT (deep venous phlebothrombosis) (after orthopedic surgery), and five of them had anamnesis about previous DVT (one with abdominal, one with vascular, one with orthopedic and two with cardiology surgery). Only one woman had DVT (after orthopedic surgery), and no one had anamnesis about previous DVT or PTE. - + 9606 Species man - + 9606 Species woman + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + - MESH:C539179 - Chemical - - PTE + OMIM:612862 + Disease + + DVT MESH:D020246 @@ -39828,18 +40009,12 @@ paragraph 6105 Patients were subjected to different types of surgical operations which are presented on Figure 1. PTE developed in different interval after operation which is presented on the Figure 2. - + 9606 Species Patients - - MESH:C539179 - Chemical - - PTE - FIG @@ -39848,24 +40023,12 @@ fig_caption 6292 Type and number of surgical operations before development of PTE. PTE developed in 15 patients with abdominal, 11with urologic, 8 with gynecologic, 15 with orthopedic, 4 with cardiologic. 2 with vascular, 3 with neurosurgical, 1 with glandular (breast) and 1 with orl operations - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE - FIG @@ -39874,36 +40037,36 @@ fig_caption 6571 The average period in days from surgery to the development of PTE. PTE developed in one to thirty days after operation, on average for abdominal operations it was 19 days, for urologic 5, for gynecologic 10, for orthopedic 15.5, for cardiologic 13, for vascular 4.5, for neurosurgical 18.5, for glandular 10 and for orl operations 10 days after operation. PPTE developed on average 10 days in women, and 14 days after operation in men. - + 9606 Species women - + 9606 Species men - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE - RESULTS paragraph 7007 Massiveness of PPTE e.g., the site or level of embolus was different-from segmental to main branches of pulmonary artery which is shown on Figure 3 and Figure 4. + + MESH:D004617 + Disease + + embolus + + + - + Chemical + + PPTE + FIG @@ -39912,25 +40075,25 @@ fig_caption 7169 The level or site of pulmonary embolus according to the type of surgery in women. The level of pulmonary embolism is different, from segmental to main branches of pulmonary artery with different number of patients according to operation. - + 9606 Species women - + 9606 Species patients - + MESH:D004617 Disease pulmonary embolus - + MESH:D011655 Disease @@ -39944,25 +40107,25 @@ fig_caption 7407 The level or site of pulmonary embolus according to the type of surgery in men. The level of pulmonary embolism is different, from segmental to main branches of pulmonary artery with different number of patients according to operation - + 9606 Species men - + 9606 Species patients - + MESH:D011655 Disease pulmonary embolism - + MESH:D004617 Disease @@ -39980,55 +40143,37 @@ paragraph 7656 There is notably increasing number of patients with pulmonary thromboembolism in recent years treated in Clinic for pulmonary diseases and TB "Podhrastovi" We are not yet sure whether it is real increasing or it is matter of better diagnostics. In three-year period 232 patients with PTE were treated in Intensive care unit of this clinic. 60 of them or 25.86% were patients where PTE developed after different types of surgical operations. We intended with this study to point out the significance and frequency of postoperative pulmonary thromboembolism, and to indicate that surgical operations of different type are significant etiologic and risk factor for the development of PTE. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - + MESH:D011655 Disease postoperative pulmonary thromboembolism - + MESH:D008171 Disease pulmonary diseases - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE - - + MESH:D011655 Disease @@ -40040,35 +40185,29 @@ paragraph 8346 This study has several limitations. First of all we did not have real data about regularity of preoperative and postoperative prophylaxis with anticoagulants, early rising from bed after surgery (mechanical prophylaxis), we did not have data about intraoperative or early postoperative deaths caused possibly by PTE. Furthermore, not small number of patients with postoperative PTE was treated in corresponding surgical clinic under the control of pulmonologist from our clinic, and released home to continue anticoagulant therapy under the control of pulmonologist in outpatient department. - + 100653365 Gene bed - + 9606 Species patients - + 9606 Species outpatient - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE + + MESH:D010149 + Disease + + postoperative deaths @@ -40076,17 +40215,11 @@ paragraph 8938 Although common risk factors for the development of PTE are well known, studied and confirmed in clinical practice there are no enough data in literature about postoperative DVT or PTE out of controlled studies. Many authors indicate the significance of preoperative anticoagulant prophylaxis. - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE + + OMIM:612862 + Disease + + DVT @@ -40094,180 +40227,192 @@ paragraph 9232 One big prospective study was done by group of authors on 75 771 patients with vascular and orthopedic operations from 1996 to 2001 in Veteran Health Administration Hospital. The average age was 65 years and 96% of patients were males. Major comorbidities included diabetes mellitus, chronic obstructive pulmonary disease, and congestive heart failure. Symptomatic PTE was diagnosed in 805 patients (0.68%) and varied significantly with procedures: 0.14% for carotid endarterectomy to 1.34% for total hip arthroplasty. In our study we dealt with patients of both sex (60% of them women), average age was 58.5 years for men, and aged 56.3 years for women, and patients were subjected to different types of surgery. We did not examine comorbidities before operation. We dealt with patients not prospectively, but only with surgical treated patients with different operations with diagnosed PPTE to indicate the importance of surgery to the development of PTE. These authors indicate the postoperative importance of pneumonia and other infections, including urinary tract infection, myocardial infarction. We did not find such factors of such importance before the development of PPTE. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species women - + 9606 Species men - + 9606 Species women - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + + MESH:D006618 + Disease + + hip arthroplasty + + MESH:D029424 Disease chronic obstructive pulmonary disease - + MESH:D014552 Disease - - tract infection + + urinary tract infection - + MESH:D009203 Disease myocardial infarction - - MESH:C539179 - Chemical - - PTE - - + MESH:D006333 Disease congestive heart failure - + + MESH:D007239 + Disease + + infections + + MESH:D011014 Disease pneumonia - + MESH:D003920 Disease diabetes mellitus - - MESH:C539179 - Chemical - - PTE - DISCUSS paragraph 10415 Only two patients in our study had DVT after operation (one man and one woman), but in all others were not be able to find the starting point of embolus. So we think that the preoperative and postoperative prophylaxis (either anticoagulant, or mechanical-early rise from bed) is of the most significance, especially in patients with risk factors for DVT or PTE, even it is only the expected lying in bed (bed-rest) longer than 4 days. There is the need for one long prospective study in different surgical clinic for the assessment of more factors (age, sex, preoperative diseases, type and site of operation, postoperative complications, duration of bed-rest, anticoagulant and mechanical prophylaxis etc.) to give the more realistic picture of postoperative pulmonary thromboembolism. - + 100653365 Gene bed - + 100653365 Gene bed - + 100653365 Gene bed - + 100653365 Gene bed - + 9606 Species patients - + 9606 Species man - + 9606 Species woman - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - + MESH:D011655 Disease postoperative pulmonary thromboembolism + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + MESH:D004617 + Disease + + embolus + CONCL @@ -40280,23 +40425,17 @@ paragraph 11216 Different surgical operations are the big risk factor for the development of PTE. There is great significance of anticoagulant prophylaxis before surgery even in patients with no anamnesis of previous DVT or PTE. - + 9606 Species patients - - MESH:C539179 - Chemical - - PTE - - - MESH:C539179 - Chemical - - PTE + + OMIM:612862 + Disease + + DVT @@ -40321,12 +40460,7 @@ 574 11477 Akutno plucno srce i plucna tromboemblija. U: Plucne bolesti - - Chemical - - Plucne - - + Disease plucna tromboemblija @@ -40345,7 +40479,7 @@ ref 11541 Acute pulmonary thromboembolism. Clinical outcomes in the International Cooperative Pulmonary Registry (ICOPER) - + MESH:D011655 Disease @@ -40364,12 +40498,18 @@ 952 11653 Is embolism risk condition by location of deep venous thrombosis? - + MESH:D020246 Disease deep venous thrombosis + + MESH:D004617 + Disease + + embolism + 100 @@ -40385,19 +40525,19 @@ 603 11719 Clinical, roendgenographic and electrocardiographic findings in patients with acute pulmonary embolism and no pre-existing cardiac or pulmonary disease - + 9606 Species patients - + MESH:D011655 Disease acute pulmonary embolism - + MESH:D008171 Disease @@ -40417,13 +40557,13 @@ 455 11871 Reproduction of chest pain by palpation: diagnostic accuracy in suspected pulmonary embolism - + MESH:D002637 Disease chest pain - + MESH:D011655 Disease @@ -40445,7 +40585,7 @@ 58 11964 Postoperative pulmonary thromoboembolism - + MESH:D001261 Disease @@ -40468,7 +40608,7 @@ surname:Kuri;given-names:SH 12005 Risk factors and clinical impact of postoperative symptomatic venous thromboembolism - + MESH:D054556 Disease @@ -40490,7 +40630,7 @@ 400s 12090 Prevention of venous thromboembolism: The Seventh ACCP Conference on Antithrombotic and Thrombolytic Therapy - + MESH:D054556 Disease @@ -40510,7 +40650,7 @@ 415 12199 A systematic review of strategies to improve prophylaxis for venous thromboembolism in hospitals - + MESH:D054556 Disease @@ -40532,31 +40672,31 @@ 800 12296 A comparison of enoxaparin with placebo for the prevention of venous thromboembolism in acutely ill medical patients. Prophylaxis in medical patients with enoxaparin study group - + 9606 Species patients - + 9606 Species patients - + MESH:D017984 Chemical enoxaparin - + MESH:D054556 Disease venous thromboembolism - + MESH:D017984 Chemical @@ -40608,30 +40748,42 @@ abstract 120 Body weight and body mass index (BMI) are considered strong predictors of osteoporotic fractures, though optimal BMI levels remain unsettled. There are several studies conducted on women about the relationship between BMI and bone mineral density (BMD), and just a few so far on men. Therefore, the objective of current study was to analyze the relationship between weight and BMI and BMD measured in lumbar spine (L1-L4), femur neck and total hip in 64 men from Kosovo. - + 9606 Species women - + 9606 Species men - + 9606 Species men - + + MESH:D020388 + Disease + + BMD + + MESH:D058866 Disease osteoporotic fractures + + MESH:D020388 + Disease + + BMD + ABSTRACT @@ -40644,13 +40796,13 @@ abstract 600 This cross-sectional study included a population of 64 men divided into three BMI groups. Dual-energy X-ray absorptiometry (DEXA) measurements were done in all the study participants. - + 9606 Species men - + 9606 Species @@ -40668,6 +40820,24 @@ abstract 794 Pearson's correlation analysis showed a significant positive correlation between weight and BMI and BMD in femur neck and in total hip, and a significant negative correlation between age and femur neck BMD. Age-adjusted linear regression analysis showed that weight and BMI had a significant positive association with BMD levels. + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + ABSTRACT @@ -40680,19 +40850,37 @@ abstract 1136 Although the results show significant relationship between BMI and BMD, the negative relationship between age and femur neck BMD may serve as guidance to initiate early assessment of the BMD in this region as well as preventive measures of osteoporosis and fractures among ageing men population - + 9606 Species men - + MESH:D050723 Disease fractures - + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + MESH:D010024 Disease @@ -40710,72 +40898,108 @@ paragraph 1447 Fractures accompanying osteoporosis have become serious health problem. There are many modifiable factors influencing bone mineral density (BMD), which is recognized as a surrogate marker of bone strength. Among them, body weight and body mass index (BMI) are considered strong predictors of osteoporotic fractures. Optimal BMI levels remain still unsettled, though a BMI of 26-28 kg/m2 has been suggested. Overweight and obesity have been considered protective to bone health, while higher BMI levels to be associated with higher BMD score. Beyond the simple mechanism of mechanical load related to body weight on bone density, there are some other assumed mechanisms not yet fully elucidated of interaction between adipose tissue and skeleton through several adipokines. However, recent studies have questioned this traditional view concluding non-linear relationship between BMI and BMD. Some studies reported no association or even lower BMD or increased risk of fractures, sometimes site dependent, in obese patients. - + 9606 Species patients - + MESH:D009765 Disease obese - - MESH:D050723 - Disease - - fractures - - + MESH:D058866 Disease osteoporotic fractures - + MESH:D010024 Disease - - Fractures accompanying osteoporosis + + osteoporosis - + + MESH:D020388 + Disease + + BMD + + MESH:D009765 Disease obesity + + MESH:D020388 + Disease + + BMD + + + MESH:D050723 + Disease + + Fractures + + + MESH:D050723 + Disease + + fractures + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + INTRO paragraph 2470 The objective of this study was to analyze the relationship between body mass index (BMI) and weight and bone mineral density (BMD) measured in lumbar spine (L1-L4), femur neck and total hip in a population of 64 men from Kosovo. The interest of several studies has been focused on women, but there are only few recent studies worldwide focused on this issue on men and, to our best knowledge, this is the first one undertaken among Kosovo men. - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species men + + MESH:D020388 + Disease + + BMD + TABLE @@ -40784,6 +41008,12 @@ table_caption 2915 Descriptive characteristics of the study subjects. BMD bone mineral density, BMI body mass index. *p <0.05, **p<0.01 compared to normal weight, p <0.05, p<0.01 compared to overweight + + MESH:D020388 + Disease + + BMD + METHODS @@ -40802,19 +41032,31 @@ paragraph 3152 The study population consisted of 64 men who were divided into three groups according to standard World Health Organization (WHO) Body Mass Index (BMI) classification: Normal Weight group (n=23), Overweight group (n=26) and Obese group (n=15). Subjects were not included in the study if they had history of diseases that might affect bone metabolism or had taken drugs that might influence BMD (glucocortiocids, bisphosphonates). Written informed consent was obtained from all participants and the study protocol was approved by local Ethical Committees. The research was conducted in accordance to guidelines in Declaration of Helsinki. - + 9606 Species men - + 9606 Species participants - + + MESH:D020388 + Disease + + BMD + + + - + Chemical + + glucocortiocids + + MESH:D004164 Chemical @@ -40832,43 +41074,55 @@ paragraph 3848 Height (m) and weight (kg) were measured in light clothing and no shoes before performing DEXA, and body mass index (BMI) was calculated as the ratio of subjects' weight to height squared (kg/m2). According to WHO criteria, participants were categorized into three BMI groups: normal weight (BMI <24.9 kg/m2), overweight (BMI=25.0-29.9 kg/m2) and obese (BMI>30 kg/m2). Dual-energy X-ray absorptiometry (DEXA; Hologic QDR-4500, USA) was used to measure bone mineral density (BMD) in lumbar vertebrae (L1-L4), femoral neck and total hip in all study participants. DEXA was calibrated daily using a phantom provided by the manufacturer. The diagnosis of osteoporosis and osteopenia was done according to WHO T-score criteria (osteoporosis if T-score <= -2.5 SD, osteopenia if T-score between -2.5 SD and -1 SD, and normal BMD if T-score > -1SD). - + 9606 Species participants - + 9606 Species participants - + MESH:D010024 Disease osteoporosis - + MESH:D010024 Disease osteoporosis - + + MESH:D020388 + Disease + + BMD + + MESH:D009765 Disease obese - + MESH:D001851 Disease osteopenia - + + MESH:D020388 + Disease + + BMD + + MESH:D001851 Disease @@ -40886,12 +41140,24 @@ paragraph 4720 SPSS version 16.0 (SPSS Inc., Chicago, IL, USA) was used for statistical analysis. All p-values in statistical analyses were two-tailed with a significance level of 0.05. Continuous variables are expressed as the means +- standard deviations (SD), while categorical variables as percentages. One-way analysis of variance (ANOVA) with Tukey test for Post Hoc Multiple Comparisons for continuous variables and Chi square test for categorical variables were used to compare characteristics of the three groups created on the basis of BMI. Person's correlation test was performed to assess correlation between age, weight, BMI and BMD. Age-adjusted linear regression analysis was calculated with BMI and weight as independent variables and BMD in L1-L4 vertebra, femoral neck and total hip as dependent variables. - + 9606 Species Person + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + RESULTS @@ -40904,19 +41170,19 @@ paragraph 5541 Descriptive characteristics of the study population in total and according to BMI groups are summarized in The recruited study subjects consisted of 64 males with a mean age of 42.17+-14.86 (SD). There were 23 subjects (36.3+-15.81 years) in normal weight group,26 subjects (42.92+-13.9 years) in overweight group and 15 subjects (49.87+-11.59 years) in obese group. Most men in this study were overweight or obese (n=41 or 64%). - + 9606 Species men - + MESH:D009765 Disease obese - + MESH:D009765 Disease @@ -40928,10 +41194,47 @@ paragraph 5971 Comparison of BMI groups by using one way ANOVA found no statistically significant difference for L1-L4 BMD, femur neck BMD and total hip BMD levels (p>0.05). Despite the fact that a statistically significant difference was not observed among BMI groups regarding BMD levels, Pearson's correlation analysis shown in table 2 reveals a significant positive correlation between weight and BMI and BMD in femur neck (r=0.445, p<0.01) and in total hip (r=0.502, p<0.01). No statistically significant correlation was shown between weight and BMI and L1-L4 BMD, while a significant negative correlation was observed between age and femur neck BMD (r=-0.302, p<0.05). - + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 Disease - - hip BMD + + BMD @@ -40941,27 +41244,66 @@ table_caption 6631 Pearson's correlation analysis between age, weight, BMI and BMD (r) *p <0.05; **p<0.01 + + MESH:D020388 + Disease + + BMD + RESULTS paragraph 6718 Age-adjusted linear regression analysis showed that weight and BMI as explanatory variables had a significant positive association with L1-L4 BMD, FN BMD and TH BMD levels (Table 3). When weight was compared to BMI, it was observed a tendency of differences in L1-L4 BMD and FN BMD (beta=0.308 vs. 0.305 and beta=0.563 vs. beta= 0.559), whereas BMI showed greater difference in total hip BMD (beta=0.577 vs. 0.540). Weight and BMI had highest squared R values for femur neck BMD, where these variables can explain 38.6% and 35.4% of the variation in FN BMD (R2=0.386 and R2=0.354, respectively). - + + MESH:D020388 Disease - - FN BMD + + BMD - + + MESH:D020388 Disease - - hip BMD + + BMD - + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 Disease FN BMD + + MESH:D020388 + Disease + + FN BMD + + + MESH:D020388 + Disease + + FN BMD + TABLE @@ -40970,6 +41312,12 @@ table_caption 7314 Age-adjusted linear regression analysis for BMD with weight and BMI as predictors.*p<0.05;**p<0.01; ***p<0.001 + + MESH:D020388 + Disease + + BMD + DISCUSS @@ -40982,44 +41330,44 @@ paragraph 7439 Body weight and body mass index as potentially modifiable factors are generally considered essential determinants of bone mineral density. Several studies demonstrated positive association of elevated body weight and/or BMI with bone mineral density or even protective effect against osteoporosis and fractures. Principal explanation of this association was heavier mechanical loading on bones with subsequent bone remodelation to resist this loading. Some of other putative mechanisms are increased estrogen synthesis (in women), increased secretion of insulin and increased plasma levels of leptin. However, there are other published studies focused on the impact of fat mass on BMD that demonstrated negative association of obesity with BMD; the assumed mechanisms are secretion of pro-inflammatory cytokines that stimulate bone resorption, decreased levels of adiponectins and increased parathormone levels. - + 3630 Gene 173 insulin - + 9606 Species women - + + MESH:D020388 + Disease + + BMD + + MESH:D050723 Disease fractures - + + MESH:D020388 + Disease + + BMD + + MESH:D010024 Disease osteoporosis - - MESH:D004967 - Chemical - - estrogen - - - MESH:D010281 - Chemical - - parathormone - - + MESH:D009765 Disease @@ -41031,33 +41379,71 @@ paragraph 8351 In accordance with results of Dogan A et al., Salamat M et al. and Lloyd J et al. results of this cross-sectional study demonstrate a positive correlation of weight and body mass index with both femur neck BMD and total hip BMD in all male subjects included in the study, but, in contrary to them, compared to normal weight, BMD levels of overweight and obese groups were not significantly different; the lack of significant difference between BMI groups might be as limitation of the small sample size. Contrary to results of our study and above mentioned studies, Greco et al. reported low lumbar BMD in obese patients, while a recent study by Emaus N et al. demonstrated a nonlinear relationship between body mass index and bone mineral density. Every unit increase in BMI was associated with an increase of 0.008 g/cm2 in L1-L4 BMD, 0.017 g/cm2 in femur neck BMD and 0.018 g/cm2 in total hip BMD. Interestingly, age was negatively correlated only with femur neck BMD in all subjects. - + 9606 Species patients - + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + MESH:D009765 Disease obese - - Disease - - hip BMD - - + MESH:D009765 Disease obese - + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 Disease - - hip BMD + + BMD @@ -41071,19 +41457,25 @@ paragraph 9353 In conclusion, we demonstrated a highly significant relationship between body mass index and bone mineral density in femur neck and total hip in this sample of men. Negative relationship between age and femur neck BMD may serve as guidance to focus the evaluation of the bone mineral density in this region and to initiate early preventive measures of osteoporosis and fractures among aging male population. - + 9606 Species men - + + MESH:D020388 + Disease + + BMD + + MESH:D050723 Disease fractures - + MESH:D010024 Disease @@ -41117,7 +41509,7 @@ 43 9809 Male osteoporosis - + MESH:D010024 Disease @@ -41139,7 +41531,7 @@ surname:Feng;given-names:X 9827 Relationship of body composition with bone mineral density in northern Chinese men by body mass index levels - + 9606 Species @@ -41165,13 +41557,13 @@ surname:Achenbach;given-names:SJ 9936 Contributions of bone density and structure to fracture risk assessment in men and women - + 9606 Species men - + MESH:D050723 Disease @@ -41207,13 +41599,13 @@ surname:Chen;given-names:YT 10123 Positive associations of bone mineral density with body mass index, physical activity, and blood triglyceride level in men over 70 years old: a TCVGHAGE study - + 9606 Species men - + MESH:D014280 Chemical @@ -41255,17 +41647,17 @@ surname:Sorge;given-names:R 10379 Relationship between body composition, body mass index and bone mineral density in a large population of normal, osteopenic and osteoporotic women - + MESH:C567172 Disease osteopenic - + MESH:D058866 Disease - - osteoporotic women + + osteoporotic @@ -41290,30 +41682,24 @@ surname:Berger;given-names:C 10526 Physical activity, body mass index and bone mineral density-associations in a prospective population-based cohort of women and men: the Canadian Multicentre Osteoporosis Study (CaMos) - + 9606 Species women - + 9606 Species men - + MESH:D010024 Disease Multicentre Osteoporosis - - MESH:D008982 - Chemical - - CaMos - surname:Janghorbani;given-names:M @@ -41328,7 +41714,7 @@ ref 10710 Relationship between Weight, Body Mass Index, and Bone Mineral Density in Men Referred for Dual-Energy X-Ray Absorptiometry Scan in Isfahan, Iran - + 9606 Species @@ -41374,11 +41760,11 @@ surname:Lee;given-names:H 11004 Visceral fat is a negative predictor of bone density measures in obese adolescent girls - - MESH:D063766 + + MESH:D009765 Disease - - obese adolescent girls + + obese @@ -41398,18 +41784,24 @@ 341 11092 Is age or the body mass index (BMI) more determinant of the bone mineral density (BMD) in geriatric women and men? - + 9606 Species women - + 9606 Species men + + MESH:D020388 + Disease + + BMD + 11 @@ -41426,13 +41818,7 @@ 14 11207 Obesity and fracture risk - - MESH:D009765 - Disease - - Obesity - - + MESH:D050723 Disease @@ -41466,13 +41852,13 @@ surname:Prossomariti;given-names:G 11233 Is obesity protective for osteoporosis? Evaluation of bone mineral density in individuals with high body mass index - + MESH:D009765 Disease obesity - + MESH:D010024 Disease @@ -41490,7 +41876,7 @@ ref 11349 Is the negative relationship between obesity and bone mineral content greater for olderwomen? - + MESH:D009765 Disease @@ -41531,19 +41917,13 @@ surname:Boonen;given-names:S 11443 Obesity is not protective against fracture in postmenopausal women: GLOW - + 9606 Species women - - MESH:D009765 - Disease - - Obesity - - + MESH:D050723 Disease @@ -41588,31 +41968,31 @@ surname:Cawthon;given-names:PM 11581 BMI and fracture risk in older men: the osteoporotic fractures in men study (MrOS) - + 8011 Gene MrOS - + 9606 Species men - + 9606 Species men - + MESH:D058866 Disease osteoporotic fractures - + MESH:D050723 Disease @@ -41641,13 +42021,13 @@ surname:Martinez-Laguna;given-names:D 11664 The association between fracture and obesity is site-dependent: a population-based study in postmenopausal women - + MESH:D050723 Disease fracture - + MESH:D009765 Disease @@ -41674,19 +42054,19 @@ surname:Nogues;given-names:X 11777 The association between fracture site and obesity in men: a population-based cohort study - + 9606 Species men - + MESH:D050723 Disease fracture - + MESH:D009765 Disease @@ -41838,101 +42218,96 @@ abstract 605 116 children with rotavirus diarrhea are included in the study. The majority boys (74.4%) and children aged 0 - 12 months (82.75%). Mean age of children in the study was 16.38 months. Almost every third child in the study was hypotrophic (29.2%). More than half of the infants (55.2%) were on mixed food, somewhat more than every third was breast feeding (36.45%), and every twelfth (8.33%) was on artificial milk (animal or formula). Apart from diarrhea, present in all patients, vomiting (97.41%) and fever (43.96%) were characteristics of the clinical presentation of the diarrhea. Two thirds of the children had mild grade dehydration (70.7%). All patients recovered with no sequels. - + 9606 Species children - + 9606 Species boys - + 9606 Species children - + 9606 Species children - + 9606 Species child - + 9606 Species infants - + 9606 Species patients - + 9606 Species children - + 9606 Species patients - + MESH:D003681 Disease dehydration - + MESH:D014839 Disease vomiting - + MESH:D003967 Disease diarrhea - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D005334 Disease fever - + MESH:D003967 Disease diarrhea - - Disease - - hypotrophic - ABSTRACT @@ -41945,13 +42320,13 @@ abstract 1307 Rotavirus continues to be responsible for a significant portion of acute diarrhea in Kosovo. Clinical features, epidemiological data and the agglutination test are safe enough to establish the diagnosis. Treated correctly rotavirus diarrhea has a favorable outcome. - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D003967 Disease @@ -41969,49 +42344,49 @@ paragraph 1587 Since 1973 when discovered by Bishop, rotavirus was found responsible for one third of the children hospitalization for acute diarrhea worldwide. Rotavirus diarrhea is a cause of 440 thousand deaths, 2 million hospitalizations, 25 million outpatients visit and 111 million episodes of domiciliary diarrhea among children. The aim of this study is to analyze the epidemiology of the acute diarrhea inflicted by rotavirus, demographics of the patients and clinical presentation of the disease in children hospitalized at the Department of Pediatric Gastroenterology of the University Clinical Center of Kosovo in Prishtina, Republic of Kosovo. - + 9606 Species children - + 9606 Species children - + 9606 Species patients - + 9606 Species children - + MESH:D012400 Disease Rotavirus diarrhea - + MESH:D003967 Disease diarrhea - + MESH:D003967 Disease diarrhea - + MESH:D003967 Disease @@ -42035,7 +42410,7 @@ paragraph 2599 This is a prospective study carried between January 1st and December 31st 2011. All data, comprising demographics, nutrition, clinical presentation, laboratory findings, management and outcome of the rotavirus diarrhea are collected on the specially designed form. - + MESH:D012400 Disease @@ -42047,7 +42422,7 @@ paragraph 2864 Retrovirus is confirmed in the stool of patients using Latex agglutination test LTA s.r.l. - Via Milan with sensitivity and specify of 96%. - + 9606 Species @@ -42065,13 +42440,13 @@ paragraph 3017 Of 1011 children admitted at the Hospital for acute diarrhea during the study period, every third is tested for rotavirus. Of 337 tested, 116 were positive on rotavirus (34.42%). - + 9606 Species children - + MESH:D003967 Disease @@ -42083,77 +42458,67 @@ paragraph 3196 The majority of the children in the study were boys (74.4) and infants (82.75%). Average age of children in the study was 16.38 months. Almost every third child in study was hypotrophic (29.2%). The hypotrophy was more frequent in girls than in boys (41% vs. 23.5%). - + 9606 Species children - + 9606 Species boys - + 9606 Species infants - + 9606 Species children - + 9606 Species child - + 9606 Species girls - + 9606 Species boys - - Disease - - hypotrophy - - - Disease - - hypotrophic - RESULTS paragraph 3463 Only 20 children were not infants (17.2%). More than half of the infants (55.2%) were on mixed food, somewhat more than every third was breast feeding (36.45%), and every twelfth (8.33%) was on artificial milk (animal or formula). - + 9606 Species children - + 9606 Species infants - + 9606 Species @@ -42173,13 +42538,13 @@ table_caption 3780 Demographics, nutritional status and mode of feeding of the patient with rotavirus diarrhea in the study - + 9606 Species patient - + MESH:D012400 Disease @@ -42191,73 +42556,79 @@ paragraph 3885 Apart from diarrhea, present in all patients, vomiting (88.8%) and fever (43.96%) were characteristics of the clinical presentation of the diarrhea. Vomiting was more frequent in boys than in girls (97.4% vs. 71.8%) as well as temperature (46.7% vs. 38.5%). Two patients in the study (1.7%), one boy and one girl have had a convulsion at the admission The frequency of the convulsions was equal (each 1.7%) - + 9606 Species patients - + 9606 Species boys - + 9606 Species girls - + 9606 Species patients - + 9606 Species boy - + 9606 Species girl - + MESH:D012640 Disease convulsions - + MESH:D003967 Disease diarrhea - + MESH:D012640 Disease convulsion - + + MESH:D014839 + Disease + + Vomiting + + MESH:D014839 Disease vomiting - + MESH:D005334 Disease fever - + MESH:D003967 Disease @@ -42269,37 +42640,37 @@ paragraph 4292 All patients in the study had moderate or severe dehydration with the later present in almost every third patient in the study (29.3%). There are no significant differences in the degree of the dehydration between girls and boys. - + 9606 Species patients - + 9606 Species patient - + 9606 Species girls - + 9606 Species boys - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease @@ -42311,13 +42682,13 @@ paragraph 4522 Clinical presentation of the rotavirus diarrhea and the degree of dehydration at the admission are presented in table 2. - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D003681 Disease @@ -42331,19 +42702,19 @@ table_caption 4643 Symptoms and degree of dehydration of the patient with rotavirus diarrhea at the admission - + 9606 Species patient - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D003681 Disease @@ -42355,43 +42726,61 @@ paragraph 4734 Only about one third of the patients with rotavirus diarrhea in our study had associated diseases (36.2%). The most frequent one was bronchopneumonia that was present in 31.9% of our patients. Less common were upper urinary tract infections (1.7%), upper respiratory tract infections (0.9%) and gastro esophageal reflux (1.7%). Both urinary tract infections in our study were in girls (5.1% of the girls). - + 9606 Species patients - + 9606 Species patients - + 9606 Species girls - + 9606 Species girls + + MESH:D012141 + Disease + + respiratory tract infections + MESH:D012141 Disease - - upper respiratory tract infections + + tract infections + MESH:D005764 + Disease + + gastro esophageal reflux + + MESH:D001996 Disease bronchopneumonia - + + MESH:D014552 + Disease + + upper urinary tract infections + + MESH:D012400 Disease @@ -42403,13 +42792,13 @@ paragraph 5140 Data on associated conditions and diseases in patients with rotavirus diarrhea are presented in table 3. - + 9606 Species patients - + MESH:D012400 Disease @@ -42423,13 +42812,13 @@ table_caption 5245 Associated conditions and diseases in patient with rotavirus diarrhea in our study - + 9606 Species patient - + MESH:D012400 Disease @@ -42441,67 +42830,67 @@ paragraph 5328 Paralytic ileus was the most common complication of the rotavirus diarrhea. In our study it occurred in 9.5% of patients or almost in every tenth patient. The incidence of this complication was for a little more frequent in girls than in boys (15.4% vs 6.5%). There was also a single case of ascites among our patients (0.86%). One patient presented with neurological symptoms (seizures). - + 9606 Species patients - + 9606 Species patient - + 9606 Species girls - + 9606 Species boys - + 9606 Species patients - + 9606 Species patient - + MESH:D007418 Disease Paralytic ileus - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D012640 Disease seizures - + MESH:D009422 Disease neurological symptoms - + MESH:D001201 Disease @@ -42513,72 +42902,54 @@ paragraph 5717 Little more than two of three patients in our study were treated without antibiotics (36.2%) and only eight patients (6.9%) were given concentrated erythrocytes or fresh frozen plasma. Ceftriaxone was the most commonly used antibiotic in our practice. The drug was given to 50% of the patients as the only antibiotic and in another 11.2% of patients combined with aminoglycosides - amikacin or gentamicin. Amikacin was given as the only drug in 2.6% of patients and Bactrim (sulfamethoxazole and trimethoprim) in 0.9% of patients. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D013420 - Chemical - - sulfamethoxazole - - - MESH:D005839 - Chemical - - gentamicin - - + MESH:D000617 Chemical aminoglycosides - + MESH:D002443 Chemical Ceftriaxone - - MESH:D000583 - Chemical - - Amikacin - MESH:D000583 Chemical @@ -42586,6 +42957,30 @@ amikacin + MESH:D005839 + Chemical + + gentamicin + + + MESH:D013420 + Chemical + + sulfamethoxazole + + + MESH:D015662 + Chemical + + Bactrim + + + MESH:D000583 + Chemical + + Amikacin + + MESH:D014295 Chemical @@ -42605,31 +43000,31 @@ table_caption 6327 The use of antibiotics, blood and blood derivates for the treatment of patients with rotavirus diarrhea in study. *Ceftriaxone in combination with amikacin or gentamicin - + 9606 Species patients - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D000583 Chemical amikacin - + MESH:D005839 Chemical gentamicin - + MESH:D002443 Chemical @@ -42641,13 +43036,13 @@ paragraph 6497 Assessment of the seasonal distribution of the rotavirus diarrhea has shown that the disease most commonly occurred in autumn, winter and early spring with peaks in September, November and January (Graph 1). All patients recovered without sequels. - + 9606 Species patients - + MESH:D012400 Disease @@ -42661,7 +43056,7 @@ fig_caption 6745 Seasonal distribution of the rotavirus diarrhea - + MESH:D012400 Disease @@ -42679,55 +43074,67 @@ paragraph 6807 Rotavirus is the common cause of diarrhea of the infant worldwide. According to Parashar and associates, who reviewed the Global mortality associated with Rotavirus disease among children, rotavirus is responsible for 527,000 deaths (95% confidence interval, 475,000-580,000 deaths) annually or 29% of all deaths due to diarrhea among children <5 years of age. This translates into approximately 1440 deaths due to rotavirus disease per day (1 of 237 children born each year would die of rotavirus disease by 5 years of age). - + 9606 Species infant - + 9606 Species children - + 9606 Species children - + 9606 Species children - + MESH:D003967 Disease diarrhea - + MESH:D012400 Disease Rotavirus disease - + + MESH:C564120 + Disease + + 000-580,000 deaths + + MESH:D012400 Disease rotavirus disease - + MESH:D003967 Disease diarrhea - + + MESH:D003643 + Disease + + 000 deaths + + MESH:D012400 Disease @@ -42739,73 +43146,79 @@ paragraph 7333 Although rotavirus is ubiquitous and the incidence of disease is similar among children in industrialized and developing countries, there is a great disparity in mortality associated with rotavirus. Whereas very few deaths due to rotavirus disease occurred in affluent countries, countries with the greatest level of child mortality accounted for approximately 86% of all deaths due to rotavirus disease. In fact, 6 countries (India, Nigeria, China, Pakistan, Ethiopia, and Democratic Republic of Congo) accounted for >50% of all deaths due to rotavirus disease, with India alone accounting for one-fourth of the deaths. - + 9606 Species children - + 9606 Species child - + MESH:D012400 Disease rotavirus disease - + MESH:D012400 Disease rotavirus disease - + MESH:D012400 Disease rotavirus disease + + MESH:D006479 + Disease + + Democratic Republic of Congo + DISCUSS paragraph 7954 Papers discussing the prevalence of diarrhea caused by rotavirus in the Republic of Kosovo are lacking. At the moment, there are only two papers, one presented by Azemi alone and another presented by Azemi with coworkers discussing this issue. According to the paper in year 1987, 24.27% of children hospitalized at the Department of Pediatric Gastroenterology of the University Clinical Center of Kosovo had acute diarrhea associated with severe dehydration (toxicosis), and, of them, 7% inflicted by rotavirus. In 2002, according to the second paper, rotavirus is found responsible for 16.8% of acute diarrhea. - + 9606 Species children - - MESH:D003967 - Disease - - acute diarrhea - - + MESH:C565846 Disease toxicosis - + + MESH:D003967 + Disease + + diarrhea + + MESH:D003681 Disease dehydration - + MESH:D003967 Disease diarrhea - + MESH:D003967 Disease @@ -42817,7 +43230,7 @@ paragraph 8567 In our study, the percentage is found to be significantly higher - 34.42%. It is rather reflection of better living standard of the Kosovars and health care provision than of the change in epidemiological behavior of the virus. Better feeding habits, better education of the mothers and better health care, no doubt, have made for other causes of the diarrhea to diminish. - + MESH:D003967 Disease @@ -42829,19 +43242,24 @@ paragraph 8942 With one third of acute diarrhea due to rotavirus (34.42%), prevalence of the disease in Kosovo is similar to the prevalence in other European countries. It is similar to the prevalence in Central and Southeastern Europe, that varies between 22% (Czech Republic) and 55.3% (Russia), with Bosnia and Hercegovina (23.9%), Hungary (27.1%), Albania (32.4%), Bulgaria (32.4%), Romania (42%) and Ukraine (42%) in between, as found by Ogilvie and coauthors in their study of burden of rotavirus gastroenteritis in the pediatric population in this region. But, also similar to the prevalence found in REVEAL study that have found prevalence ranging between 27.8% in Germany and 52% in Sweden, with Spain (31.2%), France (33.5%), Great Britain (35.9%), Italy (43.6%) and Belgium (44.7%) in between. - + MESH:D012400 Disease rotavirus gastroenteritis - + MESH:D003967 Disease diarrhea - + + Disease + + Bosnia + + Disease Romania @@ -42852,37 +43270,37 @@ paragraph 9732 The majority of children with acute diarrhea in our study were infants (48%) and boys (66%). The higher infliction of the boys with rotavirus diarrhea is noted in several other studies, both local and worldwide, but, up to day, there are no known reasons established. Furthermore, there are studies as well, stating the opposite. - + 9606 Species children - + 9606 Species infants - + 9606 Species boys - + 9606 Species boys - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D003967 Disease @@ -42894,31 +43312,37 @@ paragraph 10062 Vomiting, diarrhea and temperature usually occurred at the second day after exposition, following an exposition period of less than 48 hours. Consequently, in our study, dehydration was present in all children. In almost every third child, this dehydration was severe. - + 9606 Species children - + 9606 Species child - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease dehydration - + + MESH:D014839 + Disease + + Vomiting + + MESH:D003967 Disease @@ -42930,79 +43354,79 @@ paragraph 10331 The REVEAL study reported that the proportion of children with dehydration due to acute RVGE varied between 11.1% (Spain) and 71.4% (Sweden), and in most countries, was considerably higher than for those with rotavirus-negative disease. In Belgium and the UK, the prevalence of dehydration was comparable among children with or without RVGE, whereas the dehydration prevalence ratio of rotavirus versus non-rotavirus disease was 1.82, 5.54, 3.27, 3.47, and 2.18, respectively, in France, Germany, Italy, Spain, and Sweden. A second prospective multicenter study from France, Germany, Italy, Spain, and the UK reported that dehydration was evident in 75.7% of patients with RVGE, and was severe in 11.3% of them. In comparison, only 54.5% of children with non-RVGE were dehydrated, while 4.7% had severe dehydration (25). Another study from France confirmed this trend, showing significant differences in dehydration between rotavirus-positive and negative gastroenteritis (26.8% vs.14.7%, P < .0001) as did a study from Greece. Further, one Italian community-based study showed that dehydration at initial presentation in primary care was associated with a higher likelihood of RVGE (OR: 1.8; 95% CI, 1.1-3; P = 0.02). - + 9606 Species children - + 9606 Species children - + 9606 Species patients - + 9606 Species children - + MESH:D003681 Disease dehydration - + MESH:D012400 Disease rotavirus versus non-rotavirus disease - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease dehydration - + MESH:D005759 Disease gastroenteritis - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease dehydration - + MESH:D003681 Disease @@ -43014,19 +43438,19 @@ paragraph 11550 It is interesting to find that about every third patients in our study has bronchopneumonia as associated disease and every tenth had paralytic ileus. - + 9606 Species patients - + MESH:D007418 Disease paralytic ileus - + MESH:D001996 Disease @@ -43038,43 +43462,43 @@ paragraph 11701 Although affection of the central nervous system is not common in children with rotavirus diarrhea, it occurred in two our patients. First to report association of this disease with rotavirus diarrhea were Day and coauthors. They reported neurological symptoms and stool virology positive for rotavirus in four children, presented within 4 weeks in Barnet and Chase Farm Hospitals NHS Trust, London, UK. Similar to our patients, all four made a quick recovery and were discharged home after a few days without medication. At 6 weeks all were well, with no further neurological episodes. - + 9606 Species children - + 9606 Species patients - + 9606 Species children - + 9606 Species patients - + MESH:D009422 Disease neurological symptoms - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D012400 Disease @@ -43086,13 +43510,13 @@ paragraph 12288 Later, Fuchigami and coauthors reported a case of encephalopathy in 4 year old girl at the Department of General Pediatrics, Nihon University Nerima Hikarigaoka Hospital, Tokyo. Probably there will be more such reports in the future. - + 9606 Species girl - + MESH:D001927 Disease @@ -43104,19 +43528,19 @@ paragraph 12522 Use of antibiotics, blood and blood derivates in rotavirus diarrhea is a matter of a worldwide discussion. In out study almost two thirds of the patients were treated with antibiotics and about every twentieth was given blood or blood derivates. High association with bronchopneumonia may explain why. - + 9606 Species patients - + MESH:D012400 Disease rotavirus diarrhea - + MESH:D001996 Disease @@ -43128,7 +43552,7 @@ paragraph 12824 Rotavirus diarrhea most commonly occurred in autumn, winter and early spring with peaks in September, November and January. - + MESH:D012400 Disease @@ -43146,7 +43570,7 @@ paragraph 12962 Rotavirus diarrhea continues to be a serious health problem in Kosovo. Epidemiological and clinical features of the disease and Latex agglutination test are essential tools to establish the diagnosis at the time. - + MESH:D012400 Disease @@ -43180,13 +43604,13 @@ 1284 13223 Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis - + 9606 Species children - + MESH:D005759 Disease @@ -43217,24 +43641,18 @@ 683 13343 Interventions for the control of diarrhoeal diseases among young children: rotavirus and cholera immunisation - + 9606 Species children - - MESH:D030342 + + MESH:D004194 Disease diarrhoeal diseases - - MESH:D002771 - Disease - - cholera immunisation - REF @@ -43255,13 +43673,13 @@ S15 13481 Global Mortality Associated with Rotavirus Disease among Children in 2004 - + 9606 Species Children - + MESH:D012400 Disease @@ -43277,13 +43695,13 @@ ref 13555 The corelation of the etiological agent and acidobasic status in children with toxicosis - + 9606 Species children - + MESH:C565846 Disease @@ -43305,7 +43723,7 @@ surname:Avdiu;given-names:M 13644 Clinical and biochemical features of the acute diarrhea from rotavirus - + MESH:D003967 Disease @@ -43328,13 +43746,7 @@ 533 13715 Burden of rotavirus gastroenteritis in the pediatric population in Central and Eastern Europe. Serotype distribution and burden of illness - - MESH:D002908 - Disease - - illness - - + MESH:D012400 Disease @@ -43353,7 +43765,7 @@ S16 13854 Multicenter Prospective Study of the Burden of Rotavirus Acute Gastroenteritis in Europe 2004 - 2005: The Reveal Study - + MESH:D012400 Disease @@ -43375,7 +43787,7 @@ surname:Avdiu;given-names:M 13975 Clinical and biochemical features of the acute diarrhea from rotavirus - + MESH:D003967 Disease @@ -43392,11 +43804,17 @@ ref 14046 Impact of diseases afecting gastrointestinal system in pediatric toxicosis - + MESH:D005767 Disease - - diseases afecting gastrointestinal system in pediatric toxicosis + + diseases afecting gastrointestinal system + + + MESH:C565846 + Disease + + toxicosis @@ -43411,8 +43829,8 @@ 445 14121 Acidobasic amd electrolitic disorders in pediatric toxicosis and their role in treatment - - MESH:C565846 + + MESH:D063766 Disease pediatric toxicosis @@ -43431,17 +43849,17 @@ 148 14210 Prognosis of the children with diarrhea toxicosis.12th Congress of the pediatritian of the Yugoslavia - + 9606 Species children - + MESH:D003967 Disease - - diarrhea toxicosis.12th Congress of the pediatritian of the Yugoslavia + + diarrhea toxicosis.12th Congress @@ -43459,11 +43877,11 @@ 252 14312 The incidence and clinical presentation of infantile rotavirus diarrhoea in Sierra Leone - + MESH:D012400 Disease - - rotavirus diarrhoea + + infantile rotavirus diarrhoea @@ -43484,13 +43902,13 @@ surname:Gunduz;given-names:TS 14401 Rotavirus and adenovirus frequency among patients with acute gastroenteritis and their relationship to clinical parameters: a retrospective study in Turkey - + 9606 Species patients - + MESH:D005759 Disease @@ -43512,19 +43930,19 @@ 489 14557 Enteric adenovirus infection among infants with diarrhea in rural Bangladesh - + 9606 Species infants - + MESH:D003967 Disease diarrhea - + MESH:D000257 Disease @@ -43550,13 +43968,13 @@ surname:Gocer;given-names:S 14634 Childhood diarrhoea in Ankara, Turkey: epidemiological and clinical features of rotavirus-positive versus rotavirus-negative cases - + 9103 Species Turkey - + MESH:D003967 Disease @@ -43580,13 +43998,13 @@ surname:Htwe;given-names:TT 14765 Hospital-based surveillance for rotavirus diarrhea in children in Yangon, Myanmar - + 9606 Species children - + MESH:D012400 Disease @@ -43615,13 +44033,18 @@ surname:Gwizdek;given-names:A 14847 Epidemiological and clinical characteristics of rotaviral diarrhoea in children from Gdansk, Gdynia and Sopot - + 9606 Species children - + + Disease + + Gdynia + + MESH:D003967 Disease @@ -43650,13 +44073,13 @@ surname:Olarte;given-names:J 14958 Etiology of acute diarrhoea among children in developing countries: a multicentre study in five countries - + 9606 Species children - + MESH:D003967 Disease @@ -43673,7 +44096,7 @@ ref 15064 Rotavirus gastroenteritis - + MESH:D012400 Disease @@ -43691,11 +44114,11 @@ ref 15090 Burden of community-acquired and nosocomial rotavirus gastroenteritis in the pediatric population of Western Europe: a scoping review - + MESH:D012400 Disease - - rotavirus gastroenteritis + + nosocomial rotavirus gastroenteritis @@ -43708,7 +44131,7 @@ ref 15224 Cluster of neurological manifestations of rotavirus infection in children - + MESH:D012400 Disease @@ -43733,13 +44156,13 @@ surname:Gragjevci;given-names:V 15298 The Usefulness of Clinical and Laboratory Parameters for Predicting Severity of Dehydratation in Children with Acute Gastroenteritis - + 9606 Species Children - + MESH:D005759 Disease @@ -43785,12 +44208,6 @@ abstract 62 Elimination of stone is determined by size and its localization. Stone from the ureter in 80% of cases can be eliminated spontaneously. If the stone by its characteristics is not spontaneously eliminated, taken are further steps and therapeutic protocols to solve this problem. - - MESH:C064318 - Chemical - - Stone - ABSTRACT @@ -43803,24 +44220,19 @@ abstract 362 The study was prospective, open and comparative. It was conducted at the Urology Clinic Clinical Center of Sarajevo University in the period from 2007 to 2013. The study included 404 patients with urinary tract lithiasis treated by ESWL. ESWL treatment is performed on the machine Siemens Model Lithostar Multiline, which has a combined ultrasonographic and fluoroscopic display, large energy density in order to obtain optimum focus (without damaging surrounding tissue) and minimal pain that on rare occasions requires for mild sedation-sedation. - + 9606 Species patients - + MESH:D010146 Disease pain - - Chemical - - fluoroscopic - - + MESH:D052878 Disease @@ -43838,49 +44250,49 @@ abstract 920 From a total of 404 patients included in the study there were 234 (57.92%) male and 170 (42.08%) female patients. The most common type of stone both in female and male patients was calcium type. From a total of 262 calcium stones, 105 of them (40.07%) was present in female patients and 157 (59.92%) in male. Share of infectious type of stone in female patients was 63 (49.60%) and 64 among males (50.39%). Other stones were less abundant in both the gender groups and their total number was only 17. In women their frequency was 2 (13.33%) and 13 among males (86.67%). There was a significant difference in the frequency of different types of stones by gender (chi2 = 11.47, p = 0.009). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species women - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical @@ -43898,11 +44310,11 @@ abstract 1620 There was no statistically significant correlation between the number of treatments and localization of stones in the ureter, as well as a statistically significant correlation between the size of the stone and the localization of calculus in the ureter. - + MESH:D002137 Disease - - calculus in the ureter + + calculus @@ -43916,13 +44328,13 @@ paragraph 1891 Every year between the 1200 to 1400 persons per million develop urinary calculosis with a male to female ratio of 3:1. Urology is probably the oldest branch of surgery, knowing that one of the first operations was safe and resolve stone jammed on the outer opening of the urethra. - + 9606 Species persons - + MESH:D020347 Disease @@ -43934,132 +44346,144 @@ paragraph 2172 Creation of stones in the urinary organs (urolithiasis) is a very common disease. Urinary tract stones are sixth health problem. Epidemiological data suggest an increased prevalence of upper urinary tract calculi in developed countries. Various authors suggest that urinary calculi constitute 1-3 per thousand total hospitalized patients which makes 15-24% of patients in the urology departments. This disease have several names: nephrolithiasis-meaning kidney stone, nephrolithiasis and the word is derived from the Greek word nephros (kidney) and lithos (stone). Word of urolithiasis-urinary (urine) stone is derived from the French word urine and Greek ouron meaning urine. It is also used as renal calculus from the Latin word calculus (plural-calculi), which means gravel. Urolithiasis also affects animals: the stones were found in dogs, cats, rabbits, minks, etc., and the components are the same as in humans. Urolithiasis occurs during the most productive human age from 30 to 50 years of age, but has been described in all age groups. It was found in newborns and fetuses. However, urolithiasis in children is rare. Only 2-3% of cases occur in the pediatric age. It is believed that 12% of men and 4% of women in developed countries suffer from urinary stones. In case of a positive family history, this number doubles. One of the reasons for its frequent occurrence in men is explained by the increased endogenous production of oxalate in the liver under the influence of testosterone, while in the urine of women is on average higher concentration of citrate, otherwise, increase the solubility of calcium. Calculosa with women predominantly induced urinary infections and metabolic disorders, while in men predominate idiopathic lithiasis or calcium lithiasis and uric acid. Percentage of recurrent calculosis is 50% within 5 years and 70% over 10 years. Stones less than 2 mm easily pass through the ureter. Most calculi (90%) from 4 to 5 mm are eliminated spontaneously. Percentage decreases to 50% at stones of 4 to 6 mm and 20% at stones larger than 6 mm. - + 9606 Species patients - + 9606 Species patients - + 9615 Species dogs - + 9685 Species cats - + 9986 Species rabbits - + 9666 Species minks - + 9606 Species humans - + 9606 Species human - + 9606 Species children - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species women - + 9606 Species women - + 9606 Species men - + + MESH:D008569 + Disease + + Word + + MESH:D007669 Disease nephrolithiasis-meaning kidney stone - + MESH:D052878 Disease urolithiasis-urinary - + MESH:D052878 Disease urolithiasis - + MESH:D052878 Disease urolithiasis - + MESH:D008659 Disease metabolic disorders - + MESH:D002118 Chemical calcium + + MESH:D002137 + Disease + + plural-calculi + MESH:D053040 Disease @@ -44073,47 +44497,47 @@ oxalate + MESH:D014545 + Disease + + upper urinary tract calculi + + + MESH:D008569 + Disease + + word calculus + + MESH:D020347 Disease calculosis - + MESH:D020347 Disease lithiasis - + MESH:D001745 Disease urinary infections - + MESH:D052878 Disease Urolithiasis - + MESH:D013739 Chemical testosterone - - MESH:C102006 - Chemical - - citrate - - - MESH:D004410 - Disease - - word nephros - MESH:D007669 Disease @@ -44133,16 +44557,16 @@ uric acid - MESH:D002137 - Disease - - word calculus + MESH:D019343 + Chemical + + citrate - MESH:D002118 - Chemical - - calcium + MESH:D008569 + Disease + + word nephros MESH:D052878 @@ -44264,65 +44688,60 @@ paragraph 7000 The study was prospective, open and comparative. It was conducted at the Urology Clinic Clinical Center of Sarajevo University in the period from 2007 to 2013. The study included 404 patients with urinary tract lithiasis. All patients prior to initiation of therapy-ESWL treatments were subjected to the following diagnostic procedures: anamnesis, clinical examination of patients, laboratory tests, ultrasound examination of the urinary tract and urinary tract X-ray, from which was derived chemical qualitative analysis of morphological stone composition. Diagnostics are used in the detection of complications such as obstruction with dilatation of the renal colorectal system, reduction of the renal parenchyma and in monitoring of renal obstruction during treatment. This type of diagnosis is not suitable for stones in the ureter and can make the difference between calcified and radiolucent concrement. ESWL treatment is performed on the machine Siemens Model Lithostar Multiline, which has a combined ultrasonographic and fluoroscopic display, large energy density in order to obtain optimum focus (without damaging surrounding tissue) and minimal pain that on rare occasions requires for mild sedation-sedation. For nominal and ordinal variables chi-square test was used. In cases when the frequency was lower than expected was used the Fisher's exact test. The degree of correlation was determined by means of the Spearman. P value of <0.05 was considered statistically significant. Statistical analysis was performed using SPSS computer software for statistical analysis (SPSS Statistical Package for the Social Sciences) version 13.0. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D010146 Disease pain - + MESH:D052878 Disease urinary tract lithiasis - + MESH:D007674 Disease renal obstruction + + MESH:D007674 + Disease + + dilatation of the renal colorectal system + - MESH:D007415 + MESH:D000402 Disease obstruction - Chemical - - fluoroscopic - - - MESH:D051437 + MESH:D007674 Disease reduction of the renal parenchyma - - MESH:D015179 - Disease - - dilatation of the renal colorectal system - RESULTS @@ -44335,13 +44754,13 @@ paragraph 8660 From a total of 404 patients included in the study there were 234 (57.92%) male and 170 (42.08%) female patients. The most common age group in the sample was at age from 35 to 45 years and consisted of 110 respondents (27.09%). The minimum number of respondents had the age over 65 years 19 respondents (4.67%). - + 9606 Species patients - + 9606 Species @@ -44353,37 +44772,37 @@ paragraph 8972 The most common type of stone both in female and male patients was calcium type. From a total of 262 calcium stones, 105 of them (40.07%) was present in female patients and 157 (59.92%) in male. Share of infectious type of stone in female patients was 63 (49.60%) and 64 among males (50.39%). Other stones were less abundant in both the gender groups and their total number was only 17. In women their frequency was 2 (13.33%) and 13 among males (86.67%). There was a significant difference in the frequency of different types of stones by gender (chi2 = 11.47, p = 0.009). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species women - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical @@ -44395,55 +44814,55 @@ paragraph 9546 Due to the very low prevalence of other types of stones and inability to perform the chi-square test to a group of other stones are grouped stones which, by virtue were cystine, xanthine stones and uric acid stones. The incidence of cystine calculi was 4 (0.9%), frequency of xanthine stones 3 (0.7%) and uric acid 8 (1.9%). In the group of female respondents 74 (40.88%) had calculus size up to 10 mm, while in the group of male patients stone size up to 10 mm had 107 (59.12%). In the group of female respondents 96 of them (43.05%) had a size of stone exceeding 10 mm, while in the group of male patients stone size over 10 mm had 127 (56.95%). There was no statistically significant difference in the incidence of stones with sizes up to 10 and over 10 mm by gender (chi2 = 0.192, p = 0.661) - + 9606 Species patients - + 9606 Species patients - + MESH:D014527 Chemical uric acid - + MESH:D019820 Chemical xanthine - + MESH:D002137 Disease cystine calculi - + MESH:D002137 Disease calculus - + MESH:D003553 Chemical cystine - + MESH:D019820 Chemical xanthine - + MESH:D014527 Chemical @@ -44455,43 +44874,43 @@ paragraph 10342 Number of stones localized in the upper pole of the kidney in women was 43 (48.86%), and among men was slightly lower and amounted to 45 (51.13%). Number localized in the lower pole of the kidney in women was 22 (61.11%), and among men was slightly lower and amounted to 14 (38.88%). Number of stones localized medially among women was 38 (45.78%), and among men was slightly lower and amounted to 45 (54.21%). Number of localized in the renal pyelon in females was 27 (40.39%), and among men was slightly lower and amounted to 40 (59.70%). There was no statistically significant difference in the frequency of localization of stones by gender. - + 9606 Species women - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species @@ -44503,25 +44922,19 @@ paragraph 10987 The incidence of urolithiasis at the site of physiological narrowing of the ureter in female subjects was 22 (27.5%) and 58 in males (72.5%). The incidence of urolithiasis at the site of the physiological enlargement of the ureter in female subjects was 8 (50%), and the same among men 8 (50%). There was no statistically significant difference in the frequency of localization of calculi to physiological narrowing and widening of the ureter in relation to gender differences (p = 0.086). - + 9606 Species men - - MESH:D014516 - Disease - - enlargement of the ureter - - + MESH:D052878 Disease urolithiasis - + MESH:D052878 Disease @@ -44539,13 +44952,13 @@ paragraph 11827 Size of the calculi in women was 12 (8-15 mm), while in men it was 12 (8-15 mm). There was no statistically significant difference in the size of calculi between males and females (p = 0.557). - + 9606 Species women - + 9606 Species @@ -44563,55 +44976,55 @@ paragraph 12208 In the group of patients who had a negative family history of calculus size to 10 mm were present in 108 (59.67%), while in the group of subjects who had a family history size of stones up to 10 mm had 73 (40.33%). In the group of patients who had a negative family history of calculus size over 10 mm was registered in 160 (71.75%), while in the group of subjects who had a positive family history of calculus size over 10 mm was registered in 63 (28.25%). There was a statistically significant correlation between the size of stones and positive / negative family history (chi2 = 6.529, p = 0.011), respectively, in patients with a positive family history more often were present small stones (to 10 mm). Size of the calculi in patients who had a positive family history was 12 (9-15 mm), while the size of stones in patients who had a positive family history was 10 (8-15 cm). There was no statistically significant difference in the size of the stones between the groups of patients (p = 0.013). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D002137 Disease calculus - + MESH:D002137 Disease calculus - + MESH:D002137 Disease @@ -44623,43 +45036,43 @@ paragraph 13208 Size of calculus in patients without recurrent urolithiasis was 11 (9-15 mm), while the size of the calculus in patients who have had recurrent urolithiasis was 12 (8-15 mm). There was no statistically significant difference in the size of calculi between groups of patients (p = NS). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D002137 Disease calculus - + MESH:D052878 Disease urolithiasis - + MESH:D002137 Disease calculus - + MESH:D052878 Disease @@ -44679,7 +45092,7 @@ table_caption 13718 The dependence of the calculus size and localization in relation to poles. Fisher exact test; p=0.003 - + MESH:D002137 Disease @@ -44703,7 +45116,7 @@ paragraph 14383 Size of calculi localized medially was 14 (10-15 mm), while the localization pyelon was 15 (12-18 mm). There was a significant difference in size between the mentioned calculus localization (p = 0.022). Stones that occur in the renal pyelon are higher than the stones that are found in our study, in medial position. - + MESH:D002137 Disease @@ -44721,12 +45134,24 @@ paragraph 15140 Frequency of calculi localization at sites of physiological stricture site that had the size of 10 mm was 68 (85%), while the frequency of localization at sites of calculi physiological stricture site which had the size over 10 mm was 12 (15%). Frequency of localization at sites of calculi enlargement physiological ureter which had the size of 10 mm was 15 (93.75%), while the frequency of localization at sites of calculi enlargement physiological ureter which had the size over 10 mm, was 1 (6.25%). There was no statistically significant dependence between the size of tartar and calculus localization to physiological constriction and expansion of the ureter (p = 0.688) (Table 2). - + MESH:D002137 Disease calculus + + MESH:D002137 + Disease + + calculi enlargement + + + MESH:D002137 + Disease + + calculi enlargement + TABLE @@ -44735,43 +45160,37 @@ table_caption 15828 Dependence between the size of the stone and its localization in the ureter. Fisher exact test; p=0.688. E1- physiological narrowing of the ureter, E2 physiological enlargement of the ureter - - MESH:D014516 - Disease - - enlargement of the ureter - RESULTS paragraph 16019 In the group of patients under 35 years the incidence of calcium stones was 88 (33.58%), while in the age group of 35-55 years the incidence of calcium type of calculi was 135 (51.52%). The frequency of this type of calculi in subjects older than 55 years was 39 (14.88%). In the group of patients under 35 years the incidence of infectious stones was 26 (20.47%), and at the age of 35 to 55 years the incidence of infectious types of calculi was 63 (49.60%). The frequency of this type of calculi in subjects older than 55 years was 38 (29.92%). In the group of patients under 35 years the incidence of other types of stones was 4 (26.66%), while in the age group of 35-55 years the incidence of other types of calculi was 7 (46.66%). The incidence of other types of calculi in subjects older than 55 years was 4 (26.66%). Statistically significant dependence between age and type of calculi (chi2 = 15.170, p = 0.004) (Table 3). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical @@ -44785,7 +45204,7 @@ table_caption 16950 Dependence of age and types calculus. chi2=15.170 df=4; p=0.004 - + MESH:D002137 Disease @@ -44803,41 +45222,35 @@ paragraph 17028 Renal lithiasis is a disease in which the stones were formed in the collecting tubules, cups and pyelon. The incidence was similar in both kidneys, and about 40% of patients had bilateral stones. There is a wide range of risk factors that may be associated with the disease, including local and general factors. Of local risk factors that favor the emergence of this disease are: trail of urine, disorders of innervation, anomalies of drainage pathways, anatomical abnormalities (sponge kidney, horseshoe kidney), and recurrent infection. General risk factors are categorized as: metabolic (calciuria, cystinuria), hormonal (primary hyperparathyroidism, hyperthereodosis and hypovitaminosis) and other factors such as climatic conditions of life, feeding, pH of urine excretion of concentrated urine, prolonged immobilization, etc.. - + 9606 Species patients - + MESH:D049950 Disease primary hyperparathyroidism, hyperthereodosis and hypovitaminosis - + MESH:D020347 Disease Renal lithiasis - + MESH:D007239 Disease infection - + MESH:D003555 Disease - - calciuria, cystinuria - - - MESH:D007691 - Disease - - sponge kidney + + cystinuria @@ -44845,7 +45258,7 @@ paragraph 17861 Formation of kidney stones attempted to explain many theories but none fully explains the mechanism of occurrence. Urine is in the usual conditions supersaturated solution in which the particles are held in solution influence crystallization inhibitor and colloids. If these factors are disrupted leads to precipitation and aggregation of crystals. The organic matrix, which consists of cellular detritus, blood and bacteria may be the main factor that leads to nucleation and crystal growth. One theory in the spotlight puts deficit of crystallization inhibitors. There is a theory which states that the formation of stones occurs when the crystals, which are constantly being created in the urine oversaturated with salts, are not washed away in the urine. This may occur due to damage to the epithelial duct to which it adheres crystal. - + MESH:D007669 Disease @@ -44857,19 +45270,19 @@ paragraph 18701 Changes in the socio-economic conditions have influenced the changes in the frequency and type of urolithiasis in terms of localization and physico-chemical properties of stones. Major variations on the occurrence of urolithiasis in the world are presented to the public in terms of geographical areas. Annual incidence in England is 22 cases per 100,000 inhabitants, in Kuwait 23.9 per 100,000 population. However, in some countries there are significant differences in the incidence, such as Sweden, where the incidence of 140 per 100,000 inhabitants, Italy 168 per 100,000 inhabitants and United States with high prevalence of 277 per 100,000 population. In Europe, urinary stones are found mainly in the upper urinary tract, while the proportion of stones in the bladder does not exceed 10.0%. It is shown that the urinary bladder calculi more common in the elderly. - + MESH:D052878 Disease urolithiasis - + MESH:D001744 Disease urinary bladder calculi - + MESH:D052878 Disease @@ -44881,55 +45294,55 @@ paragraph 19572 The disease is more common in men and usually occurs after the third decade of life. It is shown that Caucasians are more likely to develop kidney stones than African Americans, and men more often than women patients. A possible explanation why men are more frequent in patients than in females lies in the fact that men in performing physical activities much sweat and lose a lot of fluid, which is also the main reason for the formation of a kidney stone or a stone in the bladder. - + 9606 Species men - + 9606 Species men - + 9606 Species women - + 9606 Species patients - + 9606 Species men - + 9606 Species patients - + 9606 Species men - + MESH:D007669 Disease kidney stones - + MESH:D007669 Disease @@ -44941,43 +45354,43 @@ paragraph 20056 Some new studies are mostly directed of change in the relationship of occurrence calculi in men and women. In the United States-and the data showed that the overall incidence of urolithiasis 10.6% for men and 7.1% for women. This is explained by balancing risks for both men and women today, compared to the past when they were significantly different. - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species women - + 9606 Species men - + 9606 Species women - + MESH:D052878 Disease @@ -44989,7 +45402,7 @@ paragraph 20411 Results of this study showed that the total number of subjects included higher percentage occupied by male respondents and 57.92%, while the remaining 42.08% are occupied by the respondents are female. The results are consistent with published data showing a higher incidence of this disease in male patients compared to female respondents. - + 9606 Species @@ -45001,37 +45414,37 @@ paragraph 20752 Greater tendency of men to concentrate urine compared to the opposite sex, may be the reason for higher incidence of this disease in men. It is shown that the higher the osmolality in men also work super saturation which is responsible for the crystallization of poorly soluble compounds. Even if the concentration of poorly soluble compounds reaches the threshold of saturation of the total 24-h urine, then it can exceed during transient episodes, the way to increase the intake of foods rich in protein, at night, or during intense exercise, especially in the summer months, or season in which men showed a significant decrease in urine volume. Although urolithiasis 2-3 times more common in men, there are no studies that clearly define the reasons for variations incidence of this disease. - + 9606 Species men - + 9606 Species men - + 9606 Species men - + 9606 Species men - + 9606 Species men - + MESH:D052878 Disease @@ -45043,49 +45456,49 @@ paragraph 21547 Daudon and colleagues show that men older than 80 years constitute 40.0% of the analyzed patients with urinary bladder calculi. Prostatic hyperplasia is more common in the elderly and causes an obstruction in the urinary tract. This may be a possible explanation for the high incidence of stones in the bladder in the elderly. Women are also at risk calculus appears in the lower parts of the urinary tract. This suggests the existence of other risk factors, such as changes in bladder function associated with relaxation of smooth muscle tone in the elderly, with an efficiency reduction bladder emptying favoring stagnation of urine and the occurrence of tartar. - + 9606 Species men - + 9606 Species patients - + 9606 Species Women - + + MESH:D000402 + Disease + + obstruction + + MESH:D001744 Disease urinary bladder calculi - + MESH:D011470 Disease Prostatic hyperplasia - + MESH:D001745 Disease efficiency reduction bladder - - MESH:D007415 - Disease - - obstruction - - + MESH:D002137 Disease @@ -45103,79 +45516,85 @@ paragraph 22716 The study by Alaya and associates, who analyzed the sample of 1301 urinary tract stones, it has been proven that the highest percentage share of 58.6% occupied stones of calcium type, that type of calcium oxalate, but they also recorded an increase in the incidence and types of stones uric acid. The authors believe that in the last 50 years there has been a change in eating habits associated with an increase in foods rich in purines (animal proteins projections and seafood), which correlates the increase in uric acid in the urine, and increasing incidence of this type of calculi. Their results show a clear increase in uric acid stones in both sexes. - + MESH:D002118 Chemical calcium - + MESH:D014527 Chemical uric acid - + MESH:D014527 Chemical uric acid - + MESH:D002129 Chemical calcium oxalate - + MESH:D014527 Chemical uric acid + + MESH:D011687 + Chemical + + purines + DISCUSS paragraph 23374 The results of our study showed that the group of patients under 35 years the incidence of calcium stones was 33.58%, the incidence of infectious stones 20.47%, while the incidence of other types of stones was 26.66%. At the age of 35 to 55 years the incidence of a calcium type of calculi was 51.52%, the frequency of the infectious type of calculi was 49.60%, and the prevalence of other types of stones was 46.66%. The frequency of this type of calcium calculi in subjects older than 55 years was 14.88%, infectious stones were 29.92%, while the incidence of other types of calculi in subjects older than 55 years was 26.66%. There was a statistically significant correlation between age and type of calculi (chi2 = 15.170, p = 0.004), i.e. among younger patients are the most common types of calcium stones, while the incidence of infectious types of stones rarer. Increasing age leads to equalization of the frequency of calcium and infectious stones. - + 9606 Species patients - + 9606 Species patients - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical @@ -45187,31 +45606,31 @@ paragraph 24331 Differences in the incidence of age and stones were analyzed through a large number of epidemiological studies. There are certain variations according to geographical areas. In 1993, Baker et al reported that in Australia peak incidence of urolithiasis calcium oxalate sampled observed in individuals between 50 and 60 years of age. In Europe, research shows that the stones which are chemically calcium oxalate more common in people between 40 and 50 years of age. In Asia, the highest prevalence of calcium oxalate stone formation occurs at an earlier age range 30 to 50 years. - + 9606 Species people - + MESH:D052878 Disease urolithiasis - + MESH:D002129 Chemical calcium oxalate - + MESH:D002129 Chemical calcium oxalate - + MESH:D002129 Chemical @@ -45223,156 +45642,156 @@ paragraph 24911 Certain hereditary disorders that run in the family increases the risk of recurrent kidney stones. A rare hereditary disease, renal tubular acidosis, increases the acidity of urine, which is favorable for the occurrence of kidney stones. Cystinuria is a hereditary disorder of the metabolism of amino acids, which results in high levels of cystine in the urine and blood, leading to frequent formation of cystine kidney stones. Second, a hereditary disorder of metabolism, hyperoxaluria, is resulting in high levels of oxalate salt in the body, which is combined with calcium in the form of kidney stones. Hypercalciuria causes high accumulation of calcium in the body, which increases the incidence of kidney stones. Hiperuricosuria increased level of uric acid in the urine, which leads to the formation of uric acid stones. - + MESH:D002118 Chemical calcium - + Disease Hiperuricosuria - + MESH:D002118 Chemical calcium - + MESH:D007669 Disease cystine kidney stones - + MESH:D007669 Disease kidney stones - + MESH:D014527 Chemical uric acid - + MESH:D007669 Disease kidney stones - + MESH:D030342 Disease hereditary disease - + MESH:D003553 Chemical cystine - + + MESH:D030342 + Disease + + hereditary disorder + + MESH:D030342 Disease hereditary disorder - + MESH:D006959 Disease hyperoxaluria - + MESH:D030342 Disease hereditary disorders - + MESH:D007669 Disease kidney stones - + MESH:D000141 Disease renal tubular acidosis - + MESH:D003555 Disease Cystinuria - + MESH:D014527 Chemical uric acid - - MESH:D030342 - Disease - - hereditary disorder of metabolism - - + MESH:D007669 Disease kidney stones - - MESH:D010070 - Chemical - - oxalate - - + MESH:D053565 Disease Hypercalciuria + + MESH:D010070 + Chemical + + oxalate salt + DISCUSS paragraph 25738 Diet, in terms of animal protein (52 g / day), sodium (50 mg / day) and the oxalate (200 mg / day) with a normal intake of calcium (1,200 mg / day), reduced recurrent stones for almost 50% and more, within five years, compared to a diet low in calcium (400 mg / day) and oxalate. - + MESH:D010070 Chemical oxalate - + MESH:D012964 Chemical sodium - + MESH:D002118 Chemical calcium - + MESH:D002118 Chemical calcium - + MESH:D010070 Chemical @@ -45384,37 +45803,37 @@ paragraph 26018 Our results show that the proportion of respondents who did not have positive personal and family history of urolithiasis was 44.94%, and patients who had a positive family history of urolithiasis, but he had a personal history was 50.05%. The share of respondents, who had a family history, and negative personal history of urolithiasis, was 42 (30.88%), while the number of respondents who had a positive to families and personal history was 136 (33.74%). There was a significant frequency dependence of family and personal history (chi2 = 7.41, p = 0.006), i.e. Higher incidence of recurrence in patients who have a positive family history of urolithiasis. - + 9606 Species patients - + 9606 Species patients - + MESH:D052878 Disease urolithiasis - + MESH:D052878 Disease urolithiasis - + MESH:D052878 Disease urolithiasis - + MESH:D052878 Disease @@ -45426,19 +45845,19 @@ paragraph 26678 Similar results were presented by the study Koyuncu and associates, which showed a significant correlation recurring calculi and positive family history. The authors also found that the time interval between the onset of recurrence was significantly shorter in patients who had a positive family history of urolithiasis. By analyzing the patients according to sex, the authors noted that the incidence of recurrence was more frequent in males than females respondents. The authors believe that the information positivity family history is very important and can provide valuable information about the possibility of future attacks as well as the severity of the disease. - + 9606 Species patients - + 9606 Species patients - + MESH:D052878 Disease @@ -45450,7 +45869,7 @@ paragraph 27349 Epidemiological and randomized studies have shown greater security ESWL treatment methods in breaking stones when it starts with applying lower energy sequences of the same, with a gradual increase energy sequences, resulting in a vasoconstriction which prevents renal damage and the difference in the fragmentation is not significant despite the fact that whether amplification is carried out or not. - + MESH:D007674 Disease @@ -45462,6 +45881,12 @@ paragraph 27751 Previous clinical and epidemiological studies have shown that as an indication of ESWL treatment of urolythiasis depends on several factors, including the size, localization, consistency and other histological characteristics of calculi. Pregnancy and specific internship and urological diseases, with an emphasis on acute urinary infection, contraindications to perform of ESWL. + + MESH:D014570 + Disease + + urological diseases + MESH:D001745 Disease @@ -45469,7 +45894,7 @@ urinary infection - MESH:D000437 + MESH:D019966 Disease urolythiasis depends @@ -45618,12 +46043,6 @@ ref 29708 Urinary Stone Disease - - MESH:D007669 - Disease - - Stone Disease - REF @@ -45640,11 +46059,6 @@ ref 29787 Hirurgija mokracnih organa - - Disease - - Hirurgija mokracnih organa - 16 @@ -45676,19 +46090,19 @@ 297 29846 Clinical analysis of 41 children's urinary calculus and acute renal failure - + 9606 Species children - + MESH:D002137 Disease calculus - + MESH:D058186 Disease @@ -45708,7 +46122,7 @@ 693 29922 Acute Renall Colic from Ureteal Calculus - + MESH:D002137 Disease @@ -45752,7 +46166,7 @@ ref 29992 Clinically relevant facets of urolithiasis - + MESH:D052878 Disease @@ -45792,7 +46206,7 @@ 165 30137 Urologic Diseases in America Project. Prevalence of kidney stones in the United States Source - + MESH:D007669 Disease @@ -45811,11 +46225,11 @@ 39 30231 Diabetes and calculations - + MESH:D003920 Disease - - Diabetes and calculations + + Diabetes @@ -45849,13 +46263,13 @@ 963 30335 Clinical practice. Calcium kidney stones - + MESH:D002118 Chemical Calcium - + MESH:D007669 Disease @@ -45878,7 +46292,7 @@ 109 30376 Family history in stone disease: how important is it for the onset of the disease and the incidence of recurrence? - + MESH:D007669 Disease @@ -45910,12 +46324,6 @@ Rustempasic N, Cvorak A, Agincic A 0 Outcome of Endovenous Laser Ablation of Varicose Veins - - MESH:C077381 - Chemical - - Varicose - ABSTRACT @@ -45934,12 +46342,35 @@ abstract 78 In Bosnia and Herzegovina according to available data, treatment of incompetent superficial lower extremity varicose veins by endovenous laser ablation (EVLA) has been introduced two years ago and so far no paper has been published regarding results of EVLA treatment of patients from our country. We wanted to present our results with EVLA treatment. - + 9606 Species patients + + - + Chemical + + EVLA + + + Disease + + Bosnia + + + - + Chemical + + EVLA + + + - + Chemical + + EVLA + ABSTRACT @@ -45958,7 +46389,7 @@ abstract_title_1 594 Patients and methods: - + 9606 Species @@ -45970,49 +46401,49 @@ abstract 616 The study was clinical and prospective. It was carried out at Clinic for vascular surgery in Sarajevo where fifty-eight (58) patients received surgical treatment for varicose veins and in Aesthetic Surgery Center "Nasa mala klinika" in Sarajevo were sixty-one (61) patients with varicose veins were treated by endovenous laser ablation. Total 119 patients (limbs) with pathologic reflux only in great saphenous vein were evaluated between 1st of January 2013 and 31st of April 2014. Following primary outcome endpoints were evaluated smean day of return to normal everyday activities, patient subjective quantification of pain during first seven days after intervention, incidence of deep venous thrombosis (DVT), incidence of wound bleeding requiring surgical intervention, incidence of peri-saphenous vein hematoma and infection rate. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + MESH:D006470 Disease bleeding - + MESH:D020246 Disease deep venous thrombosis - + MESH:D006406 Disease vein hematoma and infection - + MESH:D010146 Disease @@ -46030,49 +46461,55 @@ abstract 1466 Mean of return to normal activities (expressed in days after intervention); EVLA vs. stripping (surgery) =1.21vs12.24, T test 13,619; p=0, 000, p<0,05. T test was used for comparing Mean value of visual pain analog scale for the first 7 days between groups, for all seven days pain was significantly higher in surgical group of patients as compared to EVLA group; p<0,05. Incidence of hematoma greater than 1% of total body surface area was significantly higher in patients receiving surgical treatment; Pearson Chi Square=23,830, p<0,05; odds ratio:10,453. Incidences of infection, deep venous thrombosis and posttreatment bleeding were not statistically different between analyzed groups; EVLA vs Surgery (Pearson Chi Square =3,237; p>0,05; Pearson Chi Square=2,139, p>0,05, Pearson Chi Square=2,139, p>0,05, respectively.) - + 9606 Species patients - + 9606 Species patients - + MESH:D020246 Disease deep venous thrombosis - + + - + Chemical + + EVLA + + MESH:D014786 Disease visual pain - + MESH:D007239 Disease infection - + MESH:D010146 Disease pain - + MESH:D006406 Disease hematoma - + MESH:D006470 Disease @@ -46090,25 +46527,31 @@ abstract 2304 EVLA offers better patient recovery in terms of significantly lower post treatment pain, faster return to everyday activities and lower incidence of bruising (hematomas). - + 9606 Species patient - + MESH:D003288 Disease bruising - + MESH:D006406 Disease hematomas - + + - + Chemical + + EVLA + + MESH:D010146 Disease @@ -46126,31 +46569,31 @@ paragraph 2491 In the recent past the gold standard in the treatment of lower extremity varicose veins with incompetent great saphenous vein (GSV) or small saphenous vein (SSV) was classical surgicall method of ligation and stripping of incompetent saphenous vein along with varicectomies. However, during the last decade other minimal invasive techniques have been invented and introduced to treat the incompetent saphenous vein and varicosities. Endovenous laser ablation (EVLA) is one of them and it has been used as minimally invasive replacement to classical surgical way of treating varicose veins for many years (Figure 1, Figure 2). Avoiding surgical incisions, mechanical disruption of the sapheno-femoral junction (SFJ) as well as aggressiveness of avulsion of saphenous vein, EVLA may offer reduced postoperative pain, post intervention bleeding and peri-saphenous vein hematoma along with decreased rate of wound infection and potentially shorter periods of absenteeism from work. In addition neo-vascularisation is significantly reduced which in turn might cause decreased incidence of varicose vein recurrence. - + MESH:D006406 Disease vein hematoma - + MESH:D006470 Disease bleeding - + MESH:D010149 Disease postoperative pain - + MESH:D001523 Disease - - aggressiveness of avulsion of saphenous vein + + aggressiveness of avulsion - + MESH:D007239 Disease @@ -46164,12 +46607,6 @@ fig_caption 3601 Varicose veins before treatment - - MESH:C077381 - Chemical - - Varicose - FIG @@ -46178,24 +46615,41 @@ fig_caption 3633 Same patient after EVLA treatment - + 9606 Species patient + + - + Chemical + + EVLA + INTRO paragraph 3667 In Bosnia and Herzegovina according to available data, endovascular laser ablation of incompetent superficial lower extremity veins has been introduced two years ago and so far no paper has been published regarding EVLA treatment of patients from our country. Since many papers on this topic have certain contradictory results we wanted to present our experience with these two methods of varicose vein tretament. - + 9606 Species patients + + - + Chemical + + EVLA + + + Disease + + Bosnia + INTRO @@ -46214,7 +46668,7 @@ title_1 4246 3. PATIENTS AND METHODS - + 9606 Species @@ -46226,25 +46680,25 @@ paragraph 4270 The study was clinical and prospective. It was carried out at Clinic for vascular surgery in Sarajevo where patients received surgical treatment for varicose veins and in Aesthetic Surgery Center "Nasa mala klinika" were endovenous laser treatment of varicose vein was performed. Total 119 patients with pathologic reflux only in great saphenous vein were evaluated between 1st of January 2013 and 31st of April 2014. Sixty-one (61) patients who received endovenous laser treatment for varicose veins in Aesthetic Surgery Center "Nasa mala klinika" were assigned to group A, while fifty-eight (58) patients that received surgical treatment of varicose veins at the Clinic for Vascular surgery were assigned to Group B. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species @@ -46256,103 +46710,103 @@ paragraph 4998 Primary outcome endpoints were evaluated: mean day of return to normal everyday activities, patient subjective quantification of pain during first seven days after intervention, incidence of deep venous thrombosis (DVT), incidence of wound bleeding requiring surgical intervention, incidence of peri-saphenous vein hematoma and infection rate. Every day activities were defined as ability for self-care and all other professional, recreational, household or outdoor physical activities that used to be carried out before varicose vein treatment. Return to those activities was advised to all patients to be as soon as possible and the information about the day of return to those activities were recorded when patient resumed them without any difficulty ie pain. Infection was defined as redness of skin around wound (phlebectomy site) along with presence of pus in the wound (site of intervention) with or without patient being febrile. Hematoma along the direction of treated saphenous vein was examined on 7th day and its surface area estimated by comparing its total surface to the surface area of the palm of individual patient (1% of total body surface area)-procedure similar to the method used in assessment of burned skin surface area. Patients were then categorized either as having hematoma less than 1% or having hematoma >= 1% of total body surface area. Pain score was measured using a visual analogue scale ranging from 0 (no pain) to 5 (most severe pain) (Figure 3). - + 9606 Species patient - + 9606 Species patients - + 9606 Species patient - + 9606 Species patient - + 9606 Species patient - + 9606 Species Patients - + MESH:D006470 Disease bleeding - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - + MESH:D006406 Disease Hematoma - + MESH:D006406 Disease vein hematoma and infection - + MESH:D020246 Disease deep venous thrombosis - + MESH:D010146 Disease pain - + MESH:D010146 Disease Pain - + MESH:D006406 Disease hematoma - + MESH:D010146 Disease pain - + MESH:D006406 Disease @@ -46366,7 +46820,7 @@ fig_caption 6484 Visual analog scale pain estimation - + MESH:D010146 Disease @@ -46378,151 +46832,151 @@ paragraph 6520 In case of presence of clinical signs of lower extremity deep venous thrombosis (DVT) Color Doppler examination was performed. Any bleeding after intervention that required surgical intervention either in the form of surgical revision of bleeding or need for additional surgical suture were noted and defined as postintervention bleeding. Return to normal activities were recorded taking 30 days after intervention as maximum period. This period was sufficient for full mobilization of patient even in the case of unwanted outcome endpoints like DVT. Each patient had a pre-operative duplex ultrasound examination that was performed in order to identify the site of pathologic reflux and evaluate suitability for intervention on superficial venous system. Ultrasonography was performed by vascular surgeon using Mindray 6 apparatus for group of patients treated surgically or GE ultrasound for patients treated by endovascular laser ablation. Ultrasonographic mapping of the venous systems was performed in all patients in standing position. Incompetence of the GSV was defined as reflux > 0.5 s in duration following calf compression. Exclusion criteria were: previous surgical treatment of the GSV, GSV diameter >= 2 cm, pregnancy, allergy to lidocaine, active superficial phlebitis, presence of any dermatologic phlebostatic changes in leg like hyperpigmentation or skin ulcer, positive history of previous deep venous thrombosis, diabetes mellitus and arterial disease. Mechanical stripping of great saphenous vein was performed in general anesthesia. After a groin incision, high ligation of the GSV and ligation of all tributaries were performed. Access to distal part of GSV was achieved through a small incision at the level of knee joint. GSV stripping was performed by metalic stripper. Varicosities were removed by minimal stab avulsion technique (Klapp/Smetana method). In the EVLA group of patients, endovascular access for laser fiber introduction into GSV was ultrasonically guided and performed at the knee level. The laser fiber tip was positioned 1 cm distal to the SFJ. Before laser ablation was started tumescent anesthetic (25 ml Lidocaine 2% in 500 ml of normal saline 0,9%) was injected all around GSV under ultrasound guidance. Thermal ablation of great saphenous vein was performed by Biolitec 1470 nm radial fiber with energy 70J/cm at 10 watts. Varicosities were removed by microphlebectomy technique. Post procedurally elastic venous compression was applied to both group of patients for 30 days in surgically treated group and for only 7 days in group of patients that received laser ablation treatment. Wound dressings were changed every three days and wounds were disinfected with povidone-iodine solution and sutures removed on 10th postoperative day in surgical group of patients. Wound dressing in EVLA group of patients was left intact for total 7 days and then removed along with compressive stocking. - + 9606 Species patient - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9913 Species calf - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D004342 Disease allergy - - MESH:D003324 + + MESH:D002539 Disease arterial disease - + MESH:D020246 Disease deep venous thrombosis - + MESH:D010689 Disease phlebitis - + MESH:D006470 Disease bleeding - + MESH:D008012 Chemical Lidocaine - + MESH:D012883 Disease skin ulcer - + MESH:D017495 Disease hyperpigmentation - + MESH:D020246 Disease deep venous thrombosis - + MESH:D006470 Disease bleeding - + MESH:D011206 Chemical povidone-iodine - + MESH:D006470 Disease bleeding - + MESH:D008012 Chemical lidocaine - + MESH:D003920 Disease @@ -46540,25 +46994,25 @@ paragraph 9472 Statistical data analysis was performed using the IBM SPSS version 22.0. In this study we analyzed continuous and categorical (dichotomous) variables. Statistical data analysis was performed using mainly methods of inferential statistics. For continuous variables, parametric independent-samples t-test was performed. Non-parametric chi-square test was performed for testing hypothesis for categorical variables. Both tests were calculated on the basis of 95% confidence level. Odds ratio was calculated only for the incidence of hematoma. For other variables odds ratio calculation was not possible because of zero frequencies in the cross tabulation cells. Mean of return to normal activities was expressed in days after intervention ei EVLA vs surgery (stripping) =1.21vs12.24, T test 13,619; p=0, 000, p<0,05 (Table 1). T test was used for comparing Mean value of visual pain analog scale (figure 3) for first 7 days between groups, for all seven days pain was significantly higher in surgical group of patients as compared to EVLA group; EVLA vs Surgery: 1st day T test= -17.900, p<0.05; 2nd day T test=-19,817, p<0.05; 3rd day T test=-21.986, p<0.05; 4th day T test=-4.898, p<0.05; 5th day T test=-4.355, p<0.05; 6th day T test -3,643, p<0.05; 7th day T test=2.386, p<0.05. - + 9606 Species patients - + MESH:D006406 Disease hematoma - + MESH:D010146 Disease pain - + MESH:D014786 Disease @@ -46580,7 +47034,7 @@ table_caption 10815 Mean value of visual pain scale for first seven days after intervention - + MESH:D014786 Disease @@ -46592,13 +47046,13 @@ paragraph 10887 Incidence of hematoma greater than 1% of total body surface area was significantly higher in patients receiving surgical treatment; Pearson Chi Square=23,830, p<0,05; odds ratio:10,453 (Table 3). Incidences of infection, deep venous thrombosis and posttreatment bleeding were not statistically different between analyzed groups; EVLA vs Surgery: (Pearson Chi Square =3,237; p>0,05; Pearson Chi Square=2,139, p>0,05, Pearson Chi Square=2,139, p>0,05, respectively (Table 4, Table 5 and Table 6). - + 9606 Species patients - + MESH:D006406 Disease @@ -46612,7 +47066,7 @@ table_caption 11382 Incidence of hematoma greater than 1% of total body surface area - + MESH:D006406 Disease @@ -46626,7 +47080,7 @@ table_caption 11447 Incidence of infection - + MESH:D007239 Disease @@ -46640,7 +47094,7 @@ table_caption 11470 Incidence of wound bleeding after intervention - + MESH:D006470 Disease @@ -46654,7 +47108,7 @@ table_caption 11517 Incidence of deep venous thrombosis after intervention - + MESH:D020246 Disease @@ -46672,259 +47126,335 @@ paragraph 11586 Return to normal everyday activities in EVLA group was on first day after intervention (mean) while in surgical group it was on 12th day (Mean: 1.23 vs. 12. 24); (Table 1). The difference was statistically significant p<0.05. All patients immediately after EVLA procedure was finished, left Clinic walking without any difficulties and regular every day walking was advised as only measure for prevention of DVT in this group. - + 9606 Species patients + + - + Chemical + + EVLA + DISCUSS paragraph 12012 Pain was quantified every day (for all seven days after intervention/operation) by each patient on scale from 0-5 (Figure 1) based on mean value of visual pain scale. In EVLA group for all seven days after intervention mean value of pain intensity was significantly less than in surgical group (Table 2). Less pain is probably related to avoiding trauma to skin (groin incisions) and trauma to subcutaneous tissue caused by avulsion (mechanical stripping of great saphenous vein). These results are opposite to results published by Pronk et al. where EVLA patient had more pain in second week as compared to surgical group of patients. Underlying reason for greater pain in their EVLA group is probably the type of laser apparatus they used (older type) because new Laser systems with higher wavelengths like 1470 nm have a better absorption in water and cause less pain and bruising after the procedure because they cause less vein wall perforation. This is in accordance with recommendation that low-energy EVLA with the use of a 1,470-nm laser fiber with linear endovenous energy density (LEED) of 80 J/cm or less is advisable option for the treatment of incompetent saphenous veins. Regarding post intervention hematoma, in our study there were 5 cases of hematoma in EVLA group and 28 cases being >= 1% of total body surface area in surgical group. This difference was statistically significant, p<0.05 (Table 3). Similar results were obtained by study of Siribumrungwong et al in which they had statistically significant advantage of EVLA method over surgery in hematoma appearance. Postoperative hematomas were also significantly smaller after EVLA than those after stripping in the study of Kalteis et al.. - + 9606 Species patient - + 9606 Species patient - + 9606 Species patients - + MESH:D006406 Disease hematoma - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - + + MESH:D003288 + Disease + + bruising + + + MESH:D010149 + Disease + + Postoperative hematomas + + + MESH:D014947 + Disease + + trauma + + + - + Chemical + + EVLA + + + MESH:D010146 + Disease + + Pain + + + MESH:D014786 + Disease + + visual pain + + + MESH:D014867 + Chemical + + water + + + - + Chemical + + EVLA + + MESH:D010146 Disease pain - - MESH:D003288 - Disease - - bruising + + - + Chemical + + EVLA - + MESH:D006406 Disease hematoma - + MESH:D010146 Disease pain - + MESH:D014947 Disease trauma - - MESH:D010149 - Disease - - Postoperative hematomas - - + MESH:D006406 Disease hematoma - - MESH:D014947 - Disease - - trauma - - + MESH:D010146 Disease pain - - MESH:D010146 - Disease - - Pain - - - MESH:D014786 - Disease - - visual pain - DISCUSS paragraph 13728 Incidence of infection in this study was smaller in EVLA group (0 patients) as compared to surgical group of patients (3 patients) but this difference was not statistically significant, p>0.05 (Table 4). These findings have similar correlation with results of meta-analysis on 2245 limbs by Pan et al. which reported also fewer complications with EVLA as compared to high ligation and stripping in case of bleeding and hematoma (1.28% versus 4.83%) as well as wound infection (0.33% versus 1.91%). Striping itself is mechanically tissue destructive procedure which after removal of saphenous vein leaves avulsed tributaries that are left to bleed into subcutaneous tissue and amount of bleeding can be controlled only by subsequent compression. In that regard we had to additionally control bleeding after operations in 2 patients while there was no need to control any unwanted bleeding in patients after EVLA, (Table 5). However, this difference was not statistically relevant, p>0.05. We had no cases of DVT in EVLA group while 2 cases of crural DVT were recorded in surgical group of patients that were confirmed by color Doppler. Both of them received low molecular weight heparin therapy with switch to oral anticoagulation according to standard protocol for DVT treatment. Recovery was uneventful. This difference in incidence of DVT between groups was not statistically significant, (Table 6), p>0.05. Incidence of deep vein thrombosis (DVT) after varicose vein surgery vary between 0.4% historically and 5.7% while after EVLA from 0 to 7.7%. We administered LMWH prevention for surgical group of patients during their hospital stay although according to some authors there is no superiority of a short-term regimen of LMWH, early ambulation and compression therapy as compared with early ambulation and compression therapy alone. This decision was guided by idea that mobilization of patient after surgery due to general anesthesia was delayed as compared to EVLA group. EVLA treatment is rarely related to possible endovenous heat-induced thrombosis (EHIT) that leads to clinically significant DVT. EHIT may be caused by heat that is delivered by laser catheter placed in the saphenous vein. This brings about closure of the vein by denaturing the vessel wall with consequent thrombus formation. There are no relevant guidelines regarding advice for routine anticoagulation for EVLA patients. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 1214577 Species to 7 - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + + - + Chemical + + EVLA + + MESH:D006406 Disease hematoma - - MESH:D007239 - Disease - - infection + + - + Chemical + + EVLA - + MESH:D006470 Disease bleeding - - MESH:D007239 - Disease - - infection - - - MESH:D013927 - Disease - - thrombosis + + - + Chemical + + EVLA - + MESH:D006470 Disease bleeding - + MESH:D006470 Disease bleeding - + + - + Chemical + + EVLA + + + Disease + + EHIT + + MESH:D006470 Disease bleeding - + + - + Chemical + + EVLA + + MESH:D006495 Chemical LMWH - + MESH:D006493 Chemical heparin - - MESH:D013927 - Disease - - thrombus - - + MESH:D006495 Chemical LMWH - + + MESH:D010291 + Disease + + crural DVT + + + MESH:D007239 + Disease + + infection + + + Disease + + EHIT + + + MESH:D007239 + Disease + + infection + + + MESH:D013927 + Disease + + thrombosis + + + - + Chemical + + EVLA + + MESH:D020246 Disease @@ -46942,25 +47472,25 @@ paragraph 16145 According to obtained results this endovenous laser ablation (EVLA) enables patients after treatment of varicose veins with better recovery in terms of significantly lower post treatment pain, faster return to everyday activities and lower incidence of bruising (hematomas). - + 9606 Species patients - + MESH:D006406 Disease hematomas - + MESH:D003288 Disease bruising - + MESH:D010146 Disease @@ -46972,55 +47502,67 @@ paragraph 16420 It has also shown that EVLA had offered no advantages in terms of significantly lower incidence of post intervention bleeding, infection rate or DVT. - + MESH:D006470 Disease bleeding - + MESH:D007239 Disease infection + + - + Chemical + + EVLA + CONCL paragraph 16570 LIMITATIONS: Study was carried out on patients with varicose veins without dermatologic complications so further evaluation is required especially regarding need for inclusion of patients with C3 level of disease or higher according to CEAP classification. In addition patient expectations might have had influence on subjective pain score scale since they have received information about so far published advantages of EVLA before intervention was performed. - + 282991 Gene 45479 CEAP - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + MESH:D010146 Disease pain + + - + Chemical + + EVLA + CONCL @@ -47179,7 +47721,7 @@ surname:Thakkinstian;given-names:A 18063 A systematic review and meta-analysis of randomised controlled trials comparing endovenous ablation and surgical intervention in patients with varicose vein - + 9606 Species @@ -47241,13 +47783,18 @@ surname:Whiteley;given-names:MS 18481 Deep Vein Thrombosis (DVT) after Venous Thermoablation Techniques: Rates of Endovenous Heat-induced Thrombosis (EHIT) and Classical DVT after Radiofrequency and Endovenous Laser Ablation in a Single Centre - + MESH:D020246 Disease Deep Vein Thrombosis - + + Disease + + EHIT + + MESH:D013927 Disease @@ -47271,19 +47818,19 @@ surname:Vaquero;given-names:C 18687 Low-molecular-weight heparin for prevention of venous thromboembolism after varicose veinsurgery in moderate-risk patients: a randomized, controlled trial - + 9606 Species patients - + MESH:D054556 Disease venous thromboembolism - + MESH:D006493 Chemical @@ -47306,7 +47853,7 @@ 1585 18842 Incidence of deep vein thrombosis after varicose vein surgery - + MESH:D020246 Disease @@ -47341,16 +47888,16 @@ 0 Preoperative Kidney Tumor Embolization as Procedure for Therapy of Advanced Kidney Cancer - MESH:D007680 + MESH:D007674 Disease - - Kidney Cancer + + Kidney Tumor MESH:D007680 Disease - - Kidney Tumor + + Advanced Kidney Cancer @@ -47383,10 +47930,10 @@ kidney cancer - MESH:D009360 + MESH:D007674 Disease - - kidney tumor embolization + + kidney tumor @@ -47436,41 +47983,47 @@ abstract 677 Symptoms that are dominating among patients were haematuria and pain. Analysis of mean size of tumors based on CT evaluation showed statistically significance in between the biggest size of tumors in group from Hamburg (9.11+-3cm) and the smallest size of tumors in Sarajevo group (4.94+-1.6cm) p=0.0001. Reason for this is difference in selection of patients for treatment in Hamburg from Sarajevo. - + 9606 Species patients - + 9606 Species patients - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumors - + + MESH:D006417 + Disease + + haematuria + + MESH:D010146 Disease - - haematuria and pain + + pain @@ -47484,13 +48037,13 @@ abstract 1088 Kidney as functional finishing organ is extremely suitable for transcatheter therapeutic procedures. The gold standard in the treatment of advanced and metastatic tumor is the nephrectomy. As preparation for nephrectomy in metastatic cancer total capillary embolization is performed. After embolization, surgery is shorter, procedure can be done 24-48 hours after embolization or delayed nephrectomy done 2-3 weeks after the intervention. - + MESH:D009369 Disease tumor - + MESH:D009369 Disease @@ -47508,37 +48061,37 @@ paragraph 1543 Kidney cancer is the most common kidney tumor and the third one by the frequency in urology. Comparing to other malignancy, 2% of all tumors is the kidney cancer. In end stage kidney cancer therapy of choice is radically nephrectomy, which means and adrenalectomy with removing fat kidney capsule and proximal urethra. - + MESH:D007676 Disease end stage kidney cancer - + MESH:D007680 Disease kidney cancer - + MESH:D009369 Disease tumors - + MESH:D009369 Disease malignancy - - MESH:D007680 + + MESH:D007674 Disease kidney tumor - + MESH:D007680 Disease @@ -47550,31 +48103,31 @@ paragraph 1862 Kidney tumor embolization is the standard procedure in therapy of kidney cancer, due to embolization whole kidney or its parts should be exclude from the function. First experimental embolization was performed by lang in 1971. He used radioactive particles to induce tumor necrosis. On humans, first embolization was performed in 1973, by Almagard. From that time the technique has been changed a lot, also diagnostic procedures for detecting the tumors were fulfilled with CT and MRI. In the same time the new operative techniques were introduced laparoscopic nephrectomy, organoprotective resection and extracorporal Work-Bench procedures. - + 9606 Species humans - + + MESH:D007674 + Disease + + Kidney tumor + + MESH:D009369 Disease tumors - + MESH:D007680 Disease kidney cancer - - MESH:D009360 - Disease - - Kidney tumor embolization - - + MESH:D009336 Disease @@ -47586,17 +48139,17 @@ paragraph 2504 Preoperative tumor embolization is performed in patients with: - + 9606 Species patients - - MESH:D009360 + + MESH:D009369 Disease - - tumor embolization + + tumor @@ -47604,7 +48157,7 @@ paragraph 2567 Advanced stage of tumor - + MESH:D009369 Disease @@ -47622,32 +48175,20 @@ paragraph 2635 Infiltration of venous system - - MESH:D020246 - Disease - - venous system - INTRO paragraph 2665 Tumor in close range of kidney hillus - - MESH:D007674 - Disease - - kidney hillus - INTRO paragraph 2703 Metastatic tumors and - - MESH:D018223 + + MESH:C538445 Disease Metastatic tumors @@ -47658,7 +48199,7 @@ paragraph 2725 Patients who are not treatable to become treatable. - + 9606 Species @@ -47670,25 +48211,25 @@ paragraph 2777 Preoperative embolization has the goal to reduce intraoperative bleeding and to reduce the time of surgical procedure, by making preparation of tumor easier. This is particularly important for T3 and T4 tumors, with infiltration of renal vein and vena cava inferior. - + MESH:D009369 Disease tumor - - MESH:D059228 + + MESH:D007674 Disease infiltration of renal vein - + MESH:D016063 Disease intraoperative bleeding - + MESH:D009369 Disease @@ -47706,65 +48247,65 @@ paragraph 3069 We retrospectively observed 50 patients between 2000-2011, in which the preoperative embolization was performed. The majority of patients were treated in Eilbek Clinic in Hamburg and few in Clinical Center Sarajevo. Mean age of patients was 64 years, range between 3 and 86 years. 19 patients were female and 31 patients were male. Patients that were enrolled on study were the patients with the tumor larger than 8cm on the upper pole of left kidney, and patients wit proven bilateral kidney tumor. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species Patients - + 9606 Species patients - + 9606 Species patients - + MESH:D009369 Disease tumor - - MESH:D007680 + + MESH:D007674 Disease - - bilateral kidney tumor + + kidney tumor @@ -47772,55 +48313,55 @@ paragraph 3569 Patients that were excluded from study were the patients with tumor smaller than 8cm, if the tumor was good for surgical procedure. Embolised patient were compared with the group of 51 patients from Urology Clinic Sarajevo, which were performed surgical nephrectomy without embolization. This group has been concised from 19 female and 32 male. Mean age was 61 years, range between 32 and 80 years. The total number of patients was 101. Patients were diagnosed with CT or MRI, as outpatients and small amount of patients we diagnosed in our Institution (CT siemens Somatom duo and MRI Unit 1.5T Symphonie). - + 9606 Species Patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species Patients - + 9606 Species patients - + MESH:D009369 Disease tumor - + MESH:D009369 Disease @@ -47832,60 +48373,84 @@ paragraph 4176 Conventional angiography was performed on Siemens Polystar, 2 frames/sec. Angiography was performed by Seldinger technique, with contrast agent Xenetix 300. Angiography was performed to determine the blood supply of tumors in attacked kidney; we performed first aortography followed by selective angiography of renal arteries. Once done selective angiography, we approached embolization. Embolization is done first on the capillary level. As embolization material at the capillary level in the course of the study we used alcohol or Ivalon 150-250 mu particles. Alcohol is the cheapest and most efficient means of embolization. The disadvantage is that alcohol is the only means of embolization during one embolization must be applied in a blocked blood vessel in order to prevent reflux. We used 96% alcohol. In some patients inhibit the supply vessel is impossible, and then we performed a combined embolization with alcohol and Ivalon or just Ivalon particle embolization. The advantage in using Ivalon that supply blood vessel may not be blocked. Upon completion of the capillary embolization, we performed control angiography, and if the tumor / kidney is embolised we performed the central embolization of supply vessel with metal spirals. In the course of the study, Tornado spirals (COOK) were used. After that is done aortography to confirm complete embolization and showed situation surrounding blood vessels that were not aimed by embolization. - + 9606 Species patients - - MESH:D009369 - Disease - - tumors - - MESH:D000431 + MESH:C026699 Chemical - - Alcohol + + Ivalon - MESH:D000431 + MESH:D000438 Chemical - + alcohol - MESH:D000431 + MESH:D000438 Chemical - - alcohol + + Alcohol - MESH:D000431 + MESH:C026699 Chemical - - alcohol + + Ivalon + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 + Chemical + + alcohol + + MESH:D009369 Disease - - tumor + + tumors - - MESH:D000431 + + MESH:D000438 Chemical alcohol + + MESH:D000438 + Chemical + + alcohol + + + MESH:D009369 + Disease + + tumor + METHODS paragraph 5632 Besides angiographies findings, we were processed operating protocols and protocols of anesthesia. To test the existence of the correlation between the groups for each parameter was used Pearson correlation test. In all tests of statistical significance was considered at p <0.05 or at the level of 95%. All data were analyzed using statistical software SPSS version 13.0 for Windows, while the resulting tables and graphs subsequently processed using Microsoft Word 2003. + + MESH:D008569 + Disease + + Word + RESULTS @@ -47898,126 +48463,126 @@ paragraph 6116 Symptomatology that led the patient's physician is shown in Figure 1 and 2. A small number of patients reported the loss of body weight, 11 patients from the Hamburg group (H) and 7 patients in the Sarajevo group (S). Infiltration of the kidney hollow system has been registered with 5 (H) patients and in 7 (S) patients. Infiltration of veins, tumor thrombus was found in a total of 16 patients (10 H and 6 S) and the infiltration of adjacent organs was found in 15 patients (8 H 7 and S). AV-shunt was diagnosed in only one patient at Hamburg group. Analysis of the average size of the tumor based on CT findings showed a statistically significant difference in terms of the maximum average size of tumors in patients from Hamburg (9.11 +- 3 cm) and minimum size in patients in Sarajevo (4.94 +- 1.6 cm), p = 0.0001. The reason for this is in the selection of patients in Hamburg preoperative group compared to Sarajevo operative group. The presence of metastases in regional lymph nodes showed statistically significant differences between groups p = 0.030. It is evident that the invasion of local lymph nodes at least recorded in patients in Sarajevo (7.8%), and patients from Hamburg (22%). Distant metastases were found in 4 (H) and in 2 (S) patients. - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D013927 - Disease - - tumor thrombus - - - MESH:D009362 - Disease - - metastases - - + MESH:D009362 Disease metastases - + MESH:D009369 Disease tumors - + MESH:D009369 Disease tumor + + MESH:D013927 + Disease + + tumor thrombus + + + MESH:D009362 + Disease + + metastases + FIG @@ -48026,7 +48591,7 @@ fig_caption 7375 Number cases of Haematuria in our sample - + MESH:D006417 Disease @@ -48040,7 +48605,7 @@ fig_caption 7416 Presence of pain in our patients - + MESH:D010146 Disease @@ -48052,7 +48617,7 @@ paragraph 7449 Embolization was repeated in two patients. Patohistological classification is shown in Figure 3. - + 9606 Species @@ -48078,37 +48643,37 @@ paragraph 7742 Histological determination of tumor size confirms previous statement on CT tumor size that the same was higher in patients from Hamburg (8.2 +- 3.4 cm), compared to patients in Sarajevo (3.89 +- 1.5 cm). Analysis of variance also showed statistically significant differences p = 0.0001 in favor of more specific histopatological tumor size in patients from Hamburg. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D009369 Disease tumor - + MESH:D009369 Disease tumor - + MESH:D009369 Disease @@ -48120,30 +48685,42 @@ paragraph 8108 The average length of surgery was very similar and it was 105.6 +- 23 minutes in patients from Hamburg, and 101.4 +- 38 minutes in patients in Sarajevo. Minimum duration of the procedure was recorded in Sarajevo on 45 minutes, which was recorded as well as the maximum duration of treatment of 195 minutes. There were no statistically significant differences. The Hamburg group's surgery was done on average 23 days after embolization. Blood transfusion during and after surgery are shown in Table 1. With statistical significance, the difference can be seen that the operations carried out in Sarajevo was used more units of blood during the same. Figure 4 presents the case of preoperative embolization, which was done in so called sandwich-technique alcohol mixed with lipiodol and Ivalon particle. Central occlusion was performed with Tornado spirals. - + 9606 Species patients - + 9606 Species patients - - MESH:D000431 + + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 Chemical alcohol - + MESH:D004998 Chemical lipiodol + + MESH:D015356 + Disease + + Central occlusion + TABLE @@ -48160,8 +48737,8 @@ fig_caption 9017 a), b) present angiographic findings of kidney tumor; c) and d) present angiographic findings after embolisation. - - MESH:D007680 + + MESH:D007674 Disease kidney tumor @@ -48178,7 +48755,7 @@ paragraph 9145 Treatment of choice for kidney cancer is surgical resection. In early stage of TU and with favorable localization is sufficient to perform enucleation or partially nephrectomy, and in advanced disease should be done radical nephrectomy. - + MESH:D007680 Disease @@ -48190,37 +48767,37 @@ paragraph 9382 By age, group of our patients who were preoperatively embolised, was very heterogeneous. We performed one embolization of Wilms tumor. Wilms tumor occurs at the age of 2-4 years. After 10 years of appearing less frequently. In addition to Wilms tumor in children is possible occurrence of renal cancer. Incidence of 3- 6%. - + 9606 Species patients - + 9606 Species children - + MESH:D009396 Disease Wilms tumor - + MESH:D009396 Disease Wilms tumor - + MESH:D007680 Disease renal cancer - + MESH:D009396 Disease @@ -48232,13 +48809,13 @@ paragraph 9705 Reasons for blood loss during nephrectomy are: hypervascularisation of tumor, atypical blood vessels and hilar lymph nodes that hamper visibility of blood vessels. - + MESH:D006473 Disease blood loss - + MESH:D009369 Disease @@ -48250,187 +48827,181 @@ paragraph 9869 For decisive loss of blood atypical blood vessels are the main reason, which are often fulfilled with the blood. In violation of these vascular structures, Intraoperative bleeding is abundant because these vessels are less contractile. Another disadvantage is that these blood vessels are retroperitoneal positioned which makes preparation of large tumors very difficulty. With preoperative embolization, we have to occlude not only renal artery and capillary circulation, but must be able to occlude all atypical arteries to reduce blood loss to a minimum. - + MESH:D016063 Disease Intraoperative bleeding - + MESH:D006473 Disease blood loss - + MESH:D009369 Disease tumors - - MESH:D009383 - Disease - - decisive loss of blood atypical blood vessels - DISCUSS paragraph 10427 Craven study showed minimal blood loss in delayed nephrectomy after embolization even with advanced stages of cancer with vascularized thrombus in the inferior vena cava. Delayed nephrectomy stimulates tumor regression as well as venous thrombosis, which significantly facilitates op procedure. In our group's average delay was 23 days. Some authors recommend a delay of less than 48 hours between embolization and surgery because postinfarctus syndromes do not develop fully. The same authors as Lanigian et al. suggest that a longer interval between embolization and surgery lead to the development of collateral that hamper preparation. These data from the literature do not agree with our experience. In most of our patients, we performed delayed nephrectomy. In these patients, there was no collateral development in the periphery. - + 9606 Species patients - + 9606 Species patients - + MESH:D009369 Disease cancer - + MESH:D020246 Disease venous thrombosis - + MESH:D006473 Disease blood loss - + MESH:D009369 Disease tumor - - MESH:D013927 - Disease - - vascularized thrombus - DISCUSS paragraph 11264 Our first embolization we performed only with the alcohol, in 96%. Alcohol is very potent and very cheap embolisaton material. If it is used in combination with distal balloon occlusion, the rate of complications such as migration is very low. Alcohol is very powerful embolization material, and destruction of capillary system is very powerful, better than any other embolization material. Destruction of capillary system lead to tumor necrosis. Tolle and Cramer performed 20 embolisations with very low rate of the complications. Those authors claim that alcohol is very safe to use, and percent of recanalization is almost zero. We also performed majority of our embolisations with same techniques. The volume of alcohol that we used was determined for every patient individually. After balloon occlusion, angiography was performed. The volume of contrast material that was necessary for demarcation of whole kidney is the volume of alcohol that we used. Occlusion was performed 40 minutes after alcohol injection. In our study the rate of complication was zero. With lack of occlusion catheters we abandoned this technique. Some authors reported severe complications after alcohol embolization. Laarmann at all. reported two cases of testicular damage, as reason of anatomical variation where art. testiculariswas the branch of renal artery. Same group of authors performed alcohol embolization in 44 patients with no distal balloon occlusion. The volume of alcohol that was used was 14-22ml. Timmerman performed capillary embolization in combination of alcohol and Etiblock. - + 9606 Species patient - + 9606 Species patients - + + MESH:D000438 + Chemical + + alcohol + + MESH:D054549 Disease balloon occlusion - - MESH:D000431 + + - Chemical - - alcohol + + Etiblock - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - + + MESH:D000438 + Chemical + + Alcohol + + MESH:D054549 Disease balloon occlusion - - MESH:D000431 + + MESH:D000438 Chemical - - alcohol + + Alcohol - + MESH:D009336 Disease tumor necrosis - - MESH:D000431 + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 Chemical alcohol - + MESH:D054549 Disease balloon occlusion - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - - MESH:D000431 - Chemical - - Alcohol - - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - - MESH:D000431 - Chemical - - Alcohol - - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - + MESH:D007674 Disease @@ -48442,47 +49013,47 @@ paragraph 12845 Several publications confirmed severe complications after alcohol embolization, so the use of alcohol was diminished and Etiblock was introduced as main embolization material. Initial results with Etiblock determined vasospasm after its use, so results were incomplete embolization. After another embolization materials were introduced, such as Ivalon 150-250 microns. Ivalon embolization does not need use of occlusion balloons, and application is very simple over microcatheter. Ivalon also has very good X ray visibility. In our series we had two case of recanalization after Ivalon embolization, so embolization had to be repeated. After this we started with so called sandwich embolization technique alcohol and Ivalon particles 150-250 microns. Central embolization was performed with Tornado colils. We had no complication with these technique. - - MESH:D000431 + + MESH:C026699 Chemical - + + Ivalon + + + MESH:D000438 + Chemical + alcohol - - MESH:D020301 - Disease - - vasospasm + + MESH:D000438 + Chemical + + alcohol - - MESH:D000431 + + MESH:D000438 Chemical - + alcohol - - MESH:D004617 + + MESH:D020301 Disease - - Ivalon embolization + + vasospasm - + MESH:D054549 Disease occlusion balloons - - MESH:D000431 + + MESH:C026699 Chemical - - alcohol - - - MESH:D004617 - Disease - - Ivalon embolization + + Ivalon @@ -48490,19 +49061,19 @@ paragraph 13697 With preoperative embolization there is significant reduce of blood transfusion during surgery, comparing to patients with no embolization. Volume of blood in group of embolised patients was around 250ml, and in group of non embolised patients was around 800ml. - + 9606 Species patients - + 9606 Species patients - + 9606 Species @@ -48514,13 +49085,13 @@ paragraph 13959 Zielinski et al. in their work compare rate of surveillance of five and ten years between group of 118 patients that were embolised and group of 116 patients with no embolization prior to surgery. The both groups had same demographics. Five and ten year's surveillance was statistically significant in first group 62%, and in the second group 35%. - + 9606 Species patients - + 9606 Species @@ -48532,73 +49103,79 @@ paragraph 14307 Kauffmann in his work showed reduction of blood loss during surgery in the group of patients preoperatively embolised. Vogel et al. published the same results as Kauffmann with loss of blood of 600ml in patients that were preoperatively embolised. Embolization was performed with 2-6ml Histoacril mixed with 50 % Lipiodol. Danish group of authors with Christensen published that they had smaller loss of blood during surgery in group of patients who were embolised prior to surgery, and the time of surgery was shorter, comparing with the group of patients with no embolization prior to surgery. Embolization was performed only with coils. The surgery was performed five days after embolization. Surveillance rate of embolised patients was better than in patients with no embolization. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D007022 + + MESH:D006473 Disease reduction of blood loss - + MESH:D004998 Chemical Lipiodol + + - + Chemical + + Histoacril + DISCUSS paragraph 15093 Stosslein at all. published that blood volume loss during surgery was in average 300ml. They had 92 patients. There were no significant shortening it time of surgery between two groups of patients. Surveillance rate was much higher in group of patient with embolization than in another group. - + 9606 Species patients - + 9606 Species patients - + 9606 Species @@ -48610,13 +49187,13 @@ paragraph 15387 Latal at all published that there were no statistically significance in surveillance rate between patients who were embolised prior to surgery comparing with patients who had only surgical nephrectomy. - + 9606 Species patients - + 9606 Species @@ -48634,73 +49211,79 @@ paragraph 15603 Kidney cancer is the rare tumor considering other malignancies. Kidney as the organ is very suitable for all transcatheter procedures as for percutaneous procedures. The gold standard for treatment of kidney cancer in advanced stage is surgical procedure with nephrectomy, and for small tumor is enucleation of tumor. Full capillary embolization is method of choice to prepare surgical nephrectomy. The most potent embolization material is alcohol, but its is the most dangerous one. Because of that, alcohol should be administered over the occlusion or in small portions with conjunction of Lipiodol to be visible. The goal of this embolization is to make surgical procedure easier, thru less bleeding during the surgical procedure. On other goal is to make surgery less time consuming. The surgery can be made 24-48 h after the embolization, or it can be made as late nephrectomy after 2-3 weeks. Some percent of patients who were inoperable before the embolization, become operative after the procedure. It is a fact that the cost of treatment of those patients are bigger than with patients who are treated only with the surgery. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D006470 - Disease - - thru less bleeding - - + MESH:D007680 Disease Kidney cancer - + MESH:D009369 Disease tumor - + MESH:D007680 Disease kidney cancer - + + MESH:D006470 + Disease + + bleeding + + MESH:D058405 Disease small tumor - - MESH:D000431 + + MESH:D009369 + Disease + + malignancies + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 Chemical alcohol - + MESH:D004998 Chemical Lipiodol - - MESH:D000431 - Chemical - - alcohol - - + MESH:D009369 Disease @@ -48740,13 +49323,13 @@ 399 16796 Superselective arterial catheterization as a vehicle for delivering radioactive infarct particles to tumors - + MESH:D007238 Disease infarct - + MESH:D009369 Disease @@ -48767,17 +49350,11 @@ 479 16904 Treatment of renal adenocarcinoma by embolic occlusion of the renal circulation - - MESH:D002292 - Disease - - renal adenocarcinoma - - - MESH:D007674 + + MESH:C538614 Disease - - embolic occlusion of the renal circulation + + renal adenocarcinoma by embolic occlusion of the renal circulation @@ -48794,11 +49371,6 @@ 789 16984 Nierentumoremboisation - - Disease - - Nierentumoremboisation - 162 @@ -48814,14 +49386,14 @@ 1405 17007 Renal cell carcinoma in children: experience at a single institution in Japan - + 9606 Species children - - MESH:D002292 + + MESH:C538614 Disease Renal cell carcinoma @@ -48841,14 +49413,14 @@ 1710 17085 Renal cell carcinoma in children: the Detroit experience - + 9606 Species children - - MESH:D002292 + + MESH:C538614 Disease Renal cell carcinoma @@ -48868,11 +49440,6 @@ 587 17142 Nephroblastom - - Disease - - Nephroblastom - 42 @@ -48903,11 +49470,11 @@ 434 17194 Usefulness of preopertive transcatheter embolisaton in kidney tumors - - MESH:D007680 + + MESH:D007674 Disease - - kidney tumors + + embolisaton in kidney tumors @@ -48923,7 +49490,7 @@ 708 17263 Preopertive renal embolization as adjunct to radical Nephrectomy - + MESH:D007674 Disease @@ -48944,18 +49511,18 @@ 708 17328 Planned delayed nephrectomy after ethanol embolization of renal carcinoma - + + MESH:C538614 + Disease + + renal carcinoma + + MESH:D000431 Chemical ethanol - - MESH:D002292 - Disease - - renal carcinoma - 33 @@ -48970,8 +49537,8 @@ 170 17402 The role of arterial embolization in renal cell carcinoma - - MESH:D002292 + + MESH:C538614 Disease renal cell carcinoma @@ -48991,8 +49558,8 @@ 178 17460 The current status of embolization in renal cell carcinoma - a survey of local and national practice - - MESH:D002292 + + MESH:C538614 Disease renal cell carcinoma @@ -49011,11 +49578,17 @@ 212 17561 Klinische Erfahrungen mit der arteriellen Embolization von Nierentumoren mit Athanol - + MESH:D004617 Disease - - Embolization von Nierentumoren mit Athanol + + Embolization von + + + MESH:D000431 + Chemical + + Athanol @@ -49047,6 +49620,18 @@ 277 17729 Der absolute Alkohol, ein steuerbares Embolisationsmaterial bei Nierentumoren + + - + Chemical + + steuerbares + + + MESH:D000431 + Chemical + + Alkohol + 24 @@ -49076,7 +49661,7 @@ 204 17898 Analyse der Langzeitergebnisse nach Nephrektomie wegen Hypernephrom anhand 442 Fallen - + MESH:D014890 Disease @@ -49097,8 +49682,8 @@ 178 17985 The current status of embolization in renal cell carcinoma-a survey of local and national practice - - MESH:D002292 + + MESH:C538614 Disease renal cell carcinoma @@ -49118,13 +49703,13 @@ 731 18084 Value of preoperative renal artery embolization in Reducing blood transfusion requirements during nephrectomy for renal cell carcinoma - - MESH:D002292 + + MESH:C538614 Disease renal cell carcinoma - + MESH:D007674 Disease @@ -49161,8 +49746,8 @@ 12 18313 Comparison of preoperative embolization followed by radical nephrectomy with radical neprectomy alone for renal cell carcinoma - - MESH:D002292 + + MESH:C538614 Disease renal cell carcinoma @@ -49181,8 +49766,8 @@ 207 18440 Palliative capillary embolization renal carcinoma - - MESH:D002292 + + MESH:C538614 Disease Palliative capillary embolization renal carcinoma @@ -49215,18 +49800,12 @@ 28 18550 Prolonged survival following palliative renal tumor embolization by capillary occlusion - - MESH:D007680 + + MESH:D007674 Disease renal tumor - - OMIM:163000 - Disease - - capillary occlusion - 133 @@ -49243,8 +49822,8 @@ 193 18638 The value of transvascular embolization in the treatment of renal carcinoma - - MESH:D002292 + + MESH:C538614 Disease renal carcinoma @@ -49279,7 +49858,7 @@ 31 18805 Einfluss der Nierenocclusion auf die Uberlebensrate beim Nierenzellkarzinom: Eine retrospektive 10-Jahres-Studie - + MESH:C535733 Disease @@ -49299,8 +49878,8 @@ 535 18919 Long-term results of embolization in renal tumors - - MESH:D007680 + + MESH:D007674 Disease renal tumors @@ -49319,7 +49898,7 @@ 886 18969 Renal artery embolization: clnical indication and experience from over 100 cases - + MESH:D007674 Disease @@ -49357,8 +49936,8 @@ MESH:D009190 Disease - - Myelodysplastic Syndromes + + Myelodysplastic @@ -49387,8 +49966,8 @@ MESH:D006402 Disease - - blood cytopenia + + peripheral blood cytopenia @@ -49402,19 +49981,13 @@ abstract 437 to assess the influence of the some prognostic factors like age, gender, cytopenia, BM blast percentage, transfusion dependence, ferritin, hemoglobin (Hb), lactate dehydrogenase (LDH), albumin and specific karyotype abnormalities in myelodysplastic syndromes on overall survival (OS). - + MESH:D006402 Disease cytopenia - - MESH:D019344 - Chemical - - lactate - - + MESH:D009190 Disease @@ -49426,7 +49999,7 @@ abstract_title_1 722 Patients and methods: - + 9606 Species @@ -49438,17 +50011,17 @@ abstract 744 we retrospectively analyzed the cohort of 108 patients diagnosed between 1.1.2011 and 31.12.2013 at the University Clinic of Hematology, Ss Cyril and Methodius University, Skopje, Macedonia. They were evaluated for clinical and hematologic features at diagnosis and at leukemic transformation. - + 9606 Species patients - + MESH:D007938 Disease - - leukemic + + leukemic transformation @@ -49493,6 +50066,18 @@ patients + MESH:D007938 + Disease + + Leukemic + + + - + Chemical + + OS + + MESH:D006402 Disease @@ -49522,35 +50107,30 @@ paragraph 1987 Myelodysplastic syndromes (MDSs) are a heterogeneous group of disorders characterized by dysplasia in one or more cell lines, followed by a progressive impairment in the ability to differentiate, manifested with peripheral cytopenia and an increasing risk of evolution into acute myeloid leukemia (AML). - + MESH:D015470 Disease acute myeloid leukemia - - MESH:D015792 - Disease - - dysplasia - - Disease - - MDSs - - MESH:D009190 Disease Myelodysplastic syndromes - - MESH:D010523 + + MESH:D006402 Disease peripheral cytopenia + + MESH:D004476 + Disease + + dysplasia + MESH:D015470 Disease @@ -49563,98 +50143,104 @@ paragraph 2291 The clinical course is variable so much effort has been focused on methods for predicting prognosis. Accurately predicting the prognosis once a malignancy has been diagnosed is of great importance to both patients and their physicians alike. From a patient's perspective, the prognosis helps define the severity of disease and sets expectations as to how it is likely to impact them. In contrast, prognostic information from a physician's standpoint is essentially a means of staging the disease in a manner that can be used to help direct therapy. For both patients and physicians, the estimation of prognosis is a continual process that does not happen just at the time of diagnosis. Reevaluating the prognosis may be useful when a patient shows signs of progression or after they have become refractory to standard treatment. The vast majority of cases (80% to 90%) occur 'de novo', whereas 10% to 20% of cases are secondary. French-American British (FAB) classification for MDS is still in use, based on morphologic findings. The standard prognostic tool for prognosis in MDS is the International Prognostic Scoring System (IPSS) which classifies patients into low-, intermediate-1, intermediate-2 and high-risk categories. IPSS takes into account BM blast percentage, number of cytopenia and cytogenetic abnormalities. Beside the known prognosis factors included in the IPSS, other variables that have an impact on prognosis in MDS were studied: age, gender, FAB subtypes, degree of anemia and RBC transfusion dependence, ferritin (, LDH, albumins, mutations and comorbidities. - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + MESH:D006402 Disease cytopenia - + MESH:D009190 Disease MDS - + MESH:D009190 Disease MDS + + MESH:D009369 + Disease + + malignancy + INTRO paragraph 3878 Diagnosis requires BM examination and cytogenetic studies, and lately molecular studies. The consensual minimum criterion for diagnosis is the presence of erythroid, granulocyte or megakaryocyte dysplasia in 10% or more of informative cells. One must exclude the possibility of erythroid dysplasia associated with vitamin B12/folate/copper deficiency, viral infections, chemotherapy, or lead/arsenic poisoning. - + 4709 Gene 37630 B12 - - MESH:D014777 + + MESH:D001102 Disease viral infections - - MESH:D015792 + + MESH:D004476 Disease dysplasia - + MESH:C535468 Disease copper deficiency - + MESH:D005492 Chemical folate - + MESH:C538442 Disease erythroid dysplasia - + MESH:D020261 Disease @@ -49666,37 +50252,31 @@ paragraph 4289 The incidence of MDS increases with age (median about 70 years). MDS is rare in children Age had a significant effect on OS of the MDS population analyzed as a whole and stratified by subgroups-the older the age, the worse the prognosis. During the analysis on the subgroups, the effect of age was statistically relevant within RA and RARS patients, whereas it was not significant within and RAEB subgroup. - + 5917 Gene 68281 RARS - + 9606 Species children - + 9606 Species patients - + MESH:D009190 Disease MDS - - MESH:D009190 - Disease - - MDS increases - MESH:D009190 Disease @@ -49704,10 +50284,10 @@ MDS - MESH:D001172 + MESH:D009190 Disease - - RA + + MDS @@ -49745,31 +50325,37 @@ paragraph 5025 According to the multivariate analysis peripheral cytopenia did not reach a statistical significance on OS in MDS patients. FAB subtypes can worsen the prognosis in MDS in terms of OS, with differences among subtypes. Refractory anemia had the best and RAEB-T the worst prognosis. - + 9606 Species patients - + + MESH:D000754 + Disease + + RAEB-T + + MESH:D000740 Disease anemia - + MESH:D009190 Disease MDS - + MESH:D009190 Disease MDS - + MESH:D006402 Disease @@ -49781,55 +50367,55 @@ paragraph 5306 Blast count showed a significant predictive value on OS in MDS patients. More than 5% blasts in BM are considered as a bad prognostic factor for OS and leukemic transformation. - + 9606 Species patients - - MESH:C567932 - Disease - - OS and leukemic transformation - - + MESH:D009190 Disease MDS + + MESH:D007938 + Disease + + leukemic + INTRO paragraph 5483 Red blood cell (RBC) transfusions are a commonly used therapy to treat symptomatic anemia that affects most patients with MDS. Transfusion dependence is defined by the MDS International Study Group (2000) as requiring transfusion of at least one RBC at 8 weeks for 4 months. Transfusion-dependent patients had shorter survival rate than those who received less than 18 units of blood over a period of 36 months (37). - + 9606 Species patients - + 9606 Species patients - + MESH:D000740 Disease anemia - + MESH:D009190 Disease MDS - + MESH:D009190 Disease @@ -49841,72 +50427,66 @@ paragraph 5900 Serum ferritin level >=500 microg/l at diagnosis was a strong independent predictor of survival. Serum ferritin was significantly correlated with OS in Chinese patients. These patients suffer from comorbidities such as heart failure, diabetes, infections, disorders of the thyroid gland and liver, shortening their survival. - + 9606 Species patients - + 9606 Species patients - - MESH:D013959 - Disease - - disorders of the thyroid gland - - + MESH:D003920 Disease diabetes - + MESH:D006333 Disease heart failure + + MESH:D007239 + Disease + + infections + INTRO paragraph 6225 Lactate dehydrogenase (LDH) is a parameter that should be recognized as a prognostic factor in MDS. One German study pointed out the correlation between LDH level and OS. Serum albumin is an independent prognostic factor that influences OS in patients with MDS. Hypoalbuminemia is a marker for shorter OS in MDS. - + 9606 Species patients - + MESH:D009190 Disease MDS - + MESH:D009190 Disease MDS - + MESH:D009190 Disease MDS - - MESH:D019344 - Chemical - - Lactate - MESH:D034141 Disease @@ -49919,19 +50499,31 @@ paragraph 6538 Recurrent chromosomal abnormalities have been identified in 40-70% of the 'de novo' MDS and 95% of secondary MDS. These chromosomal aberrations include 5q-, 7q-/-7, +8, 20q-, 12p-, abnormalities in 17p, 11q23 and chromosome 3. Favorable prognostic markers according to IPSS include: a normal karyotype, 5q-as an isolated anomaly, 20q-as an isolated anomaly and -Y chromosome. Karyotype findings associated with poor prognosis include complex karyotype and abnormalities of chromosome 7. Other cytogenetic abnormalities confer an intermediate prognosis. Abnormal karyotype correlates with poor prognosis and shorter OS. According to the multivariate analysis, cytogenetics showed a significant predictive value on OS. - + MESH:D009190 Disease MDS - + MESH:D009190 Disease MDS - + + MESH:D000014 + Disease + + anomaly + + + MESH:D000014 + Disease + + anomaly + + MESH:D002869 Disease @@ -49943,19 +50535,19 @@ paragraph 7263 Somatic mutations are identified in more than 70% of patients with MDS, including more of the patients with normal karyotype. These mutations are major predictors of the clinical phenotype, and could also be predictors of prognosis. - + 9606 Species patients - + 9606 Species patients - + MESH:D009190 Disease @@ -49967,127 +50559,127 @@ paragraph 7496 The incidence of MDS increases with the age, so does the prevalence of comorbidities. About 50% of MDS patients have one or more comorbidities. Congestive heart disease, hypertension, lung diseases, diabetes, liver failure, bleeding and solid tumors are cited as the most often reasons for non-leukemic dead. Comorbidities are significant and independent predictors in MDS. Congestive heart disease and chronic obstructive lung disease are associated with shorter OS, while diabetes and cerebro-vascular diseases do not change prognosis in MDS. - + 9606 Species patients - + + MESH:D009369 + Disease + + solid tumors + + + MESH:D009190 + Disease + + MDS + + MESH:D029424 Disease chronic obstructive lung disease - + MESH:D006973 Disease hypertension - + MESH:D009190 Disease MDS - - MESH:D014652 - Disease - - cerebro-vascular diseases - - + MESH:D003920 Disease diabetes - + MESH:D006333 Disease Congestive heart disease - + MESH:D017093 Disease liver failure - + + MESH:D000783 + Disease + + cerebro-vascular diseases + + MESH:D009190 Disease MDS - + + MESH:D007938 + Disease + + non-leukemic + + MESH:D006333 Disease Congestive heart disease - + MESH:D006470 Disease bleeding - + MESH:D008171 Disease lung diseases - + MESH:D009190 Disease MDS - - MESH:D007938 - Disease - - leukemic - - - MESH:D009369 - Disease - - tumors - - + MESH:D003920 Disease diabetes - - MESH:D009190 - Disease - - MDS increases - INTRO paragraph 8041 The median OS in MDS is 2.5 years. Prognosis is poor for patients with MDS, with 3-year survival rates estimated at less than 50%. The median survival rates according to IPSS are estimated at 8, 5.3, 2.2, and 0.9 years, respectfully. - + 9606 Species patients - + MESH:D009190 Disease MDS - + MESH:D009190 Disease @@ -50105,31 +50697,25 @@ paragraph 8282 The aim of this study was to assess the influence of the some prognostic factors in myelodysplastic syndromes (age, gender, cytopenia, BM blast percentage, transfusion dependence, serum levels of hemoglobin, ferritin, lactate dehydrogenase and albumin, and specific karyotype abnormalities) on OS. - + MESH:D006402 Disease cytopenia - + MESH:D009190 Disease myelodysplastic syndromes - - MESH:D019344 - Chemical - - lactate - METHODS title_1 8580 3. PATIENTS AND METHODS - + 9606 Species @@ -50141,132 +50727,138 @@ paragraph 8604 We retrospectively analyzed the cohort of 108 patients (62 male and 46 female) diagnosed between 1.1.2011 and 31.12.2013 at the University Clinic of Hematology, Ss Cyril and Methodius University, Skopje, Macedonia. They were evaluated for clinical and hematologic features at diagnosis and leukemic transformation. Observation time was 36 months. Diagnosis was made upon dysplastic changes in peripheral blood smear and bone marrow aspirate. Confirmation analysis included bone marrow aspirate, cytochemical stain, immunohistochemical tests, cytochemical stain of medullar iron and karyotype analysis in some cases. We evaluated some parameters that could influence OS: age, gender, cytopenia, BM blast percentage, transfusion dependence, serum levels of Hb, ferritin, LDH and albumin as well as specific karyotype abnormalities. Chromosomal analysis was performed only in 5 patients using BM aspirate, according to laboratory procedures. IPSS was not calculated in the vast majority of patients, because we could not perform cytogenetic analysis in all patients. Most patients (pts) received supportive care: transfusion of RBC and platelets, red cell and granulocyte growth factors (21 pts), vitamins, corticosteroids, iron chelatation therapy (7 pts), some received chemotherapy (4pts) and few patients allogeneic transplantation (3 pts). OS was estimated in months including the period from the date of diagnosis to the time of death / time of last visit. Leukemic transformation was noticed in 10 (9,3%) patients. Mortality was 36,1%. Infectious and hemorrhagic complications as well as BM failure were considered causes of MDS-related deaths. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species pts - + 9606 Species pts - + 9606 Species pts - + 9606 Species pts - + 9606 Species pts - + MESH:D007501 Chemical iron - + MESH:D007501 Chemical iron - + MESH:D007938 Disease leukemic transformation - - MESH:D000305 - Chemical - - corticosteroids - - + MESH:D009190 Disease MDS - - MESH:C059630 - Chemical - - vitamins + + MESH:D007938 + Disease + + Leukemic - + MESH:D006470 Disease hemorrhagic complications - + + MESH:D017093 + Disease + + BM failure + + MESH:D006402 Disease cytopenia - + MESH:D003643 Disease death + + MESH:D004416 + Disease + + dysplastic + RESULTS @@ -50279,90 +50871,72 @@ paragraph 10264 Mean age at diagnosis was 66,6 years (range 18-89) (SD 14.2). Mean OS depending on age was 16,4 months, and it was not statistically significant (p= .46375). In our study, 62 (57.4%) patients were men, 46 (42.6%) women. Male to female ratio was 1.35 to 1. Women had better OS than men (p = .00819). (Figure 1). According to the FAB classification patients were classified as follows: 75 patients as having refractory anemia (RA; 69.4%), 1-RA with ringed sideroblasts (RARS; 0.9%), 21-RA with excess of blasts (RAEB; 19.4%), 3-RAEB in transformation (RAEB-T 2.8%), and 8 as having chronic myelomonocytic leukemia (CMML 8.2%). (Table 1) - + 5917 Gene 68281 RARS - + 9606 Species patients - + 9606 Species men - + 9606 Species women - + 9606 Species Women - + 9606 Species men - + 9606 Species patients - + 9606 Species patients - - MESH:D000753 - Disease - - refractory anemia - - MESH:C039620 - Chemical - - 3-RAEB + MESH:D054429 + Disease + + myelomonocytic leukemia - MESH:D001172 + MESH:D000754 Disease - - RA + + RAEB-T - MESH:D001172 - Disease - - RA - - - MESH:D015477 - Disease - - chronic myelomonocytic leukemia - - - MESH:D001172 + MESH:D000740 Disease - - RA + + anemia @@ -50380,7 +50954,7 @@ table_caption 10928 Distribution of patients according to the FAB classification - + 9606 Species @@ -50392,17 +50966,17 @@ paragraph 10989 Statistically significant differences in OS were found among patients with RA, RAEB, RAEB-T and CMML (p = .03015). (Figure 2) - + 9606 Species patients - - MESH:D001172 + + MESH:D000754 Disease - - RA + + RAEB-T @@ -50418,54 +50992,54 @@ paragraph 11144 Cytopenia had no influence on OS (p=.09340). In our group 23 patients (21.3%) presented with one cytopenia, 46 patients (42.6) with bicytopenia and 39 patients (36.1%) with pancytopenia. There was no statistically significant difference in OS among patients with different cytopenia (p = .33755). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D006402 Disease Cytopenia - - MESH:D006402 - Disease - - cytopenia - - + MESH:D010198 Disease pancytopenia - + + MESH:D006402 + Disease + + different cytopenia + + Disease bicytopenia - + MESH:D006402 Disease @@ -50483,49 +51057,49 @@ paragraph 11750 Mean Hb level in our cohort was 85,1 g/L (range 41-150). Hb had no impact on OS (16.4 months) (p=.12142). The patients were grouped in three categories according to the severity of anemia: severe anemia (Hb <70 g/L), moderate anemia (Hb 70-100 g/L) and patients with Hb >100 g/L. OS was highest in patients with Hb >100 g/L-21.6 months, in moderate anemia-14,8 months and in severe anemia-15.2 months, being not statistically significant (p=0.07535). - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D000740 Disease anemia - + MESH:D000740 Disease anemia-15 - + MESH:D000740 Disease anemia-14 - + MESH:D000740 Disease anemia - + MESH:D000740 Disease @@ -50545,6 +51119,12 @@ fig_caption 12472 OS in groups with different serum ferritin levels + + - + Chemical + + OS + RESULTS @@ -50629,18 +51209,24 @@ paragraph 14141 The statistical analysis was performed using statistical package SSPS 16.0. For the parameters or variables used in this paper, the range, mean and standard deviation were presented. Differences among variables were evaluated by the Chi-square test. The determinations of correlations between different variables was based on the Pearson correlation coefficient in all cases where the variables had a normal distribution, and a certain correlation was considered statistically significant if p <0.05. Overall survival was defined as the time interval between diagnosis date and death date. Patients who were alive were censored at the last follow-up date. The probabilities of OS were estimated using the method of Kaplan and Meier. Cox proportional hazards regression models were used to assess the association between prognostic factors and OS. - + 9606 Species Patients - + MESH:D003643 Disease death + + - + Chemical + + OS + DISCUSS @@ -50653,56 +51239,68 @@ paragraph 15002 Our results considering age (66.6 years) correspond with those in the literature. Age in this group did not affect OS neither as a whole nor stratified by subgroups, similar with some studies. Considering gender, men had worse OS than women, corresponding with the data in the literature. FAB subtypes had a significant effect on OS. Considering subtypes, the worst results were observed in RAEB-T, than in CMML, RAEB and RA subtype, similar with those in the literature. There was only one patient with RARS and he was not taken in consideration. Cytopenia had no impact on OS, corresponding with some data in the literature. But, others showed opposite results. OS correlated inversely with BM blast percentage, showed in other studies. Hemoglobin as a whole and stratified into subgroups did not show any impact on OS. Transfusion dependence could be an independent prognostic factor for bad prognosis, although in our study group the influence on OS was not significant. Serum ferritin level >=500 microg/L at presentation is an independent poor predictor for OS. The group with ferritin <500 microg/L had better OS. LDH is a factor sited as having predictive value, although we could not prove that. Hypoalbuminemia is an independent poor prognostic factor in MDS patients, but in our study group it was not significant. - + 5917 Gene 68281 RARS - + 9606 Species men - + 9606 Species women - + 9606 Species patient - + 9606 Species patients - - MESH:D001172 + + MESH:D000754 Disease - - RA + + RAEB-T - + + - + Chemical + + OS + + MESH:D009190 Disease MDS - + + - + Chemical + + OS + + MESH:D034141 Disease Hypoalbuminemia - + MESH:D006402 Disease @@ -50714,13 +51312,13 @@ paragraph 16328 Cytogenetics in other studies showed a significant predictive value on OS and patients were accordingly stratified in groups with different IPSS score. Only 5 patients were analyzed for cytogenetic abnormalities, three of them with good prognosis, and two of them (+8, inv16) with intermediate prognosis. - + 9606 Species patients - + 9606 Species @@ -50732,12 +51330,24 @@ paragraph 16633 Limitations in our study include retrospective analysis, insufficient data on cytogenetic analysis and impossibility to stratify patients according to IPSS score, lack of data on comorbidities, insufficient data on ferritin, LDH and albumins, that probably reflected on the results making some of them different than those cited in the literature. - + 9606 Species patients + + MESH:D000309 + Disease + + insufficient + + + MESH:D000309 + Disease + + insufficient + DISCUSS @@ -50763,17 +51373,23 @@ 351 17029 Risk assessment in myelodysplastic syndromes and myelodysplastic/ myeloproliferative neoplasms - + + MESH:D009190 + Disease + + myelodysplastic + + MESH:D009196 Disease myeloproliferative neoplasms - + MESH:D009190 Disease - - myelodysplastic syndromes and myelodysplastic + + myelodysplastic syndromes @@ -50788,13 +51404,13 @@ 538 17124 Myelodysplastic syndromes-coping with ineffective hematopoiesis - + MESH:C536227 Disease hematopoiesis - + MESH:D009190 Disease @@ -50813,11 +51429,11 @@ 964 17188 Clinical and Genetic Predictors of Prognosis in Myelodysplastic Syndromes - + MESH:D009190 Disease - - Myelodysplastic Syndromes + + Myelodysplastic @@ -50834,7 +51450,7 @@ 199 17262 Proposals for the classification of the myelodysplastic syndromes - + MESH:D009190 Disease @@ -50855,7 +51471,7 @@ 2088 17328 International scoring system for evaluating prognosis in myelodysplastic syndromes - + MESH:D009190 Disease @@ -50876,19 +51492,19 @@ 673 17411 A European survey on the detection and management of iron overload in transfusion-dependent patients with MDS - + 9606 Species patients - + MESH:D007501 Chemical iron - + MESH:D009190 Disease @@ -50908,23 +51524,23 @@ 9 17521 Prognostic relevance of anemia and transfusion dependency in myelodysplastic syndromes and primary myelofibrosis - + MESH:D000740 Disease anemia - + MESH:D055728 Disease myelofibrosis - + MESH:D009190 Disease - - myelodysplastic syndromes + + transfusion dependency in myelodysplastic syndromes @@ -50940,13 +51556,13 @@ 448 17634 Myelodysplastic syndrome in elderly patients: correlation of CBC with cytogenetic and FISH analysis - + 9606 Species patients - + MESH:D009190 Disease @@ -50965,7 +51581,7 @@ 358 17734 Myelodysplastic syndromes - + MESH:D009190 Disease @@ -50985,7 +51601,7 @@ 368 17760 Prognostic factors and scoring systems in myelodysplastic syndromes - + MESH:D009190 Disease @@ -51010,19 +51626,19 @@ surname:Kuendgen;given-names:A 17828 Impact of the degree of anemia on the outcome of patients with myelodysplastic syndromes and its integration into the WHO Classification-based Prognostic Scoring System (WPSS) - + 9606 Species patients - + MESH:D009190 Disease myelodysplastic syndromes - + MESH:D000740 Disease @@ -51044,7 +51660,7 @@ 92 18004 Myelodysplastic syndromes: a study of 101 cases according to the FAB classification - + MESH:D009190 Disease @@ -51068,7 +51684,7 @@ surname:Zhang;given-names:Y 18088 Serum Ferritin Is an Independent Prognostic Factor in Chinese with Myelodysplastic Syndromes Classified as IPSS Intermediate-1 - + MESH:D009190 Disease @@ -51088,13 +51704,13 @@ 5255 18215 Objectives of iron chelation in myelodysplastic syndromes: more than meets the eye? - + MESH:D009190 Disease myelodysplastic syndromes - + MESH:D007501 Chemical @@ -51113,11 +51729,11 @@ 2852 18299 Incidence and Clinical Complications of Myelodysplastic Syndromes among United States Medicare Beneficiaries - + MESH:D009190 Disease - - Complications of Myelodysplastic + + Myelodysplastic Syndromes @@ -51131,25 +51747,25 @@ 494 18408 Retrospective nationwide surv1077y of Japanese patients with transfusion-dependent MDS and aplastic anemia highlights the negative impact of iron overload on morbidity/mortality - + 9606 Species patients - + MESH:D000741 Disease aplastic anemia - + MESH:D009190 Disease MDS - + MESH:D007501 Chemical @@ -51176,19 +51792,19 @@ surname:Pivkova;given-names:A 18586 Iron overload in patients with transfusion dependent myelodysplastic syndrome - + 9606 Species patients - + MESH:D009190 Disease myelodysplastic syndrome - + MESH:D007501 Chemical @@ -51209,24 +51825,18 @@ 976 18664 Prognostic significance of serial determinations of lactate dehydrogenase (LDH) in the foolow-up of patients with myelodysplastic syndromes - + 9606 Species patients - + MESH:D009190 Disease myelodysplastic syndromes - - MESH:D019344 - Chemical - - lactate - 87 @@ -51239,13 +51849,13 @@ 1009 18804 Hypoalbuminenemia is an independent prognostic factor for overall survival in myelodysplastic syndromes - + MESH:D009190 Disease myelodysplastic syndromes - + Disease Hypoalbuminenemia @@ -51263,7 +51873,7 @@ 964 18908 Clinical and genetic predictors of prognosis in myelodysplastic syndromes - + MESH:D009190 Disease @@ -51284,101 +51894,101 @@ 408 18982 Combined mutations of ASXL1, CBL, FLT3, IDH1,IDH2, JAK2, KRAS, NPM1, NRAS, RUNX1, TET2 and WT1 genes in myelodysplastic syndromes and acute myeloid leukemias - + 171023 Gene 9098 ASXL1 - + 867 Gene 3802 CBL - + 2322 Gene 3040 FLT3 - + 3417 Gene 21195 IDH1 - + 3418 Gene 37590 IDH2 - + 3717 Gene 21033 JAK2 - + 3845 Gene 37990 KRAS - + 4869 Gene 81697 NPM1 - + 4893 Gene 55661 NRAS - + 861 Gene 1331 RUNX1 - + 54790 Gene 49498 TET2 - + 7490 Gene 11536 WT1 - + MESH:D009190 Disease myelodysplastic syndromes - - MESH:D007951 + + MESH:D015470 Disease - - myeloid leukemias + + acute myeloid leukemias @@ -51393,11 +52003,11 @@ 885 19140 The Multifaceted Nature of Myelodysplastic Syndromes: Clinical, Molecular and Biological Prognostic Features - + MESH:D009190 Disease - - Myelodysplastic Syndromes + + Myelodysplastic @@ -51421,7 +52031,7 @@ surname:Vigil;given-names:CE 19249 Association of comorbidities with overall survival in myelodysplastic syndrome: development of a prognostic model - + MESH:D009190 Disease @@ -51442,7 +52052,7 @@ 951 19363 The 2008 revision of the World Health Organization (WHO) classification of myeloid neoplasms and acute leukemia: rationale and important changes - + MESH:D015470 Disease @@ -51462,7 +52072,7 @@ 1045 19508 Myelodysplastic syndromes - Many New Drugs, Little Therapeutic Progress - + MESH:D009190 Disease @@ -51497,23 +52107,23 @@ surname:Abruzzo;given-names:L 19582 Acquisition of cytogenetic abnormalities in patients with IPSS defined lower-risk myelodysplastic syndrome is associated with poor prognosis and transformation to acute myelogenous leukemia - + 9606 Species patients - + MESH:D009190 Disease myelodysplastic syndrome - - MESH:D007951 + + MESH:D015470 Disease - - myelogenous leukemia + + acute myelogenous leukemia @@ -51528,7 +52138,7 @@ 526 19772 Cytogenetic features in myelodysplastic syndromes - + MESH:D009190 Disease @@ -51549,7 +52159,7 @@ 515 19822 Unraveling the molecular pathophysiology of myelodysplastic syndromes - + MESH:D009190 Disease @@ -51570,13 +52180,13 @@ 52 19892 Epidemiology of myelodysplastic syndromes and chronic myeloproliferative disorders in the United States 2001-2004, using data from the NAACCR and SEER programs - + MESH:D009196 Disease myeloproliferative disorders - + MESH:D009190 Disease @@ -51597,19 +52207,19 @@ 2231 20052 Refinement of the international prognostic scoring system (IPSS) by including LDH as an additional prognostic variable to improve risk assessment in patients with primary myelodysplastic syndromes (MDS) - + 9606 Species patients - + MESH:D009190 Disease MDS - + MESH:D009190 Disease @@ -51638,13 +52248,13 @@ surname:Krieger;given-names:O 20255 Prognostic impact of age and gender in 897 untreated patients with primary myelodysplastic syndromes - + 9606 Species patients - + MESH:D009190 Disease @@ -51677,19 +52287,19 @@ surname:Vallespi;given-names:T 20356 Spanish Myelodysplastic Syndrome Registry, Prognostic impact of severe thrombocytopenia in low-risk myelodysplastic syndrome - + MESH:D009190 Disease myelodysplastic syndrome - + MESH:D009190 Disease Myelodysplastic - + MESH:D013921 Disease @@ -51709,11 +52319,11 @@ 39 20481 The Myelodysplastic Syndromes - + MESH:D009190 Disease - - Myelodysplastic Syndromes + + Myelodysplastic @@ -51734,19 +52344,19 @@ surname:Beyne-Rauzy;given-names:O 20511 Prognostic factors for response and overall survival in 282 patients with higher-risk myelodysplastic syndromes treated with azacitidine - + 9606 Species patients - + MESH:D009190 Disease - - myelodysplastic + + myelodysplastic syndromes - + MESH:D001374 Chemical @@ -51776,7 +52386,7 @@ surname:Boni;given-names:M 20648 Prognostic Factors and Life Expectancy in Myelodysplastic Syndromes Classified According to WHO Criteria: A Basis for Clinical Decision Making - + MESH:D009190 Disease @@ -51799,7 +52409,7 @@ 302 20791 Prognostic factors in Myelodysplastic syndromes - + MESH:D009190 Disease @@ -51872,6 +52482,11 @@ abstract 825 Students recognized 10 main medical professionalism dimensions (empathy, respect, responsibility, autonomy, trust, communication, difference between professional and private life, team work, partnership) and two dimensions associated with it (physician's characteristics, external factors). Slight change of the attitudes towards a more self-centred future physicians' figure was observed in the fifth-year medical students. + + Disease + + empathy + ABSTRACT @@ -51896,7 +52511,7 @@ paragraph 1570 In the era of sophisticated medical technology, family physicians should use technological advances but continue to practice comprehensive medicine in spite of health policy demands for more focus on public than individual outcomes. Consequently, the medical professionalism is slowly finding its way back to medical education. It is widely acknowledged that medical professionalism can be learned and taught. However, usually it is a part of a so called hidden curriculum: it is incorporated into teaching different subjects without setting clear learning objectives. There are voices that medical students besides communication skills, team work, practice organization, and knowledge management also should be taught in humanism, accountability and altruism. However, not much is known about the role of medical education in building a future physician who will possess the appropriate professional values, attitudes and behaviors. Even a unifying theoretical or practical model to use as a format for developing a professionalism teaching curriculum does not exist yet. - + Disease altruism @@ -51907,24 +52522,24 @@ paragraph 2643 There is neither an unified definition nor theoretical model of medical professionalism at the moment. The American Board of Internal Medicine suggested that professionalism has six components: altruism, accountability, excellence, duty, honour/integrity and respect. A model from USA and Europe declared that professionalism consists of three fundamental principles (patient welfare, patient autonomy and social justice) with ten professional responsibilities. The model from UK suggests that medical professionalism consists of integrity, compassion, altruism, continuous improvement, excellence and working in partnership with team members. - + 9606 Species patient - + 9606 Species patient - + Disease altruism - + Disease altruism @@ -51935,12 +52550,17 @@ paragraph 3287 So far, several studies have tried to determine the views of undergraduate medical student regarding the professionalism. They showed that undergraduate students in general have a clear perception of what medical professionalism was. On the other hand, a recent qualitative study from Australia showed that undergraduate students have had some difficulties with perceiving what a professional physician was; they differentiated between the term "good" doctor and professional doctor as they perceived professionalism as an external and imposed construct. It also seems that some differences exist regarding the perceptions of medical professionalism in undergraduate students of different study years. Students in higher years of education reported the decline of professionalism in terms of idealism, empathy, the perception of physicians' status and income and the rise of the negative views on primary care and the importance of lifestyle and family. This change of attitudes seems to be largely dependent on the curriculum and the teaching method applied. The shift from informal to formal teaching of professionalism resulted in higher sensitivity to professionalism and the early patient contact and small groups in professionalism enhanced the understanding of professionalism. - + 9606 Species patient + + Disease + + empathy + INTRO @@ -51977,7 +52597,7 @@ paragraph 5148 This was a qualitative study, based on focus groups. It took part on the Faculty of Medicine in Maribor, Slovenia and was performed in June 2014. The study was approved by the National Ethics Board (No. 226/09/13). - + Disease Slovenia @@ -51988,7 +52608,7 @@ title_2 5363 3.2. Participants and focus groups - + 9606 Species @@ -52006,31 +52626,31 @@ paragraph 5653 The students were included in two focus groups. The first one consisted of the five first-year medical students (three women and two men), each 19 years old. The second one consisted of seven fifth-year medical students (two men and five women), each 23 years old. Both focus groups were moderated by one of the researchers (HV) and observed by the second researcher (ZKK). The focus groups participants discussed upon the several questions (Box 1). - + 9606 Species women - + 9606 Species men - + 9606 Species men - + 9606 Species women - + 9606 Species @@ -52050,7 +52670,7 @@ paragraph 6143 Focus groups were audio-recorded. Oral informed consent was sought from the participants prior to the session. - + 9606 Species @@ -52092,7 +52712,7 @@ paragraph 6837 Students of both study years expressed the importance of setting a clear line between objectivity and subjectivity in order not to let the subjective feelings influence the management of the patients. - + 9606 Species @@ -52104,7 +52724,7 @@ paragraph 7038 "The sympathy towards a patient should not blur your professional objectivity." (Fifth-year female student) - + 9606 Species @@ -52116,7 +52736,7 @@ paragraph 7150 "...exactly here...in the attitude towards the patient...to set the line between subjective and objective." (First-year female student) - + 9606 Species @@ -52128,7 +52748,7 @@ title_2 7290 4.2. Empathy - + Disease Empathy @@ -52139,31 +52759,36 @@ paragraph 7303 First-year students perceived empathy as something you could learn but not in order to really become an empathic person but in to make an impression that you are an empathic person. - + 9606 Species person - + 9606 Species person + + Disease + + empathy + RESULTS paragraph 7485 "I think it is important that the physicians do not feel sorry for the patient as their primary goal is to cure the patients" (First-year female student) - + 9606 Species patient - + 9606 Species @@ -52175,7 +52800,7 @@ paragraph 7643 "I think it is important to ask the patients how they are feeling. Not because you are really interested but to show them that you care. (First-year female student) - + 9606 Species @@ -52187,6 +52812,11 @@ paragraph 7810 "...(empathy) is learned not innate...you can act that you care for someone but you really don't." (First-year male student) + + Disease + + empathy + RESULTS @@ -52199,7 +52829,7 @@ paragraph 8063 "It is necessary to find a healthy line between taking the emotional stuff home and not being able to sleep and between being a misanthrope and a cold person." (Fifth-year female student) - + 9606 Species @@ -52211,7 +52841,7 @@ paragraph 8255 "Not only that we write in the patient's record that we will keep him for another week in the hospital because he has nowhere to go but also to understand his situation and to adjust the management and communication." (Fifth-year male student) - + 9606 Species @@ -52229,7 +52859,7 @@ paragraph 8516 Students of both study years recognized the importance of a respectful relationship towards coworkers and patients. - + 9606 Species @@ -52241,7 +52871,7 @@ paragraph 8632 "I cannot imagine the relationship my colleague will have towards patients if he is not able to say hi to me when we meet even though he has been my classmate for the last five years. " (Fifth-year female student) - + 9606 Species @@ -52253,7 +52883,7 @@ paragraph 8850 "To show respect to the patients by addressing them with sir or madam." (First-year female student) - + 9606 Species @@ -52283,7 +52913,7 @@ paragraph 9234 "...being responsible to your patients in a way to be able to help them as much as you can." (Fifth-year female student) - + 9606 Species @@ -52307,7 +52937,7 @@ paragraph 9496 The first-year students perceived autonomy as the right of patients to make their own decisions and also as their obligation for taking more responsibility. The fifth-year students referred also to the autonomy of the physicians. - + 9606 Species @@ -52319,7 +52949,7 @@ paragraph 9726 "I think that nowadays patients have way to much autonomy and very little responsibility and this is not right." (First-year female student) - + 9606 Species @@ -52331,7 +52961,7 @@ paragraph 9871 "...that you do not let the patient to control you i.e. to tell you which drug to prescribe, to set this line." (Fifth-year female student) - + 9606 Species @@ -52343,7 +52973,7 @@ paragraph 10015 "That the explanatory duty/informed consent does not become some kind of a blackmail and a form of persuasion that your truth is the only truth for this patient but that you only try to explain and present different treatment options." (Fifth-year female student) - + 9606 Species @@ -52367,7 +52997,7 @@ paragraph 10394 "I perceive professionalism as the relationship where patient's problems stay between you and him. That you do not talk about them outside your office." (First-year male student) - + 9606 Species @@ -52391,13 +53021,13 @@ paragraph 10750 Students perceived communication as an essential part of professionalism practice. They considered that patient should understand physician and also physician should understand patient. - + 9606 Species patient - + 9606 Species @@ -52409,7 +53039,7 @@ paragraph 10936 "It is important to now that nowadays patients read a lot and then they can mislead their physician that they understand something but they really don't. They just talk about something they've read somewhere and do not understand it." (Fifth-year female student) - + 9606 Species @@ -52421,7 +53051,7 @@ paragraph 11207 "I think that medicine has become a service activity ad people seek service when they go to the doctor. They sometimes forget how good it felt when physicians took the time and listened to them." (Fifth-year male student) - + 9606 Species @@ -52433,7 +53063,7 @@ paragraph 11433 "Professionalism is being able to communicate in such a way that the patient understands you. If he comes from low social class you communicate in a more lay language and if he comes from a high class you can communicate in a more sophisticate way." (First-year male student) - + 9606 Species @@ -52457,7 +53087,7 @@ paragraph 12005 "...if you take this home as a luggage then this can limit your ability to perform your work. Which also affects the patients." (Fifth-year female student) - + 9606 Species @@ -52487,7 +53117,7 @@ paragraph 12373 "...meaning that there is not any gossiping and malicious imputation in the team so you can focus on work. This is what patients deserve." (Fifth-year male student) - + 9606 Species @@ -52499,7 +53129,7 @@ paragraph 12542 "For example, the patients can see if there are some disputes within the team and this is a sign for them that they will not receive a quality management." (Fifth-year female student) - + 9606 Species @@ -52517,7 +53147,7 @@ paragraph 12747 Partner relationship with patients was only recognized by fifth-year students. - + 9606 Species @@ -52529,7 +53159,7 @@ paragraph 12826 "That you do not give the patient the feeling that he has no clue and that you do not place yourself above him." (Fifth-year female student) - + 9606 Species @@ -52541,7 +53171,7 @@ paragraph 12971 "I think that the patient has to feel as he has the possibility to participate in the decisions but that is our duty to present out professional opinion in such a way that he will agree with it and still think that it was solely his decision." (Fifth-year female student) - + 9606 Species @@ -52571,7 +53201,7 @@ paragraph 13466 "I think that this is a thing that people bring it from their childhood, from their families." (Fifth-year female student) - + 9606 Species @@ -52583,7 +53213,7 @@ paragraph 13593 "Some people always ignore those who for example fall on the street while others always stop and help. This is not something you could learn." (Fifth-year female student) - + 9606 Species @@ -52595,13 +53225,13 @@ paragraph 13768 "Professionalism largely depends on the personality. Some are more collegial and help their colleagues even now and lend them their notes while others keep the things to themselves. You know that they will be like that with their patients : they will not be willing to listen to anybody or to recognize their mistakes." (First-year male student) - + 651272 Species the things - + 9606 Species @@ -52643,7 +53273,7 @@ paragraph 14505 "Someone can be a great expert but has a very low professional relationship towards patients." (Fifth-year female student) - + 9606 Species @@ -52655,13 +53285,13 @@ paragraph 14632 "I think that there are also doctors who are not very professional but are great experts. They can diagnose a patient very good but are not considered to be good doctors by patients." (First-year female student) - + 9606 Species patient - + 9606 Species @@ -52697,13 +53327,13 @@ paragraph 15121 "I think that the medical profession can soon become inhuman. So, the humanistic part of it should be constantly a part of education. Otherwise, we can quickly start regarding patients as objects and not humans." (Fifth-year male student) - + 9606 Species patients - + 9606 Species @@ -52715,7 +53345,7 @@ paragraph 15364 "I think that you change a lot during your study. You start to hate people." (Fifth-year female student) - + 9606 Species @@ -52745,7 +53375,7 @@ paragraph 15722 "I still think that the health care system made the people to perceive the health care workers as their slaves or as salesmen in the store - they expect to get some kind of drug each time they go to the doctor." (First-year female student) - + 9606 Species @@ -52769,21 +53399,37 @@ paragraph 16576 As previously stated, there is no unified theoretical model of medical professionalism and it is therefore difficult to relate our results to the theoretical concepts. If we compare the medical professionalism categories recognized by the undergraduate students to the aforementioned theoretical models (Table 1), we can see that the main professionalism components have been recognized: altruism, autonomy, integrity, relationship, empathy and confidentiality. Of course, the wording and the terms of some components are sometimes different but the meaning is similar, i.e. objectivity and responsibility relate to altruism and autonomy relates to integrity. However, the students failed to recognize some professionalism components, i.e. continuous improvement, quality of care, access to care, distribution of finite resources and scientific knowledge. If we take a look at some studies that dealt with professionalism attitudes in undergraduate students, we can see that students recognized a very wide range of professionalism components and some of them are comparable to the categories in our study. For example, five out of seven components reported by Blackall et al were recognized in our study: accountability, altruism, duty, honour and integrity and respect. In another study, similar to our study, respect, relationship and partnership were recognized. Yet another study put forward confidentiality - similar to our study. However, the students in our study also recognized empathy, communication, team work and difference between professional and private life which other studies failed to recognize. - + + Disease + + empathy + + Disease altruism - + Disease altruism - + Disease altruism + + Disease + + empathy + + + MESH:D008569 + Disease + + wording + DISCUSS @@ -52796,12 +53442,37 @@ paragraph 19385 In this study, some differences were detected between the first-and fifth-year students' attitudes towards professionalism. First of all, some categories were recognized only by the fifth-year students: teamwork, partnership, role modelling and education. It seems that students do learn about professionalism through their medical studies. Also, they seem to grasp the importance of education in developing a figure of a professional physician. Additionally, there were some qualitative differences in perception of several professionalism components. Although the students of both study years recognized empathy as one of the components, first-year students expressed a very pragmatic attitude towards it whereas fifth-year students were concerned about how to cope with too much empathy. Similar findings were reported regarding drawing the line between personal and professional life. Several studies reported on the decline of empathy through the medical studies. Although our study was qualitative and hence the differences are hard to detect, our results suggest that the students become more sensible to empathy as they move towards the end of their studies. Based on the results of our study, we cannot claim that they become more or less empathic; we can only say that they become aware of potential harm empathy could have on their personal lives. This finding relates also to the finding about the autonomy. Namely, the fifth-year students also recognized the autonomy of the physician not only of the patient. - + 9606 Species patient + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + DISCUSS @@ -52866,7 +53537,7 @@ 143 22691 Incorporating professionalism into medical education: the Mayo Clinic experience - + 162683 Species @@ -52915,6 +53586,11 @@ 100 22921 Validation of Slovenian version of Jefferson scale of empathy for students + + Disease + + empathy + surname:Omona;given-names:K @@ -53008,12 +53684,18 @@ 241 23456 Improving the quality of care for patients with chronic diseases: what research and education in family medicine can contribute - + 9606 Species patients + + MESH:D002908 + Disease + + chronic diseases + surname:Meador;given-names:KG @@ -53230,11 +53912,10 @@ 41 24756 An empirical study of decline in empathy in medical school - - MESH:D029424 + Disease - - decline in empathy + + empathy @@ -53349,6 +54030,12 @@ abstract 134 The main aim of this research was to determine the influence of socioeconomic status and residence/living conditions on the status of oral health (e.g. health of mouth and teeth) in primary school students residing in Canton Central Bosnia. + + MESH:D012607 + Disease + + Canton Central Bosnia + ABSTRACT @@ -53361,30 +54048,24 @@ abstract 384 The study was designed as a cross-sectional study. Our research included two-phased stratified random sample of 804 participants. The quantitative research method and newly designed survey instrument were utilized in order to provide data on the oral health of the examined children. The alternate hypothesis foresaw that "there were significant statistical differences between the levels of incidence of dental caries in comparison to the incidence in children of different socioeconomic status. - + 9606 Species participants - + 9606 Species children - + 9606 Species children - - MESH:D003731 - Disease - - dental caries - ABSTRACT @@ -53397,18 +54078,24 @@ abstract 892 The Chi square of 22.814, degree of freedom (Df) = 8, coefficient of contingency of 0.163 and T-test (Stat) of-0.18334 showed that there were no significant statistical differences at p < 0.05 level between the primary school children from urban and rural areas. The obtained results showed that the caries indexes in elementary schools in Central Bosnia Canton were fairly uniform. Research showed that there were a difference in the attitudes towards a regular dental visits, which correlated with social-educational structure of the children's' families. - + 9606 Species children - + 9606 Species children + + MESH:D012607 + Disease + + Central Bosnia Canton + ABSTRACT @@ -53421,24 +54108,18 @@ abstract 1469 According to the results, we can see that the socioeconomic status of patients had an effect on the occurrence of dental caries and oral hygiene in patients in relation to the rural and urban areas, because we can see that by the number of respondents, the greater unemployment of parents in both, rural and urban areas, caused a host of other factors, which were, either, directly or indirectly connected with the development of caries. - + 9606 Species patients - + 9606 Species patients - - MESH:D003731 - Disease - - dental caries - INTRO @@ -53451,71 +54132,81 @@ paragraph 1923 Oral health (i.e. health of mouth and teeth) of school children represents a significant social and economic value of the modern world. Retrospective research data show that there is a difference in attitudes of examined children regarding the regular control of oral health, which is in correlation with social and educational structure of their families. The opinion that by practicing a regular preventive controls diseases of mouth and teeth can be prevented, obviously and unfortunately is not prevalent as life practice in many of primary school children's families Even though the children are exposed to the oral health at the fairly young age and although the parents are increasingly aware of the importance of the oral health for the entire well-being of their children, in many instances the role of parents and children in maintaining health of mouth and teeth is not sufficient, so the necessity arises to seek the professional help. It is best to seek help of dental professionals preventively, even then when everything seems to be well with the health of mouth and teeth. - + 9606 Species children - + 9606 Species children - + 9606 Species children - + 9606 Species children - + 9606 Species children - + 9606 Species children + + MESH:D009059 + Disease + + diseases of mouth + INTRO paragraph 3012 In 2005, as an answer to the rather poor state of primary school children's oral health in Federation entity of Bosnia and Herzegovina (F.B&H), within all of its ten Cantons, a promotional-preventive activities have been carried out with the main objective to familiarize the matriculated first grade children with the importance of proper oral hygiene and its correct application. This ambitious program was carried out in cooperation with Federal Ministry of Health, Federal Institute for Public Health, Federal Institute for Insurance and Re-insurance and primary health care institutions in F.B&H. In the academic school year 2004/2005 this program included 99.5 percent of all primary schools in F.B&H (i.e. 398 of central primary schools and majority of regional schools), while, at the same time education 89.2 percent of first grade children. In the academic school year 2006/2007 this program was additionally carried out to the first and second grade primary school pupils in Federation of Bosnia and Herzegovina. - + 9606 Species children - + 9606 Species children - + 9606 Species children - - MESH:D012211 - Chemical - - Re + + Disease + + Federation of Bosnia + + + Disease + + Bosnia @@ -53523,13 +54214,13 @@ paragraph 4036 The progress of the infection process is exemplified by the penetration into the tissue around the tooth's root and development of periodontitis. Each hole in the tooth represents an additional place for keeping food, plaque formation and propagation of a large number of bacteria. - + MESH:D007239 Disease infection - + MESH:D010518 Disease @@ -53541,7 +54232,7 @@ paragraph 4318 The treatment actually consists in detecting caries damage and removing softened dentin, followed by the appropriate treatment and disinfection of the oral cavity and eventually sealing. - + MESH:D003731 Disease @@ -53553,12 +54244,6 @@ paragraph 4505 While there are many theories about the origin of caries none of them give a complete answer to all the questions that come to mind when it comes to the etiopathogenesis of dental caries. According to contemporary views, some factors have greater importance in its origination, namely: - - MESH:D003731 - Disease - - dental caries - INTRO @@ -53583,7 +54268,7 @@ paragraph 4880 d) Carbohydrates. - + MESH:D002241 Chemical @@ -53595,13 +54280,13 @@ paragraph 4898 Dental plaque is of great importance, both in the development of caries, as well as in the pathogenesis of parondotopathy. It consists of desquamated epithelium of the oral mucosa, microorganisms and mucin (constituents of saliva). - + 100508689 Gene mucin - + Disease parondotopathy @@ -53612,13 +54297,25 @@ paragraph 5130 The accumulation of plaque on the tooth surfaces on which the physiological self-cleaning and cleaning with a brush is rather inefficient, is preceded and followed by caries. A special attention should be paid to microorganisms that are found in plaque, especially cariogenic strains of streptococci (Streptococcus mutans and sangus). Their biochemical activities carry out decomposition of carbohydrates to simple acids and synthesize different polysaccharides (i.e. dextran, levan and amylopectin). - + 1309 Species Streptococcus mutans - + + MESH:D003911 + Chemical + + dextran + + + MESH:D011134 + Chemical + + polysaccharides + + MESH:D002241 Chemical @@ -53636,7 +54333,7 @@ paragraph 5656 The study was designed as a cross-sectional study. The survey instrument was used for the survey interview and clinical examination of the mouth and teeth to provide data on oral health of the examined children. The survey was conducted in all units of observation, in selected schools in the stage of choice within these departments-selected groups of students. This also meant that two phases stratified sample of compatible groups, where 804 students were surveyed in elementary schools and where 355 students were from urban and 449 from rural schools. - + 9606 Species @@ -53660,12 +54357,6 @@ paragraph 6305 Members were familiarized with the available data on oral hygiene habits and their impact on oral health, various techniques and tools for proper oral hygiene, and tools and techniques for the determination of OHI (e.g. index hygiene oral). - - MESH:C489255 - Chemical - - OHI - METHODS @@ -53696,7 +54387,7 @@ paragraph 7150 Assessment of the socioeconomic status in children from urban and rural areas was carried out on the basis of the group questions in the application form, which were related to employment and qualifications of father / mother, the place of residence, and on the basis of their financial status. The Chi squared had the value of 22.814, the Level of Significance of P = 0.0001 and Contingency Coefficient of 0.163. - + 9606 Species @@ -53722,7 +54413,7 @@ paragraph 7745 No statistical significance was observed at p < 0.05, and therefore there is no statistically significant difference between urban and rural primary school children according to the socioeconomic status. - + 9606 Species @@ -53734,30 +54425,24 @@ paragraph 7949 From total amount of 72.37 percent of surveyed children listed that their father was employed, (i.e. 44.00 percent in urban and 28.37 percent in rural areas), 25.75 listed that their father was unemployed (i.e. 14.00 percent from urban and 11.37% from rural areas). Situation with mothers' employment status was quite different; whereas 52.5 percent of surveyed children listed (i.e. 22.25 percent in urban and 30.25 percent from rural areas) listed that their mother was employed, while 66.62 percent (i.e. 18.5 percent from urban and 48.125 percent from rural areas) listed that their mother was unemployed. 86.5 percent of examinees listed to reside in their own house or apartment (e.g. 34.37 percent in urban and 52.12 percent in rural areas). 40.50 percent of surveyed answered to have at disposal its own room, while 53.75 percent listed to share the room with brother or sister. By taking into consideration the answers regarding employment and socioeconomic status of examined children, where are clearly visible changed in rural and urban areas, we can ascertain that socioeconomic status (as an independent variable) was in causal relationship with the causation and development of dental caries and adverse changes in oral hygiene of examinees. Poorer socioeconomic status seems to have caused the incidence of the increasing number of examinees from rural areas who have visited dentist(s) more than four times; (i.e. 19.75 percent) in comparison to the examinees from the urban areas measured at 16.5 percent. At the same time there were not a single examinee residing in the urban areas who had reported to have visited a dentist more than ten times, while there were 1.5 percent of surveyed from the rural areas who visited a dentist more than ten times. - + 9606 Species children - + 9606 Species children - + 9606 Species children - - MESH:D003731 - Disease - - dental caries - FIG @@ -53802,7 +54487,7 @@ paragraph 11827 It is important to note that this factor had an impact on the status of oral hygiene in our patients, and that our sample consisted of a very large number of respondents who consumed sweets and snacks, each day and several times a day. Encouraging responses were in the forms of answers where the fruits were consumed more than three times in the last three days, although it was directly linked with socioeconomic status of respondents. - + 9606 Species @@ -53820,18 +54505,6 @@ children - - MESH:D003731 - Disease - - tooth decay - - - MESH:D003731 - Disease - - dental caries - DISCUSS @@ -53856,7 +54529,7 @@ paragraph 13208 Conditions of oral hygiene habits' among school children was on a satisfactory level and no large differences were observed between respondents in urban and rural areas; - + 9606 Species @@ -53874,48 +54547,30 @@ paragraph 13518 Factors that were predominant in relation to residence and caries were: eating poor quality food during the big break at school, and in urban areas more than in rural areas, one of the essential factors of consuming sweets and snacks was equally represented in all areas. Regular brushing was the factor that had an impact on the formation of cavities, but it could not be correlated with the geographic distribution. Taking diet that was supposed to improve tooth nutrition, such as drinking glasses of milk, gave no statistically significant results in both groups, but, rather favored the rural areas. - - MESH:D044342 - Disease - - tooth nutrition - CONCL paragraph 14127 According to the results, we can see that the socioeconomic status of patients was affected by the occurrence of dental caries and oral hygiene, in patients, in relation to their rural and urban residence areas, because we could observe that by the number of respondents and greater unemployment in both parents in rural areas than in the city, which caused a host of other factors that were directly and indirectly connected with the development of caries. - + 9606 Species patients - + 9606 Species patients - - MESH:D003731 - Disease - - dental caries - CONCL paragraph 14585 According to the type of food consumed we could see that in urban and rural areas the responses were fairly balanced, with the city where the larger number of respondents ate foods that posed a risk factors for tooth decay and deterioration of an oral hygiene than in rural areas, while the answers to the sample size and the use of a healthy diet were conditioned with the geographical position, (i.e. housing in rural areas where fruits and vegetables more available than in the city center). - - MESH:D003731 - Disease - - tooth decay - CONCL @@ -53944,7 +54599,7 @@ 3029 15128 Dental caries in an ecological perspective - + MESH:D003731 Disease @@ -53979,7 +54634,7 @@ surname:Higuera;given-names:C 15245 Plaque caries level and oral hygiene habits in young patients receiving orthodontic treatment - + 9606 Species @@ -54000,7 +54655,7 @@ 185 15339 Dental caries in children - + MESH:D003731 Disease @@ -54022,12 +54677,6 @@ 539 15365 A systematic review of dental recall intervals and incidence of dental caries - - MESH:D003731 - Disease - - dental caries - 44 @@ -54061,12 +54710,6 @@ surname:Longbotttom;given-names:C 15518 Defining dental caries for 2010 and beyond - - MESH:D003731 - Disease - - dental caries - 208 @@ -54129,12 +54772,6 @@ 367 15816 Nutritional advice of great importance to prevent dental caries - - MESH:D003731 - Disease - - dental caries - 141 @@ -54163,18 +54800,12 @@ 321 15899 Effective use of fluorides for the preventions of dental caries in the 21st century: the WHO approach - + MESH:D005459 Chemical fluorides - - MESH:D003731 - Disease - - dental caries - surname:Markovic;given-names:N @@ -54193,6 +54824,11 @@ surname:Ganibegovic;given-names:M 16001 The relationship between socioeconomic backgrounds, caries associated micro flora and caries experience in 12-year-olds in Bosnia and Herzegovina in 2004 + + Disease + + Bosnia + 117 @@ -54207,12 +54843,6 @@ 367 16155 Nutritional advice of great importance to prevent dental caries - - MESH:D003731 - Disease - - dental caries - 57 @@ -54228,12 +54858,6 @@ 414 16219 Social factors affecting health (2) Dental diseases - - MESH:D009057 - Disease - - Dental diseases - 114 @@ -54249,7 +54873,7 @@ 546 16271 Epidemiological aspects of frontal edentation of child and adolescent - + 9606 Species @@ -54306,12 +54930,6 @@ 84 16536 Nutritional status and prevalence of dental caries amon 12-year-old childer at public schools: a case - control study - - MESH:D003731 - Disease - - dental caries - 141 @@ -54328,12 +54946,6 @@ 539 16656 A systematix review of dental recall intervals and incidence of dental caries - - MESH:D003731 - Disease - - dental caries - 141 @@ -54465,12 +55077,6 @@ paragraph 1813 With the modernization of radiology departments, along with using electronic information systems such as PACS, RIS, speech recognition systems, preliminary report systems, hospital information systems, and electronic health records, workflow and informatics infrastructure supporting them became very complex. This is because it requires combining many computerized systems with different user environments to provide quality efficient services in less time. - - MESH:C508168 - Chemical - - PACS - INTRO @@ -54483,12 +55089,18 @@ paragraph 2675 Dashboards is an intellectual and visual tool for monitoring of KPI which can capture key data from different systems and represent them summarized and useful in real-time with easier reading and interpretation for users. This business intelligence tool can applied in radiology department as used in other sections such as operation rooms, emergency rooms, diabetes, and hospital infection control. - + MESH:D003920 Disease diabetes + + MESH:D056987 + Disease + + operation rooms + MESH:D003428 Disease @@ -54755,11 +55367,6 @@ paragraph 15963 About dashboards implementation, for achieve to a comprehensive set of dashboards that contain managerial and clinical operations of the radiology department So as to be able to create the accountability and strategic value in all aspects of financial, quality and clinical, the following technical infrastructure is necessary to be considered: - - Disease - - dashboards implementation - DISCUSS @@ -54802,7 +55409,7 @@ paragraph 16984 Utilizing interoperability standards such as HL7 messaging standards and DICOM to make connections and data exchange among different systems - + CVCL:2492 CellLine @@ -54893,12 +55500,6 @@ 21 18641 The radiology dashboards: A user's guide to a "high-performance" PACS - - MESH:C508168 - Chemical - - PACS - surname:Richardson;given-names:JS @@ -55004,17 +55605,17 @@ surname:Wakefield;given-names:DS 19090 A diabetes dashboards and physician efficiency and accuracy in accessing data needed for high-quality diabetes care - + MESH:D003920 Disease - - diabetes + + A diabetes dashboards - + MESH:D003920 Disease - - diabetes dashboards + + diabetes @@ -55032,12 +55633,6 @@ 167 19206 TOCSIN: a proposed dashboards of indicators to control healthcare-associated infections - - MESH:D007239 - Disease - - infections - REF @@ -55172,12 +55767,6 @@ 75 20010 Flying blind: using a digital dashboards to navigate a complex PACS environment - - MESH:C508168 - Chemical - - PACS - surname:Martin;given-names:DP @@ -55306,6 +55895,12 @@ 54 20612 Your business dashboards: Knowing when to change the oil + + MESH:D009821 + Chemical + + oil + surname:Gravel;given-names:JW;suffix:Jr @@ -55394,7 +55989,7 @@ 25 21066 Real-time analytics increase early discharges. Dashboards lowers cost per case, improves patient satisfaction and minimizes staff frustration - + 9606 Species @@ -55452,12 +56047,6 @@ ref 21445 A visual dashboards for moving health technologies from "lab to village" - - MESH:D014786 - Disease - - visual dashboards - REF @@ -56084,145 +56673,150 @@ paragraph 10141 The parent is the first and most important link in the upbringing of the child. For the child's emotional stability and proper development it is most important that parents establish proper communication with the child from its earliest years. A warm family environment in the home, harmonious parental relations, relationships among siblings and other family members (grandparents) contributes to the proper development of the child, without the stress and frustration. The aforementioned departure of a child to kindergarten is the first stress to the child's psyche, which is why parents need to be prepared to that milestone in a child's life. In the first place, it is essential that their positive attitude towards kindergarten and confidence in educators, shortening the stay of a child in kindergarten the first day, parent involvement in all activities related to kindergarten, establishing a precise rhythm of arrival and departure from the kindergarten, and in the interim period adaptations allow the child to the brings his/hers toys, pacifier, bottle, etc.., which remind the child to home. Period of adjustment of the child is different and depends on the age group, of course, to quickly adapt to children older groups. A sudden separation from home child interprets as treason by parents and often reacts with anger, disapproval and aggression, often regressive behavior (crawling, bedwetting, etc.). For all these options parent should be ready to answer them with stronger affection, conversation and rationalization, of course, to the extent that to the child is understandable. Before kindergarten, a child needs to prepare and get used to this act, an occasional visit to a kindergarten, observing children at play, talking with staff, to be able to eat independently and possibly self-dresser. It is useful to occasionally leave the child with her grandparents or people who kept the diet for a few hours or a whole day to get the child used to the occasional separation from parents. Also, it is very important to build a relationship of trust and mutual respect with educators, proper communication and collaborative partnership because they are for parents and educators directed towards a common goal-successful adaptation of the child in the new circumstances. - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species people - + 9606 Species child - + 9606 Species child - + MESH:D001523 Disease aggression + + Disease + + psyche + INTRO paragraph 12432 Educator is the next link in the educational process of the child. To educator belongs a fundamental and very responsible role, "surrogate parent" for parents employed because they due to the workload have less time to socialize with the child, which are increasingly leaving the work in kindergarten (full day, 10-hour programs). - + 9606 Species child - + 9606 Species @@ -56234,19 +56828,19 @@ paragraph 12767 Certainly in the educational process in the kindergarten equally affect the other participants: psychologist, pedagogue, special education teacher, speech therapist, and others who are important to the upbringing of the child (assistant, cooks, other staff), therefore all complementary to achieve complex missions in child-rearing. - + 9606 Species participants - + 9606 Species child - + 9606 Species @@ -56264,49 +56858,49 @@ paragraph 13151 As stated above word, verbal communication is the foundation of every communication, including with pre-school children, some of which are still learning to speak. Proper verbal communication seeks calm approach to child, educator's equidistance towards other children in kindergarten (not favoring one child, so the other would not feel neglected) and finally individual approach, if necessary (e.g., children with special needs). Apart from this, regular communication, according to psychological divisions, there are also non-verbal communication ("body language"), and paraverbal communication (mode of speech, facial expressions, tone of voice). Like adults, children are very well understood by all three forms of communication, so that the educator in her work has extremely careful in all three forms (in) direct communication with the child. Child communication seeks educator's attention, equal relationship and the need for him to tutor his full attention to the conversation, i.e. that it is not in any way neglected. - + 9606 Species children - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species children - + 9606 Species children - + 9606 Species child - + 9606 Species @@ -56318,13 +56912,13 @@ paragraph 14185 So, if the educator is occupied some other business (e.g. preparation of workbooks, or other activities) and the child spoke, educator should stop working and devote its full attention. This means, not with some other work, it is necessary to look into the eyes of a child, to patiently hear its question or comment. - + 9606 Species child - + 9606 Species @@ -56336,13 +56930,13 @@ paragraph 14502 Also, body language-gestures that betray impatience (tapping a pencil, interrupting the conversation, watching over the child, non-listening...) and the desire to talk or request as soon as possible, lead to poor communication between educators and child. - + 9606 Species child - + 9606 Species @@ -56354,13 +56948,13 @@ paragraph 14760 Finally, the raised voices in conversation with the child, threatening expression (paraverbal communication) at the start eliminate child's desire to communicate, to which particular must watch every educator. - + 9606 Species child - + 9606 Species @@ -56372,19 +56966,19 @@ paragraph 14970 It should be noted that the child during the entire stay in the kindergarten assess teacher's person, personality, way of communicating, and vice-versa, in a similar way to educator evaluate children in behavior and adoption of knowledge. - + 9606 Species child - + 9606 Species person - + 9606 Species @@ -56396,49 +56990,49 @@ paragraph 15209 The most important part of a successful adaptation of the child to kindergarten makes teachers. During the period of adjustment child educator needs to know, as a group, parents and children, recognize the individual needs of each child. Also, as a token of good communication relations educators need to be accepted by the child, through their activities, providing support to the child and parents. Educators monitor, observe the child during his stay in kindergarten take notes, plan activities and design child room to learn, play and other activities. - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species @@ -56450,61 +57044,61 @@ paragraph 15766 What is the key moment in relation educator-child is that the everything that child is going through in kindergarten transmits to parents, or independently of its, spontaneous narratives, parents questioned in detail about (every)day stay in kindergarten and events (the ratio of educators to child, other children, food hygiene, etc.). Follow every move, every educator's raised voice, or some other form of punishment of the child (rough handling, pushing, pulling hair, etc..) Which may lead to very poor communication form educator-child. From such, disturbed, relations subsequently followed poor communication parent-educator, because any change in the relationship between child communications signals to parents. Of course, the relationship between educators and parents (the child's testimony parents often exaggerates!) Two vital components in preschool, thereby permanently disrupted. According to the findings of misfortunate events in kindergarten, and after talking with the child, the parents immediately, ultimately the director of the institution seeking punishment or removal of educators for the sake of the child's testimony. - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species @@ -56516,13 +57110,13 @@ paragraph 16914 The above triad is the subject of numerous studies, psychological, pedagogical, as well as in the field of communication sciences, and they all agree with the fundamental postulates of communication, which in relation to the child has its specificities. So, parents and educators, spending time with your child should adhere to the following postulates: - + 9606 Species child - + 9606 Species @@ -56534,13 +57128,13 @@ paragraph 17268 Talk with child-teach introvert child to talk, encourage it to conversation, avoid conversation with the answers yes and no. - + 9606 Species child - + 9606 Species @@ -56552,13 +57146,13 @@ paragraph 17397 Call child by name-if you call them by the name, the child interprets this intimacy with a sense of responsibility, contrary to anonymous communication which means indifference. - + 9606 Species child - + 9606 Species @@ -56570,7 +57164,7 @@ paragraph 17581 Look child in the eyes: a direct sight means that you are interested in it (not too long, because fixed look achieve the opposite effect, watching manner means indifference or its absence). - + 9606 Species @@ -56582,7 +57176,7 @@ paragraph 17775 Touch of a hand, touch child's hair: intimacy, commitment, "reward" for a completed task. - + 9606 Species @@ -56624,7 +57218,7 @@ paragraph 18362 Point out errors, teach the child conversation. - + 9606 Species @@ -56636,7 +57230,7 @@ paragraph 18414 If you mad tell him that you are sorrowful and sad because he did, do not allow yourself to be provoked or shout at a child. - + 9606 Species @@ -56654,29 +57248,29 @@ paragraph 18618 Although Chinese proverb (quoted at the beginning of the text) says that children are telling the truth, we know from experience that it cannot be true. Reason false accusations may be because of relations between parents, (the so-called. Said syndrome sexual allegations in divorce) but also false allegations that the child expresses (invent). What happened was that children of parents accuse educators for (not) work that the same is not really so. - + 9606 Species children - + 9606 Species child - + 9606 Species children - - MESH:D012735 + + MESH:D016097 Disease - - syndrome sexual allegations + + Said syndrome sexual allegations @@ -56684,25 +57278,25 @@ paragraph 19071 Example. (Statement of an educator). "Peter was lively and happy child often would come in my lap. He was not always willing to fulfill the tasks, but with a little persuasion he gets them done. One morning he came with his father, corpulent worker from shipyard and very rudely attacked me, because if I had yesterday beaten child. I told him it was not true, and that such a thing has never happened in our kindergarten, but the father was insistent, saying that his Peter never lies. Immediately I called Peter and asked him in front of his father: "Did I beat you yesterday?" He embarrassingly said that I did not, and that he invented it so he does not have to do a task that for him was so hard. Parent immediately pounced the child, but now with a story that is a little liar and that his house does not believe anything. " - + 7939 Gene lap - + 9606 Species child - + 9606 Species child - + 9606 Species @@ -56714,73 +57308,85 @@ paragraph 19910 In some cases the child, out of fear, hides a traumatic event in kindergarten fearing retaliation of the educators. Example: (testimony of parents) ... My Igor, a boy with signs of mild MCD, which required a special, individual approach because it is often obstinately persisting in their demands. Educator obviously had no patience with him, because the lady who earlier came by my child said that he is an educator led him to the bathroom and well beaten because he in a loud voice yelled. I have noticed that since that day my son did not want to go to kindergarten, he was afraid, he was crying but he would not say why, I guess out of fear. Of course ... I'm also reported the director of the kindergarten "... - + 9606 Species child - + 9606 Species boy - + 9606 Species child - + MESH:D012514 Disease MCD + + MESH:D014947 + Disease + + traumatic + INTRO paragraph 20630 These cases are sporadic, rare, but should be assiduously investigated, because the truth always eventually came to light. For example after talks with parents who report changes in child behavior (frustration, fear, anxiety ...) followed by psychological testing (processing, drawing, etc..) That reveal educators / woman who got lost in the water for their upbringing and that systematically punish children in kindergarten. These cases are rare, but there are some, an example of a case where educators are parents set up a hidden camera that has discovered what all the kids did and how they were punished. - + 9606 Species child - + 9606 Species woman - + 9606 Species children - + MESH:D001008 Disease anxiety + + MESH:D014867 + Chemical + + water + INTRO paragraph 21242 However, in most cases by children honestly convey to parents all events, was pleasant and unpleasant, and the knowledge and application relevant to kindergarten every case non pedagogic work with the child should be thoroughly investigated and, if necessary, sanctioned. - + 9606 Species children - + 9606 Species @@ -56792,31 +57398,31 @@ paragraph 21514 Without a doubt, these situations are extreme, because the vast majority of educators do their job responsibly, and finally themselves have chosen the honorable invitation educators. Therefore, any case report violations of the child's integrity and personality (login parents or third parties, public or anonymously) should be accepted, checked thoroughly investigated and, if necessary, conduct psychological testing or other actions that would not have occurred cases which, needless, throw filth the honorable job of educators. A child who is developing normally, during his stay in kindergarten, collective, rapidly improves your vocabulary, speech impediments, talking with other children and with the teacher. However, the unconscious child can take on some negative qualities of speech and facial expressions (drawl, mispronounced words, tics) of children with these problems, imitating them. - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species @@ -56828,25 +57434,25 @@ paragraph 22415 Example: Mirko child from highly educated family, whose parents are alternately brought and taken away from the nursery. (testimony of parents)... "In the beginning was staying in kindergarten flowed really well, Mirko quickly adapted and played with children. The problem arose when we noticed that Mirko weird talking, delaying and modulating voice in a special way, which has never before been done. I'm a little bothered, questioned and found that Mirko most endeared and hung out with the boy John, (a boy with Down syndrome), which, when we checked spoke in the same manner as is now saying our Mirko! " - + 9606 Species child - + 9606 Species children - + 9606 Species boy - + 9606 Species @@ -56858,13 +57464,13 @@ paragraph 23031 All these are the moments that must be observed educator who, as a corrective, talking with parents, seeks to correct the child's behavior, to direct it and to give special attention to such children. Because once mistakenly learned, it is difficult to correct. Kids play, learn unencumbered through the game, absorbing every word of educators, even if they do not know the true meaning of the word, but according to the sense and context are decrypted. - + 9606 Species child - + 9606 Species @@ -56876,37 +57482,37 @@ paragraph 23485 Every word spoken, remembered the child may return like a boomerang, if a parent mechanically in a situation or someone send a curse, the child will remember and reproduce in approximately similar situation (anxiety, anger, resentment). Child resonates: If dad (or mom) so saying it is wrong and can also say in front of other people. So can it happen in a completely unmanageable situation juicy cursed child, even if you do not know the meaning of swearing, because it is a situation where it occurs associated with parental cursing in a similar situation. - + 9606 Species child - + 9606 Species child - + 9606 Species Child - + 9606 Species people - + 9606 Species child - + MESH:D001008 Disease @@ -56918,14 +57524,14 @@ paragraph 24044 Example 1: (testimony of parents) "The car journey with us was a little Ida, in her chair tied in last place. While driving in front of us carelessly, high speed from the side street crashed driver who almost hit us. I admit I slipped a juicy curse, but it all ended well. Next time driving, when I slammed on the brakes, Ida repeated cursing in the same way as I did I previously spoke". - + 3425 Gene 170 Ida - + 3425 Gene 170 @@ -56938,13 +57544,13 @@ paragraph 24437 Example 2 (testimony of parents). "We looked at (wife, little Emma) TV with an entertainment show in which he participated in one of our show biz stars be rampant. I quietly (not to hear me daughter!) Told the woman to switch the channel because I cannot watch that bitch..... Ok, a woman had done a few days back on TV appeared the same stars from the entertainment industry. Emma immediately cried loudly: There goes that son of a ... immediately shut down the TV." - + 9606 Species woman - + 9606 Species @@ -56956,13 +57562,13 @@ paragraph 24909 This will also happen in relation to the teacher: child will house use phrases that he heard from educators, was in communication with him or with other children. - + 9606 Species child - + 9606 Species @@ -56974,182 +57580,194 @@ title_1 25072 6. IMPORTANCE OF PROPER COMMUNICATION WITHIN THE FAMILY + + MESH:D001929 + Disease + + WITHIN + INTRO paragraph 25128 Immeasurable importance of proper communication relation child and parents. Famous aphorism Earl of Rochester's (I had six theories about raising children when I had no child, now I have six of them and no theories ...) confirms this. Parents with a debut child learn communication and education on the forms that they experienced in their youth when they were raised by their parents. Parents who grew up as an only child, unconsciously accepted patterns of his youth, and almost identical to convey the paradigm of education to their child. Parents who grew up in a large family with different forms of education and communication with the child. What is now clear, the economic situation in the society, parents are reluctant to have another child, let alone more: partly out of conformism, but more for economic reasons. In families with one child, a kindergarten is a surrogate for a large family, and as such is necessary in the education of an individual, especially as parents occupied a job cannot be adequately measured participate in the upbringing of the child. Of course it is necessary nursery with large families in which the parents work, and the parents were all agree that education is the first child toughest. This is partly from inexperience and partly because of a family with one child unconsciously, exaggerated care, such children spared all possible situations in which a child could possibly be the endangered (e.g. Child is not sent to the nursery or day care due to the risk of transmission of infections, keeping the cold, rain, presvalacenje several times a day, especially feeding, mixed foods, avoiding the society of other children in the building where they live or work outside of a possible infection, avoidance of vaccination for the possible consequences, avoid rooms which housed more people, etc ...). 14 Because over care and isolation of the child, in such families are left only for parents, lonely, eager to socialize with other children. Company other kids did milieu in which the child is developing normally, realizes communication relationships with children their own age, and through the game with the children a feeling of satisfaction and happiness. Children who are only children in the family, "condemned" the only parents that if he wanted to, I cannot provide what the child wants, subconsciously are lonely and dissatisfied, because it is the day in the house monotonous and uneventful, until now the prevailing mood state. All knowledge about the life of children from their parents and in the best case of the second generation (today's increasingly rare!) Grandparents, who take the role of the nursery. So evident is the discrepancy between the generations, the generation gap, it is questionable as a good solution in education of a child. That is why the kindergarten indeed the solution for their situation, the postulation of kindergarten (kindergarten), which was established in 1840 by Frobel and remains very topical today, of course, in the new, modern conditions. - + 54922 Gene 9847 rain - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species Child - + 9606 Species children - + 9606 Species people - + 9606 Species child - + 9606 Species children - + 9606 Species child - + 9606 Species children - + 9606 Species children - + 9606 Species Children - + 9606 Species children - + 9606 Species child - + 9606 Species children - + 9606 Species child - + + MESH:D007239 + Disease + + infections + + MESH:D007239 Disease @@ -57222,7 +57840,7 @@ ref 28465 The ecology of human development: Experiments by design and nature - + 9606 Species @@ -57243,7 +57861,7 @@ surname:Moorehouse;given-names:M 28532 Interacting systems in human development. Research paradigms: Present and future - + 9606 Species @@ -57263,7 +57881,7 @@ 7 28613 Including Parents in Evaluation of a Child Development Program: Relevance of Parental Involvement - + 9606 Species @@ -57318,13 +57936,13 @@ 481 28910 Is the child victim of sexual abuse telling the truth? - + 9606 Species child - + MESH:D012735 Disease @@ -57341,7 +57959,7 @@ 23 28965 True and False Allegations of Child Sexual Abuse: Assessment and Case Management - + 9606 Species @@ -57375,7 +57993,7 @@ 830 29064 A 12-Year Prospective Study of the Long-term Effects of Early Child Physical Maltreatment on Psychological, Behavioral, and Academic Problems in Adolescence - + 9606 Species @@ -57456,16 +58074,16 @@ Mouse - MESH:D012090 + MESH:D010146 Disease - - Occupational Overuse Syndrome + + Pain - MESH:D010146 + MESH:D012090 Disease - - Pain Syndrome + + Occupational Overuse Syndrome @@ -57515,79 +58133,73 @@ paragraph 1398 Computers are considered as an integral part of everyday life in today's work and life. They are used in a wide variety of professions from banking, health, communication to entertainment and leisure time. "Thanks to the" great use of computers, occurs the diseases of the modern era, which one of the authors of this article named "Technological diseases" - Carpal Tunnel Syndrome (CTS), a Mouse Shoulder (MS) and Cervical Pain Syndrome (CPS). Within past a few decades rapidly increased automation of offices and very few authors in the scientific literature have reported about positive association between computer use and musculoskeletal symptoms as consequences. Under term of"Technological diseases" we mean the diseases caused by the harmful influence of the job in the first place, the working position in which it is involved excessive work on a computer, such as the banking sector, the health sector and many others who are directly or indirectly connected with the work on the computer and overuse of keyboard and computer mice, which are in large number of cases of non-ergonomic, and inappropriate for a hand. It is therefore recommended to use ergonomic computer mice. Using them can prevent diseases of the modern era, technological diseases associated with prolonged sitting and working on a computer such as Carpal Tunnel Syndrome, Cervical syndrome, and certainly the most interesting for the many unknown, but not less important disease, mouse shoulder an increasingly common diagnosis in clinics of family medicine. These diseases are described in more detail in this paper. In Bosnia and Herzegovina to overuse injuries has not paid enough attention from the medical nor the social and economic aspects, although it is evident that large amounts of funds are allocated for treatment and rehabilitation of these diseases. In Clinical Center University of Sarajevo in the cabinet for EMNG of the Neurology clinic, monthly are diagnosed 10 cases with carpal tunnel syndrome, while cases of cervical syndrome is in a significant expansion during last 5 years which more and more represent not only a medical but also social economic problem. - + 7276 Gene 317 CTS - + 10090 Species Mouse - + 10090 Species mice - + 10090 Species mice - + 10090 Species mouse - + MESH:D002575 Disease cervical syndrome - + MESH:D020165 Disease CPS - + MESH:D019547 Disease Cervical Pain - + MESH:D002349 Disease carpal tunnel syndrome - + MESH:D012090 Disease Bosnia and Herzegovina to overuse injuries - + MESH:D002575 Disease Cervical syndrome - - MESH:D009140 - Disease - - musculoskeletal - INTRO @@ -57600,46 +58212,46 @@ paragraph 3599 Carpal tunnel syndrome (CTS) is the most common canaliculus syndrome resulting from pressure on the central nerve (median nerve) in the carpal tunnel (lat. canalis carpi) and reflected in reduction of sensation, pain, paresthesia (numbness, tingling), and muscle weakness in the hands and forearms. - + 27040 Gene 7811 lat - + 7276 Gene 317 CTS - + MESH:D018908 Disease muscle weakness - + MESH:D006987 Disease numbness - + MESH:D010146 Disease pain - + MESH:D002349 Disease Carpal tunnel syndrome - - MESH:D061325 + + MESH:D013577 Disease canaliculus syndrome @@ -57650,26 +58262,20 @@ paragraph 3898 The median nerve in its course passes through the carpal canal (Figure 1). The bones of the hand (lat. ossa carpi) make gutter channels, and rectangular connection (lat. retinaculum flexorum) covers the channel. In the channel along with the nerve are located tendon flexor muscle of fingers, m. pronator teres, m. pronator quadrates, m. flexor carpi radialis, m. palmaris longus, m. flexor carpi ulnaris, m. flexor digitorum superficialis et profundus and m. flexor policis longus. All these muscles supplying the median nerve. Carpi ulnaris M.flexor the inner half, m. Flexor digitorum profundus is innervated by n. ulnaris. - + 27040 Gene 7811 lat - + 27040 Gene 7811 lat - - MESH:D007645 - Disease - - palmaris longus - FIG @@ -57684,50 +58290,55 @@ paragraph 4553 Carpal tunnel syndrome (CTS) was first described in the mid-1800 by James Paget's (Figure 1). The world's best known as peripheral compressive neuropathy and one of the fastest growing technology diseases in the United States. At least 1 out of 10 people develop this disorder or suffering from the symptoms of this syndrome. This is one of the most common causes of absenteeism in the workplace and disability in the United States - + 7276 Gene 317 CTS - + 9606 Species people - - MESH:D009422 + + MESH:D010523 Disease - - neuropathy + + peripheral compressive neuropathy - + MESH:D002349 Disease Carpal tunnel syndrome + + Disease + + absenteeism + INTRO paragraph 4986 Signs and symptoms are: a) pain in the palm of the hand, especially near the thumb and first two fingers; b) numbness and tingling often occur in the thumb and first two fingers; and c) weakness, usually thenar eminence (muscles at the base of the thumb), can occur when a severe clinical picture (Figure 2). - + MESH:D006987 Disease numbness - + MESH:D018908 Disease weakness - + MESH:D010146 Disease @@ -57747,25 +58358,25 @@ paragraph 5345 Compression of the nerve in the carpal tunnel (carpal tunnel syndrome) is recognized as the impact of trauma and attributed to tenosynovitis of the rheumatoid arthritis. Even since 1954, Albert and his colleagues concluded that there are numerous reports describing the spontaneous development of carpal tunnel syndrome of unknown pathogenesis. - + MESH:D014947 Disease trauma - + MESH:D001172 Disease tenosynovitis of the rheumatoid arthritis - + MESH:D002349 Disease carpal tunnel syndrome - + MESH:D002349 Disease @@ -57777,25 +58388,25 @@ paragraph 5690 In case of a longer duration of illness and long-term nerve compression, there is an unnecessary prolongation of the patient discomfort, severe nerve damage, prolonged recovery time after surgery, and generally reduced chances for successful surgical treatment. Thereby reducing the working ability of the patient, leading both to the individual and to the socioeconomic consequences. - + 9606 Species patient - + 9606 Species patient - - MESH:D003389 + + MESH:D004194 Disease nerve damage - + MESH:D002908 Disease @@ -57807,38 +58418,38 @@ paragraph 6075 At the Department of Orthopedic Surgery, Faculty of Medicine, University of Zagreb, and Zagreb University Hospital Center is conducted a retrospective study on 114 patients and 154 hands with CTS that were surgically treated between 1999 and 2004. In this study, it was found that patients in only 52% of cases within one year since reporting to the physician were referred to an orthopedist. In the period prior to surgical treatment, 96.1% of patients were treated by physical therapy, although all eventually underwent a surgical procedure, and 42% of patients in the same time period it was on sick leave, usually from three to six months. The results demonstrate that the timeliness of surgical treatment is essential to the success of the outcome of the procedure, and that the time which elapses from entering the patient's health care system to the surgical treatment of unnecessary cost. - + 7276 Gene 317 CTS - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species @@ -57850,24 +58461,29 @@ paragraph 6972 This situation, besides being detrimental for the patient, and leads to unnecessary economic losses include the expense of physical therapy and absenteeism. The patients involved in the study in question is estimated that these economic losses, in the period prior to surgical treatment, for several hundred thousand Euros higher than in the postoperative period. - + 9606 Species patient - + 9606 Species patients - + 9319 Species Euros + + Disease + + absenteeism + INTRO @@ -57892,54 +58508,44 @@ paragraph 7697 In the diagnosis of CTS we also use provoking tests of which we will mention four that are suitable for use in general practice. These are the Tinel, Phalen, Bilic and Tourniquet test. Tinel's test is positive if the percussion in the carpal ligament is followed with occurrence of pain and/or paresthesia. Phalen's test is performed so that the wrist is placed in the second forced palmar flexion for 60 seconds, and is positive if within this time span occurs pain and/or parasthesias (Figure 2). Compression of median nerve in this position with carpal tunnel syndrome causes the characteristic symptoms of burning and tingling. - + 7276 Gene 317 CTS - + MESH:D010146 Disease pain - - Chemical - - Phalen - - + MESH:D002349 Disease carpal tunnel syndrome - + MESH:D010146 Disease pain - - Chemical - - Phalen - INTRO paragraph 8329 Bilic test is performed in palmar flexion of the wrist by 45 degrees by pressing above the median nerve in the distal ridges and mid wrist for 30 seconds. The test is positive if within this time span occurs pain and/or paresthesia. Tourniquet test is performed so that the cuff pressure gauge is mounted on the upper arm and inflated above the systolic pressure value. The test is positive during 60 seconds occurs pain and/or paresthesia in the innervation area of median nerve. - + MESH:D010146 Disease pain - + MESH:D010146 Disease @@ -57951,7 +58557,7 @@ paragraph 8810 Alongside this symptom, electrical neurophysiological diagnostic is the"gold standard" in the diagnosis of carpal tunnel syndrome, in order to determine the exact location of nerve compression, and in order to set the indication for surgical treatment (Figure 3, 4, 5). - + MESH:D002349 Disease @@ -57987,19 +58593,13 @@ paragraph 9226 In the case of known primary disease that caused Carpal tunnel syndrome the primary disease is treated first. The conservative methods of treatment include immobilization, local application of corticosteroids and avoidance of chronic, repeated trauma. If the cause of the syndrome is mechanical compression, the treatment is surgical (cutting of retinaculum flexorum). - - MESH:D000305 - Chemical - - corticosteroids - - + MESH:D014947 Disease trauma - + MESH:D002349 Disease @@ -58011,37 +58611,37 @@ paragraph 9595 In Turkey was conducted a study that suggests that treatment with vitamin B6 improves clinical symptoms as well as sensory and electro diagnostic results in patients with carpal tunnel syndrome, and therefore it is advisable to treat carpal tunnel syndrome with vitamin B6. - + 9103 Species Turkey - + 9606 Species patients - + MESH:D002349 Disease carpal tunnel syndrome - + MESH:D002349 Disease carpal tunnel syndrome - + MESH:D025101 Chemical vitamin B6 - + MESH:D025101 Chemical @@ -58053,19 +58653,19 @@ paragraph 9869 Another study in Turkey was carried out at the Institute of Neurology in cooperation with the Clinic for Physiotherapy in which patients were included with carpal tunnel syndrome. They are divided into two groups, one in the treatment arm used immobilization and massages, while the other control group used only immobilized arm. - + 9103 Species Turkey - + 9606 Species patients - + MESH:D002349 Disease @@ -58077,13 +58677,13 @@ paragraph 10199 The study results showed that the massage of the painful area only in patients with carpal tunnel syndrome is sufficient for mild cases. - + 9606 Species patients - + MESH:D002349 Disease @@ -58095,19 +58695,19 @@ paragraph 10336 This was also one of the biggest studies with massage treatment in patients with carpal tunnel syndrome. The researchers recommend self-administration of massage methods, and this would reduce the time to go to the doctor and rehabilitation centers, and thereby save money, both for health funds, and patients. - + 9606 Species patients - + 9606 Species patients - + MESH:D002349 Disease @@ -58125,41 +58725,47 @@ title_1 10834 3. CERVICAL PAIN SYNDROME - - MESH:D009477 + + MESH:D000699 Disease PAIN + + MESH:D002575 + Disease + + CERVICAL + INTRO paragraph 10860 Cervical pain syndrome (CPS) refers to a range of disorders caused by changes in the cervical spine and soft tissue surrounding it, with pain as the predominant symptom. Neck pain is a common problem for a large portion of today's population. - + MESH:D019547 Disease Neck pain - + MESH:D020165 Disease CPS - + MESH:D010146 Disease - - pain + + pain syndrome - - MESH:D019547 + + MESH:D010146 Disease - - Cervical pain syndrome + + pain @@ -58173,23 +58779,35 @@ paragraph 11366 Anatomical specificity of the cervical spinal column are: a small side vertebral joints that cervical spine provide great mobility, transversaria foramina through which the vertebral artery (C6-C2) and the spinal cord enters in the vertebral canal. - - MESH:C023714 - Chemical + + OMIM:217000 + Disease C6-C2 + + MESH:D007592 + Disease + + small side vertebral joints + INTRO paragraph 11615 Cervical radiculopathy causes symptoms that radiate from the neck. Although the problem is in the spine, the symptoms can be felt in the shoulder or arm. Symptoms will be felt in the area where the nerve is in its path. - + MESH:D011843 Disease - - Cervical radiculopathy + + radiculopathy + + + MESH:D004194 + Disease + + radiate @@ -58198,17 +58816,17 @@ 11835 By clinical examination of the spine the specialist can usually determine which nerve is involved. Symptoms include pain, stiffness and weakness. Reflexes on the upper arm may be affected.. - MESH:D018908 - Disease - - weakness - - MESH:D010146 Disease pain + + MESH:D018908 + Disease + + stiffness and weakness + INTRO @@ -58271,25 +58889,19 @@ paragraph 12482 In the younger population, cervical radiculopathy is the result of disc herniation or acute injury. In the older population, cervical radiculopathy is often the result of foramen narrowing and the formation of osteophytes then reduced the amount of disk and degenerative changes in the joints. - + MESH:D058186 Disease acute injury - + MESH:D011843 Disease - - radiculopathy - - - MESH:D054850 - Disease - - osteophytes + + cervical radiculopathy - + MESH:D011843 Disease @@ -58301,7 +58913,7 @@ paragraph 12776 Cervical syndrome classification: - + MESH:D002575 Disease @@ -58313,43 +58925,43 @@ paragraph 12810 CERVICAL SYNDROME patients complain of pain in the neck that occurs gradually or abruptly, after taking a certain position, rapid or uncontrolled movements of the neck or cooling. - + 9606 Species patients - + MESH:D010146 Disease pain + + MESH:D002575 + Disease + + CERVICAL + INTRO paragraph 12990 CERVICOCEPHALIC SYNDROME usually develops as a result of irritation or compression of the nerve roots C1-C3. The main symptom is headache of varying character and intensity. Headache is localized in the neck with a tendency to spread to the shoulders, the face, orbital or auricular region. - - MESH:D004428 - Disease - - auricular - - + MESH:D006261 Disease Headache - + MESH:D006261 Disease headache - + MESH:D001523 Disease @@ -58361,14 +58973,19 @@ paragraph 13281 CERVOCOBRACHIAL SYNDROME result is irritation or compression of the nerve roots C4-C8, mostly due to prolapse of the intervertebral discus, severe degenerative changes or injury. - + MESH:D001523 Disease irritation + + Disease + + CERVOCOBRACHIAL + - MESH:D014947 + MESH:D058186 Disease injury @@ -58386,7 +59003,7 @@ irritation - MESH:D014652 + MESH:D000783 Disease vascular disorders @@ -58397,17 +59014,28 @@ paragraph 13654 Irritation commonly caused by degenerative changes in the cervical spine: spondylosis (osteophytes), uncarthrosis, spondylosis. Atherosclerosis can worsen symptoms. - + MESH:D050197 Disease Atherosclerosis - + MESH:D055009 Disease - - spondylosis (osteophytes), uncarthrosis, spondylosis + + spondylosis + + + Disease + + uncarthrosis + + + MESH:D055009 + Disease + + spondylosis @@ -58415,31 +59043,31 @@ paragraph 13819 The typical clinical presentation of patients with cervical syndrome is characterized by the presence of pain and sensitivity in the muscles back of the neck with the spread of pain in the back of the head, shoulders or scapular region (Figure 6). The sensitivity of the muscle can occur during execution of one or more movements, and the headache is a common symptom. - + 9606 Species patients - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - + MESH:D006261 Disease headache - + MESH:D002575 Disease @@ -58451,66 +59079,66 @@ paragraph 14188 Symptoms and signs are: a) pain in the neck, which may be sharp or blunt; b) the tension in the neck; c) painful and limited mobility; d) inability to perform daily duties due to stiffness in the neck; e) pain in the shoulders and arms associated with pain in the neck; f) fainting; g) dizziness; h) tinnitus; i) blurred vision; j) headache; k) diplopia; l) weakness; m) feeling of heaviness, tenderness and paresthesia in the upper extremities; n) impaired concentration and memory, etc.. - + MESH:D010146 Disease pain - + Disease tenderness - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - - MESH:D009207 + + MESH:D008595 Disease - - upper extremities + + heaviness - + MESH:D004244 Disease dizziness - + MESH:D004172 Disease diplopia - + MESH:D014786 Disease blurred vision - + MESH:D018908 Disease weakness - + MESH:D014012 Disease tinnitus - + MESH:D006261 Disease @@ -58522,19 +59150,19 @@ paragraph 14678 Diagnostic imaging such as magnetic resonance imaging, computerized tomography, or myelography should be used as a strategy for assessment. Electromyography is useful in differentiating between the various entities when distinguishing clinical diagnosis. Treatment of this disorder has not been studied systematically in a controlled manner. However, using a variety of treatments, radiculopathy usually improves without the need for surgery. Indications for surgery were persistent pain, increased weakness, or new or progressive process. Future studies evaluating different treatment options will be helpful in guiding practitioners towards optimal economic evaluation. - + MESH:D018908 Disease weakness - + MESH:D011843 Disease radiculopathy - + MESH:D010146 Disease @@ -58546,19 +59174,19 @@ paragraph 15350 The goal of treatment is absolutely the same as for any other illness. Always seeks the same goal, or cure, and if this is not possible, at least alleviate symptoms and apply palliative care. Since we are talking about a technological disease that affects more and more to older and younger people should therefore work on the prevention of disease. It is very important to advise patients about lifestyle, how to maintain mobility and that despite the difficult situation improve the quality of life. - + 9606 Species people - + 9606 Species patients - + MESH:D002908 Disease @@ -58570,7 +59198,7 @@ paragraph 15852 Treatment without medicationshould be: a) massage hot/cold; b) rest; c) exercises with light stretching (gently stretch your neck to one, then the other aside and keep 30 seconds); d) exercises for neck recommended by physiologists; e) transcutaneous electrical stimulation (TENS); f) short-term immobilization; g) surgical treatment; h) acupuncture; i) traction (enlargement of the intervertebral space), j) medicamentous treatment: - + 137872 Gene 5865 @@ -58583,49 +59211,43 @@ paragraph 16286 Early and adequate treatment with analgesics, anti-inflammatory drugs, muscle relaxants and help with sleep problems are carriers of pharmacological treatment in patients with neck pain. - + 9606 Species patients - + MESH:D019547 Disease neck pain - - MESH:D012893 - Disease - - sleep problems - INTRO paragraph 16473 Performed is a study on how the steroid injections affect the reduction of pain in patients with cervical syndrome, who are waiting for surgery discus, and whether the possible suspension of surgical treatment. - + 9606 Species patients - + MESH:D002575 Disease cervical syndrome - + MESH:D013256 Chemical steroid - + MESH:D010146 Disease @@ -58637,25 +59259,25 @@ paragraph 16684 Transforaminal injection of steroids has gained popularity with the explanation that the inflammation of nerve roots causing radicular pain, and because steroids are placed on the local level should relieve symptoms. - - MESH:D011842 - Disease - - radicular pain - - + MESH:D013256 Chemical steroids - + MESH:D007249 Disease inflammation - + + MESH:D010146 + Disease + + radicular pain + + MESH:D013256 Chemical @@ -58667,13 +59289,13 @@ paragraph 16901 Prospective-cohort study indicates a decrease in the need for surgical treatment because of treatment-injection steroid injections. The clinical effect is measurable and statistically significant improvement registered radicular pain. - + MESH:D010146 Disease pain - + MESH:D013256 Chemical @@ -58685,19 +59307,19 @@ paragraph 17138 Another in a series of studies on the treatment of cervical syndrome confirmed the effectiveness of acupuncture in the cervical region of patients with radicular symptoms. Favorable results have been seen in almost 90% of cases. These results indicate that treatment of acupuncture cervical region can be effective as the conservative therapy for the treatment of cervical radiculopathy. - + 9606 Species patients - + MESH:D002575 Disease cervical syndrome - + MESH:D011843 Disease @@ -58709,7 +59331,7 @@ title_1 17526 4. MOUSE SHOULDER - + 10090 Species @@ -58721,7 +59343,7 @@ paragraph 17544 Computers are considered as an integral part of everyday life. They are used in education, health and medicine, science, banking, recreation and entertainment, politically engaged people. Approximately 75% of jobs are dependent on the work on the computer. - + 9606 Species @@ -58733,19 +59355,19 @@ paragraph 17801 Chen and colleagues examined the effect of five computer mice made at different angles in 12 respondents employed in the business sector in Taiwan. They concluded that too frequent use of non ergonomic computer mice causes extreme discomfort in the muscle and tendon system of the hand along with various manifest symptoms in the shoulder and forearm. The use of ergonomic mice custom hand shape, thanks to the different corners of the obliquity have less adverse effect on the activity of the forearm and shoulder. - + 10090 Species mice - + 10090 Species mice - + 10090 Species @@ -58765,13 +59387,13 @@ paragraph 18407 The shoulder belt is common, even among the most frequent localization of musculoskeletal pain after pain in the back and knees. - + MESH:D059352 Disease musculoskeletal pain - + MESH:D010146 Disease @@ -58783,19 +59405,25 @@ paragraph 18536 The long-term work at the computer leads to inflammation of tendons and exchanges shoulder joint, and in severe cases can lead to tearing of tendons and muscles "rotator cuff". If inflammation persists, the capsule and ligaments of the shoulder joint becomes stiffer and limited mobility. Inappropriate placement of computers may have adverse effects on posture in children. - + 9606 Species children - + + MESH:D052256 + Disease + + tendons + + MESH:D007249 Disease inflammation - + MESH:D007249 Disease @@ -58807,13 +59435,13 @@ paragraph 18915 Limited and painful mobility is especially pronounced when raising your arms above your head or behind your back. Long-term pain becomes stronger and leads to the development of muscle weakness areas of the shoulder belt through a series of hands, and preventing further activities. - + MESH:D010146 Disease pain - + MESH:D018908 Disease @@ -58825,13 +59453,13 @@ paragraph 19198 Symptoms of pain in the shoulder differently vary from intense that it can spread to other parts of the body, to moderate that lingers on the shoulder belt, but still hinder the function of the hinge, and can even lead to sleep disorders. - + MESH:D010146 Disease pain - + MESH:D012893 Disease @@ -58843,13 +59471,19 @@ paragraph 19437 The anatomy of the shoulder is a specific because the shoulder joint consists of primary, secondary and ancillary wrist joints. The anatomical structure makes the most complex joint of the body. Joints shoulders: glenohumeral joint (shoulder joint primary), Scapulothoracic joint (secondary shoulder joint), and sternoclavicular joint (extra joints). + + MESH:D007592 + Disease + + sternoclavicular joint + INTRO paragraph 19788 Muscle groups that may be a potential source of pain are rotator cuff (responsible for balance of the glenohumeral joint), stabilizing the scapula (responsible for the position of the scapula) the primary drivers (responsible for the strong movements). - + MESH:D010146 Disease @@ -58861,32 +59495,32 @@ paragraph 20041 For the diagnosis of mouse shoulder, the most important is detailed history with regard to occupation and profession of the patient and physical review. We should not forget to examine whether the patient has in the past had a shoulder injury, or some localized inflammation in the same. - + 10090 Species mouse - + 9606 Species patient - + 9606 Species patient - + MESH:D007249 Disease inflammation - - MESH:D014947 + + MESH:D058186 Disease injury @@ -58897,19 +59531,19 @@ paragraph 20329 Information about the character, pain intensity and directions of its expansion are of great importance. In fact it is necessary to establish that the primary cause of shoulder pain, and that the anatomical structure of the shoulder is affected. It is known that the pain can be transmitted from the adjacent region to the shoulders, for example. The cervical region, or by visceral fibers of the heart. - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - + MESH:D010146 Disease @@ -58921,13 +59555,13 @@ paragraph 20733 In equivocal cases, x-rays of the shoulder joint may help in differentiating existing inflammation of the joints and structure surrounding joint of a possible slight shoulder dislocation. - + 255564 Species rays - + MESH:D007249 Disease @@ -58939,13 +59573,13 @@ paragraph 20921 The primary method for the treatment of mouse shoulder in a timely manner to reduce the strain on shoulders, mostly working with computer mouse. - + 10090 Species mouse - + 10090 Species @@ -58957,19 +59591,19 @@ paragraph 21066 If the patient is in question, and cannot make a break, they definitely need the to replace old with new ergonomic computer mouse that is adapted to grip and allows the natural position of the hand during mouse use. - + 9606 Species patient - + 10090 Species mouse - + 10090 Species @@ -58981,13 +59615,13 @@ paragraph 21282 NSAIDs are the first drugs of choice. In the long-term intractable pain states and may prescribe therapy application of corticosteroid injections in the shoulder joint. An indispensable aspect of therapy is physical therapy. Physical therapy is appropriate in the chronic phase of the disease, which aims to stabilize and strengthen the muscles of the shoulder belt, and the resultant reduction in pain. - + MESH:D010146 Disease pain - + MESH:D010146 Disease @@ -58999,13 +59633,13 @@ paragraph 21686 Jan C. Winters and colleagues came to the conclusion that only 50% of patients with newly diagnosed painful condition experience a full recovery within the first 6 months. This percentage rises to 60% in the first year of the onset of pain. They also proved that speaking to the long-term treatment, at the end of treatment, there was no difference in effect between the corticosteroid and physical therapy. - + 9606 Species patients - + MESH:D010146 Disease @@ -59017,47 +59651,36 @@ paragraph 22094 Onyebeke LC investigated the effects of props during the use of the mouse on the palm, forearm and upper extremities, and came to the result that 90% of respondents who have used the support arm had less pain in the shoulder girdles of those who are not used to. The use of any support results in less tension, and less applied force to a computer mouse during operation, and therefore less hassle and pain in the shoulder. - + 10090 Species mouse - + 10090 Species mouse - + MESH:D010146 Disease pain - + MESH:D010146 Disease pain - - MESH:D009207 - Disease - - upper extremities - INTRO title_1 22518 5. PHYSICAL REHABILITATION AND PREVENTION OF TECHNOLOGICAL DISEASES - - Chemical - - TECHNOLOGICAL - INTRO @@ -59070,13 +59693,13 @@ paragraph 22666 If the patient required outpatient treatment, family doctor referred him Physiotherapy in CBR (Community based rehabilitation) i.e. the center for rehabilitation in the community. CBR's are organized within the framework of primary health care and are located within the health centers. CBR-rehabilitation in the community-the definition of a Joint Paper, ILO, UNESCO, WHO, in 1994. - + 9606 Species patient - + 9606 Species @@ -59088,60 +59711,42 @@ paragraph 23053 Community rehabilitation strategy based on community development for the rehabilitation, equalization of opportunities and social integration of people with disabilities, their families and communities and the appropriate health of educational, professional and social services. - + 9606 Species people - - MESH:D009069 - Disease - - disabilities - INTRO paragraph 23332 Community rehabilitation allows people with disabilities to take initiative and improve their own lives, and to contribute to the community, and not just use the funds and resources that are available. Thus, the entire community and all its members are winners. - + 9606 Species people - - MESH:D009069 - Disease - - disabilities - INTRO paragraph 23594 CBR became the backbone of the carrier non-institutional organization of the program in physical medicine and rehabilitation of persons with disabilities as well as a large number of chronically ill patients in the prevention of disability. - + 9606 Species persons - + 9606 Species patients - - MESH:D009069 - Disease - - disabilities - INTRO @@ -59160,25 +59765,19 @@ paragraph 24061 Treatment in the community increases the accessibility and quality of services for people with disabilities and their families. - + 9606 Species people - - MESH:D009069 - Disease - - disabilities - INTRO paragraph 24189 Services include primary treatment damage and disability in the center and at home, as well as patient education, health promotion and prevention of disease and disability. - + 9606 Species @@ -59196,7 +59795,7 @@ title_2 24667 5.3. The most common reasons for referral of patients in the CBR - + 9606 Species @@ -59208,7 +59807,7 @@ paragraph 24732 The most common reasons due to which family physician refers patients to the ambulance treatment in rehabilitation centers in the community are: - + 9606 Species @@ -59220,54 +59819,54 @@ paragraph 24877 Adult patients: a) Cervical pain syndrome; b) Lumbar pain syndrome; c) Rheumatic diseases; d) Rehabilitation after stroke. Children and young people: a) Neurorisk children; b) Deformities of the spinal column; c) Deformities of the lower extremities; d) Rehabilitation after injuries of the locomotor apparatus - + 9606 Species patients - + 9606 Species Children - + 9606 Species people - + 9606 Species children - + + MESH:D010146 + Disease + + pain syndrome + + MESH:D020521 Disease stroke - - MESH:D019547 + + MESH:D010146 Disease - - Cervical pain + + pain - + MESH:D012216 Disease Rheumatic diseases - - MESH:D010146 - Disease - - Lumbar pain - INTRO @@ -59286,18 +59885,12 @@ paragraph 25306 Health Center promotes the concept of active participation of disabled people and their families in identifying needs and resources in rehabilitation develop a shared vision of their life in society, the implementation of the vision and the monitoring and evaluation of implementation. - + 9606 Species people - - MESH:D014786 - Disease - - implementation of the vision - INTRO @@ -59310,7 +59903,7 @@ paragraph 25601 5I.295 Within the health center there is a center for the rehabilitation of disabled persons (CBR) with support groups and self-help. - + 9606 Species @@ -59358,7 +59951,7 @@ paragraph 26524 5I.302 There are dated, documented criteria for referring patients to physical therapy. The criteria are written/reviewed in the past three years. - + 9606 Species @@ -59376,7 +59969,7 @@ paragraph 27026 5I.304 There are documented procedures on the regulation of proper completion of the treatment of patients having implemented the rehabilitation plan. The procedure is written/rewritten in the past three years. - + 9606 Species @@ -59388,7 +59981,7 @@ paragraph 27237 5I.305 estimated needs of the patient/user for the physical treatment by physical examination which receives measurable data for evaluation. - + 9606 Species @@ -59400,7 +59993,7 @@ paragraph 27378 5I.306 Each patient was referred to physical therapy has an individual rehabilitation plan for the implementation of physical therapy. - + 9606 Species @@ -59412,7 +60005,7 @@ paragraph 27513 5I.307 Before each procedure/treatment is carried out risk assessments. Note: This includes risk assessment, contraindications to treatment and precautions. Also, include the verification of hazards such as wet floors, and the provision of suitable clothing and shoes worn by therapists and patients/users. - + 9606 Species @@ -59436,7 +60029,7 @@ paragraph 28005 5I.310 Before using the apparatus and its application to a patient, is made visual and physical verification of security equipment. - + 9606 Species @@ -59448,7 +60041,7 @@ paragraph 28137 5I.311 There is plenty of space and are available partitions/curtains in the room/rooms for physiotherapy treatment that patients provide visual privacy when using the equipment. - + 9606 Species @@ -59466,135 +60059,111 @@ paragraph 28443 5I.313 Health Centre organizes regular meetings for people with disabilities and their family members, performs basic assessment of their needs and implements early education and basic rehabilitation programs under the guidance of an appropriate therapist. - + 9606 Species people - - MESH:D009069 - Disease - - disabilities - INTRO paragraph 28700 5I.314 health center, or CBR, is developing a multi-sectoral cooperation and contracts with appropriate institutions for labor and social policy, education, institutes Employment, sports institutions, etc. Panel evidence-based clinical practice guidelines on musculoskeletal rehabilitation interventions, - + MESH:D048949 Disease labor - - MESH:D009140 - Disease - - musculoskeletal - DISCUSS title_1 29005 7. DISCUSSION AND RECCOMENDATIONS ON THERAPEUTIC MODALITIES - - MESH:D045506 - Chemical - - THERAPEUTIC - DISCUSS paragraph 29065 Several studies have reported possitive or negative association between computer use and musculosceletal symptoms. The prevalence of Computer Carpal Syndrome (CTS) among computer professionals based on clinical signs and sympoms is approximately 13,1% and almost 1 out of every computer professional suffers from this condition. Very few data published about this topic - studies about CTS prevalence based on clinical signs and symptoms, according to Ali KM and Sathiyasekaran BWC, ranging from 3 to 6 %b only, but we think real situation is much worse. Postural stress due to inadeqauate workstation ergonomics (inappropriate location of monitor, keyboard or mouse) discussed as cause of all mentioned technological diseases in this text (Figure 7). Aydan Oral et al. Had written paper about Evidence Based Physical Medicine and Rehabilitation strategies for patients with cervical radiculopathy due to disc herniation. They reported that neck pain affected 4,82% of the world population in 2010, ranks second after low back pain among musculoskeletal disorders as one of the leading causes of years lived with disability, with contribution of 33,6 million years. Physical medicine and rehabilitation interventions in radicular neck pain and their evidence based are: a) educational interventions; b) exercise; c) workplace interventions/ergonomics; d) phyasical agents (TENS, therapeutic ultrasound, low-level laser therapy) pulsed electromagnetic field therapy, non-invasive brain stimulation techniques, etc); e) injection therapy (epidural steroids, botulinum toxin, ozone); f) cervical collars; g) traction; manual therapies (massage, manipulation and mobilization (commonly used in the management of discogenic neck pain); complementary and alternative medicine treatments (acupuncture, herbal medicine, etc.). Much research efforts have been spent and several risk factors such as heavy lifting, lifestyle, psichosocial factors identified, but the etiology of technological diseases are still or not enough unclear. Recurrences and functional limitations can be minimized limitations with appropriate conservative management, including medications, physical therapy, exercise and patient education. But, these subgroup of developing chronic and disabling symptoms generating large social costs. - + 7276 Gene 317 CTS - + 7276 Gene 317 CTS - + 10090 Species mouse - + 9606 Species patients - + 1407750 Species herbal medicine - + 9606 Species patient - + MESH:D010146 Disease pain - - MESH:D019547 + + MESH:D002349 Disease - - neck pain + + Computer Carpal Syndrome - + MESH:D009140 Disease musculoskeletal disorders - - MESH:D010126 - Chemical - - ozone + + MESH:D019547 + Disease + + neck pain - - MESH:D017116 + + MESH:D001416 Disease - - low back pain + + back pain - + MESH:D019547 Disease radicular neck pain - - MESH:D013256 - Chemical - - steroids - - + MESH:D011843 Disease @@ -59606,12 +60175,6 @@ paragraph 31371 According to the Panel evidence-based clinical practice guidelines on musculoskeletal rehabilitation interventions the key points to clinicians are (Table 1 and 2): - - MESH:D009140 - Disease - - musculoskeletal - TABLE @@ -59620,11 +60183,17 @@ table_caption 31536 Summary grid of neck pain guidelines. *Adapted from the Philadelphia Panel Members and Ottawa Methods Group.2. A, benefit demonstrated; C, no benefit demonstrated; EMG, electromyographic; ID, insufficient or no data; TENS, transcutaneous electrical nerve stimulation - - MESH:D019547 + + MESH:D000309 Disease - - neck pain + + insufficient + + + MESH:D010146 + Disease + + pain @@ -59634,7 +60203,13 @@ table_caption 31803 Summary grid of shoulder pain guidelines. *Adapted from the Philadelphia Panel Members and Ottawa Methods Group.1A, benefit demonstrated; C, no benefit demonstrated; EMG, electromyographic; ID, insufficient or no data; TENS, transcutaneous electrical nerve stimulation. - + + MESH:D000309 + Disease + + insufficient + + MESH:D010146 Disease @@ -59646,13 +60221,13 @@ paragraph 32073 The Philadelphia Panel recommends continued normal activities for acute, uncomplicated low back pain and therapeutic exercise for chronic, subacute, and postsurgical low back pain. - + MESH:D017116 Disease low back pain - + MESH:D017116 Disease @@ -59670,7 +60245,7 @@ paragraph 32341 exercise for knee osteoarthritis. - + MESH:D010003 Disease @@ -59682,7 +60257,7 @@ paragraph 32375 For chronic neck pain, the Philadelphia Panel recommends proprioceptive and thera-peutic exercise. - + MESH:D019547 Disease @@ -59694,7 +60269,7 @@ paragraph 32474 The Philadelphia Panel found evidence to sup-port the use of therapeutic ultrasound in thetreatment of calcific tendonitis of the shoulder. - + MESH:D052256 Disease @@ -59712,55 +60287,55 @@ paragraph 32775 Acute neck pain is often associated with injury or accident, whereas chronic neck pain is related to repetitive injury. Neck pain is commonly managed with analgesics and rest, but referrals to rehabilitation are increasing. The Philadelphia Panel sought to improve the appropriate use of rehabilitation interventions for neck pain by providing evidence-based guidelines. A summary of the Panel's recommendations can be found in Table 2. - + MESH:D019547 Disease Neck pain - + MESH:D019547 Disease neck pain - + + MESH:D000081084 + Disease + + injury or accident + + MESH:D019547 Disease neck pain - + MESH:D012090 Disease repetitive injury - + MESH:D019547 Disease neck pain - - MESH:D020521 - Disease - - injury or accident - DISCUSS paragraph 33212 Rehabilitation specialists offer several conservative interventions for the management of shoulder pain. There are few published guidelines for the management of shoulder pain. - + MESH:D010146 Disease pain - + MESH:D010146 Disease @@ -59778,43 +60353,37 @@ paragraph 33403 Thanks to the high use of computers, resulting diseases of the modern era, known as the"Technological disease." Under this term we mean the diseases caused by the harmful influence of the job in the first place, the working position in which it is involved excessive work for a computer, such as the banking sector, the health sector and many others who are directly or indirectly connected with the work on the computer and overuse keyboards and computer mice, which are in a number of cases of non-ergonomic, and inappropriate for a handful. Establishing the diagnosis of diseases of technology is an interdisciplinary process that requires special knowledge in medicine and related fields related to health and safety at work. Determining the causes and diagnosis of occupational diseases is carried out according to the criteria of modern medicine work. - + 10090 Species mice - + MESH:D009784 Disease occupational diseases - - MESH:D012090 - Disease - - overuse keyboards - CONCL paragraph 34265 First, we identify the clinical picture of the disease on the one hand and identification in the working process on the other side, and their immediate connections. Medical history is the gold standard, because without it, it would be possible to find information on working conditions and the duration and intensity of exposure to a particular hazard from the workplace. The intensity and length of exposure to harmful factor must be that level that is known and proven scientific research that can damage health. Prevention would involve, educate employees by computers to ensure better and more regular position of the spine and shoulder while working at the computer, as well as the use of stylish ergonomic mice in hand. Of course, adequate work breaks are required, in order to avoid fatigue and exhaustion syndrome, but today it is difficult to appreciate the extent and speed of work. No less important and efficient are exercises to strengthen the shoulder belt, as well as exercises for posture. - + 10090 Species mice - + MESH:D005221 Disease fatigue - + MESH:D006359 Disease @@ -59866,7 +60435,7 @@ surname:Ditmars;given-names:D 35394 Diagnosis of carpal tunnel syndrome - + MESH:D002349 Disease @@ -59902,7 +60471,7 @@ 637 35453 Sistemic disease and the carpal tunnel syndrome - + MESH:D002349 Disease @@ -59951,13 +60520,13 @@ surname:Aghili;given-names:A 35565 Effect of vitamin B6 on clinical symptoms and electrodiagnostic results of patients with carpal tunnel sndrome - + 9606 Species patients - + MESH:D025101 Chemical @@ -59981,13 +60550,13 @@ surname:Gur;given-names:A 35676 Reliability and efficacy of the new massage technique on the treatment in the patients with carpal tunnel syndrome - + 9606 Species patients - + MESH:D002349 Disease @@ -60041,7 +60610,7 @@ surname:Pandza;given-names:H 35928 Cervical Pain Syndrome as Consequence of Computer Use in Daily Practice - + MESH:D010146 Disease @@ -60069,7 +60638,7 @@ 2899 36002 Cervical radiculopathy - + MESH:D011843 Disease @@ -60090,13 +60659,13 @@ 1070 36025 Transforaminal steroid injections in the treatment of cervical radiculopathy - + MESH:D011843 Disease - - cervical radiculopathy + + radiculopathy - + MESH:D013256 Chemical @@ -60118,7 +60687,7 @@ 367 36102 Clinical effect of acupuncture on cervical spondylotic radiculopathy: results of a case series - + MESH:D011843 Disease @@ -60167,7 +60736,13 @@ 178 36292 Musculoskeletal pain in the Netherlands: prevalences, consequences and risk groups, the DMC(3)-study - + + MESH:C023025 + Chemical + + DMC + + MESH:D059352 Disease @@ -60204,7 +60779,7 @@ surname:Chung;given-names:CC 36550 Issues about home computer workstations and primary school children in Hong Kong: A pilot study - + 9606 Species @@ -60225,24 +60800,18 @@ 53 36646 Evidence-Based Physical Medicine and Rehabilitatio Strategies for Patients with Cervical Radiculopathy Due to Disc Herniation - + 9606 Species Patients - + MESH:D011843 Disease Cervical Radiculopathy - - MESH:D007405 - Disease - - Disc Herniation - 12 @@ -60258,7 +60827,7 @@ 325 36772 Computer Professionals and Carpal Tunel Syndrome (CTS) - + 7276 Gene 317 @@ -60283,13 +60852,13 @@ surname:Martinaj;given-names:M 36827 Prevalence and Pharmacologic Treatment of Patients with Low Back Pain Treated at Kosovo Energetic Corporation - + 9606 Species Patients - + MESH:D017116 Disease @@ -60340,108 +60909,96 @@ abstract 68 Multiple myeloma is still an incurable disease with pattern of regression and remission followed by multiple relapses raising from the residual myeloma cells surviving even in the patients who achieve complete clinical response to treatment. In recent years there is a huge improvement in treatment of patients with multiple myeloma. The milestones of these improvement are: autologous transplantation and high-dose melphalan, imunomodulating drugs (thalidomide, lenalidomide), proteosom inhibitors (bortesomib, carfilzomib). The most significant improvement in overall survival has been achieved in the patients younger than 65 years. So, the major challenge for hematologist is to translate this improvement in the elderly patients with multiple myeloma. Today, physicians are able to offer wider variety of treatment options for elderly patients with multiple myeloma. Therapeutic options should be tailored and personalized according to patient's characteristics by balancing efficacy and toxicity of each drug which is especially important for elderly patients. In the mode of sequencing treatment for elderly patients with multiple myeloma, our goal is to achieve and maintain maximal response while limiting treatment -related toxicities as much as possible. Second-generation novel agent, such as carfilzomib, pomalidomide, elotuzumab, bendamustine are currently being evaluated as an option to improve treatment outcome in elderly patients. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:C001758 + + - Chemical - - bendamustine + + bortesomib - + MESH:D009101 Disease multiple myeloma - - MESH:C467566 - Chemical - - pomalidomide - - + MESH:D008558 Chemical melphalan - - MESH:D013792 + + MESH:D000069461 Chemical - - thalidomide + + bendamustine - + MESH:D064420 Disease toxicity - + MESH:D009101 Disease Multiple myeloma - - MESH:D064420 - Disease - - toxicities - MESH:D009101 Disease @@ -60449,10 +61006,10 @@ multiple myeloma - MESH:D009101 - Disease - - multiple myeloma + MESH:D000077269 + Chemical + + lenalidomide MESH:D009101 @@ -60461,24 +61018,48 @@ multiple myeloma - MESH:C467567 + MESH:C524865 Chemical - - lenalidomide + + carfilzomib + MESH:C467566 + Chemical + + pomalidomide + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D064420 + Disease + + toxicities + + + MESH:D009101 + Disease + + multiple myeloma + + MESH:D009101 Disease myeloma - - MESH:C524865 + + MESH:C546027 Chemical - - carfilzomib + + elotuzumab - + MESH:C524865 Chemical @@ -60490,25 +61071,25 @@ paragraph 1518 Myeloma multiplex is a malignant disorder first recognized in the 19th century and at that time point was described as "mollities ossium" accompanied by the presence of Bence Jones protein in urine. At that time there was no effective treatment and median overall survival was only short: a few months. Over the time, especially over the past decade, many advances in myeloma treatment have been made and improvement of the median overall survival have been achieved. But, myeloma multiplex is still considered incurable disease. - + MESH:D009101 Disease Myeloma multiplex - + MESH:D009101 Disease - - myeloma multiplex + + myeloma - + MESH:D009101 Disease - + myeloma - + MESH:D009369 Disease @@ -60520,87 +61101,87 @@ paragraph 2052 Myeloma multiplex is a malignant disorder that arises from the malignant proliferation of plasma cell and is characterized by the presence of at least 10% clonal bone marrow plasma cells and serum and/or urinary monoclonal protein. Myeloma multiplex accounts for 1% of all types of cancers and is the second most common hematologic neoplasm, approximately 13% for all hematologic malignances. Myeloma multiplex is considered as a disease of elderly reflected by the average age at diagnosis 70 years, with 37% of patients younger than 65 years, 26% aged 65 to 74 years, and 37% older than 75years. - + 9606 Species patients - + MESH:D009369 Disease cancers - + MESH:D009101 Disease - - Myeloma multiplex + + Myeloma - + MESH:D019337 Disease hematologic neoplasm - - MESH:D009101 - Disease - - Myeloma multiplex - - + MESH:D009369 Disease malignant disorder - + MESH:D009101 Disease Myeloma multiplex + + MESH:D009101 + Disease + + Myeloma + INTRO paragraph 2650 Patient with myeloma multiplex could be diagnosed as symptomatic or asymptomatic disease. Patients with a symptomatic disease should be treated immediately, and the mainstay of asymptomatic disease is still an observation. Symptomatic disease could be defined with so called CRAB features: C-hypercalcemia (>11.5mg/dl (2.65mmol/l); R-renal failure (serum creatinin >mg/dl; 1.73mmol/l); A-anemia (hemoglobin <10g/dl; 12.5 mmol/l) or 2 g/dl; 1.25mmol/l below the lower limit of normal; and B-bone disease (lytic lesions, severe osteopenia, or pathologic fractures). Patients are stratified into three risk groups according to the International Staging system (ISS). This system defines three risk groups based on serum beta 2 micro globulin and albumin levels at diagnosis. High-risk disease and poor prognosis are defined with the presence of high levels of serum beta2 micro globulin (stage III). - + 10242 Gene 4257 beta 2 - + 10242 Gene 4257 beta2 - + 9606 Species Patient - + 9606 Species Patients - + 9606 Species Patients - + MESH:D009101 Disease @@ -60612,13 +61193,13 @@ paragraph 3547 Nowadays, chromosomal abnormalities t(4;14), t(14;16) and t(14;20) chomosome 1 abnormalities and del17 detected by fluorescent in situ hybridization (FISH) are associated with poor prognosis. Hiperdiplpdy, t (11;14), t, are associated with good prognosis and can be considered "standard risk". - + MESH:D002869 Disease chromosomal abnormalities - + Disease Hiperdiplpdy @@ -60629,109 +61210,103 @@ paragraph 3845 Initial therapy for multiple myeloma depends to a certain extent on patients characteristics such as: eligibility for autologous stem cell transplantation per se; age and co-morbidities. The role of induction therapy is to induce remission, but patient's characteristics have a significant role in the initial treatment approach. Goals of treatment are: to eradicate the tumor clone, including cancer stem cell, to search for an appropriate balance between efficacy and toxicity with three different but complementary aims: quality of life, survival prolongation and eventually the dream of cure. This can be achieved if we use appropriate tools to evaluate treatment efficacy. Achieving the lowest level of minimal residual disease can be an important goal of therapy, a step in the path to cure. - + 9606 Species patients - + 9606 Species patient - + MESH:D009369 Disease tumor - + MESH:D009369 Disease cancer - + MESH:D064420 Disease toxicity - + MESH:D009101 Disease multiple myeloma depends - - MESH:D008133 - Disease - - prolongation - INTRO paragraph 4643 Today, treatment goals in myeloma patients are shifting and the goal of therapy in elderly patients is to achieve and maintain maximal response. Many believe that multiple myeloma can be converted into a chronic disease and that a functional cure maybe a realistic goal. Attainment of complete remission at any time point during treatment is associated with improvement outcome; so it is likely to be established as a goal of therapy. The increasing number of treatment possibilities as an optimal therapeutic strategy improve patient outcome. Physicians have the opportunity to choose the best treatment regimen according to patient characteristics, while limiting treatment - related toxicities as much as possible. In elderly patients optimal treatment should be always balance efficacy and toxicity. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patient - + 9606 Species patients - + MESH:D064420 Disease toxicities - + MESH:D064420 Disease toxicity - + MESH:D002908 Disease chronic disease - + MESH:D009101 Disease multiple myeloma - + MESH:D009101 Disease @@ -60743,112 +61318,137 @@ paragraph 5449 Initial therapy for multiple myeloma depends on eligibility for high-dose therapy and autologous stem cell transplant (HDT-ASCT). In many European countries, elderly patients (older than 65) are generally considered ineligible for autologous stem cell transplantation (ASCT). In past times in practice, patients were stratified to those who are transplant eligible and consolidated with high-dose melphalan/SCT and those who were transplant ineligible and received oral melphalan with prednisolone (MP). This stratification has been made in the era of conventional therapy where response rates to the induction therapy were poor and the goal was achievement of partial response and prevention of organ damage. So, in contrast, today we considered that biological age and chronological age do not always correspond, and a greater emphasis should be placed on the former rather than the latter. CR is a good surrogate end point for survival in transplant-ineligible patients and novel drugs have been incorporated into the treatment of non-transplant candidates. Patients are generally considered eligible for ASCT if they have good performance status, no-comorbidities, and normal cardiac, pulmonary, liver and renal function. Patients older than 75 years or vulnerable ones are more susceptible to adverse events and in this setting we are looking for less toxic regimens and appropriate dose reductions should be adopted. Even, ASCT with a reduced melpahlan conditioning dose is well tolerated by the selected population of patients up to the age of 75 in good clinical conditions. Until novel agents were introduced, for more than 40 years the combination of melpahalan and prednisone (MP) was considered the standard approach for transplant -ineligible patients, with PFS of approximately 18 months and 2-3 years (at best) overall survival in the population treated with MP. A meta-analysis including 27 randomized studies compared MP with other chemotherapy-containing regimens. The introduction of novel agents has challenged this combination and new and more effective combination is available. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species Patients - + 9606 Species Patients - + 9606 Species patients - + 9606 Species patients - + MESH:D008558 Chemical melphalan - + MESH:D011241 Chemical prednisone - - MESH:D009101 - Disease - - multiple myeloma + + - + Chemical + + MP - + MESH:D011239 Chemical prednisolone - - MESH:D008558 - Chemical - - melphalan - - + + - Chemical melpahalan + + MESH:D009101 + Disease + + myeloma depends + + + - + Chemical + + MP + INTRO paragraph 7552 Six randomized phase III studies have shown that the combination melphalan-prednisone-thalidomide (MPT) is superior to MP in terms of response and progression-free survival, but it was translated into improved survival in the 2 IFM studies. An efficacy meta-analysis of the six MPT trials including 1685 patients was conducted and has showed that the addition of thalidomide to MP significantly prolonged both PFS and extended OS by 20%-39 months compared to MP-33 months. The meta-analysis further confirmed the progression-free survival advantage achieved with MPT. So, MPT is therefore regarded as a new standard of care in transplant ineligible patients. - + 9606 Species patients - + 9606 Species patients - + MESH:D013792 Chemical thalidomide - + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + MESH:D013792 Chemical - - melphalan-prednisone-thalidomide + + thalidomide + + + - + Chemical + + MPT @@ -60856,43 +61456,55 @@ paragraph 8215 For selected elderly patients, particularly for standard risk patients with favorable FISH, CTD is a feasible approach. A phase III assessed the role of thalidomide in combination with different alkylating agent and steroid than those commonly used, respectively cyclophosphamide and dexamethasone with an attenuated schedule (CTDa). Despite a deeper response, no differences were noted in median PFS and OS between patients treated with CTDa and MP. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D013792 Chemical thalidomide - + + - + Chemical + + CTDa + + MESH:D013256 Chemical steroid - + + - + Chemical + + CTDa + + MESH:D003907 Chemical dexamethasone - + MESH:D003520 Chemical @@ -60904,137 +61516,139 @@ paragraph 8666 Lenalidomide is an immunomodulatory drug with higher potency than its analogue thalidomide and without sedative or neurotoxic adverse effects. Lenalidomide showed to be safe and effective in relapsed/refractory multiple myeloma patients, as well as in newly diagnosed multiple myeloma patients. - + 9606 Species patients - + 9606 Species patients - - MESH:C467567 + + MESH:D000077269 Chemical - + Lenalidomide - + MESH:D013792 Chemical thalidomide - + MESH:D009101 Disease myeloma - + MESH:D009101 Disease multiple myeloma - - MESH:D020258 - Disease - - neurotoxic adverse - - - MESH:C467567 + + MESH:D000077269 Chemical - + Lenalidomide + + MESH:D020258 + Disease + + neurotoxic + INTRO paragraph 8961 The phase III trial RD (lenalidomide plus high-dose dexamethason) versus Rd (lenalidomide plus low-dose dexamethason) included newly diagnosed multiple myeloma patients both eligible and ineligible for autologus stem cell transplantation. Rd seems preferable for elderly patients because has significantly longer 1-year overall survival, particularly evident in patients older than 65 years of age and can be considered a valid therapeutic option for elderly newly diagnosed myeloma patients. RD remains a good option for patients with renal failure, hypercalcemia, pain or spinal cord compression because more adverse events occurred when RD was given compared with Rd. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + + MESH:D000077269 + Chemical + + lenalidomide + + MESH:D006934 Disease hypercalcemia - + MESH:D009101 Disease myeloma - - Chemical - - dexamethason - - + MESH:D010146 Disease pain - + MESH:D051437 Disease renal failure - - MESH:C467567 - Chemical - - lenalidomide - - + + MESH:D003907 Chemical dexamethason - - MESH:C467567 + + MESH:D000077269 Chemical - + lenalidomide - + + MESH:D003907 + Chemical + + dexamethason + + MESH:D009101 Disease @@ -61046,41 +61660,53 @@ paragraph 9632 The role of lenalidomide was assessed in a recent phase III study that compared melphalan-prednisone-lenalidomide followed by lenalidomide maintenance (MPR-R), with MPR and MP. MPR-R significantly improved median progression-free survival compared with MPR and MP (31 months versus 14 months vs. 13 months; p<0.001), which means that MPR-R reduced the risk of progression by 51% and 60% compared with MPR and MP. Lenalidomide maintenance was well tolerated with few reported incidence of grade 3/4 adverse events and second cancer. However the benefit associated with MPR-R outweigh the increased risk of second primary malignancies. - - MESH:C467567 + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D000077269 Chemical lenalidomide - - MESH:C467567 + + MESH:D000077269 Chemical - - melphalan-prednisone-lenalidomide + + lenalidomide - + MESH:D009369 Disease cancer - - MESH:D009369 - Disease - - malignancies + + MESH:D011241 + Chemical + + prednisone - - MESH:C467567 + + MESH:D008558 + Chemical + + melphalan + + + MESH:D000077269 Chemical Lenalidomide - - MESH:C467567 - Chemical - - lenalidomide + + MESH:D009369 + Disease + + malignancies @@ -61088,29 +61714,15 @@ paragraph 10266 VISTA study compared MP versus VMP and proved that addition of bortezomib to MP is rational, because responses, time to progression and survival were significantly higher with VMP. - - 140767 - Gene - 7597 - - VMP - - - 140767 - Gene - 7597 - - VMP - - + 64115 Gene 81923 VISTA - - MESH:C400082 + + MESH:D000069286 Chemical bortezomib @@ -61121,85 +61733,94 @@ paragraph 10447 In the Spanish PETHEMA trial a cohort of 260 patients older than 65, transplant ineligible, were randomized to receive induction treatment with 6 cycles of bortezomib-thalidomeide-prednisone VTP, or VMP considered as more gentle approach where melphalan is used instead of thalidomide. Both regimens led to high ORR rate (81%and 80%), but there were more adverse events among patients with VDT. VMP with once-weekly schedule of botezomib should be preferred to VTP as induction for elderly multiple myeloma patients. One-weekly bortezomib was scheduled instead of standard twice-weekly administration to reduce adverse events, especially neuropathy associated with bortezomib administration. - - 140767 - Gene - 7597 - - VMP - - - 140767 - Gene - 7597 - - VMP - - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + + MESH:D011241 + Chemical + + prednisone + + + MESH:C562515 + Disease + + especially neuropathy + + MESH:D013792 Chemical thalidomide - - MESH:D009422 - Disease - - neuropathy + + - + Chemical + + botezomib - + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D000069286 + Chemical + + bortezomib + + MESH:D008558 Chemical melphalan - - MESH:C400082 + + - Chemical - - bortezomib + + VTP - + MESH:D009101 Disease multiple myeloma - - MESH:C037167 + + MESH:D000069286 Chemical - - PETHEMA + + bortezomib - - MESH:C400082 + + - Chemical - - bortezomib-thalidomeide-prednisone VTP + + thalidomeide - - MESH:C400082 - Chemical - - bortezomib + + Disease + + VDT @@ -61207,173 +61828,146 @@ paragraph 11139 The combination of bortezomib-lenalidomide-dexamethasone (VRD) was evaluated in both young and elderly patients with multiple myeloma. - + 9606 Species patients - + MESH:D009101 Disease multiple myeloma - - MESH:C400082 + + - Chemical - - bortezomib-lenalidomide-dexamethasone - - - Disease VRD + + - + Chemical + + bortezomib-lenalidomide-dexamethasone + INTRO paragraph 11274 The addition of thalidomide to the new treatment standard with VMP followed by bortezomib-thalidomide maintenance is valid alternative with 3-years overall survival 565 with VMP-VT compared with 41% with VMP. VMP-VT with once weekly bortezomib seems to be a valid alternative for elderly patients particularly those younger than 75 years, where there is the same efficacy, but without additional toxicity, particular peripheral neuropathy. - - 140767 - Gene - 7597 - - VMP - - - 140767 - Gene - 7597 - - VMP - - + 9606 Species patients - - 140767 - Gene - 7597 - - VMP + + MESH:D010523 + Disease + + particular peripheral neuropathy - - 140767 - Gene - 7597 - - VMP + + MESH:D017180 + Disease + + VT - - MESH:C400082 + + MESH:D000069286 Chemical bortezomib - + + - + Chemical + + bortezomib-thalidomide + + MESH:D064420 Disease toxicity - + MESH:D017180 Disease VT - - MESH:C400082 - Chemical - - bortezomib-thalidomide - - - MESH:D010523 - Disease - - peripheral neuropathy - - + MESH:D013792 Chemical thalidomide - - MESH:D017180 - Disease - - VT - INTRO paragraph 11714 A sequential approach consisting of an induction regimen associated with a high rate of complete response, followed by consolidation, maintenance therapy, induce a profound cytoreduction and delays relapse, thus improving survival as a therapy approach that minimized toxicity maximized quality of life and emphasized patient's preference. This choice of treatment strategy improve outcome and prevent the occurrence of relapse with a continuous treatment keeping residual disease under control. The international MM015 phase III study assessed the role of lenalidomide given in maintenance. Having this in mind MPR followed by lenalidomide maintenance emerges as a new standard treatment option for elderly multiple myeloma patients. Another treatment option for elderly patients with impressive result is more intensive regimen VMPT followed by VT maintenance. Bortezomib showed same efficacy with decreased toxicity administrated in a new schedule from twice to once weekly. - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + MESH:D009101 Disease multiple myeloma - - MESH:C467567 + + MESH:D000077269 Chemical lenalidomide - + MESH:D064420 Disease toxicity - - MESH:C400082 + + MESH:D000069286 Chemical Bortezomib - + MESH:D017180 Disease VT - + MESH:D064420 Disease toxicity - - MESH:C467567 + + MESH:D000077269 Chemical lenalidomide @@ -61384,198 +61978,205 @@ paragraph 12692 As we already emphasized, the choice among these regimens should be based on the patient's characteristics. For elderly patients with renal impairment bortezomib or thalidomide containing regiment is considered the best treatment option, because lenalidomide is excreted by the kidneys. In such a case dose of lenalidomide should be reduced according to creatinin clearance. Despite that, for patients with peripheral neuropathy lenalidomide is considered as the best treatment option because of the reduced neurological toxicity. In elderly patients there is an increased risk of treatment-related adverse events in response to novel agent-containing regimens. Clinicians should be aware and proposed guidelines for dose reduction of novel agents and protocol modification in elderly should be followed in clinical practice for elderly patients with multiple myeloma to optimize outcome. So, there is an attempt to personalize the therapy in multiple myeloma according to patient age and vulnerability proposed by European Myeloma Network (EMN). - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + + MESH:D000077269 + Chemical + + lenalidomide + + MESH:D013792 Chemical thalidomide - - MESH:C467567 + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000077269 Chemical lenalidomide - + MESH:D009101 Disease - - Myeloma Network + + Myeloma - - MESH:D009101 + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D010523 Disease - - multiple myeloma + + peripheral neuropathy - - MESH:D009422 + + MESH:D020258 Disease neurological toxicity - - MESH:C467567 - Chemical - - lenalidomide - - + MESH:D007674 Disease renal impairment bortezomib - - MESH:D010523 - Disease - - peripheral neuropathy lenalidomide - - + MESH:D009101 Disease multiple myeloma - - Chemical - - EMN - INTRO paragraph 13739 Multiple myeloma is still an incurable disease with pattern of regression and remission followed by multiple relapses raising from the residual myeloma cells surviving even in the patients who achieve complete clinical response to treatment. New anti-myeloma drugs change treatment paradigm providing both tumor reduction and tumor suppression. There is so much progress, but still many unsolved questions. Today, physicians are able to offer wider variety of treatment options for both young and elderly patients with multiple myeloma. Therapeutic options should be tailored and personalized according to patient's characteristics by balancing efficacy and toxicity of each drug which is especially important for elderly patients. In the mode of sequencing treatment for elderly patients with multiple myeloma, our goal is to achieve and maintain maximal response while limiting treatment -related toxicities as much as possible. Second-generation novel agent, such as carfilzomib, pomalidomide, elotuzumab, bendamustine are currently being evaluated as an option to improve treatment outcome in elderly pateints. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + MESH:D064420 Disease toxicities - + MESH:D064420 Disease toxicity - + MESH:C467566 Chemical pomalidomide - + MESH:D009101 Disease multiple myeloma - + MESH:D009101 Disease myeloma - + + MESH:D000069461 + Chemical + + bendamustine + + MESH:D009101 Disease Multiple myeloma - + + MESH:D009101 + Disease + + multiple myeloma + + MESH:C524865 Chemical carfilzomib - + MESH:D009369 Disease tumor reduction and tumor - - MESH:D009101 - Disease - - multiple myeloma - - - MESH:C001758 + + MESH:C546027 Chemical - - bendamustine + + elotuzumab - + MESH:D009101 Disease @@ -61607,7 +62208,7 @@ 2972 14902 Multiple myeloma - + MESH:D009101 Disease @@ -61629,7 +62230,7 @@ 170 14919 New approaches to management of multiple myeloma - + MESH:D009101 Disease @@ -61649,7 +62250,7 @@ 1060 14968 Multiple myeloma - + MESH:D009101 Disease @@ -61669,7 +62270,7 @@ 3420 14985 International staging system for multiple myeloma - + MESH:D009101 Disease @@ -61688,7 +62289,7 @@ 9 15035 Criteria for diagnosis, staging, risk stratification and response assessment of multiple myeloma - + MESH:D009101 Disease @@ -61708,7 +62309,7 @@ 124 15132 Current Threpautic Starategy for Multiple myeloma - + MESH:D009101 Disease @@ -61729,13 +62330,13 @@ 2221 15182 International Myeloma Working Group molecular classification of multiple myeloma: spotlight review - + MESH:D009101 Disease multiple myeloma - + MESH:D009101 Disease @@ -61756,13 +62357,13 @@ 717 15281 Combining fluorescent in situ hybridization data with ISS staging improves risk assessment in myeloma: an International Myeloma Working Group collaborative project - + MESH:D009101 Disease myeloma - + MESH:D009101 Disease @@ -61783,19 +62384,19 @@ 600 15445 International Myeloma Working Group Consensus Stateet for the Managmnent, treatment, and Supportive Care of Patients with myeloma not eligible for standard Autologous Stem-Cell transplantation - + 9606 Species Patients - + MESH:D009101 Disease myeloma - + MESH:D009101 Disease @@ -61817,19 +62418,19 @@ 242 15638 European myeloma network recommendations on the evaluation and treatment of newly diagnosed patients with multiple myeloma - + 9606 Species patients - + MESH:D009101 Disease multiple myeloma - + MESH:D009101 Disease @@ -61848,17 +62449,17 @@ 495 15762 How should we treat newly diagnosed multple myeloma patients? - + 9606 Species patients - + MESH:D009101 Disease - - multple myeloma + + myeloma @@ -61873,13 +62474,13 @@ 577 15824 How to treat elderly patients with multiple myeloma: combination of therapy or sequencing - + 9606 Species patients - + MESH:D009101 Disease @@ -61899,11 +62500,11 @@ 273 15914 Diagnosis and therapy of multiple myeloma - + MESH:D009101 Disease - - multiple myeloma + + myeloma @@ -61919,11 +62520,11 @@ 268 15956 Association of response endpoints with survival outomes in multiple myeloma - + MESH:D009101 Disease - - multiple myeloma + + myeloma @@ -61940,7 +62541,7 @@ 2520 16032 Improved survival in multiple myeloma and the impact of novel therapies - + MESH:D009101 Disease @@ -61962,22 +62563,17 @@ 4529 16104 Personalized therapy in multiple myeloma according to p[atient age and vulnerability: a report of the Eouropean Myeloma Network (EMN) - - Chemical - - EMN - - + MESH:D009101 Disease multiple myeloma - + MESH:D009101 Disease - - Myeloma Network + + Myeloma @@ -61995,12 +62591,18 @@ 1137 16238 Age and aging in blood disoreders: multiple myeloma - + MESH:D009101 Disease myeloma + + MESH:D006402 + Disease + + blood disoreders + 25 @@ -62015,13 +62617,13 @@ 73 16290 Management of older patients with multiple myeloma - + 9606 Species patients - + MESH:D009101 Disease @@ -62042,13 +62644,13 @@ 890 16341 How we manage autologous stem cell transplantation for patients with multiple myeloma - + 9606 Species patients - + MESH:D009101 Disease @@ -62070,7 +62672,7 @@ 601 16427 Evolving Starategies in the Initial Treatment of multiple myeloma - + MESH:D009101 Disease @@ -62091,13 +62693,13 @@ 584 16493 Initial treatment of nontransplant patients With Multiple myeloma - + 9606 Species patients - + MESH:D009101 Disease @@ -62115,31 +62717,31 @@ 3842 16559 Myeloma trialist Collaborative Group. Combination chemotherapy versus melphalan plus prednisone as treatment for myltiple myeloma: an overview of 6,633 patients from 27 randomized trials - + 9606 Species patients - + MESH:D009101 Disease myltiple myeloma - + MESH:D009101 Disease Myeloma - + MESH:D011241 Chemical prednisone - + MESH:D008558 Chemical @@ -62160,49 +62762,43 @@ 1218 16746 Melphalan and prednisone plus thalidomide versus melphalan and prednisone alone or reduced-intensity autologous stem cell transplantation in elderly patients with multiple myeloma (IFM 99-06): a randomized trial - + 9606 Species patients - + MESH:D008558 Chemical melphalan - + MESH:D011241 Chemical prednisone - + MESH:D008558 Chemical Melphalan - + MESH:D009101 Disease multiple myeloma - - MESH:C000595265 - Chemical - - IFM 99-06 - - + MESH:D011241 Chemical prednisone - + MESH:D013792 Chemical @@ -62223,31 +62819,31 @@ 3670 16958 Efficacy of melphalan and prednisone plus thalidomide in patients older than 75 years with newly diagnosed multiple myeloma: IFM01/01 trial - + 9606 Species patients - + MESH:D008558 Chemical melphalan - + MESH:D013792 Chemical thalidomide - + MESH:D009101 Disease multiple myeloma - + MESH:D011241 Chemical @@ -62267,37 +62863,37 @@ 22 17098 Addition of thalidomide to oral melphalan/prednisone in patients with multiple myeloma not eligible for transplantation: result of a randomized trial from tha Turkish Myeloma Study Group - + 9606 Species patients - + MESH:D009101 Disease multiple myeloma - + MESH:D011241 Chemical prednisone - + MESH:D009101 Disease Myeloma - + MESH:D013792 Chemical thalidomide - + MESH:D008558 Chemical @@ -62318,31 +62914,31 @@ 1412 17285 Melphalan and prednisone plus thalidomide or placebo in elderly patients with multiple myeloma - + 9606 Species patients - + MESH:D013792 Chemical thalidomide - + MESH:D008558 Chemical Melphalan - + MESH:D009101 Disease multiple myeloma - + MESH:D011241 Chemical @@ -62363,31 +62959,31 @@ 3166 17380 Phase III study of the value of thalidomide added to melphalan plus prednisone in elderly patients with newly diagnosed multiple myeloma: the HOVON 49 Study - + 9606 Species patients - + MESH:D011241 Chemical prednisone - + MESH:D009101 Disease multiple myeloma - + MESH:D008558 Chemical melphalan - + MESH:D013792 Chemical @@ -62408,31 +63004,31 @@ 3114 17537 Oral melphalan, prednisone, and thalidomide in elderly patients with multiple myeloma: update results oa a randomized controlled trial - + 9606 Species patients - + MESH:D008558 Chemical melphalan - + MESH:D011241 Chemical prednisone - + MESH:D009101 Disease multiple myeloma - + MESH:D013792 Chemical @@ -62452,25 +63048,25 @@ 94 17672 Safety of thalidomide in newly diagnosed elderly myeloma patients: a mata-analysis of data from individual patients in six randomized trials - + 9606 Species patients - + 9606 Species patients - + MESH:D009101 Disease myeloma - + MESH:D013792 Chemical @@ -62491,31 +63087,38 @@ 1238 17813 Cyclophosphamide, thalidomide and dexamathasone (CTD) as initial therapy for patients with multiple myeloma unsuitable for autologous transplantation - + 9606 Species patients - + + - + Chemical + + CTD + + MESH:D003520 Chemical Cyclophosphamide - + MESH:D013792 Chemical thalidomide - + MESH:D009101 Disease multiple myeloma - + + - Chemical dexamathasone @@ -62535,31 +63138,32 @@ 37 17963 lenalidomide plus high-dose dexamathasone versus lenalidomide plus low-dose dexamethasone as initial therapy for newly diagnosed multiple myeloma : an open - lablel randomized controlled trial - + MESH:D009101 Disease multiple myeloma - + MESH:D003907 Chemical dexamethasone - - MESH:C467567 - Chemical - - lenalidomide - - + + - Chemical dexamathasone - - MESH:C467567 + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D000077269 Chemical lenalidomide @@ -62580,18 +63184,18 @@ 1769 18158 Continuos lenalidomide treatment for newly diagnosed multiple myeloma - - MESH:D009101 - Disease - - multiple myeloma - - - MESH:C467567 + + MESH:D000077269 Chemical lenalidomide + + MESH:D009101 + Disease + + myeloma + 371 @@ -62607,7 +63211,7 @@ 905 18228 Autologous Transplantation and Maintenance Therapy in Multiple Myeloma - + MESH:D009101 Disease @@ -62628,29 +63232,23 @@ 917 18299 Bortezomib plus melphalan and prednisone for initial treatment of multiple myeloma - - MESH:D008558 - Chemical - - melphalan - - + MESH:D009101 Disease multiple myeloma - + MESH:D011241 Chemical prednisone - - MESH:C400082 - Chemical - - Bortezomib + + MESH:D007625 + Disease + + Bortezomib plus melphalan @@ -62667,44 +63265,38 @@ 2266 18382 Bortezomib plus melphalan and prednisone compared with melphalan and prednisone in previously untretated multiple myeloma: update follow-up and impact of subsequent therapy in phase III VISTA trial - + 64115 Gene 81923 VISTA - + MESH:D011241 Chemical prednisone - - MESH:D009101 + + MESH:D007625 Disease - - myeloma + + Bortezomib plus melphalan - - MESH:D008558 - Chemical - - melphalan - - + MESH:D011241 Chemical prednisone - - MESH:C400082 - Chemical - - Bortezomib + + MESH:D009101 + Disease + + untretated multiple myeloma - + MESH:D008558 Chemical @@ -62725,78 +63317,78 @@ 941 18580 Bortezomib, melphalan, and prednisone versuis bortezomib, thalidomide, and prednisone as induction therapy followed by maintenance treatment with bortezomib and thalidomide versus bortezomib and prednisone in elderly patients with untreated multiple myeloma: a randomized trial - + 9606 Species patients - - MESH:C400082 + + MESH:D000069286 Chemical Bortezomib - - MESH:C400082 + + MESH:D000069286 Chemical - + bortezomib - + MESH:D011241 Chemical prednisone - + MESH:D013792 Chemical thalidomide - + MESH:D011241 Chemical prednisone - + MESH:D008558 Chemical melphalan - + MESH:D009101 Disease multiple myeloma - + + MESH:D000069286 + Chemical + + bortezomib + + MESH:D013792 Chemical thalidomide - - MESH:C400082 + + MESH:D000069286 Chemical - + bortezomib - + MESH:D011241 Chemical prednisone - - MESH:C400082 - Chemical - - bortezomib - 28 @@ -62812,24 +63404,48 @@ 5109 18858 Bortezomib-melphalan-prednisone-thalidomide followed by maintenance with bortezomib-thalidomide compared with bortezomib-melphalan-prednisone for initial treatment of multiple myeloma: a randomized controlled trial - - MESH:C400082 + + - + Chemical + + melphalan-prednisone-thalidomide + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + - Chemical bortezomib-thalidomide - - MESH:C400082 + + MESH:D000069286 Chemical - - bortezomib-melphalan-prednisone + + Bortezomib - + MESH:D009101 Disease multiple myeloma + + MESH:D000069286 + Chemical + + bortezomib + 116 @@ -62845,14 +63461,14 @@ 4753 19073 Efficacy and safety of once- weekly bortezomib in multiple myeloma patients - + MESH:D009101 Disease multiple myeloma - - MESH:C400082 + + MESH:D000069286 Chemical bortezomib @@ -62870,19 +63486,19 @@ ref 19149 Age and organ damage correlate with poor survival in myeloma patients: meta - analysis of 1435 individual patient data from 4 randomiuzed trials - + 9606 Species patients - + 9606 Species patient - + MESH:D009101 Disease @@ -62929,25 +63545,20 @@ paragraph 458 Dr. Morris Collen has had a profound influence, not only on the creation of the field of informatics, but also on health-care delivery and the creation of new models of payment and prevention. Dr. Collen's remarkable career began in 1942 when he was selected by Dr. Sidney Garfield, a surgeon, to join him as an internist in a California group practice. Drs Garfield and Collen subsequently worked with the industrialist Henry Kaiser, who is credited with creating one of the first comprehensive prepaid health plans for both office and hospital care. This led to the establishment of Kaiser Perma-nente in the post-World War II period plus a comprehensive infrastructure of hospitals in the Bay Area near San Francisco and near Portland, Oregon. In the subsequent decades, the Kaiser organization grew to become a nationwide healthcare provider with millions of enrollees. - + 8406 Gene 4598 Drs - - Disease - - infrastructure - INTRO paragraph 1332 Collen became a nationally recognized authority on the treatment of pneumonia during World War II. His gift for research showed early in his published studies in The Permanente Foundation Medical Bulletin of which he was long-time editor. After two decades as an internist with Kaiser Permanente, his career took a turn into early medical information technology. Morris Collen and his team set to work to automate the 10-year-old multiphasic health screening exam to develop a prototype electronic health record. Within a decade, Dr. Collen accumulated several millions of health checkup data sets on more than a million subjects, creating in the process not only a prototype electronic health record, but also a phenomenal and unique basis for research, and this despite the immaturity of the technology available in the fifties and sixties. For the pursuit of the scientific aspects of his work, Dr. Collen founded the Medical Methods Research Division within Kaiser Permanente in Oakland, to which he added the Division of Technology Assessment in 1979 that he directed until his retirement in 1983, at age 70. - + MESH:D011014 Disease @@ -63003,7 +63614,7 @@ 373 3233 Collen at 100: A Trib ute to "The Father of Medical Informatics" - + 26150 Gene 9230 @@ -63086,15 +63697,15 @@ 246 This retrospective study was performed at department of infertility and sexual medicine from September 2007 to February 2014. 307 couples with male factor infertility were included and 672 IUI cycles were analyzed. - MESH:D007246 + MESH:D007247 Disease - + infertility - MESH:D007246 + MESH:D007247 Disease - + infertility @@ -63121,18 +63732,6 @@ abstract 935 The initial total progressive sperm count lower than 5*106 means the poor outcome of IUI in the infertile couples with male factor. If the infertile couples with male factor don't get pregnancy after three IUI cycles, the couples should receive re-assessment or other artificial reproductive technology. - - MESH:D007246 - Disease - - infertile - - - MESH:D007246 - Disease - - infertile - INTRO @@ -63145,46 +63744,40 @@ paragraph 1257 Intrauterine insemination (IUI) is a widely used fertility treatment for couples with infertility and is a simple, non-invasive, and a cost-effective technique. IUI is often suggested to infertile couples in which the woman has at least one permeable fallopian tube. This method is indicated in cases of cervical infertility, male factor infertility, anovulation, endometriosis, and unexplained infertility. - + 9606 Species woman - - MESH:D007246 - Disease - - infertility - - - MESH:D007246 + + MESH:D007247 Disease cervical infertility - + MESH:D004715 Disease endometriosis - - MESH:D007246 + + MESH:D007247 Disease - - infertile + + unexplained infertility - - MESH:D007246 + + MESH:D007247 Disease infertility - - MESH:D007246 + + MESH:D007247 Disease - + infertility @@ -63193,43 +63786,37 @@ paragraph 1665 There are certain variables that are currently known to be predictive of IUI success, most of which relate to the female partner including follicle number, endometrial thickness, duration of infertility, and sperm motility. Pregnancy rate after IUI differ between studies according to patient selection criteria, the presence of various infertility factors, ovarian stimulation methods, number of cycles performed, different sperm parameters and preparation technique. - + 9606 Species patient - - MESH:D007246 - Disease - - infertility factors - - - MESH:D016889 - Disease - - endometrial thickness - - - MESH:D007246 + + MESH:D007247 Disease infertility + + MESH:D007247 + Disease + + infertility factors + INTRO paragraph 2134 Semen analysis is the first step to accurately diagnose male infertility. Sperm count, sperm motility and the percentage of sperm with normal morphology are the main criteria for the quality of semen and number of motile sperm and normal morphology have been shown to correlate with IUI outcome. Others found that male factors were not found to correlate with the treatment outcome. The aim of this retrospective study is to exam the use of semen parameters in predicting IUI outcomes in couples diagnosed with male factor infertility. - - MESH:D007246 + + MESH:D007247 Disease infertility - + MESH:D007248 Disease @@ -63247,112 +63834,112 @@ paragraph 2694 In this retrospective cohort study, all couples undergoing intrauterine insemination at department of infertility and sexual medicine from September 2007 to February 2014 were retrospectively enrolled into our database for evaluation of a diagnosis of male factor infertility. Before each course of treatment, infertile couples underwent the following tests: hysterosalpingography, hormone concentrations, semen analysis and postcoital test. Male factor infertility was diagnosed if the male had at least two pre-treatment semen analyses which were abnormal. In this analysis 672 IUI were performed in couples which fit the criteria listed above for male factor infertility. Clinical features of couples, pre and post processing semen analysis parameters at the time of IUI were included in the analyses. - + + MESH:D007247 + Disease + + infertility + + MESH:D007248 Disease male factor infertility - - MESH:D007246 - Disease - - infertility - - - MESH:D007246 + + MESH:D007247 Disease infertility - - MESH:D007246 + + MESH:D007247 Disease infertility - - MESH:D007246 - Disease - - infertile - METHODS paragraph 3499 Patients who underwent natural cycles have regular menstrual cycle and ovulation. Clomiphene citrate (50 or 100 mg daily) and letrozole (2.5 mg or 5 mg daily) were administered orally for 5 days stating on cycle day 3 to 5. Gonadotropin injections were performed daily starting on cycle day 3 to 5 and titrated to develop 2 to 3 mature follicles. When the lead follicle reached at least 16 mm (usually 18 mm) in diameter 10000IU of human chorionic gonadotropin (hCG, livzon pharmaceutical Co.Ltd, China) subcutaneously to induce ovulation. Washed sperm was inseminated 24-36 hours after hCG injection according the LH level on HCG day. All patients were monitored with either ultrasound or both serum LH, estradiol and progesterone levels. - + 93659 Gene 37338 hCG - + 93659 Gene 37338 HCG - + 93659 Gene 37338 hCG - + 9606 Species Patients - + 9606 Species human - + 9606 Species patients - + MESH:D011374 Chemical progesterone - + MESH:D002996 Chemical Clomiphene citrate - - MESH:C067431 + + MESH:D000077289 Chemical letrozole - + MESH:D004958 Chemical estradiol + + MESH:D007986 + Chemical + + LH + METHODS paragraph 4239 For the purpose of semen collection, individuals were asked to refrain from ejaculating for two to four days prior to collection of the specimen. Specimens were produced with masturbation in a collection room adjacent to the laboratory. Freshly ejaculated sperm was allowed to liquefy before initial semen analysis. Liquefied semen was thoroughly mixed before an aliquot was placed on a standard count slide for the pre-processing analysis. The samples were assessed by CASA system (Sperm Class Analyzer, V4.0.0, Spain) and at least four random fields were evaluated for each analysis. - + 1446 Gene 48054 @@ -63365,13 +63952,13 @@ paragraph 4825 After place semen sample in a discontinuous density (40%-80%) gradient column (SpermGrad TM, SpermRinse, Vitrolife Sweden AB, Sweden), the gradient was centrifuged for 20 min at 400g and subsequently, the 40% layer and the seminal plasma fraction were removed from the test tube, leaving the 80% layer undisturbed. Approximately 6-8 ml of sperm-washing medium was added to the 80% layer and centrifuged for 5 min at 400g. The sperm pellet was then reconstituted to approximately 0.5 ml. The analysis of an aliquot of the processed sample was performed as previously. The insemination was performed in a sterile fashion, using a flexible plastic catheter with the patient in the dorsal lithotomy position. The patient did not assume a prone position for at least thirty minutes after the end of the insemination. - + 9606 Species patient - + 9606 Species @@ -63383,13 +63970,13 @@ paragraph 5639 Serum beta-human chorionic gonadotropin (beta-hCG) levels were analyzed 14 to 17 days after IUI to determine pregnancy status. A level greater than 5 mIU/ml was considered positive for pregnancy including biochemical pregnancy and clinical pregnancy. Clinical pregnancies were defined as those with a gestational sac on ultrasound. In this article, we use clinical pregnancy rate to analyze. - + 9606 Species human - + 93659 Gene 37338 @@ -63420,29 +64007,23 @@ paragraph 6300 From 672 IUI cycles performed on 307 couples, there are 27.36% couples get pregnancy (84 out of 307) and the overall pregnancy rate was 12.95% (87 out of 672) of inseminations. At the primary infertile group pregnancy rate was found as 14.29% (68 out of 475). In a group of patients with secondary infertility pregnancy rate was observed as 9.69% (19 out of 197). There were more dominant preovulatory follicles (>= 16mm) in pregnant group than that in not pregnant groups (1.72+-0.89 vs 1.50+-0.77, P<0.05). The mean age of couple, duration of infertility were compared between two groups. When comparing the semen analysis parameters between the two outcome groups, we found that the initial motile and progressive sperm count and post motile and progressive sperm count was higher in the pregnant and not pregnant groups, whereas the post motile sperm count has no significant difference between two groups (Table 1). - + 9606 Species patients - - MESH:D007246 - Disease - - infertility - - - MESH:D007246 + + MESH:D007247 Disease infertility - - MESH:D007246 + + MESH:D007247 Disease - - infertile + + infertility @@ -63486,11 +64067,11 @@ paragraph 7863 Intrauterine insemination using the husband's sperm is commonly performed to overcome male factor problems, as well as to enhance the probability of conception in various other infertility conditions. The pregnancy rate depends on sperm parameters, female factors, and the number of dominant preovulatory follicles. In Stephanie M. Luco's study, the total pregnancy rate observed is 5.3% in 356 IUI cycles. In Ahmed Badawy's study, Seventy-nine clinical pregnancies followed 714 IUI cycles, for a clinical pregnancy rate per cycle of 11.06% and a clinical pregnancy rate per couple of 20.1%. The total clinical pregnancy rate observed in this study, 12.95% per cycle, is considerably higher than Stephanie M. Luco's study and is comparable with Ahmed Badawy's study. - - MESH:D007246 + + MESH:D007247 Disease - - infertility + + infertility conditions @@ -63498,25 +64079,49 @@ paragraph 8630 Miller et al. reported a pregnancy rate per cycle of 12.4% when the total number of collected motile spermatozoa (TMS) is over 20 million, compared with 7.4% when it is between 10 and 20 million. For a value under 10 million, he suggests steering the couple towards IVF. In the same sense, Dickey et al. and Van Voorhis et al. reported the best pregnancy rates when more than 10 million motile spermatozoa were selected. It has been suggested that the number of motile spermatozoa inseminated is a potential predictive factor. In our study, when comparing the semen analysis parameters between the two outcome groups, we found that the initial and post progressive sperm count and the initial motile sperm count was higher in the pregnant group than not pregnant group. These results mean that the semen parameters were related to the outcome of IUI in the current study. In this study, when initial total progressive sperm count was >=5x106, the pregnancy rate was higher than in the group with initial total progressive sperm count <5x106 and the pregnancy rate was compared among the groups with initial total progressive sperm count 5-9.9x106, 10-19.9x106 and >=20x106 (12.73%, 11.11%, and 13.68%, respectively (Table 2). There was no pregnant in the cycles which the initial total progressive sperm count was lower than 5*106, so we should told these infertile couples the poor outcome of IUI and suggested them receive in vitro fertilization (IVF). - + + MESH:C537182 + Disease + + IVF + + Disease TMS + + MESH:D000072660 + Disease + + motile spermatozoa + + + MESH:D000072660 + Disease + + motile spermatozoa + + + MESH:D000072660 + Disease + + motile spermatozoa + DISCUSS paragraph 10096 The findings in Stephanie Marticle indicated that post processing semen analysis parameters are not more predictive of pregnancy than are preprocessing semen analysis results in couples with male factor infertility. Tan et al. found that post washed total progressively motile sperm count (TPMSC) is an independent predictor of pregnancy test result and TPMSC of half million or greater is adequate to achieve statistically similar pregnancy test results after non-donor IUI cycles. The predicted odd of positive pregnancy result is statistically significantly higher when TPMSC is >0.51 million compared to the TPMSC of <0.51 million (OR = 1.68, 95 % CI: 1.04-2.71). Ok EK found that an average TPMSC of 10x106 may be a useful threshold value for IUI success. In this study, there was no pregnancy occurred when TPMSC was <4 million (0/20). Our results also showed that with the increase of post total progressive sperm count, the clinical pregnancy rate increased, similar with the results of those articles (Table 2). - + 9606 Species donor - - MESH:D007246 + + MESH:D007247 Disease infertility @@ -63528,28 +64133,28 @@ 11118 In the present study, 85 clinical pregnancies had been achieved (97.70%) at the end of the third cycle (Table 3). Plosker and Amato advised infertile couples to received IVF after three unsuccessful IUI. Nuoja-Huttunen et al noted that the highest pregnancy rate in 811 cycles (18%) was seen during the first cycle and that 97% of all pregnancies result from the first four cycles. In cases of unexplained infertility, Aboulghar et al found a cumulative pregnancy rate of 39.2% after three cycles and 48.5% after six cycles in a study of 1112 IUI cycles (16.4% per cycle). At present, it is generally admitted that IUI should be limited to four or six cycles and that IVF should be performed. Our results supported the opinion that three inseminations prior to IVF treatments have been usually performed in couples where the cause of infertility was unexplained, due to unilateral tubal factor, mild endometriosis or mild male factor infertility. - MESH:D007246 + MESH:D007247 Disease - - infertile + + infertility - MESH:D007246 + MESH:C537182 Disease - - infertility + + IVF - MESH:D007246 + MESH:D007247 Disease - + infertility - MESH:D007246 + MESH:C537182 Disease - - infertility + + IVF MESH:D004715 @@ -63557,6 +64162,18 @@ endometriosis + + MESH:D007247 + Disease + + unexplained infertility + + + MESH:C537182 + Disease + + IVF + CONCL @@ -63570,23 +64187,11 @@ 12079 IUI is a powerful method to deal with infertile couple with male factors. The initial total progressive sperm count lower than 5*106 means the poor outcome of IUI in the infertile couples with male factor. If the couples with male factor infertility don't get pregnancy after three IUI cycles, the couple should receive re-assessment or other artificial reproductive technology. - MESH:D007246 - Disease - - infertile - - - MESH:D007246 + MESH:D007247 Disease infertility - - MESH:D007246 - Disease - - infertile - CONCL @@ -63612,8 +64217,14 @@ ref 12508 Intrauterine insemination versus fallopian tube sperm perfusion for non tubal infertility + + MESH:D005184 + Disease + + Intrauterine insemination versus fallopian tube + - MESH:D007246 + MESH:D007247 Disease tubal infertility @@ -63654,6 +64265,12 @@ surname:Konidaris;given-names:S 12691 Impact of isolated teratozoospermia on the outcome of intrauterine insemination + + MESH:D000072660 + Disease + + teratozoospermia + 91 @@ -63699,7 +64316,7 @@ 106 12979 Analysis of prognostic factors for successful outcome in patients undergoing intrauterine insemination - + 9606 Species @@ -63720,6 +64337,12 @@ 2065 13082 Influence of the number of motile spermatozoa inseminated and of their morphology on the success of intrauterine insemination + + MESH:D000072660 + Disease + + motile spermatozoa + surname:Barry;given-names:Behr @@ -63736,11 +64359,11 @@ 162 13208 The evaluation of pre and post processing semen analysis parameters at the time of intrauterine insemination in couples diagnosed with male factor infertility and pregnancy rates based on stimulation agent. A retrospective cohort study - - MESH:D007248 + + MESH:D007247 Disease - - male factor infertility + + infertility @@ -63787,6 +64410,12 @@ 668 13692 Effect of the total motile sperm count on the efficacy and cost-effectiveness of intrauterine insemination and in vitro fertilization + + MESH:D005317 + Disease + + cost-effectiveness of intrauterine insemination + 31 @@ -63803,14 +64432,14 @@ 1153 13826 Predictive value of postwashed total progressively motile sperm count using CASA estimates in 6871 non-donor intrauterine insemination cycles - + 1446 Gene 48054 CASA - + 9606 Species @@ -63878,17 +64507,11 @@ 91 14295 Controlled ovarian hyperstimulation and intrauterine insemination for treatment of unexplained infertility should be limited to a maximum of three trials - - MESH:D007246 - Disease - - infertility - - - MESH:D016471 + + MESH:D007247 Disease - - ovarian hyperstimulation + + unexplained infertility @@ -64168,31 +64791,19 @@ paragraph 9302 This research shows that there is a significant and meaningful relationship between the sexes of users and using computer systems; thus, female middle manager show more utilization of the computer systems than male managers and this finding is consistent with the finding of Al-Gahtani, 2004 and Meade, et al., 2009. In this research between the age and the education level of the participants with computer usage system did not show any meaningful relationship, while in the studies of Marchewka and Al-Gahtani it showed a significant relationship. - + 9606 Species participants - - MESH:D000535 - Chemical - - Al - - - MESH:D000535 - Chemical - - Al-Gahtani - DISCUSS paragraph 9852 In a Level of %95 between organizational resources and IT implementation there is a significant relationship which showed a correlation intensity of r= 0.297. Regarding the relationship between these two variables it can be said that due to some facts like accessible financial and human resources, enough budget for the application of new technologies, strategic planning of the organizations' resources that can lead the managers to realize the mission and targets of the organizational resources, the attitude of the users might be influenced and it is concluded that the more available resources is provided for the users, the more tendency they show in application of Information technology. - + 9606 Species @@ -64204,7 +64815,7 @@ paragraph 10551 The above theory is confirmed due to a significant relationship between the users' attitude toward IT application and organizational knowledge with a level of %99 and a correlation coefficient of (r= 0.297). Therefore, it can be inferred however the organizational knowledge is not in a technology oriented category, information technology application is a necessity for knowledge management projects to succeed. Hence some organizations seek knowledge management systems for executing their organizational knowledge management projects. In this way they might become confused because of lack of deep and correct understanding of capabilities and abilities of the organizations and susceptible areas for knowledge technology products. Considering the main three factors of knowledge management (individuals, contents, processes) an organization should identify the proper technology for itself. Accordingly, the tools for knowledge management should be determined based on the needs of the organization. Thus, it can be generally stated that when organizational knowledge in an organizations is used properly, people tend to use more of information technology because it facilitates the access and management of this knowledge. These findings are in agreement with previous researches like Armenakis&Bernerth in 2007 and Bartunek& Rousseau in 2006. - + 9606 Species @@ -64216,7 +64827,7 @@ paragraph 11902 The relationship between processes and the attitude toward application of information technology is confirmed with a level of %99 and a correlation coefficient of (r= 0.409) that approves the above theory. Therefore, it can be concluded that in organizations in which processes are shorter and easier beside shorter organizational communication and fewer organizational levels, they show more flexibility in case of facing with environmental and technological changes. In other words, staff in organizations with horizontal structure can communicate with managers easier and they participate in decision makings more effectively. So, organizations with the least division of responsibilities and informal processes have an organic structure in which people tend to use information technology more. This finding is also in consistent with Armenakis Schraeder & Tsiknakis &Kouroubali, 2009 and Yu li H, P, 2009. - + 9606 Species @@ -64234,7 +64845,7 @@ paragraph 13613 The relationship between values and goals and the attitude toward application of information technology is confirmed with a level of %95 and a correlation coefficient of (r= 0.246) in this regard, it can be stated that goals and strategies are as the main framework of planning. Any plan requires a goal which explains the philosophy of existence and it also elaborates the needs, the objectives and the final results in order to support the subordinate goals of the organization. In fact, the goals create a kind of network or a hierarchy system. In addition, just like the need to know the tradition for a better understanding of organizational behavior, noting the values would be another aspect that can affect people's behavior in the organizations. Values have great influence on career decision makings, directions, behaviors, relationships and the attitudes of the staffs. According to the findings in this research, when implementation and application of information technology becomes a priority in the organizations' goals, there should be a strategy and it also ought to be valued in order to be relatively effective in application of information technology. This is in accordance with Simpson & Dansereau, 2007. - + 9606 Species @@ -64246,7 +64857,7 @@ paragraph 14840 Among the variables included in the equation, managerial structure (beta= 0.760) and then human resources (beta=0.153) have the strongest affect on information technology application, so it is concluded that senior managers should focus more attention to these variables since they are the main important variables. - + 9606 Species @@ -64300,7 +64911,7 @@ 52 15794 Organizational factors affecting successful adoption of innovative eHealth services: A case study employing the FITT framework - + MESH:C043685 Chemical @@ -64445,7 +65056,7 @@ 558 16728 What factors affect the use of electronic patient records by Irish GPs? - + 9606 Species @@ -64724,54 +65335,48 @@ abstract 511 A cross sectional study was conducted on thirty (30) good quality lateral cephalograms. Only those lateral cephalograms were selected where Point A was easily identified. Landmarks: Sella (S), Nasion (N), Point A and three substitution points Y, L, X were traced. Angles formed by SN with Point A (Angle SNA) and three substitution points (Angle SNY, SNX, SNL) were measured. Correlation of angle SNA with angles SNY, SNX and SNL were derived. Statistical analysis used - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - - MESH:D009584 - Chemical - - N - ABSTRACT @@ -64784,105 +65389,105 @@ abstract 990 Mean and standard deviation for Angles SNA, SNY, SNL and SNX were calculated individually for males and females. 'T' Test was applied to determine statistical significance for all the parameters i.e Age, Angles SNA, SNY, SNL and SNX respectively. Karl Pearson correlation coefficient was carried out to determine the statistical significant correlation for Angle SNA with SNY, SNL and SNX. Results: A mean value of 82.8 +-1.9 , 83.1 +-1.8 , 78.3 +-2.9 and 78.7 +-2.7 for Angle's SNA, SNY, SNL and SNX respectively was observed. A statistically significant correlation was observed between angles SNA & SNY, SNL, SNX & strong positive correlation was observed with angle SNY. Conclusions: We conclude that Point Y is the most nearing maxillary apical base landmark to Point A. Hence maxillary apical base landmark can be substituted by Point Y where identification of point A is not obvious. - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -64901,7 +65506,7 @@ paragraph 1915 Cephalometric assessment of patients is an essential adjunct to achieve an accurate orthodontic diagnosis aiding for comprehensive orthodontic treatment planning. The extreme range or radiolucency between bone and soft tissues makes it impossible to locate consistently all landmarks on routine radiographs. Studies regarding the reliability of cephalometric landmarks have been differentiated by (a) differences between two films of the same subject, (b) observed differences in locating the points, and (c) variations in measuring the distance between two marked points. The factors influencing accurate identification were quoted as distinctness of structural detail, noise from adjacent structures due to superimposition of conflicting anatomic details, and conceptual judgment, a factor which is largely based on the past experience and radiological knowledge of the observer. In spite of improved techniques, occasionally certain landmarks are still difficult to locate, among them Point A or Subspinale is one such landmark. Point A is a midline point whose relationship to the anterior teeth in a lateral head film may be influenced by head position. Almost all cephalometric analysis such as Steiner's, Down's, Wit's Appraisal, Mc Namara to name a few use point A or the NA plane as a reference point or plane. Because of difficulty in locating point A various authors i.e Van der Linden, Jarabak and Fizzel, Jacobson R. and Jacobson A have given different substitutions for Point A. The aim of this study was to identify the most nearing alternate maxillary apical base landmark from pre-existing cephalometric points given by different authors. - + 9606 Species @@ -64919,49 +65524,49 @@ paragraph 3595 The study was started after obtaining the approval from the Ethical Committee. A cross-sectional study was conducted on 30 (12 males; 18 females) pretreatment good quality lateral cephalograms from patients visiting to Department of Orthodontics and Dentofacial Orthopedics. The lateral cephalograms were selected such that the Point A could be accurately located. A purposive convenience sampling technique was utilized for our study. Patients of age more than 16 yrs only were included in the sample. Patients with tooth agenesis or supernumeraries, any developmental anomalies, traumatic injuries or fractured upper and lower incisors and molars, complex craniofacial deformities or syndromes, Patients who have undergone any orthodontic treatment were excluded from our study. The lateral cephalograms (Kodak 8000C Digital Panoramic and Cephalometric Systems) utilized in our study were of true size (1:1) and any faulty radiographs with image distortion were excluded. The exposure time ranged from 12.8 to 13.9 seconds with kV 69-71 and m/A 10-12. All the cephalograms were manually traced for landmark by one investigator. - + 9606 Species patients - + 9606 Species Patients - + 9606 Species Patients - + 9606 Species Patients - + MESH:D050723 Disease traumatic injuries or fractured upper - + MESH:D000848 Disease tooth agenesis - + MESH:D019465 Disease craniofacial deformities - + MESH:D000014 Disease @@ -64979,34 +65584,28 @@ paragraph 4753 After categorization of the cephalograms, on an acetate tracing paper the following landmarks were traced, Sella (S), Nasion (N), Point A, Point B, Point Y, Point L and Point X (Table 1, Figure 1) were located and the angles SNA, SNY, SNL and SNX were measured (Figure 2). - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - - MESH:D009584 - Chemical - - N - - + MESH:D000085 Chemical @@ -65036,14 +65635,14 @@ fig_caption 5117 Various angles formed a: Angle SNA, b: Angle SNY, c: Angle SNL, d: Angle SNX - + 6624 Gene 48164 SNL - + 6615 Gene 4363 @@ -65062,63 +65661,63 @@ paragraph 5214 All the statistical tests were conducted using SPSS (version 19.0) Mean and standard deviation for Angles SNA, SNY, SNL and SNX were calculated individually for males and females. 'T' Test was applied to determine statistical significance for all the parameters i.e Age, Angles SNA, SNY, SNL and SNX respectively. Karl Pearson correlation coefficient was carried out to determine the statistical significant correlation for Angle SNA with SNY, SNL and SNX. - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -65137,21 +65736,21 @@ paragraph 5686 Males: A mean value of 22.1 +-4.3 years for Age was observed. A mean value of 82.80 +-1.950 for Angle SNA was observed. A mean value of 83.10 +-1.80 was observed with Angle SNY. A mean value of 78.30 +-2.90 was observed with Angle SNL. A mean value of 78.70 +-2.70 was observed with Angle SNX (Table 2). - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 @@ -65166,21 +65765,21 @@ table_caption 5990 Measured angles of SNA, SNY, SNL and SNX in our study - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -65193,49 +65792,49 @@ paragraph 6044 Females: A mean value of 19.8 +-3.2 years of Age was observed. A mean value of 80.90 +-2.40 was observed for Angle SNA. A mean value of 81.10 +-2.50 was observed with Angle SNY. A mean value of 77.30 +-3.40 was observed with Angle SNL. A mean value of 77.90 +-3.40 was observed with Angle SNX (Table 2). T test was carried out to determine significance amongst all the parameter Age, SNA, SNY, SNL and SNX. Angles SNA (P<0.033) and SNY (P<0.023) revealed statistical significance (Table 3). - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -65250,21 +65849,21 @@ table_caption 6535 Determination of statistical signifi cance for parameters SNA, SNY, SNL, SNX and Age - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -65277,42 +65876,42 @@ paragraph 6620 A correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. SNA revealed a strong statistically significant correlation with SNY>SNX>SNL (Table 4). - + 6615 Gene 4363 SNA - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - + 310 Gene 36149 @@ -65327,21 +65926,21 @@ table_caption 6853 Determination of overall correlation coeffi cient for parameters SNA, SNY, SNL, SNX and Age - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 6615 Gene 4363 @@ -65354,28 +65953,28 @@ paragraph 6945 In males correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. Only SNA revealed a strong statistically significant correlation with SNY only. (Table 5). - + 6615 Gene 4363 SNA - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 @@ -65390,21 +65989,21 @@ table_caption 7188 Determination of correlation coefficient for parameters SNA, SNY, SNL, SNX and Age for females. - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 @@ -65417,42 +66016,42 @@ paragraph 7284 In females correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. SNA revealed a strong statistically significant correlation with SNY, SNL and SNX (Table 6). - + 6615 Gene 4363 SNA - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 SNX - + 310 Gene 36149 @@ -65467,21 +66066,21 @@ table_caption 7531 Determination of correlation coefficient for parameters SNA, SNY, SNL, SNX and Age for males - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 310 Gene 36149 @@ -65500,168 +66099,168 @@ paragraph 7638 Apical base of maxilla and mandible help in determining the spatial relation of both maxilla and mandible to the cranial base. It also determines the limit of placement of incisors in the anteroposterior position. Numerous controversies exist in landmarks which are difficult to identify. Among those points, Point A is the most common point which encounters difficulty in identification. The cephalometric landmark, Point A, was investigated with regard to definition, location and usefulness in cephalometric analysis. Point A or Subspinale represents the maxillary apical base; the projection of cheeks frequently obscures this landmark in lateral cephalogram. Due to shortcomings of Point A various substitute landmarks have been sought by different authors by keeping the root apex of maxillary central incisor as a stable landmark. Van der Linden, suggested the use of point L, which is located on the anterior surface of the image of the labial lamella at the region of the apex of the maxillary incisors Jarabak and Fizzel identified a Point X 2mm ahead of the root apex as a redefinition of point A. Another Point Y plotted 3mm labial to a point between upper and lower two-thirds of the long axis of the root of the maxillary central incisor as suggested by Jacobson R. and Jacobson A. Comparison of mean values of Angle SNA with Angles SNY, SNL and SNX When the mean values of angles SNA, SNL, SNX and SNY were compared we observed the mean value of SNY was more in comparison to SNA, SNL and SNX. The mean values of SNL and SNX was less in comparison to SNA. Probable cause for such an observation could be attributed the variation in the definition of those cephalometric landmarks. When T test was carried out to determine significance amongst all the parameters individually like Age, SNA, SNY, SNL and SNX, Angles SNA and SNY revealed statistical significance (Table 3). This suggested that angles SNA and SNY were more specific for all the samples included in the study. On an overall SNA and SNY; SNX and SNL revealed strong statistical correlation between each other. Clear visibility of maxillary central incisor would act as a guide for marking the landmarks as require for Point Y could be attributed for such an observation. In males there was high correlation of angle SNA with angle SNY. In females, angle SNA correlated with Angle SNY, SNL and SNX, as we had observed that all the angles were less in comparison to males, which resulted in such an observation. However a further study has to be conducted on females to ascertain the probable cause for such an observation. - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6615 Gene 4363 SNA - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6624 Gene 48164 SNL - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 310 Gene 36149 SNX - + 6615 Gene 4363 SNA - + 6615 Gene 4363 SNA - + 6615 Gene 4363 @@ -65674,14 +66273,14 @@ paragraph 10237 In almost all cases the angles SNA and SNY were equal and showed high statistical significant correlation. Abdwani et al stated that the effects of incisal inclination changes, due to orthodontic treatment, are of no clinical relevance to the position of Point A and B, even though they may be statistically significant. However Kazem et al reported that the position of Point A is affected by local bone remodeling associated with proclination of the upper incisor in Class II division 2 malocclusion, but this minor change does not significantly affect the SNA angle. According to Jacobson a point closer to the center of the root of a tooth is less vulnerable to displacement than, say, a point close to the root apex during crown tipping procedures. Point Y represents closer to center of the tooth root hence point Y can be used more precisely as a substitute for point A. Whereas Point L and Point X are located in relation to the root apex, which can change if the tooth is proclined or retroclined. - + 6615 Gene 4363 SNA - + 6615 Gene 4363 @@ -65821,12 +66420,6 @@ 884 12262 Maxillary Incisor Proclination Effect on the Position of Point A in Class II division 2 Malocclusion - - MESH:D008310 - Disease - - Malocclusion - @@ -65853,18 +66446,12 @@ Jafari F, Mobasheri M, Mirzaeian R 0 Effect of Diet Education on Blood Pressure Changes and Interdialytic Weight in Hemodialysis Patients Admitted in Hajar Hospital in Shahrekord - + 9606 Species Patients - - MESH:D015431 - Disease - - Weight in Hemodialysis - ABSTRACT @@ -65877,23 +66464,29 @@ abstract 162 Nutrition is a key factor in the treatment of patients with chronic kidney disease because kidney burden decrease causes uremic reduction and its side effects. The aim of this research is to examine the effect of diet education on blood pressure changes and interdialytic weight in Hemodialysis patients admitted to Hemodialysis ward of Hajar hospital in Shahrekord. - + 9606 Species patients - + 9606 Species patients + + MESH:D051437 + Disease + + kidney burden decrease causes uremic reduction + - MESH:D007676 + MESH:D051436 Disease - - chronic kidney disease because kidney burden decrease causes uremic reduction + + chronic kidney disease @@ -65985,239 +66578,227 @@ paragraph 1769 Advanced chronic renal failure is a progressive and irreversible impairment where the kidney's ability to filter metabolic waste products from the blood, and maintain water and electrolyte balance disappears, hence resulting in uremia. In patients with advanced chronic renal failure, it is impossible for them to live without adequate alternative treatment. Nowadays, the best alternative treatment is hemodialysis. Renal failure is highly prevalent in the world and in Iran. Nowadays, 242 out of 1000 people in the world suffer from renal failure; besides, about 8% are added to this figure annually. Although hemodialyses has improved renal disease and prolong patients' lives, without the proper diet and medication, it is not highly efficient. Following diet, fluid restriction and drugs are essential in care and health maintenance of renal dialysis patients. For these patients, the purpose of diet is the reduction of kidney burden, prevention of renal complications, maintaining proper nutrition, and prevention of uremia and its complications. Evidence indicates that many patients undergoing hemodialysis do not adhere to proper diet and fluid intake restrictions. It seems that many patients are not aware of diet and fluid restriction. Dialysis alone cannot provide the health and survival to the patients in long term. Besides hemodialysis, therefore, diet, fluid restriction, and drug therapies are essential treatments for chronic renal failure. Poor diet, and lack of fluid restriction can have serious consequences for patients, including impaired physical abilities, depression, acute pulmonary edema, congestive heart failure, and premature death. A poor dietary habit is associated with low quality of life, and morbidity and mortality of dialysis patients. Further, malnutrition and inflammation increase cardiovascular risk and mortality in hemodialysis patients, while following the diet and restricting fluid intake decrease the risk of complications and increase quality of life. Yet, many patients do not follow their diet. Previous studies on the role of nurses in the renal failure patients' diet plan show that the most important objectives in hemodialysis patient care are preventing malnutrition and maintaining good nutrition, edema control, delaying renal osteodystrophy by controlling phosphorus and calcium levels, and finally encouraging the patient to consume tolerable foods which are in accordance with lifestyle changes. Lack of awareness regarding the effects of diet on blood pressure made hemodialysis patients always suffer from its complications. Since this matter is of vital importance, the present study aims to determine the effect of diet on the interdialytic blood pressure and weight in patients undergoing hemodialysis treatment. - + 9606 Species patients - + 9606 Species people - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - - MESH:D007674 - Disease - - renal dialysis + + MESH:D014867 + Chemical + + water - - MESH:D051437 + + MESH:D000275 Disease - - reduction of kidney burden + + depression - + MESH:D014511 Disease uremia - + MESH:D051437 Disease renal failure - + MESH:D051437 Disease Renal failure - + MESH:D007676 Disease chronic renal failure - + MESH:D051437 Disease renal failure - + MESH:D007676 Disease chronic renal failure - + MESH:D004487 Disease edema - - MESH:D011654 - Disease - - pulmonary edema - - - MESH:D059445 - Disease - - impaired physical abilities - - + MESH:D007674 Disease renal complications - + MESH:D006333 Disease congestive heart failure - + MESH:D010758 Chemical phosphorus - + MESH:D012080 Disease delaying renal osteodystrophy - + MESH:D002118 Chemical calcium - + MESH:D007676 Disease chronic renal failure - + MESH:D014511 Disease uremia - + MESH:D003643 Disease premature death - + MESH:D002318 Disease inflammation increase cardiovascular - + MESH:D007674 Disease renal disease - - MESH:D003866 + + MESH:D011654 Disease - - depression + + acute pulmonary edema @@ -66231,61 +66812,61 @@ paragraph 4580 This quasi-experimental and interventional study of 100 dialysis patients admitted in Hemodialysis ward of Hajar hospital was conducted in 2011 where a pretest as well as a post test took place. Patients were enrolled after giving informed consent. The patients who intended to undergo a kidney transplant during the educational program, or leave the place of dialysis, or those who are frequently ill and unable to afford a full time hemodialysis or those who suffer from another known disease besides advanced chronic kidney failure were excluded from the study. The instrument used for data collection was a two-part researcher-designed checklist which consisted of questions on demographic information, knowledge of consumption of food, systolic and diastolic hypertension control, as well as the weight control of the patients. The reliability of this checklist was determined by nutritionists. Before starting the educational program, data on weight, blood pressure and the results of Cr and BUN tests were recorded. The educational program was conducted during two months for the patients. Carried out through face to face interview, bulletins, pamphlets and educational CD, this program was concerned with the control of salt and fluid intake, and the right way of food consumption. Two weeks after beginning of the educational program, weight and blood pressure changes were measured, then at the end of the first month of program, a questionnaire was given to patients through which demographic data were recorded. Their knowledge of proper diet and nutrition was evaluated, as well. Two weeks later and at the end of the second month, weight and blood pressure were measured, and again the patients were given the questionnaire, then data obtained from the questionnaires, including weight, blood pressure and other variables were recorded. The data were entered into SPSS 16, and then the results were analyzed by using descriptive statistics, paired t-test and ANOVA. - + 9606 Species patients - + 9606 Species Patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + + MESH:D012492 + Chemical + + salt + + MESH:C563897 Disease diastolic hypertension - - MESH:D002857 - Chemical - - Cr - - + MESH:D007676 Disease @@ -66303,7 +66884,7 @@ paragraph 6572 Results showed that patients were between 13-80 years old with a mean age of 56.2+- 15.14. Of them, 69% were male and the rest 31% were female. In terms of education, the majority of the subjects (81.4%) were illiterate or educated below the 9th grade, and 18.6% were educated beyond this grade. The mean duration of dialysis was 40.7+- 36.40. The maximum and minimum of duration for the subjects undergoing dialysis were 20 years and 4 month, respectively. Further, the mean frequency of dialysis was 22.40+- 2.9 times a week. - + 9606 Species @@ -66315,68 +66896,56 @@ paragraph 7100 According to the patient's medical record, which was filled by the physician regarding muscle mass and cardiovascular problems each month, the initial mean weight of the patients was 66.15 +- 15.10. The lightest patient was 31.5 kg and the heaviest one 105 kg. After the first stage of educational program, the mean of weight become 64.43 +- 14.67. - + 9606 Species patient - + 9606 Species patients - + 9606 Species patient - - MESH:D002318 - Disease - - cardiovascular problems - RESULTS paragraph 7449 Results showed that the mean of BUN and Cr for patients before training were 77.98 +- 21.74 and 9.47 +- 2.80 respectively, which become 66.14 +- 12.13 and 8.48 +- 2.36 after the first stage of training. There was a significant difference between the Cr of the patients before and after training (P <= 0/01). However, the BUN of the patients were not significantly different before and after the training (P <= 0/31). - + 5714 Gene 37686 P <= 0/31 - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D002857 - Chemical - - Cr - - + MESH:D002857 Chemical @@ -66388,14 +66957,14 @@ paragraph 7868 Moreover, the findings represented that the mean systolic and diastolic blood pressure in the three stages of the research reduced to 6.65 +- 1.51 mmHg and 2.24 +- 1.82 mmHg, respectively. The mean systolic blood pressure before and after the test was significantly different (P <= 0/05). However, there was no significant difference between the mean diastolic blood pressure before and after the test (P <= 0/20) (Table 1). - + 51673 Gene 32659 P <= 0/20 - + 106479023 Gene @@ -66409,13 +66978,13 @@ table_caption 8295 Weight, creatinine, urea, and systolic and diastolic blood pressure in stages of study - + MESH:D003404 Chemical creatinine - + MESH:D014508 Chemical @@ -66435,42 +67004,36 @@ paragraph 8430 Findings on the determination and comparison of the problems of the patients undergoing hemodialysis (with regard to the systolic and diastolic blood pressure, weight gain, and the changes before and after training including the changes in Urea, creatinine, blood pressure and weight) are important indicators and have a huge role in creating problems and reducing the quality of life. These findings indicated that there was no significant difference in the Cr means of males and females after diet education in the experimental investigation (P=0.2). The relationship between gender and BUN showed a difference in males and females after training (P=0.026). Further, males and females significantly differed from each other in terms of interdialytic weight gain and systolic blood pressure. (P=0. 003), but no significant difference was observed between diastolic blood pressure (P=0.4). - + 9606 Species patients - + MESH:D015430 Disease interdialytic weight gain - + MESH:D015430 Disease weight gain - + MESH:D003404 Chemical creatinine - + MESH:D014508 Chemical Urea - - MESH:D002857 - Chemical - - Cr - DISCUSS @@ -66483,35 +67046,35 @@ paragraph 9334 The findings represented that educating patients with a special emphasis on diet can have a positive effect on their attitude. Interdialytic weight loss indicated that training had an effect on patients and enabled them to make adjustments in their diet. Toolabi has investigated the effect of diet education on the knowledge and practice of Hemodialysis Patients in Lorestan, which indicated a significant increase in knowledge of patients after training. - + 9606 Species patients - + 9606 Species patients - + 9606 Species Patients - + 9606 Species patients - + MESH:D015431 Disease - - Interdialytic weight loss + + weight loss @@ -66519,162 +67082,174 @@ paragraph 9791 One of the common problems to these patients is Uremia that can cause poor appetite, memory loss, drowsiness, misjudgment and poor concentration. Interdialytic weight gain increases blood pressure by 3 mm Hg per kg of body weight. Results of nutritional evaluation of 99 hemodialysis patients in Tehran represented a significant positive correlation between protein intake through the correct way of nutrition, and body weight. Therefore, nutrition education is an important variable in increasing awareness and health status of hemodialysis patients. Previous research showed that it is necessary to educate patients with high blood pressure to follow a good diet and restrict the use of fat and sodium. Lack of attention to the patient education regarding drug use, and food and fluid restriction has adverse consequences. Considering the persistent problems as well as their kinds, inefficiency of dialysis in complete resolution of these problems, and the need for continuous care, education can provide an informed and active participation of the patient to care for themselves. The subjects in this study had a low level of awareness before training. This was due to the insufficient diet education which is consistent with the study carried out by Toulabi entitled Impact of education on knowledge and practice of dialysis patients in Lorestan. In this study, there was a significant relationship between education and weight after training which was consistent with Alipour's findings on knowledge, attitude and practice of renal dialysis patients' nutrition. In the study entitled the effect of diet education on dialysis efficacy which is conducted in Zanjan, Talebipour in three stages of training showed that after training, there was an increase in the tendency of these patients to follow an efficient diet. The results of Talebipour's study is similar to those of the current research. Shab Bidar, et al. in their study of dialysis adequacy and prevalence of malnutrition in hemodialysis patients have suggested that in 23% of patients albumin level was below 3 (severe malnutrition), and in 32% of them albumin was in the range of 3-3.5 (medium malnutrition). Moreover, 75 to 80% of hemodialysis patients have high blood pressure, and despite using various drugs, the major cause of hypertension in these patients is the increase in blood volume, which is due to water and salt retention in the body; therefore, withdrawing fluid in a short period from these patients has had a significant effect on blood pressure. Durose, et al have shown that educating patients about diet and alternative consumption can lead to the fluid intake restriction and subsequently to weight loss. Based on the findings, there was a significant association between the weight of the subjects and their level of education that is consistent with the findings of the study conducted by Alipour representing a positive correlation between the knowledge and practice of the subjects, and their level of education. The main achievement in this study was that education is positively effective since the goal of education is to enable us to establish an assuring relationship between patients and nurses:considering education and support of the patients. This is implemented if once patients are allowed to actively participate in decision-making and planning for themselves. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patient - + 9606 Species patient - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - - MESH:D044342 + + MESH:D000309 Disease - - malnutrition in hemodialysis + + insufficient - - MESH:D007674 + + MESH:D012492 + Chemical + + salt + + + MESH:D015430 Disease - - renal dialysis + + weight gain increases blood - + MESH:D014511 Disease Uremia - + + MESH:D015431 + Disease + + weight loss + + MESH:D008569 Disease memory loss - + + MESH:D014867 + Chemical + + water + + MESH:D012964 Chemical sodium - + MESH:D006970 Disease drowsiness - + MESH:D006973 Disease hypertension - - MESH:D015430 - Disease - - Interdialytic weight gain increases blood - CONCL @@ -66687,25 +67262,25 @@ paragraph 13179 Diet and drug therapy besides hemodialysis can improve health condition and therapeutic effectiveness for patients with renal failure. However, it seems that many patients are not fully aware of diet and fluid intake restriction. This study showed that periodic educational programs can play a significant role in the health of patients and their attitude and knowledge of nutrition. - + 9606 Species patients - + 9606 Species patients - + 9606 Species patients - + MESH:D051437 Disease @@ -66735,17 +67310,6 @@ 1334 13611 Brunner and Suddarth S - - MESH:C563156 - Disease - - Brunner - - - Chemical - - Suddarth - surname:Bare;given-names:BG @@ -66758,17 +67322,6 @@ 1150 13634 Brunner and Suddarth S - - MESH:C563156 - Disease - - Brunner - - - Chemical - - Suddarth - surname:Braunwald;given-names:E @@ -66788,7 +67341,7 @@ surname:Fauci;given-names:AS 13657 Chronic renal failure - + MESH:D007676 Disease @@ -66807,12 +67360,6 @@ 163 13679 Intervention on Strategies for improving fluid restriction compliance in chronic hemodialysis patients - - MESH:D002908 - Disease - - chronic hemodialysis - surname:Stump;given-names:SE @@ -66845,7 +67392,7 @@ 68 13814 Are hemodialysis patient population at risk - + 9606 Species @@ -66865,13 +67412,13 @@ 646 13858 Optimizing nutrition in patients with chronic kidney disease - + 9606 Species patients - + MESH:D051436 Disease @@ -66894,13 +67441,13 @@ surname:Phipps;given-names:W 13919 Management of persons with renal failure - + 9606 Species persons - + MESH:D051437 Disease @@ -66924,7 +67471,7 @@ surname:Galetta;given-names:F 13960 Nutritional status and dietary manipulation in predialysis chronic renal failure patients - + MESH:D007676 Disease @@ -66949,7 +67496,7 @@ surname:Stevens;given-names:P 14050 Quality of life in patients on chronic dialysis: self-assessment 3 months after the start of treatment - + 9606 Species @@ -66980,7 +67527,7 @@ 47 14173 Response of patients undergoing Hemodialysis - + 9606 Species @@ -67001,23 +67548,18 @@ 288 14218 Risk factors for hypertension in chronichemodialysis patients: baseline data from the HEMO study - + 9606 Species patients - + MESH:D006973 Disease hypertension - - Chemical - - HEMO - REF @@ -67026,7 +67568,7 @@ ref 14315 Evaluating the hemodialysis patients based on nutrition - + 9606 Species @@ -67045,13 +67587,13 @@ 16 14371 Evaluatin the relationship between the weight during the hemodialys session in hemodialysis patients and hypertension increasing in Booali hospital - + 9606 Species patients - + MESH:D006973 Disease @@ -67072,7 +67614,7 @@ 62 14519 Study of knowledge, attitude and practice of nutrition for kidney patients undergoing hemodialysis - + 9606 Species @@ -67091,7 +67633,7 @@ 36 14618 Establishment of dietary education in hemodialysis efficacy in dialysis patients in Dr. Reisi hospital - + 9606 Species @@ -67104,7 +67646,7 @@ ref 14721 Estimating the efficience of dialysis and malnutrition prevalence in hemodialysis patients. Razi training and treatment center of Rasht - + 9606 Species @@ -67126,7 +67668,7 @@ 42 14857 Knowledge of dietary restrictions and the medical consequences of noncompliance by patients on Hemodialysis are not predictive of dietary compliance - + 9606 Species @@ -67210,7 +67752,7 @@ children - MESH:C029045 + - Chemical Technituim-99m-Methyle diphosphonate @@ -67282,85 +67824,91 @@ abstract 648 Bone scintigraphy revealed 17 (25%) abnormal scans in 11 boys and 6 girls. Scans findings were suggestive of spondylolysis (n=4); malignancy including primary tumors and metastases (n=3); infection including osteomyelitis and discitis (n=3); sacroiliitis (n=2); benign tumors (n=2); pseudo fractures in ribs (n=1); necrosis in femoral head epiphysis(n=1) and nonskeletal-renal retention due to hydronephrosis (n=1). Sensitivity, specificity and accuracy of bone scan in detecting gross skeletal abnormality as a cause for back pain were 94% and 100% and 99% respectively. - + 9606 Species boys - + 9606 Species girls - + MESH:D007239 Disease infection - - MESH:D060048 + + MESH:C538496 Disease - - femoral head epiphysis(n=1) and nonskeletal-renal retention + + skeletal abnormality - + MESH:D009362 Disease metastases - + MESH:D001416 Disease back pain - + + MESH:D007674 + Disease + + nonskeletal-renal retention + + MESH:D015299 Disease osteomyelitis and discitis - + MESH:D009369 Disease malignancy - + MESH:D058566 Disease sacroiliitis - + MESH:D009369 Disease benign tumors - + MESH:D009369 Disease tumors - + MESH:D006869 Disease hydronephrosis - + MESH:D050723 Disease fractures - + MESH:D009336 Disease @@ -67378,13 +67926,13 @@ abstract 1232 Bone isotope scan is a sensitive imaging modality in the assessment of pediatric patients with back pain. It is a reliable modality to detect and role out most benign and aggressive serious etiologies. - + 9606 Species patients - + MESH:D001416 Disease @@ -67402,31 +67950,31 @@ paragraph 1450 Back pain in relatively less common in children than in adults. Most cases of back pain in children clear up in a few days without medical attention. In few cases the symptoms will persist for a longer time which may indicate underlying serious problem more than just a simple backache. Causes can range from serious malignant causes to just benign lesions. Therefore, it is important to start with sensitive and cost effective imaging modality that is capable of distinguishing simple back pain from serious underlying diseases. - + 9606 Species children - + 9606 Species children - + MESH:D010146 Disease pain - + MESH:D001416 Disease back pain - + MESH:D001416 Disease @@ -67438,13 +67986,13 @@ paragraph 1980 A variety of imaging techniques has been used to confirm the cause of back pain. A conventional x-ray is often the first imaging technique which has been used. Unfortunately; this technique suffers from low sensitivity, and can miss most causes of back pain including serious pathologies. On the other hand, advanced imaging modalities like computerized tomography (CT scan) and magnetic resonance imaging (MRI) can be ordered. However, such modalities are not cost effective, and not used routinely to image the whole body. - + MESH:D001416 Disease back pain - + MESH:D001416 Disease @@ -67456,13 +68004,13 @@ paragraph 2505 Bone scintigraphy (BS) has been used in the diagnostic work-up of unexplained bone pain. The technique is sensitive and allows visualization of the whole skeleton in a short time. This modality is more sensitive than x-rays, since early or non-aggressive lesions generally cannot be seen on radiographs until significant bone mineral matrix has been lost. - + 255564 Species rays - + MESH:D010146 Disease @@ -67474,13 +68022,13 @@ paragraph 2861 Our study will assess the diagnostic value of BS in children presented with back pain. We will try to emphasize on efficiency of this modality in localizing many pathologies that would have been missed by conventional x-ray study. Also, we will make an overview for scan findings in some lesions encountered during our study. - + 9606 Species children - + MESH:D001416 Disease @@ -67492,7 +68040,7 @@ title_1 3187 2. PATIENTS AND METHODS - + 9606 Species @@ -67504,55 +68052,49 @@ paragraph 3211 Included in this retrospective study were 68 children with ages ranging from 4 to 15 years (mean age of 11+- 3). There were 45 boys and 23 girls. All children presented with back pain of 7 to 88 days duration with a mean value of 18+-5 days. All children underwent BS for assessment of back pain. There initial laboratories and conventional radiological investigation failed to define or localize any possible etiology. All patients underwent 99mTc MDP BS which has been carried out two hours after intravenous administration of radioisotope tracer with calculated dose according to body weights by the formula: Pediatric dose = age (in years)/age+12 x adult dose (20 mCi). - + 9606 Species children - + 9606 Species boys - + 9606 Species girls - + 9606 Species children - + 9606 Species children - + 9606 Species patients - + 1800 Gene 80192 MDP - - MESH:C107828 - Chemical - - 99mTc - MESH:D001416 Disease @@ -67613,67 +68155,61 @@ paragraph 4807 Bone scintigraphy revealed 17 (25%) abnormal scans in 11 boys and 6 girls. Abnormal scans were described according to anatomical sites which were congruent with the site of pathology in all cases. Table 1 shows anatomical distribution of scan abnormalities in 17 abnormal scans. Scans findings were suggestive of spondylolysis (n=4); malignancy including primary tumors and metastases (n=3); sacroiliitis (n=2); infection including osteomyelitis and discitis (n=3); benign tumors (n=2); pseudofracture (ribs) (n=1); necrosis in femoral head epiphysis (n=1) and nonskeletal (renal) (n=1). Table 2 shows BS findings and diagnosis in 17 abnormal scans encountered during our study. BS failed to detect one case of disc herniation. - + 9606 Species boys - + 9606 Species girls - + MESH:D009336 Disease necrosis - + MESH:D009369 Disease benign tumors - + MESH:D007239 Disease infection - + MESH:D058566 Disease sacroiliitis - + MESH:D009369 Disease tumors - - MESH:D060048 - Disease - - femoral head epiphysis - - + MESH:D009362 Disease metastases - + MESH:D009369 Disease malignancy - + MESH:D015299 Disease @@ -67687,12 +68223,6 @@ table_caption 5535 Anatomical distribution of bone scans abnormalities - - MESH:D004401 - Disease - - bone scans abnormalities - TABLE @@ -67707,7 +68237,13 @@ paragraph 5650 Sensitivity, specificity and accuracy of bone scan in detecting gross skeletal abnormality as a cause for back pain were 94% and 100% and 99% respectively. Positive predictive value (PPV) and negative predictive value (NPV) were found to be 100% and 98% respectively. - + + MESH:C538496 + Disease + + skeletal abnormality + + MESH:D001416 Disease @@ -67725,67 +68261,67 @@ paragraph 5932 Back pain is uncommon in children younger than 3 years, increasing its frequency with age until reaching a plateau during early adolescence. Most often, back pain is benign and self-limited. However, it is occasionally the presenting symptom of systemic diseases as cancer or infection. Thus, the major diagnostic task is to distinguish the patients with simple back pain from those with serious underlying diseases. - + 9606 Species children - + 9606 Species patients - + + MESH:D001416 + Disease + + back pain + + MESH:D034721 Disease systemic diseases - + MESH:D007239 Disease infection - + MESH:D001416 Disease back pain - + MESH:D010146 Disease pain - + MESH:D009369 Disease cancer - - MESH:D001416 - Disease - - simple back pain - DISCUSS paragraph 6349 Low cost and availability make plain radiography the most common spinal imaging test. Conventional x-ray is often the first imaging technique which has been used. Unfortunately, the interpretation of spine X-ray in children is more difficult than in adults due to the presence of epiphyseal and apophyseal growth center. Also conventional x-ray is lack the sensitivity and would miss most causes of back pain including serious pathologies. The limitation of plain radiography was obvious in our study group, in which most x-ray studies were negative or inconclusive. - + 9606 Species children - + MESH:D001416 Disease @@ -67797,14 +68333,14 @@ paragraph 6916 In our study, bone scintigraphy was able to detect many benign and malignant conditions in 25% of children (Table 2). Our results for scintigraphy with 99mTc-MDP showed an overall sensitivity, specificity values of 94%, 100% respectively. - + 1800 Gene 80192 MDP - + 9606 Species @@ -67816,55 +68352,49 @@ paragraph 7155 Skeletal scintigraphy is highly sensitive for detecting stress-induced changes in bone and is an important part in the evaluation of the young athlete with low back pain. Spondylolysis is an osseous defect of the pars interarticularis, thought to be a developmental or acquired stress fracture secondary to chronic low-grade trauma. Most patients with spondylolysis present with either acute or chronic pain. It can be found in as many as 47% of adolescent athletes with low back pain. Most of pars defects (90%) occur at the L5 level and unilateral defect found in 30% of cases. Spondylolysis will show focal tracer accumulation in the region of the pars interarticularis during delayed BS (Figure 1). SPECT BS is more sensitive than planar images and can detect abnormalities in about one third of individuals with normal planar exams. - + 9606 Species patients - + MESH:D015775 Disease stress fracture - + MESH:D017116 Disease low back pain - + MESH:D015868 Disease pars defects - - MESH:D013169 + + MESH:D014947 Disease - - Spondylolysis + + trauma - + + MESH:D015868 + Disease + + osseous defect of the pars interarticularis + + MESH:D010146 Disease pain - - MESH:D013169 - Disease - - Spondylolysis - - - MESH:D014947 - Disease - - trauma - - + MESH:D017116 Disease @@ -67878,14 +68408,14 @@ fig_caption 7993 Whole body scan in 14 years old male boy with LBP of 3 weeks duration; scan showed focus with abnormal activity at L5 level (arrow); finding was suggestive of spondylolysis. - + 3929 Gene 3055 LBP - + 9606 Species @@ -67897,31 +68427,25 @@ paragraph 8167 Also, BS is a very sensitive exam for the detection of acute bone fractures, and it does have increased sensitivity for the detection of rib fractures compared to plain radiographs. BS can be the modality of choice in the assessment of suspected child abuse cases. Also, findings can sometimes draw attention to possible underlying metabolic bone disease (Figure 2). - + 9606 Species child - + MESH:D001851 Disease metabolic bone disease - + MESH:D050723 Disease fractures - - MESH:C535569 - Disease - - abuse - - + MESH:D050723 Disease @@ -67935,7 +68459,7 @@ fig_caption 8534 10 years old male child with multiple pseudofractures invoving multiple ribs and lower tibii. - + 9606 Species @@ -67947,83 +68471,59 @@ paragraph 8628 Other causes of low back pain in children like Spinal process apophysitis, Lumbosacral transitional vertebra, Facet Syndrome, Schuermann's Disease and disc herniation can have nonspecific findings during BS. For instant; spinal process apophysitis produces uptake of tracer along the lumbar spinous process and is due to a hyperextension overuse injury. A lumbosacral transitional vertebra is a common anomaly characterized by a large transverse process that follows the contour of the sacral ala and forms a non-ossified articulation with the sacrum. Bertolotti syndrome refers to increased tracer uptake along the transverse process pseudoarthrosis of a transitional vertebrae which is indicative of stress change. This finding can be seen in about 80% of patients with a transitional vertebra and low back pain. - + 9606 Species children - + 9606 Species patients - - MESH:D016301 - Disease - - Spinal process apophysitis - - - MESH:D061325 - Disease - - Bertolotti syndrome - - - MESH:C562952 - Disease - - vertebra - - + MESH:D010300 Disease Schuermann's Disease - + MESH:D000014 Disease anomaly - - MESH:D017116 - Disease - - low back pain - - + MESH:D012090 Disease hyperextension overuse injury - + + MESH:D017116 + Disease + + low back pain + + MESH:D017116 Disease low back pain - + MESH:D011542 Disease pseudoarthrosis - - MESH:C536344 - Disease - - Lumbosacral transitional vertebra - - - MESH:C562952 + + MESH:D013577 Disease - - vertebra + + Bertolotti syndrome @@ -68031,36 +68531,36 @@ paragraph 9443 Facet Syndrome can produce both local and radiating pain. SPECT BS will usually detect an abnormality (sensitivity 100%), but with poor specificity (70%). However, a normal exam can rule out facet syndrome. - + MESH:D010146 Disease pain - - MESH:D061325 - Disease - - facet syndrome - DISCUSS paragraph 9650 Schuermann's Disease is relatively uncommon disorder, occurring in 3 to 5% of adolescents. The diagnosis is established when the radiographs demonstrate anterior wedging greater than 5 degrees with involvement of 3 or more contiguous vertebral bodies and a structural kyphosis of greater than 40 degrees plate. - + MESH:D007738 Disease kyphosis - + MESH:D010300 Disease Schuermann's Disease + + MESH:D025063 + Disease + + contiguous vertebral bodies + DISCUSS @@ -68073,56 +68573,50 @@ paragraph 10307 Early stage of Perth's disease has a characteristic appearance during BS with photopenic defect within femoral epiphysis. On the other hand; slipped Capital Femoral Epiphysis in the absence of avascular necrosis the bone scan findings is nonspecific and consist of mildly increased activity about the hip and widening and blurring of the growth plate activity. - - MESH:D060048 - Disease - - Femoral Epiphysis - - - MESH:D060048 - Disease - - photopenic defect within femoral epiphysis - - + MESH:D010020 Disease avascular necrosis - + MESH:D007873 Disease Perth's disease + + MESH:D001929 + Disease + + photopenic defect within femoral epiphysis + DISCUSS paragraph 10668 99mTc-MDP BS can detect osteomyelitis one to two weeks before radiologic changes. For the detection of osteomyelitis in non-violated bone, the 3 phase bone scan has a reported sensitivity of 90-100% and a specificity of 70-95%. Osteomyelitis has characteristic appearance in 3 phase BS with abnormal increase in flow, blood pool and delayed radiotracer uptake. - + 1800 Gene 80192 MDP - + MESH:D010019 Disease Osteomyelitis - + MESH:D010019 Disease osteomyelitis - + MESH:D010019 Disease @@ -68134,31 +68628,37 @@ paragraph 11029 There is a higher incidence of discitis in children with peaks seen between the ages of 1 to 4, and 12 to 15 years. In young children, the higher incidence is felt to be related to the vascular supply of the spine. In children subchondral vessels are present until age of 8 and supply the disc, and may persist until teenage years. During BS delayed images; discitis will show increased tracer activity in the disc space and the contiguous vertebral bodies (Figure 3). - + 9606 Species children - + 9606 Species children - + 9606 Species children - + + MESH:D025063 + Disease + + contiguous vertebral bodies + + MESH:D015299 Disease discitis - + MESH:D015299 Disease @@ -68172,19 +68672,19 @@ fig_caption 11498 14 years old girl with back pain; bone scan revealed intervertebral disc activity at level of T12-L1 extending to adjacent vertebral end plates (arrow). Finding was consistent with discitis. - + 9606 Species girl - + MESH:D001416 Disease back pain - + MESH:D015299 Disease @@ -68196,38 +68696,55 @@ paragraph 11689 99mTc-MDP BS can detect synovial inflammations of rheumatoid arthritis and ankylosing spondylitis. Sacroiliitis can have prominent unilateral or bilateral uptake within sacroiliac joints (Figure 4). However; bilateral sacroiliitis, can be difficult to detect due to the normal increased activity within the SI joints. SI joint to sacrum (SIJ index) of greater than 1.35 can have value in this case. - + 1800 Gene 80192 MDP - + MESH:C563037 Disease sacroiliac joints - + MESH:D001172 Disease inflammations of rheumatoid arthritis - + MESH:D013167 Disease ankylosing spondylitis - + + Disease + + SIJ + + + MESH:D007592 + Disease + + SI joint + + MESH:D058566 Disease Sacroiliitis - + + MESH:D007592 + Disease + + SI joints + + MESH:D058566 Disease @@ -68241,104 +68758,121 @@ fig_caption 12088 15 years old boy with low back pain of one month duration. Bone scan revealed prominent activity in both SIJs which is more prominent in the left side. Finding was consistent with sacroiliitis. - + 9606 Species boy - + MESH:D058566 Disease sacroiliitis - + MESH:D017116 Disease low back pain + + Disease + + SIJs + DISCUSS paragraph 12282 99mTc-MDP BS is 50 to 80% more sensitive than radiographs in detecting skeletal metastases. This is probably because about 30-75% of the bone mineral content must be lost before a metastasis is evident on a radiograph. In contrast, as little as a 5-10% change is required to detect an abnormality on BS. The sensitivity for BS in detecting bone metastases is between 62%-100%. About 90% of patients with skeletal metastases present with multiple lesions which increase its specificity of this modality. Nearly 80% of all metastatic lesions are in the axial skeleton. Multiplicity of active lesions can help to define metastatic bone lesions and tracer uptake involving both the vertebral body and pedicles is usually indicative of metastatic disease. - + 9606 Species patients - + 1800 Gene 80192 MDP - + MESH:D009362 Disease - - metastases + + bone metastases - + MESH:D009362 Disease - + metastases - + MESH:D009362 Disease metastases + + MESH:C538445 + Disease + + metastatic disease + + + MESH:C536543 + Disease + + vertebral body + DISCUSS paragraph 13035 Primary bone tumors also can present with back pain in children since more than 80% of the Ewing's sarcomas occur in patients under 20 years of age. The tumor usually arises in the diaphysis of a long bone, but 20% arise in the pelvis. Bone scan can show gross abnormality during the three phases of bone scan; and hardly differentiated from osteomyelitis in some cases (Figure 5). - + 9606 Species children - + 9606 Species patients - + MESH:D010019 Disease osteomyelitis - + + MESH:C563168 + Disease + + Ewing's sarcomas + + MESH:D009369 Disease tumor - - MESH:D012512 - Disease - - Ewing's sarcomas - - + MESH:D001859 Disease Primary bone tumors - + MESH:D001416 Disease @@ -68352,19 +68886,19 @@ fig_caption 13417 13 years old child with low back pain of 2 months duration; bone scan revealed prominent activity within left iliac bone suggestive of aggressive bone pathology. Biopsy revealed primary malignant Ewing sarcoma. - + 9606 Species child - - MESH:D012512 + + MESH:C563168 Disease - - Ewing sarcoma + + malignant Ewing sarcoma - + MESH:D017116 Disease @@ -68376,47 +68910,53 @@ paragraph 13628 Benign lesions like osteoblastomas and osteoid osteoma can involve the posterior elements of the spine and they have characteristic appearance during 3 phase bone scan, osteoblastomas and ostioid osteomas will be hot on both blood pool and delayed images. Osteoblastomas are typically larger than 2 cm and possess both sclerotic and lytic features. Osteoid osteoma may occur in the spine (10-20% of cases), it most commonly involves the lumbar spine, nearly always occurs in the posterior elements (50%); only 10% of lesions involve the vertebral body, and is frequently associated with a painful scoliosis with the convexity of the curve oriented away from the side of the lesion. + + MESH:D010016 + Disease + + ostioid osteomas + - MESH:D010017 + MESH:C538213 Disease - - Osteoid osteoma + + sclerotic - MESH:D010017 + MESH:D018215 Disease - - osteoid osteoma + + osteoblastomas MESH:D018215 Disease - - Osteoblastomas + + Benign lesions like osteoblastomas and osteoid osteoma - MESH:D018215 + MESH:D010017 Disease - - osteoblastomas and ostioid osteomas + + Osteoid osteoma - MESH:D018215 + MESH:C536543 Disease - - Benign lesions like osteoblastomas + + vertebral body - MESH:D012600 + MESH:D018215 Disease - - painful scoliosis + + Osteoblastomas - MESH:C538213 + MESH:D010146 Disease - - sclerotic + + painful scoliosis @@ -68424,26 +68964,20 @@ paragraph 14310 99mTc-MDP B.S showed nonskeletal abnormalities that would have been missed by conventional imaging. Retention of tracer in the kidney due to Hydronephrosis is frequently encountered, which can draw attention to presence of obstructive uropathy as a possible etiology (Figure 6). - + 1800 Gene 80192 MDP - - MESH:C107828 - Chemical - - 99mTc - - + MESH:D006869 Disease Hydronephrosis - + MESH:C536483 Disease @@ -68457,19 +68991,19 @@ fig_caption 14589 6 years old girl with back pain; scan revealed radiotracer retention within left kidney. Finding was suggestive of hydronephrosis-PUJ stenosis. - + 9606 Species girl - + MESH:D001416 Disease back pain - + MESH:D006869 Disease @@ -68481,61 +69015,61 @@ paragraph 14733 Finally we believe that plain x-ray can miss serious pathologies in pediatric patients with back pain, causing delay and unnecessary radiation exposure to this group of patients. Bone isotope scan has high negative predictive value in this study (98%), and consequently it can be safe to proceed into clinical follow up and maybe symptomatic management without the need to do any advanced investigations. We do not recommend the use bone isotope scan in each child with back pain, but when we have clinical data suggestive of an underlying bony pathology, bone isotope scan should be the first line of radiological investigation, before proceeding to CT or MRI. Also, Bone scintigraphy SPECT/CT which includes optimized CT as part of the study, provide improved diagnostic accuracy for back pain and oncology compared to routine BS, which may decrease the number of studies an individual child may need. This imaging modality, however, must be tailored on an individual basis as the addition of the CT study can increase exposure to the child and should only be performed after appropriate justification and with adherence to optimized low dose pediatric protocols. - + 9606 Species patients - + 9606 Species patients - + 9606 Species child - + 9606 Species child - + 9606 Species child - + MESH:D001416 Disease back pain - + MESH:D001847 Disease bone isotope - + MESH:D001416 Disease back pain - + MESH:D001416 Disease back pain - + MESH:D001847 Disease @@ -68553,13 +69087,13 @@ paragraph 15913 Bone isotope scan is a non-invasive, highly sensitive and reliable imaging modality in the assessment of pediatric patients with back pain and in detecting most possible bony pathologies. We recommend this investigation as first imaging procedure to avoid any delay in the diagnosis and to avoid unnecessary investigations. - + 9606 Species patients - + MESH:D001416 Disease @@ -68591,13 +69125,13 @@ 264 16285 The epidemiology of pain in children and adolescents: a review - + 9606 Species children - + MESH:D010146 Disease @@ -68618,13 +69152,13 @@ 179 16348 Low back pain in school children. An epidemiological study - + 9606 Species children - + MESH:D017116 Disease @@ -68645,7 +69179,7 @@ 814 16407 Back pain in children - + MESH:D010146 Disease @@ -68669,19 +69203,19 @@ surname:McDonald;given-names:JV 16429 Disk-caused nerve compression in patients with acute low-back pain: diagnosis with MR, CT myelography, and plain CT - + 9606 Species patients - + MESH:D017116 Disease low-back pain - + MESH:C564570 Disease @@ -68705,7 +69239,7 @@ surname:Kormano;given-names:M 16545 Magnetic Resonance Study of Disc Degeneration in Young Low Back Pain Patients - + MESH:D017116 Disease @@ -68725,13 +69259,13 @@ 592 16623 Bone scintigraphy in the evaluation of children with obscure skeletal pain - + 9606 Species children - + MESH:D010146 Disease @@ -68753,18 +69287,18 @@ 512 16698 Low-back pain in adolescent athletes: detection of stress injury to the pars interarticularis with SPECT - - MESH:D040701 - Disease - - stress injury - - + MESH:D017116 Disease Low-back pain + + MESH:D015775 + Disease + + stress injury + 21 @@ -68780,7 +69314,7 @@ 62 16803 Applications of bone scintigraphy in ankylosing spondylitis - + MESH:D013167 Disease @@ -68802,14 +69336,14 @@ 211 16863 Diagnosis of discitis by SPECT technetium-99m MDP scintigram. A case report - + 1800 Gene 80192 MDP - + MESH:D015299 Disease @@ -68842,19 +69376,13 @@ 914 16994 Skeletal scintigraphy of young patients with low-back pain and a lumbrosacral transitional vertebra - + 9606 Species patients - - MESH:C562952 - Disease - - lumbrosacral transitional vertebra - - + MESH:D017116 Disease @@ -68890,7 +69418,7 @@ ref 17125 Low back pain in the paediatric athlete - + MESH:D017116 Disease @@ -68909,6 +69437,12 @@ 555 17165 Taking the stress out of evaluating stress injuries in children + + MESH:D015775 + Disease + + stress injuries + 23 @@ -68922,7 +69456,7 @@ 320 17229 Spine pain in children - + MESH:D010146 Disease @@ -68941,7 +69475,7 @@ 457 17252 Interpretation and classification of bone scintigraphic findings in stress fractures - + MESH:D015775 Disease @@ -68961,18 +69495,12 @@ 333 17337 The role of bone scintigraphy in detecting child abuse - + 9606 Species child - - MESH:C535569 - Disease - - abuse - 44 @@ -68986,19 +69514,13 @@ 914 17392 Skeletal scintigraphy of young patients with low-back pain and a lumbrosacral transitional vertebra - + 9606 Species patients - - MESH:C562952 - Disease - - lumbrosacral transitional vertebra - - + MESH:D017116 Disease @@ -69018,12 +69540,6 @@ 44 17492 Planar and high-resolution SPECT bone imaging in the diagnosis of facet syndrome - - MESH:D061325 - Disease - - facet syndrome - 22 @@ -69037,19 +69553,19 @@ 417 17573 Diagnostic imaging of musculoskeletal infection. Roentgenography;gallium, indium-labeled white blood cell, gammaglobulin, bone scintigraphy;and MRI - + MESH:D007204 Chemical indium - + MESH:D005708 Chemical gallium - + MESH:D009140 Disease @@ -69068,25 +69584,25 @@ 506 17721 Subacute and chronic bone infections: Diagnosis using In-111, Ga-67 and Tc-99m MDP bone scintigraphy, and radiography - + 1800 Gene 80192 MDP - - MESH:C048993 - Chemical - - Ga-67 - - + MESH:D001847 Disease bone infections + + MESH:D005708 + Chemical + + Ga + 21 @@ -69102,7 +69618,7 @@ 62 17839 Applications of bone scintigraphy in ankylosing spondylitis - + MESH:D013167 Disease @@ -69147,14 +69663,14 @@ 831 18040 From the radiologic pathology archives. Ewing sarcoma family of tumors: radiologic-pathologic correlation - + MESH:D009369 Disease tumors - - MESH:D012512 + + MESH:C563168 Disease Ewing sarcoma @@ -69174,17 +69690,17 @@ 38 18146 Osteoid osteomas and osteoblastoma of the spine - + MESH:D010017 Disease Osteoid osteomas - + MESH:D018215 Disease - - osteoblastoma of the spine + + osteoblastoma @@ -69199,7 +69715,7 @@ 1052 18194 Pediatric bone imaging: diagnostic imaging of osteoid osteoma - + MESH:D010017 Disease @@ -69221,7 +69737,7 @@ 1785 18256 Detection of obstructive uropathy by bone scintigraphy - + MESH:C536483 Disease @@ -69255,10 +69771,10 @@ 0 Telemetric Detection of Chronic Obstructive Pulmonary Disease and Investigation of Quality of Life for People Working in Shipbuilding Industry - 9606 - Species - - People + MESH:D006212 + Disease + + People Working MESH:D029424 @@ -69590,60 +70106,59 @@ paragraph 4203 A group of one hundred people working in the Shipbuilding Industry (SI) was studied as well as a control group of one hundred people of the general population. All were middle aged men and constituted a convenience sample. - + 9606 Species people - + 9606 Species people - + 9606 Species men + + Disease + + SI + METHODS paragraph 4426 Data were collected on demographic characteristics, education, marital status, smoking dependence and personal medical history. Additionally, data were collected on employment status, self-assessment of health and use of health services. Independence in everyday life activities and quality of life were assessed by using St. George's Respiratory Questionnaire and smoking habit by using Fagerstrom test. St. George's Respiratory Questionnaire is a questionnaire designed to assess the relationship of pulmonary disease with quality of life associated with health and wellbeing. The average time to fill it out ranges from 10 to 15 minutes depending on age and educational level of the individual. The questionnaire provides a sum total and scores for individual symptoms (cough, sputum, wheezing, dyspnea, their frequency and duration), activities (which cause or are limited by dyspnea) and impacts (in a wide range of social functioning and mental disorders as a result of airway obstruction). The lowest score represents a better quality of life. - + MESH:D003371 Disease cough - + MESH:D004417 Disease dyspnea - + MESH:D001523 Disease mental disorders - + MESH:D012135 Disease wheezing - - MESH:D000402 - Disease - - airway obstruction - MESH:D004417 Disease @@ -69692,30 +70207,18 @@ paragraph 6112 Data were stored in an electronic page which was accessed by the specialist physician who recorded the COPD diagnosis. Diagnosis was based on symptoms and spirometry findings and airway obstruction was staged according to GOLD guidelines. Participants were asked to make at least three attempts and the highest values of FEV1 and FVC were used to classify the severity of the degree of airway obstruction. - + 260431 Gene COPD - + 9606 Species Participants - - MESH:D000402 - Disease - - airway obstruction - - - MESH:D000402 - Disease - - airway obstruction - METHODS @@ -69728,6 +70231,12 @@ paragraph 6539 Descriptive and inferential analysis was performed Depending on indications, Student's t-test or Mann-Whitney nonparametric test was used to compare quantitative variables between two groups. Respectively, parametric analysis of variance (ANOVA) or Kruskal-Wallis nonparametric test was used to compare quantitative variables between more than two groups. Bonferroni correction was used to counteract type I error Pearson's chi-squared test or Fisher's exact test was used to compare proportions where necessary. Spearman's or Pearson's (r) correlation coefficient was used to control the relationship between two quantitative variables. The level of statistical significance was set at 0.05 and SPSS 17.0 statistical software was used in the analysis. + + MESH:C536353 + Disease + + type I error Pearson + RESULTS @@ -69746,7 +70255,7 @@ paragraph 7335 The mean age of workers and general population group was 51.8 +- 8.2 and 51.1 +- 6.4 years with 25.1 +- 11.5 and 25.46 +- 10 years duration of smoking, respectively. There was a significantly higher percentage of workers in the general population (75%) compared with the corresponding percentage in the group of people who worked in the shipbuilding zone (62%). The working years in the two groups were 10 +- 3 and 25.4 +- 8.7 respectively. - + 9606 Species @@ -69758,7 +70267,7 @@ paragraph 7776 Fagerstrom score of participants did not differ between the two groups, as well as their smoking dependence level. - + 9606 Species @@ -69770,49 +70279,49 @@ paragraph 7891 The following diseases were found in the general population in order of frequency: arterial hypertension 12%, COPD 10%, heart diseases 9%, orthopedic problems 2%, diabetes mellitus 6% and other 7%. Respectively, in the shipbuilding industry workers were found: arterial hypertension 7%, COPD 13%, heart diseases 4%, orthopedic problems 7%, diabetes mellitus 4% and other 7%. - + 544442 Gene COPD 10 - + 544417 Gene COPD 13 - + MESH:D006331 Disease heart diseases - + MESH:D006331 Disease heart diseases - + MESH:D006973 Disease hypertension - + MESH:D003920 Disease diabetes mellitus - + MESH:D003920 Disease diabetes mellitus - + MESH:D006973 Disease @@ -69824,43 +70333,43 @@ paragraph 8266 No participant was found in stage IV in the general population group and 3 persons respectively were found in the group of people working in the shipbuilding industry, according to GOLD guidelines. The workers in the shipbuilding zone had significantly lower values (p < 0.001) in FVC, FEV1 and FEV1/FVC compared with the participants of the general population. There was no significant difference in COPD stage of participants depending on their workplace, although most workers in the zone had advanced-stage COPD compared with the general population (Table 1). - + 260431 Gene COPD - + 260431 Gene COPD - + 9606 Species participant - + 9606 Species persons - + 9606 Species people - + 9606 Species participants - + 9606 Species @@ -69886,49 +70395,49 @@ paragraph 8948 Significantly higher score in "Symptoms and Activity" dimensions was found in the workers in the shipbuilding industry, those with basic education and non-actively working during this period. Health evaluation as moderate/poor and comorbidity are also associated with high score in these particular dimensions of life quality (p < 0.001). Also, people who were hospitalized during the last year and those diagnosed with III-IV COPD stage had significantly higher values in Symptoms and Activity score, compared to participants with I-II stage. There was a significant difference in Symptoms score depending on smoking dependence. Participants with low smoking dependence had significantly lower scores compared with participants with high smoking dependence (p = 0.016). Participants who visited a general practitioner more than 3 times during the last year had significantly higher values in Activity score compared to participants who visited him/her 1-2 times and to participants who did not visit him/her at all (p = 0.002 and p = 0.008 respectively) (Table 2). - + 260431 Gene COPD - + 9606 Species people - + 9606 Species participants - + 9606 Species Participants - + 9606 Species participants - + 9606 Species Participants - + 9606 Species participants - + 9606 Species @@ -69948,31 +70457,31 @@ paragraph 10131 In "Impact score" dimension, a significantly higher score (p < 0.001), corresponding to more problems of social conciliation, was exhibited by those who considered their health as moderate/poor, suffered from a chronic disease and were diagnosed with III-IV COPD stage. In "Total score", participants who had visited a general practitioner more than three times during the last year had significantly higher values in Total score compared both to participants who had visited him/her 1-2 times and to those who had not visited him/her at all (p = 0.004 and p = 0.001 respectively) (Table 3). A significant positive correlation was observed between the score of participants in dimensions under study of quality of life with age, years of smoking and values of FVC, FEV1, FEV1/FVC and Fagerstrom score (p < 0.001) (Table 4). - + 260431 Gene COPD - + 9606 Species participants - + 9606 Species participants - + 9606 Species participants - + MESH:D002908 Disease @@ -70006,13 +70515,13 @@ paragraph 11125 Health professionals have recognized the need for early detection of COPD and introduced new ways to facilitate its diagnosis by using modern telehealth methods such as telemetric data transfer. Masa et al. argue that, if high quality spirometry could be performed online through a connection of the specialist to the Pulmonary Laboratory and the patient in a PHC structure by qualified personnel and high quality spirometer, possibly the problem of performing a reliable and qualitative spirometry would be resolved. - + 260431 Gene COPD - + 9606 Species @@ -70024,25 +70533,25 @@ paragraph 11643 Spirometry with telemetric data transmission method was used in the present study and it was found that the prevalence of COPD in the general population was 10%. Similarly, in their study Tzanakis et al., found a prevalence of 11.6% in men and in an epidemiological study conducted by Sichleridis et al the prevalence of COPD was also estimated at 8.2% among men. - + 260431 Gene COPD - + 260431 Gene COPD - + 9606 Species men - + 9606 Species @@ -70054,79 +70563,73 @@ paragraph 12007 The results of an epidemiological study conducted in Ireland from 1999 to 2001 on a sample of the general population aged 40-69 years showed that the prevalence of obstructive lung disease was 6.3%. In order to estimate the prevalence of COPD in this study, medical history, spirometry, a special respiratory symptoms questionnaire as in ours and dermal tests were used. Also, the BOLD (Burden of Obstructive Lung Disease) study that attempted to identify the prevalence of COPD by using population sample from 12 different countries found that the prevalence of COPD was 11.8 +- 7.9% in men. It is noted that in our study the prevalence of COPD was estimated at 10% in the general population and 13% in people working in the shipbuilding industry and no statistically significant difference was observed between the two groups. It is well known that there are many factors responsible for COPD such as environmental pollution, occupation, place of residence and respiratory infections during childhood. In this study, it was not possible to study the effects of each of these factors, but probably the highest prevalence of COPD in the group of people working in the shipbuilding industry is due to overburdened environment because of occupational exposure. Although it is known that air pollution has a serious impact on respiratory symptoms, the relationship between pollution and COPD remains to be elucidated. - + 260431 Gene COPD - + 260431 Gene COPD - + 260431 Gene COPD - + 260431 Gene COPD - + 260431 Gene COPD - + 260431 Gene COPD - + 260431 Gene COPD - + 9606 Species men - + 9606 Species people - + 9606 Species people - - MESH:D008173 - Disease - - Burden of Obstructive Lung Disease - - - MESH:D012141 + + MESH:D012131 Disease respiratory infections - + MESH:D008173 Disease @@ -70138,19 +70641,19 @@ paragraph 13422 The results of our research showed that there was a significant positive correlation of the participants' score in the SGRQ scales with age, years of smoking and Fagerstrom Score. Regarding the presence of respiratory symptoms, limitation of activities and their impacts which affect the field of social functioning, results were similar in a research of Xydeas-Kikemenis et al. in a sample of 203 people working in the construction industry. It was observed that age and duration of working activity were strong determinants of mental problems incidence affected by the workload and the negative characteristics of the workplace. Pierrakos et al., in their work concerning the quality of life related to smokers and non-smokers health, find that high smoking dependence negatively affects the quality of life which is exacerbated when tobacco consumption increases. - + 9606 Species participants - + 9606 Species people - + 4097 Species @@ -70162,19 +70665,19 @@ paragraph 14291 In contrast, SGRQ scales showed a high negative correlation with spirometric results (FEV1%, FVC% and FEV1/FVC) and thus, as values of the above measurements of the participants increase, the respective score implying better quality of life decreases. The studies of Stahl . and Ketelaars CA., in patients with COPD also demonstrate that the quality of life is positively correlated with good respiratory function while it is burdened with advancing age and disease. A moderate correlation of FEV1 values with health level and life quality emerges from the studies of Aibar Arregui et al., and Pereira et al.. - + 260431 Gene COPD - + 9606 Species participants - + 9606 Species @@ -70186,55 +70689,55 @@ paragraph 14905 In our study, comorbidities showed a significant positive correlation with all subscales of quality of life, as supported by many researchers, to the extent that the disease has physical and psychosocial impacts on the overall situation of the individual and chronic patient is observed living under conditions acting bindingly as to his/her quality of life. Overall, the coexistence of chronic diseases burdens the person mainly with organic and functional problems, frequent hospitalizations, continuous medical visits and examinations and secondarily with social, psychological and occupational problems that affect his/her quality of life. - + 9606 Species patient - + 9606 Species person - - MESH:C535569 - Disease - - psychosocial impacts - - + MESH:D002908 Disease chronic diseases burdens + + MESH:C535569 + Disease + + psychosocial + DISCUSS paragraph 15549 Educational level and occupational status have significant positive correlation with all SGRQ scales. Participants with basic educational level had higher scores in scales of symptoms, impacts, activities and overall, as in the study of Dimitropoulos et al., in which people with lower educational level had a more aggravated symptoms scale compared to the more educated participants. Based on that, people with low educational level tend to occupy lower-paid jobs in the labor market and often those associated with exposure to harmful for health factors. - + 9606 Species Participants - + 9606 Species people - + 9606 Species participants - + 9606 Species @@ -70246,7 +70749,7 @@ paragraph 16106 In our study, admission to hospital during the last year is significantly correlated with the quality of life scales and is partially consistent with the studies of Dimitropoulos et al., and Miravitlles et al., where patients with more than two admissions to hospital during the last year had worse scores in all scales, but without statistical correlation. - + 9606 Species @@ -70258,7 +70761,7 @@ paragraph 16464 The contact frequency of participants (> 3 times) with a general practitioner in the last 6 to 12 months was positively correlated with the quality of life scales and is consistent with the results of the study of Tountas et al., of whom 37% of the sample under study had visited a doctor >1 time in the previous month. - + 9606 Species @@ -70282,24 +70785,24 @@ paragraph 17167 Educational level, occupational status and presence of chronic diseases affect the quality of life of individuals, while "low" self-assessment of health, increased frequency of visits to a general practitioner and repeated admissions to hospital are associated with social dysfunction and reduced quality of life. Telemetric diagnosis of COPD can become a method of choice in a community environment, but requires enhancement of education and training of health professionals that are going to perform it. - + 260431 Gene COPD - + + MESH:D000067404 + Disease + + social dysfunction + + MESH:D002908 Disease chronic diseases - - OMIM:300082 - Disease - - social dysfunction - CONCL @@ -70330,7 +70833,7 @@ surname:Caeser;given-names:M 17725 Epidemiology and health economics of COPD across Europe: a critical analysis - + 260431 Gene @@ -70343,7 +70846,7 @@ ref 17802 GOLD, Global Initiative for Chronic Obstructive Lung Disease. Global Strategy for the diagnosis, management and prevention of COPD - + MESH:D029424 Disease @@ -70364,13 +70867,13 @@ 128 17933 Health status deterioration in patients with chronic obstructive pulmonary disease - + 9606 Species patients - + MESH:D029424 Disease @@ -70392,7 +70895,7 @@ 746 18016 Association between COPD and Employment by Industry and Occupation in the US Population: A Study of Data from the Third National Health and Nutrition Examination Survey - + 260431 Gene @@ -70412,7 +70915,7 @@ ref 18185 Telehealth: A Useful Strategy in the Management of COPD? - + 260431 Gene @@ -70475,7 +70978,7 @@ 1127 18381 The Fagerstrom Test for Nicotine Dependence: a revision of the Fagerstrom Tolerance Questionnaire - + MESH:D009538 Chemical @@ -70528,7 +71031,7 @@ 900 18588 Prevalence of COPD in Greece - + 260431 Gene @@ -70574,7 +71077,7 @@ surname:Kee;given-names:F 18663 Prevalence of obstructive lung disease in a general population sample: the NICECOPD study - + MESH:D008173 Disease @@ -70589,11 +71092,11 @@ ref 18753 Fishman's Pulmonary Diseases and Disorders - - MESH:D008171 + + MESH:C535736 Disease - - Pulmonary Diseases and Disorders + + Fishman's Pulmonary Diseases @@ -70617,7 +71120,7 @@ 253 18807 Building a national strategy for the prevention and management of and research in chronic obstructive pulmonary disease: National Heart, Lung, and Blood Institute workshop summary - + MESH:D029424 Disease @@ -70670,7 +71173,7 @@ surname:Svensson;given-names:K 19093 Health-related quality of life is related to COPD disease severity - + MESH:D029424 Disease @@ -70694,13 +71197,13 @@ surname:Halfens;given-names:RJ 19160 Determinants of health-related quality of life in patients with chronic obstructive pulmonary disease - + 9606 Species patients - + MESH:D029424 Disease @@ -70721,7 +71224,7 @@ 477 19262 Relationship between objective data and health-related quality of life in COPD patients - + 260431 Gene @@ -70744,7 +71247,7 @@ surname:Mota;given-names:RM 19350 Influence of respiratory function parameters on the quality of life of COPD patients - + 260431 Gene @@ -70778,19 +71281,19 @@ 269 19489 Mood disorders and handling of daily routine in patients with diabetes mellitus type II - + 9606 Species patients - + MESH:D003924 Disease diabetes mellitus type II - + MESH:D019964 Disease @@ -70815,7 +71318,7 @@ surname:Tsapas;given-names:C 19577 The health related quality of life of Greekpatients with chronic obstructive pulmonary disease - + MESH:D029424 Disease @@ -70849,13 +71352,13 @@ 395 19700 Effect of exacerbations on quality of life in patients with chronic obstructive pulmonary disease: a 2 year follow up study - + 9606 Species patients - + MESH:D029424 Disease @@ -70918,71 +71421,59 @@ abstract 69 Conditions have been defined for promoting growth and differentiation of hypertrophic chondrocytes obtained in culture starting from chick embryo tibiae. Hypertrophic chondrocytes, grown in suspension culture as described (Castagnola P., G. Moro, F. Descalzi Cancedda, and R. Cancedda. 1986. J. Cell Biol. 102:2310-2317), when they reached the stage of single cells, were transferred to substrate-dependent culture conditions in the presence of ascorbic acid. Cells showed a change in morphology, became more elongated and flattened, expressed alkaline phosphatase, and eventually mineralized. Type II and X collagen synthesis was halted and replaced by type I collagen synthesis. In addition the cells started to produce and to secrete in large amount a protein with an apparent molecular mass of 82 KD in reducing conditions and 63 KD in unreducing conditions. This protein is soluble in acidic solutions, does not contain collagenous domains, and is glycosylated. The Ch21 protein, a marker of hypertrophic chondrocytes and bone cells, was synthesized throughout the culture. We have defined this additional differentiation stage as an osteoblast-like stage. Calcium deposition in the extracellular matrix occurred regardless of the addition of beta glycerophosphate to the culture medium. Comparable results were obtained both when the cells were plated at low density and when they were already at confluence and maintained in culture without passaging up to 50 d. When retinoic acid was added to the hypertrophic chondrocyte culture between day 1 and day 5 the maturation of the cells to the osteoblast-like stage was highly accelerated. The switch in the collagen secretion was already observed after 2 d and the production of the 63-kD protein after 3 d. Mineralization was observed after 15-20 d. - + 9031 Species chick - + MESH:D006984 Disease hypertrophic - + MESH:D014212 Chemical retinoic acid - + MESH:C031463 Chemical beta glycerophosphate - - MESH:C537014 - Disease - - KD - - + MESH:D006984 Disease - - Hypertrophic chondrocytes + + Hypertrophic - + MESH:D002118 Chemical Calcium - + MESH:D001205 Chemical ascorbic acid - - MESH:C537014 - Disease - - KD - - + MESH:D006984 Disease hypertrophic - + MESH:D006984 Disease - - hypertrophic chondrocytes + + hypertrophic @@ -71017,112 +71508,124 @@ abstract 120 Lactating mammary epithelial cells secrete high levels of caseins and other milk proteins. The extent to which protein secretion from these cells occurs in a regulated fashion was examined in experiments on secretory acini isolated from the mammary glands of lactating mice at 10 d postpartum. Protein synthesis and secretion were assayed by following the incorporation or release, respectively, of [35S]methionine-labeled TCA-precipitable protein. The isolated cells incorporated [35S]methionine into protein linearly for at least 5 h with no discernible lag period. In contrast, protein secretion was only detectable after a lag of approximately 1 h, consistent with exocytotic secretion of proteins immediately after passage through the secretory pathway and package into secretory vesicles. The extent of protein secretion was unaffected by the phorbol ester PMA, 8-bromo-cAMP, or 8- bromo-cGMP but was doubled by the Ca2+ ionophore ionomycin. In a pulse- label protocol in which proteins were prelabeled for 1 h before a chase period, constitutive secretion was unaffected by depletion of cytosolic Ca2+ but ionomycin was found to give a twofold stimulation of the secretion of presynthesized protein in a Ca(2+)-dependent manner. Ionomycin was still able to stimulate protein secretion after constitutive secretion had terminated. These results suggest that lactating mammary cells possess both a Ca(2+)-independent constitutive pathway and a Ca(2+)-activated regulatory pathway for protein secretion. The same proteins were secreted by both pathways. No ultrastructural evidence for apocrine secretion was seen in response to ionomycin and so it appears that regulated casein release involves exocytosis. Ionomycin was unlikely to be acting by disassembling the cortical actin network since cytochalasin D did not mimic its effects on secretion. The regulated pathway may be controlled by Ca2+ acting at a late step such as exocytotic membrane fusion. - + 10090 Species mice - - MESH:D002118 + + MESH:D000069285 Chemical - + Ca2+ - - MESH:D002118 + + MESH:C000615320 Chemical - - Ca2+ + + 35S - + + MESH:D014238 + Chemical + + TCA + + + MESH:D008715 + Chemical + + methionine + + MESH:C016276 Chemical 8- bromo-cGMP - - MESH:C013111 + + MESH:D010703 Chemical - - [35S]methionine + + phorbol ester - + MESH:D015759 Chemical Ionomycin - + + MESH:D000069285 + Chemical + + Ca2+ + + MESH:D015124 Chemical 8-bromo-cAMP - - MESH:D010703 + + MESH:D000069285 Chemical - - phorbol ester PMA + + Ca(2+) - + MESH:D015759 Chemical Ionomycin - - MESH:D002118 - Chemical - - Ca(2+) - - - MESH:C013111 + + MESH:D000069285 Chemical - - [35S]methionine + + Ca2+ - + MESH:D015759 Chemical ionomycin - + MESH:D015759 Chemical ionomycin - + MESH:D015759 Chemical ionomycin - - MESH:C000589078 + + MESH:C000615320 Chemical - - TCA + + 35S - - MESH:D002118 + + MESH:D008715 Chemical - - Ca(2+) + + methionine - - MESH:D002118 + + MESH:D000069285 Chemical - - Ca2+ + + Ca(2+) - - MESH:D002118 + + MESH:D000069285 Chemical - + Ca(2+) @@ -71158,46 +71661,65 @@ abstract 128 We have used stably transfected CHO cell lines to characterize the pathway of intracellular transport of the lgp120 (lgp-A) to lysosomes. Using several surface labeling and internalization assays, our results suggest that lgp120 can reach its final destination with or without prior appearance on the plasma membrane. The extent to which lgp120 was transported via the cell surface was determined by two factors: expression level and the presence of a conserved glycine-tyrosine motif in the cytoplasmic tail. In cells expressing low levels of wild-type lgp120, the majority of newly synthesized molecules reached lysosomes without becoming accessible to antibody or biotinylation reagents added extracellularly at 4 degrees C. With increased expression levels, however, an increased fraction of transfected lgp120, as well as some endogenous lgp-B, appeared on the plasma membrane. The fraction of newly synthesized lgp120 reaching the cell surface was also increased by mutations affecting the cytoplasmic domain tyrosine or glycine residues. A substantial fraction of both mutants reached the surface even at low expression levels. However, only the lgp120G----A7 mutant was rapidly internalized and delivered from the plasma membrane to lysosomes. Taken together, our results show that the majority of newly synthesized wild-type lgp120 does not appear to pass through the cell surface en route to lysosomes. Instead, it is likely that lysosomal targeting involves a saturable intracellular sorting site whose affinity for lgp's is dependent on a glycine-tyrosine motif in the lgp120 cytoplasmic tail. - + 100689406 Gene lgp-A - + 100689316 Gene lgp-B - + 10029 Species CHO cell lines - - MESH:D005998 + + MESH:D014443 Chemical - - glycine-tyrosine + + tyrosine - + + MESH:D005998 Chemical - - tyrosine + + glycine - + MESH:D005998 Chemical glycine - + MESH:D005998 Chemical - - glycine-tyrosine + + glycine + + + MESH:D014443 + Chemical + + tyrosine + + + - + Chemical + + lgp120 + + + MESH:D014443 + Chemical + + tyrosine @@ -71350,35 +71872,17 @@ Wong SH, Low SH, Hong W 0 The 17-residue transmembrane domain of beta-galactoside alpha 2,6- sialyltransferase is sufficient for Golgi retention - - MESH:C007816 - Chemical - - beta-galactoside - - - MESH:D016055 - Disease - - Golgi retention - ABSTRACT abstract 119 beta-Galactoside alpha 2,6-sialyltransferase (ST) is a type II integral membrane protein of the Golgi apparatus involved in the sialylation of N-linked glycans. A series of experiments has shown that the 17-residue transmembrane domain of ST is sufficient to confer localization to the Golgi apparatus when transferred to the corresponding region of a cell surface type II integral membrane protein. Lectin affinity chromatography of chimeric proteins bearing this 17-residue sequence suggests that these chimeric proteins are localized in the trans-Golgi cisternae and/or trans-Golgi network. Further experiments suggest that this 17-residue sequence functions as a retention signal for the Golgi apparatus. - - MESH:D009584 - Chemical - - N - - - MESH:C007816 + + - Chemical - - beta-Galactoside + + N-linked glycans @@ -71401,13 +71905,7 @@ Eveleth DD, Bradshaw RA 0 Nerve growth factor nonresponsive pheochromocytoma cells: altered internalization results in signaling dysfunction - - MESH:C566796 - Disease - - signaling dysfunction - - + MESH:D010673 Disease @@ -71419,76 +71917,76 @@ abstract 115 Variant rat pheochromocytoma (PC12) cells which fail to respond to nerve growth factor (NGF) (PC12nnr5) (Green, S. H., R. E. Rydel, J. L. Connoly, and L. A. Greene. 1986. J. Cell Biol. 102:830-843) bind NGF at both high and low affinity sites. Although still undefined at the molecular level, these have been referred to as type I (high) and type II (low) receptors. They are apparently composed of two membrane-bound proteins, p75 and the protooncogene trk, both of which bind NGF, and apparently contribute singularly or in concert to the two observed affinities, and to the promotion of the NGF effects. In native PC12 cells, only the high affinity receptors are apparently capable of mediating internalization and degradation. PC12nnr5 cells also display type I binding, but the subsequent internalization is not the same fashion as in the parental cell line, nor is it subjected to lysosomal degradation. Rather it is initially sequestered during the first 15 min, and is eventually released intact into the medium. In contrast, EGF is bound, internalized, and degraded by PC12nnr5 cells, albeit less efficiently than in the parent cells. These observations argue that the defect(s) preventing the PC12nnr5 variants from responding to NGF prevents competent internalization, which in the case of NGF, may be required for the full expression of activity. The absence of trk, as one alteration in PC12nnr5 cells (Loeb, D. M., J. Maragos, D. Martin- Zanca, M. V. Chao, L. F. Parada, and L. A. Greene. 1991. Cell. 66:961- 966), is consistent with this conclusion. - + 24596 Gene 1877 p75 - + 59109 Gene 1898 trk - + 59109 Gene 1898 trk - + 10116 Species rat - + CVCL:0481 CellLine PC12 - + MESH:D010673 Disease pheochromocytoma - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine @@ -71521,61 +72019,67 @@ abstract 127 Investigating the regulation of very late antigen (VLA)-mediated functions, we found that TS2/16, a mAb directed against the beta chain of the VLA group of integrins, can induce binding of resting peripheral blood lymphocytes, cloned T lymphocytes, and Epstein Barr virus- transformed B cells to extracellular matrix components, fibronectin, laminin, and collagen, but not to fibrinogen. The antibody stimulates VLA-4-, VLA-5-, and VLA-6-mediated binding. Furthermore, it induces VLA- 4-mediated binding to vascular cell adhesion molecule-1 expressed by rTNF-alpha-stimulated endothelial cells, but it does not stimulate homotypic aggregation of cells as described for a number of anti-VLA-4 alpha antibodies (Bednarczyk, J.L., and B. W. McIntyre. 1990. J. Immunol. 144: 777-784; Campanero, M. R., R. Pulido, M. A. Ursa, M. Rodriguez-Moya, M. O. de Landazuri, and F. Sanchez-Madrid. 1990. J. Cell Biol. 110:2157-2165). Therefore, the stimulating activity of this anti-beta 1 antibody clearly contrasts with that of the anti-VLA-4 alpha antibodies, which induce homotypic cell aggregation, but not binding of cells to extracellular matrix components or endothelial cells, indicating that TS2/16 may generate different signals. The observation that also F(ab')2 or Fab fragments of this anti-beta 1 antibody stimulate binding to extracellular matrix components and endothelial cells excludes the possibility that binding requires receptor crosslinking, or is Fc receptor mediated. Induction of this adhesion is cation and energy dependent and requires an intact cytoskeleton. Although changes in the conformation of VLA integrins induced by this antibody may regulate their functional activity, the dependence on metabolic energy indicates that intracellular processes may also play a role. - + 3678 Gene 20508 VLA-5 - + 3655 Gene 20091 VLA-6 - + 7412 Gene 838 vascular cell adhesion molecule-1 - + 24835 Gene 496 rTNF-alpha - + 3779 Gene 3054 beta 1 - + 2187 Gene 51880 Fab - + 3779 Gene 3054 beta 1 - + 10376 Species Epstein Barr virus + + - + Chemical + + Campanero + @@ -71604,11 +72108,10 @@ mammalian - 342538 - Gene - 133972 - - Na(+)-Ca2 + MESH:D000069285 + Chemical + + Ca2+ @@ -71669,26 +72172,25 @@ guinea pig - 342538 - Gene - 133972 - - Na(+)-Ca2 + MESH:D000069285 + Chemical + + Ca2+ + MESH:D000069285 + Chemical + + Ca2+ + + MESH:D009101 Disease myeloma - - MESH:D002118 - Chemical - - Ca2+ - - MESH:D002118 + MESH:D000069285 Chemical Ca2+ @@ -71720,221 +72222,251 @@ abstract 58 Storage triacylglycerols (TAG) in plant seeds are present in small discrete intracellular organelles called oil bodies. An oil body has a matrix of TAG, which is surrounded by phospholipids (PL) and alkaline proteins, termed oleosins. Oil bodies isolated from mature maize (Zea mays) embryos maintained their discreteness, but coalesced after treatment with trypsin but not with phospholipase A2 or C. Phospholipase A2 or C exerted its activity on oil bodies only after the exposed portion of oleosins had been removed by trypsin. Attempts were made to reconstitute oil bodies from their constituents. TAG, either extracted from oil bodies or of a 1:2 molar mixture of triolein and trilinolein, in a dilute buffer were sonicated to produce droplets of sizes similar to those of oil bodies; these droplets were unstable and coalesced rapidly. Addition of oil body PL or dioleoyl phosphatidylcholine, with or without charged stearylamine/stearic acid, or oleosins, to the medium before sonication provided limited stabilization effects to the TAG droplets. High stability was achieved only when the TAG were sonicated with both oil body PL (or dioleoyl phosphatidylcholine) and oleosins of proportions similar to or higher than those in the native oil bodies. These stabilized droplets were similar to the isolated oil bodies in chemical properties, and can be considered as reconstituted oil bodies. Reconstituted oil bodies were also produced from TAG of a 1:2 molar mixture of triolein and trilinolein, dioleoyl phosphatidylcholine, and oleosins from rice (Oryza sativa), wheat (Triticum aestivum), rapeseed (Brassica napus), soybean (Glycine max), or jojoba (Simmondsia chinensis). It is concluded that both oleosins and PL are required to stabilize the oil bodies and that oleosins prevent oil bodies from coalescing by providing steric hindrance. A structural model of an oil body is presented. The current findings on seed oil bodies could be extended to the intracellular storage lipid particles present in diverse organisms. - + 100284535 Gene phospholipase A2 - + 100284535 Gene Phospholipase A2 - + 4577 Species maize - + 4577 Species Zea mays - + 4530 Species rice - + 4530 Species Oryza sativa - + 4565 Species wheat - + 4565 Species Triticum aestivum - + 3708 Species Brassica napus - + 3847 Species soybean - + 3847 Species Glycine max - + 3999 Species jojoba - + 3999 Species Simmondsia chinensis - - MESH:C069631 - Chemical - - TAG - - - MESH:D014304 - Chemical - - triolein - - - MESH:C510904 + + - Chemical - + oleosins - - MESH:C510904 + + MESH:D009821 Chemical - - oleosins + + oil - - MESH:C510904 + + MESH:D009821 Chemical - - oleosins + + oil - - MESH:C510904 + + - Chemical - + oleosins - + MESH:C017251 Chemical dioleoyl phosphatidylcholine - + MESH:C017251 Chemical dioleoyl phosphatidylcholine - - MESH:C510904 + + MESH:D010743 Chemical - - oleosins + + PL - + + - + Chemical + + seed oil + + + MESH:D009821 + Chemical + + Oil + + MESH:D014280 Chemical triacylglycerols - - MESH:D014304 - Chemical - - triolein - - - MESH:C069631 + + MESH:D010743 Chemical - - TAG + + phospholipids - - MESH:C510904 + + - Chemical oleosins - + + MESH:D009821 + Chemical + + oil + + MESH:C009317 Chemical stearylamine - - MESH:C069631 + + - Chemical - - TAG + + oleosins - - MESH:C069631 + + MESH:D010743 Chemical - - TAG + + PL - - MESH:D013229 + + MESH:D009821 Chemical - - stearic acid + + oil - - MESH:C069631 + + MESH:D009821 Chemical - - TAG + + oil - - MESH:C069631 + + MESH:D010743 Chemical - - TAG + + PL - + + MESH:D008055 + Chemical + + lipid + + + MESH:D009821 + Chemical + + oil + + + MESH:D009821 + Chemical + + oil + + MESH:C017251 Chemical dioleoyl phosphatidylcholine - - MESH:C510904 + + MESH:D009821 Chemical - + + oil + + + - + Chemical + oleosins - - MESH:C009564 + + MESH:C031183 Chemical - - trilinolein + + stearic acid - - MESH:C009564 + + - Chemical - - trilinolein + + oleosins + + + - + Chemical + + oleosins @@ -72118,41 +72650,41 @@ fibronectin + MESH:D008545 + Disease + + melanoma + + CVCL:0007 CellLine U-937 - + + MESH:D010703 + Chemical + + phorbol ester + + CVCL:0004 CellLine K-562 - + CVCL:0007 CellLine U-937 - + MESH:D002118 Chemical calcium - - MESH:D008545 - Disease - - melanoma - - - MESH:D010703 - Chemical - - phorbol ester - @@ -72174,175 +72706,217 @@ Rapiejko PJ, Gilmore R 0 Protein translocation across the ER requires a functional GTP binding site in the alpha subunit of the signal recognition particle receptor + + MESH:D006160 + Chemical + + GTP + ABSTRACT abstract 140 The signal recognition particle (SRP)-mediated translocation of proteins across the RER is a GTP dependent process. Analysis of the primary amino acid sequence of one protein subunit of SRP (SRP54), as well as the alpha subunit of the SRP receptor (SR alpha), has indicated that these proteins contain predicted GTP binding sites. Several point mutations confined to the GTP binding consensus elements of SR alpha were constructed by site specific mutagenesis to define a role for the GTP binding site in SR alpha during protein translocation. The SR alpha mutants were analyzed using an in vitro system wherein SR alpha- deficient microsomal membranes were repopulated with SR alpha by in vitro translation of wild-type or mutant mRNA transcripts. SRP receptors containing SR alpha point mutants were analyzed for their ability to function in protein translocation and to form guanylyl-5'- imidodiphosphate (Gpp[NH]p) stabilized complexes with the SRP. Mutations in SR alpha produced SRP receptors that were either impaired or inactive in protein translocation. These SRP receptors were likewise unable to form Gpp(NH)p stabilized complexes with the SRP. One SR alpha point mutant, Thr 588 to Asn 588, required 50- to 100-fold higher concentrations of GTP relative to the wild-type SR alpha to function in protein translocation. This mutant has provided information on the reaction step in protein translocation that involves the GTP binding site in the alpha subunit of the SRP receptor. - + 6729 Gene 2362 SRP54 - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 6729 Gene 2362 SRP - + 4481 Gene 12822 SR alpha - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - + 4481 Gene 12822 SR alpha - + 4481 Gene 12822 SR alpha - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - + 6729 Gene 2362 SRP - - MESH:D000641 + + MESH:D006160 Chemical - - NH + + GTP - + MESH:D006165 Chemical + + Gpp(NH)p + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006160 + Chemical + + GTP + + + - + Chemical guanylyl-5'- imidodiphosphate - + p.T588N ProteinMutation Thr 588 to Asn - - MESH:D006165 + + MESH:D009422 + Disease + + impaired + + + MESH:D006160 Chemical - - Gpp[NH]p + + GTP + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006160 + Chemical + + GTP @@ -72583,157 +73157,91 @@ Duronio RJ, Knoll LJ, Gordon JI 0 Isolation of a Saccharomyces cerevisiae long chain fatty acyl:CoA synthetase gene (FAA1) and assessment of its role in protein N- myristoylation - + 854495 Gene 137160 FAA1 - + 4932 Species Saccharomyces cerevisiae - - MESH:D000214 - Chemical - - fatty acyl - ABSTRACT abstract 145 Regulation of myristoylCoA pools in Saccharomyces cerevisiae plays an important role in modulating the activity of myristoylCoA:protein N- myristoyltransferase (NMT), an essential enzyme with an ordered Bi Bi reaction that catalyzes the transfer of myristate from myristoylCoA to greater than or equal to 12 cellular proteins. At least two pathways are available for generating myristoylCoA: de novo synthesis by the multifunctional, multisubunit fatty acid synthetase complex (FAS) and activation of exogenous myristate by acylCoA synthetase. The FAA1 (fatty acid activation) gene has been isolated by genetic complementation of a faal mutant. This single copy gene, which maps to the right arm of chromosome XV, specifies a long chain acylCoA synthetase of 700 amino acids. Analyses of strains containing NMT1 and a faal null mutation indicated that FAA1 is not essential for vegetative growth when an active de novo pathway for fatty acid synthesis is present. The role of FAA1 in cellular lipid metabolism and protein N-myristoylation was therefore assessed in strains subjected to biochemical or genetic blockade of FAS. At 36 degrees C, FAA1 is required for the utilization of exogenous myristate by NMT and for the synthesis of several phospholipid species. This requirement is not apparent at 24 or 30 degrees C, suggesting that S. cerevisiae contains another acylCoA synthetase activity whose chain length and/or temperature optima may differ from Faalp. - + 854495 Gene 137160 FAA1 - + 850892 Gene 101539 NMT1 - + 854495 Gene 137160 FAA1 - + 854495 Gene 137160 FAA1 - + 854495 Gene 137160 FAA1 - + 4932 Species Saccharomyces cerevisiae - + 4932 Species S. cerevisiae - - MESH:D009226 - Chemical - - myristate - - - MESH:D000214 - Chemical - - acylCoA - - - MESH:C040349 - Chemical - - myristoylCoA - - - MESH:D009226 - Chemical - - myristate - - - MESH:C040349 - Chemical - - myristoylCoA - - - MESH:D000214 - Chemical - - acylCoA - - - MESH:D005227 - Chemical - - fatty acid - - - MESH:D001729 - Chemical - - Bi Bi - - - MESH:D000214 + + MESH:D008055 Chemical - - acylCoA + + lipid - + MESH:D005227 Chemical fatty acid - - MESH:C040349 - Chemical - - myristoylCoA - - - MESH:D009226 - Chemical - - myristate - - - MESH:C040349 + + MESH:D010743 Chemical - - myristoylCoA + + phospholipid - + MESH:D005227 Chemical @@ -72760,55 +73268,61 @@ Chen Q, Linsenmayer C, Gu H, Schmid TM, Linsenmayer TF 0 Domains of type X collagen: alteration of cartilage matrix by fibril association and proteoglycan accumulation + + MESH:D002357 + Disease + + cartilage + ABSTRACT abstract 111 During endochondral bone formation, hypertrophic cartilage is replaced by bone or by a marrow cavity. The matrix of hypertrophic cartilage contains at least one tissue-specific component, type X collagen. Structurally type X collagen contains both a collagenous domain and a COOH-terminal non-collagenous one. However, the function(s) of this molecule have remained largely speculative. To examine the behavior and functions of type X collagen within hypertrophic cartilage, we (Chen, Q., E. Gibney, J. M. Fitch, C. Linsenmayer, T. M. Schmid, and T. F. Linsenmayer. 1990. Proc. Natl. Acad. Sci. USA. 87:8046-8050) recently devised an in vitro system in which exogenous type X collagen rapidly (15 min to several hours) moves into non-hypertrophic cartilage. There the molecule becomes associated with preexisting cartilage collagen fibrils. In the present investigation, we find that the isolated collagenous domain of type X collagen is sufficient for its association with fibrils. Furthermore, when non-hypertrophic cartilage is incubated for a longer time (overnight) with "intact" type X collagen, the molecule is found both in the matrix and inside of the chondrocytes. The properties of the matrix of such type X collagen-infiltrated cartilage become altered. Such changes include: (a) antigenic masking of type X collagen by proteoglycans; (b) loss of the permissiveness for further infiltration by type X collagen; and (c) enhanced accumulation of proteoglycans. Some of these changes are dependent on the presence of the COOH-terminal non-collagenous domain of the molecule. In fact, the isolated collagenous domain of type X collagen appears to exert an opposite effect on proteoglycan accumulation, producing a net decrease in their accumulation, particularly of the light form(s) of proteoglycans. Certain of these matrix alterations are similar to ones that have been observed to occur in vivo. This suggests that within hypertrophic cartilage type X collagen has regulatory as well as structural functions, and that these functions are achieved specifically by its two different domains. - + MESH:D002357 Disease hypertrophic cartilage - + MESH:D002357 Disease hypertrophic cartilage - - MESH:C102887 - Chemical - - COOH + + MESH:D002357 + Disease + + hypertrophic cartilage type X - - MESH:C535964 + + MESH:D002357 Disease - - hypertrophic cartilage type X collagen + + cartilage - - MESH:C102887 - Chemical - - COOH + + MESH:C535964 + Disease + + cartilage collagen fibrils - + MESH:D002357 Disease hypertrophic cartilage - + MESH:D002357 Disease hypertrophic cartilage - + MESH:D002357 Disease @@ -72835,124 +73349,154 @@ Ferreira A, Niclas J, Vale RD, Banker G, Kosik KS 0 Suppression of kinesin expression in cultured hippocampal neurons using antisense oligonucleotides + + MESH:D009841 + Chemical + + oligonucleotides + ABSTRACT abstract 99 Kinesin, a microtubule-based force-generating molecule, is thought to translocate organelles along microtubules. To examine the function of kinesin in neurons, we sought to suppress kinesin heavy chain (KHC) expression in cultured hippocampal neurons using antisense oligonucleotides and study the phenotype of these KHC "null" cells. Two different antisense oligonucleotides complementary to the KHC sequence reduced the protein levels of the heavy chain by greater than 95% within 24 h after application and produced identical phenotypes. After inhibition of KHC expression for 24 or 48 h, neurons extended an array of neurites often with one neurite longer than the others; however, the length of all these neurites was significantly reduced. Inhibition of KHC expression also altered the distribution of GAP-43 and synapsin I, two proteins thought to be transported in association with membranous organelles. These proteins, which are normally localized at the tips of growing neurites, were confined to the cell body in antisense-treated cells. Treatment of the cells with the corresponding sense oligonucleotides affected neither the distribution of GAP-43 and synapsin I, nor the length of neurites. A full recovery of neurite length occurred after removal of the antisense oligonucleotides from the medium. These data indicate that KHC plays a role in the anterograde translocation of vesicles containing GAP-43 and synapsin I. A deficiency in vesicle delivery may also explain the inhibition of neurite outgrowth. Despite the inhibition of KHC and the failure of GAP- 43 and synapsin I to move out of the cell body, hippocampal neurons can extend processes and acquire as asymmetric morphology. - + 3799 Gene 55829 kinesin heavy chain - + 3799 Gene 55829 KHC - + 3799 Gene 55829 KHC - + 3799 Gene 55829 KHC - + 3799 Gene 55829 KHC - + 3799 Gene 55829 KHC - + 2596 Gene 1545 GAP-43 - + 6853 Gene 48483 synapsin I - + 2596 Gene 1545 GAP-43 - + 6853 Gene 48483 synapsin I - + 3799 Gene 55829 KHC - + 2596 Gene 1545 GAP-43 - + 6853 Gene 48483 synapsin I - + 3799 Gene 55829 KHC - + 2596 Gene 1545 GAP- 43 - + 6853 Gene 48483 synapsin I + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:C567751 + Disease + + A deficiency in vesicle delivery + @@ -73081,34 +73625,34 @@ mouse - MESH:C032259 + MESH:D012967 Chemical - + SDS - MESH:C032259 + MESH:D012967 Chemical SDS - MESH:C032259 + MESH:D012967 Chemical - + SDS - MESH:C032259 + MESH:D012967 Chemical - + SDS - MESH:C032259 + MESH:D012967 Chemical - - 2-D SDS + + SDS @@ -73151,125 +73695,131 @@ abstract 83 A 40-kD protein kinase C (PKC)epsilon related activity was found to associate with human epithelial specific cytokeratin (CK) polypeptides 8 and 18. The kinase activity coimmunoprecipitated with CK8 and 18 and phosphorylated immunoprecipitates of the CK. Immunoblot analysis of CK8/18 immunoprecipitates using an anti-PKC epsilon specific antibody showed that the 40-kD species, and not native PKC epsilon (90 kD) associated with the cytokeratins. Reconstitution experiments demonstrated that purified CK8 or CK18 associated with a 40-kD tryptic fragment of purified PKC epsilon, or with a similar species obtained from cells that express the fragment constitutively but do not express CK8/18. A peptide pseudosubstrate specific for PKC epsilon inhibited phosphorylation of CK8/18 in intact cells or in a kinase assay with CK8/18 immunoprecipitates. Tryptic peptide map analysis of the cytokeratins that were phosphorylated by purified rat brain PKC epsilon or as immunoprecipitates by the associated kinase showed similar phosphopeptides. Furthermore, PKC epsilon immunoreactive species and CK8/18 colocalized using immunofluorescent double staining. We propose that a kinase related to the catalytic fragment of PKC epsilon physically associates with and phosphorylates cytokeratins 8 and 18. - + 5581 Gene 48343 PKC)epsilon - + 3856 Gene 55643 CK8 - + 3856 Gene 55643 CK8 - + 5581 Gene 48343 PKC epsilon - + 5581 Gene 48343 PKC epsilon - + 3856 Gene 55643 CK8 - + 3875 Gene 55448 CK18 - + 5581 Gene 48343 PKC epsilon - + 3856 Gene 55643 CK8 - + 5581 Gene 48343 PKC epsilon - + 3856 Gene 55643 CK8 - + 3856 Gene 55643 CK8 - + 5581 Gene 48343 PKC epsilon - + 3856 Gene 55643 CK8 - + 5581 Gene 48343 PKC epsilon - + 9606 Species human - + 10116 Species rat - - MESH:C566847 + + MESH:D010455 + Chemical + + peptide + + + MESH:C537180 Disease brain PKC epsilon @@ -73308,18 +73858,24 @@ abstract 101 The Balbiani Rings (BR) in the polytene chromosomes of Chironomus salivary glands are intense sites of transcription. The nascent RNPs fold during transcription into 40-50-nm granules, containing in the mature transcript approximately 37-kb RNA. Using a new nucleic acid specific stain, osmium ammine B on Lowicryl sections, in combination with electron energy filtered imaging of sections containing BR granules, we demonstrate a RNA-rich particulate substructure (10-nm particle diameter; 10-12 particles per BR granule). Elemental imaging supports that these particles are enriched in phosphorus. The possible relationship of these RNA-rich particles to ribonucleosomes is discussed, as well as models for their arrangement in the mature BR granules. - + MESH:D010758 Chemical phosphorus - + MESH:C058639 Chemical osmium ammine B + + - + Chemical + + Lowicryl + @@ -73354,108 +73910,102 @@ abstract 131 In a previous study we observed that neutrophils respond with a rapid rise in [Ca2+]i during adherence to cytokine-activated endothelial cells (EC), caused by EC membrane-associated platelet-activating factor (PAF). In the present study, we investigated whether this form of PAF was important in neutrophil adherence and migration across monolayers of rIL-1 beta- or rTNF alpha-prestimulated EC. PAF receptor antagonists prevented neutrophil migration across cytokine-pretreated EC by approximately 60% (P less than 0.005) without interfering with the process of adherence. The antagonists WEB 2086 and L-652,731 had no effect on neutrophil migration across resting EC induced by formylmethionyl-leucyl-phenylalanine (FMLP). A murine anti-IL-8 antiserum was found to also partially inhibit the neutrophil transmigration across cytokine-activated EC. When the anti-IL-8 antiserum was used in combination with a PAF receptor antagonist, neutrophil migration across cytokine-pretreated monolayers of EC was completely prevented. During transmigration, LAM-1 and CD44 on the neutrophils were down-modulated; both WEB 2086 and anti-IL-8 antiserum partially prevented this down-modulation caused by cytokine- prestimulated EC. Our results indicate that human neutrophils are activated and guided by EC-associated PAF and EC-derived IL-8 during the in vitro diapedesis in between cytokine-stimulated EC. - + 9768 Gene 8829 membrane-associated platelet-activating factor (PAF) - + 9768 Gene 8829 PAF - + 3576 Gene 47937 IL-8 - + 3576 Gene 47937 IL-8 - + 6402 Gene 539 LAM-1 - + 960 Gene 508 CD44 - + 3576 Gene 47937 IL-8 - + 3576 Gene 47937 IL-8 - + 9768 Gene 8829 PAF - + 10090 Species murine - + 9606 Species human - + 9768 Gene 8829 PAF - + 9768 Gene 8829 PAF - - MESH:C047504 - Chemical - - L-652,731 - - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ - + MESH:D009240 Chemical @@ -73488,46 +74038,52 @@ abstract 87 We have surveyed the proteins expressed at the surface of different primary neurons as a first step in elucidating how axons regulate their ensheathment by glial cells. We characterized the surface proteins of dorsal root ganglion neurons, superior cervical ganglion neurons, and cerebellar granule cells which are myelinated, ensheathed but unmyelinated, and unensheathed, respectively. We found that the most abundant proteins are common to all three types of neurons. Reproducible differences in the composition of the integral membrane proteins (enriched by partitioning into a Triton X-114 detergent phase) were detected. These differences were most striking when the expression of glycosylphosphatidyl-inositol (GPI)-anchored membrane proteins by these different neurons was compared. Variations in the relative abundance and degree of glycosylation of several well known GPI- anchored proteins, including Thy-1, F3/F11, and the 120-kD form of the neural cell adhesion molecule (N-CAM), and an abundant 60-kD GPI-linked protein were observed. In addition, we have identified several potentially novel GPI-anchored glycoproteins on each class of neurons. These include a protein that is present only on superior cervical ganglion neurons and is 90 kD; an abundant protein of 69 kD that is essentially restricted in its expression to dorsal root ganglion neurons; and proteins of 38 and 31 kD that are expressed only on granule cell neurons. Finally, the relative abundance of the three major isoforms of N-CAM was found to vary significantly between these different primary neurons. These results are the first demonstration that nerve fibers with diverse ensheathment fates differ significantly in the composition of their surface proteins and suggest an important role for GPI-anchored proteins in generating diversity of the neuronal cell surface. - + 7070 Gene 4580 Thy-1 - + 4684 Gene 40754 neural cell adhesion molecule - + 4684 Gene 40754 N-CAM - + 4684 Gene 40754 N-CAM - - MESH:D011092 - Chemical - - Triton - - MESH:D007294 + MESH:D017261 Chemical glycosylphosphatidyl-inositol + + MESH:C010615 + Chemical + + Triton X-114 + + + MESH:D017261 + Chemical + + GPI + @@ -73555,25 +74111,19 @@ abstract 108 Regulated secretory cells have two pathways that transport secreted proteins from the Golgi complex to the cell surface. To identify carrier vesicles involved in regulated and constitutive secretion, PC12 pheochromocytoma cells were labeled with [35S]sulfate to identify markers for the two secretory pathways, then mechanically permeabilized and incubated in vitro. Small constitutive secretory vesicles, containing mostly sulfated proteoglycans, accumulated during an in vitro incubation with ATP. In the presence of GTP gamma S, the constitutive vesicles became significantly more dense, suggesting that a coated intermediate was stabilized. Larger immature regulated secretory granules, enriched in sulfated secretogranin II, also escaped from the permeabilized cells in vitro. During granule maturation, their density increased and the amount of cofractionating proteoglycans diminished. The data suggest that sorting continues during secretory granule maturation. - + 24765 Gene 2591 secretogranin II - + MESH:D010673 Disease pheochromocytoma - - MESH:C031512 - Chemical - - [35S]sulfate - MESH:D016244 Chemical @@ -73581,12 +74131,24 @@ GTP gamma S + MESH:C000615320 + Chemical + + 35S + + MESH:D000255 Chemical ATP - + + MESH:D013431 + Chemical + + sulfate + + CVCL:0481 CellLine @@ -73766,58 +74328,70 @@ abstract 50 In this study, the putative laminin receptor function of the alpha 6 beta 4 integrin was assessed. For this purpose, we used a human cell line, referred to as clone A, that was derived from a highly invasive, colon adenocarcinoma. This cell line, which expresses the alpha 6 beta 4 integrin, adheres to the E8 and not to the P1 fragment of laminin. The adhesion of clone A cells to laminin is extremely rapid with half- maximal adhesion observed at 5 min after plating. Adhesion to laminin is blocked by GoH3, and alpha 6 specific antibody (60% inhibition), as well as by A9, a beta 4 specific antibody (30% inhibition). Most importantly, we demonstrate that alpha 6 beta 4 binds specifically to laminin-Sepharose columns in the presence of either Mg2+ or Mn2+ and it is eluted from these columns with EDTA but not with NaCl. The alpha 6 beta 4 integrin does not bind to collagen-Sepharose, but the alpha 2 beta 1 integrin does bind. Clone A cells do not express alpha 6 beta 1 as evidenced by the following observations: (a) no beta 1 integrin is detected in beta 1 immunoblots of GoH3 immunoprecipitates; and (b) no alpha 6 beta 1 integrin is seen in GoH3 immunoprecipitates of clone A extracts that had been immunodepleted of all beta 4 containing integrin using the A9 antibody. These data establish that laminin is a ligand for the alpha 6 beta 4 integrin and that this integrin can function as a laminin receptor independently of alpha 6 beta 1. - + 3688 Gene 22999 beta 1 integrin - + 9606 Species human - + 3688 Gene 22999 beta 1 integrin - + 3688 Gene 22999 beta 1 integrin - - MESH:D015179 + + - + Chemical + + Mg2+ + + + - + Chemical + + Mn2+ + + + MESH:D003110 Disease colon adenocarcinoma - - MESH:D008345 + + MESH:D012685 Chemical - - Mn2+ + + Sepharose - - MESH:D008274 + + MESH:D012685 Chemical - - Mg2+ + + Sepharose - + MESH:D004492 Chemical EDTA - + MESH:D012965 Chemical @@ -73850,32 +74424,25 @@ abstract 54 The N-ethylmaleimide sensitive fusion protein (NSF) is required for fusion of lipid bilayers at many locations within eukaryotic cells. Binding of NSF to Golgi membranes is known to require an integral membrane receptor and one or more members of a family of related soluble NSF attachment proteins (alpha-, beta-, and gamma-SNAPs). Here we demonstrate the direct interaction of NSF, SNAPs and an integral membrane component in a detergent solubilized system. We show that NSF only binds to SNAPs in the presence of the integral receptor, resulting in the formation of a multisubunit protein complex with a sedimentation coefficient of 20S. Particle assembly reveals striking differences between members of the SNAP protein family; gamma-SNAP associates with the complex via a binding site distinct from that used by alpha- and beta-SNAPs, which are themselves equivalent, alternative subunits of the particle. Once formed, the 20S particle is subsequently able to disassemble in a process coupled to the hydrolysis of ATP. We suggest how cycles of complex assembly and disassembly could help confer specificity to the generalized NSF-dependent fusion apparatus. - + 4905 Gene 4502 N-ethylmaleimide sensitive fusion protein - - 4905 - Gene - 4502 - - NSF - 4905 Gene 4502 - + NSF 4905 Gene 4502 - + NSF @@ -73905,6 +74472,18 @@ ATP + + MESH:D019962 + Disease + + NSF attachment proteins + + + MESH:D008055 + Chemical + + lipid + @@ -73932,18 +74511,24 @@ abstract 98 The inner row of dynein arms contains three dynein subforms. Each is distinct in composition and location in flagellar axonemes. To begin investigating the specificity of inner dynein arm assembly, we assessed the capability of isolated inner arm dynein subforms to rebind to their appropriate positions on axonemal doublet microtubules by recombining them with either mutant or extracted axonemes missing some or all dyneins. Densitometry of Coomassie blue-stained polyacrylamide gels revealed that for each inner dynein arm subform, binding to axonemes was saturable and stoichiometric. Using structural markers of position and polarity, electron microscopy confirmed that subforms bound to the correct inner arm position. Inner arms did not bind to outer arm or inappropriate inner arm positions despite the availability of sites. These and previous observations implicate specialized tubulin isoforms or nontubulin proteins in designation of specific inner dynein arm binding sites. Further, microtubule sliding velocities were restored to dynein-depleted axonemes upon rebinding of the missing inner arm subtypes as evaluated by an ATP-induced microtubule sliding disintegration assay. Therefore, not only were the inner arm dynein subforms able to identify and bind to the correct location on doublet microtubules but they bound in a functionally active conformation. - + MESH:D000255 Chemical ATP - - MESH:C040388 + + MESH:C016679 Chemical polyacrylamide + + MESH:C048139 + Chemical + + Coomassie blue + @@ -73971,65 +74556,59 @@ abstract 175 We have purified proteins of 70 kD from Drosophila, HeLa cells, and Z. mays that specifically bind nuclear localization sequences (NLSs). These proteins are recognized by antibodies raised against a previously identified NLS-binding protein (NBP) from the yeast S. cerevisiae. All NBPs are associated with nuclei and also present in the cytosol. NBPs are phosphorylated and phosphatase treatment abolished NLS binding. The requirement for NBPs in nuclear protein uptake is demonstrated in semipermeabilized Drosophila melanogaster tissue culture cells. Proper import of a fluorescent protein containing the large T antigen NLS requires cytosol and ATP. In the absence of cytosol and/or ATP, NLS- containing proteins are bound to cytosolic structures and the nuclear envelope. Addition of cytosol and ATP results in movement of this bound intermediate into the nucleus. Anti-NBP antibodies specifically inhibited the binding part of this import reaction. These results indicate that a phosphoprotein common to several eukaryotes acts as a receptor that recognizes NLSs before their uptake into the nucleus. - + 4577 Species Z. mays - + 4932 Species yeast - + 4932 Species S. cerevisiae - + 7227 Species Drosophila melanogaster + + 7227 + Species + + Drosophila + MESH:D000255 Chemical - + ATP - MESH:C000592574 - Chemical - - NBP - - MESH:D000255 Chemical - - ATP - - - MESH:D000255 - Chemical - + ATP - + CVCL:0030 CellLine HeLa - - MESH:C000592574 + + MESH:D000255 Chemical - - NBP + + ATP @@ -74219,11 +74798,17 @@ Foxall C, Watson SR, Dowbenko D, Fennie C, Lasky LA, Kiso M, Hasegawa A, Asa D, Brandley BK 0 The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide - - MESH:D002241 + + MESH:D009844 Chemical - - carbohydrate + + oligosaccharide + + + MESH:D000082 + Chemical + + x @@ -74231,124 +74816,130 @@ abstract 127 The selectins (lectin-EGF-complement binding-cell adhesion molecules [LEC-CAMs]) are a family of mammalian receptors implicated in the initial interactions between leukocytes and vascular endothelia, leading to lymphocyte homing, platelet binding, and neutrophil extravasation. The three known selectins, L-selectin (leukocyte adhesion molecule-1 [LECAM-1]), E-selectin (endothelial-leukocyte adhesion molecule-1 [ELAM-1]), and P-selectin (GMP-140) share structural features that include a calcium-dependent lectin domain. The sialyl Lewis(x) carbohydrate epitope has been reported as a ligand for both E- and P-selectins. Although L-selectin has been demonstrated to bind to carbohydrates, structural features of potential mammalian carbohydrate ligand(s) have not been well defined. Using an ELISA developed with a sialyl Lewis(x)-containing glycolipid and an E- selectin-IgG chimera, we have demonstrated the direct binding of the L- selectin-IgG chimera to sialyl Lewis(x). This recognition was calcium dependent, and could be blocked by Mel-14 antibody but not by other antibodies. Recognition was confirmed by the ability of cells expressing the native L-selectin to adhere to immobilized sialyl Lewis(x). These data suggest that the sialyl Lewis(x) oligosaccharide may form the basis of a recognition domain common to all three selectins. - + 6402 Gene 539 L-selectin - + 6402 Gene 539 leukocyte adhesion molecule-1 - + 6402 Gene 539 LECAM-1 - + 6401 Gene 389 E-selectin - + 6401 Gene 389 endothelial-leukocyte adhesion molecule-1 - + 6401 Gene 389 ELAM-1 - + 6403 Gene 2260 P-selectin - + 6403 Gene 2260 GMP-140 - + 6402 Gene 539 L-selectin - + 6402 Gene 539 L- selectin - + 6402 Gene 539 L-selectin - + 9606 Species mammalian - + 9606 Species mammalian - + + MESH:D006017 + Chemical + + glycolipid + + MESH:D002118 Chemical calcium - + MESH:D002241 Chemical carbohydrate - + MESH:D002241 Chemical carbohydrates - + MESH:D002118 Chemical calcium - - MESH:D002241 + + - Chemical - - carbohydrate + + (x) oligosaccharide @@ -74371,73 +74962,43 @@ Paschke KA, Lottspeich F, Stuermer CA 0 Neurolin, a cell surface glycoprotein on growing retinal axons in the goldfish visual system, is reexpressed during retinal axonal regeneration - + 7957 Species goldfish - - MESH:D020410 - Chemical - - Neurolin - ABSTRACT abstract 144 The mAb E 21 recognizes a cell surface glycoprotein selectively associated with fish retinal ganglion cell axons that are in a state of growth. All retinal axons and ganglion cells in goldfish embryos stained for E 21. In adult fish, however, E 21 immunoreactivity exhibited a patterned distribution in ganglion cells in the marginal growth zone of the continuously enlarging fish retina and the new axons emerging from these cells in the retina, optic nerve, and optic tract. The E 21 antigen was absent from older axons, except the terminal arbor layer in the tectum, the Stratum fibrosum et griseum superficiale where it was uniformly distributed. Upon optic nerve transection, the previously unlabeled axons reacquired E 21 positivity as they regenerated throughout their path to the tectum. Several months after ONS, however, E 21 staining disappeared from the regenerated axons over most of their lengths but reappeared as in normal fish in the terminal arbor layer. The immunoaffinity-purified E 21 antigen, called Neurolin, has an apparent molecular mass of 86 kD and contains the HNK1/L2 carbohydrate moiety, like several members of the class of cell adhesion molecules of the Ig superfamily. The NH2-terminal amino acid sequence has homologies to the cell adhesion molecule DM-Grasp recently described in the chicken. Thus, retinal ganglion cell axons express Neurolin during their development and are able to reexpress this candidate cell adhesion molecule during axonal regeneration, suggesting that Neurolin is functionally important for fish retinal axon growth. - + 396092 Gene 1229 DM-Grasp - + 7957 Species goldfish - + 9031 Species chicken - - MESH:D020410 - Chemical - - Neurolin - - + MESH:D002241 Chemical carbohydrate - - MESH:D020410 - Chemical - - Neurolin - - - MESH:D000641 - Chemical - - NH2 - - - MESH:D020410 - Chemical - - Neurolin - @@ -74478,10 +75039,10 @@ mouse - MESH:C418805 + MESH:D019297 Chemical - - radioiodinated + + DNP @@ -74504,225 +75065,189 @@ Piper RC, Tai C, Slot JW, Hahn CS, Rice CM, Huang H, James DE 0 The efficient intracellular sequestration of the insulin-regulatable glucose transporter (GLUT-4) is conferred by the NH2 terminus - + 6517 Gene 74381 GLUT-4 - + 3630 Gene 173 insulin - - MESH:D000641 - Chemical - - NH2 - - - MESH:D005947 - Chemical - - glucose - ABSTRACT abstract 131 GLUT-4 is the major facilitative glucose transporter isoform in tissues that exhibit insulin-stimulated glucose transport. Insulin regulates glucose transport by the rapid translocation of GLUT-4 from an intracellular compartment to the plasma membrane. A critical feature of this process is the efficient exclusion of GLUT-4 from the plasma membrane in the absence of insulin. To identify the amino acid domains of GLUT-4 which confer intracellular sequestration, we analyzed the subcellular distribution of chimeric glucose transporters comprised of GLUT-4 and a homologous isoform, GLUT-1, which is found predominantly at the cell surface. These chimeric transporters were transiently expressed in CHO cells using a double subgenomic recombinant Sindbis virus vector. We have found that wild-type GLUT-4 is targeted to an intracellular compartment in CHO cells which is morphologically similar to that observed in adipocytes and muscle cells. Sindbis virus-produced GLUT-1 was predominantly expressed at the cell surface. Substitution of the GLUT-4 amino-terminal region with that of GLUT-1 abolished the efficient intracellular sequestration of GLUT-4. Conversely, substitution of the NH2 terminus of GLUT-1 with that of GLUT-4 resulted in marked intracellular sequestration of GLUT-1. These data indicate that the NH2-terminus of GLUT-4 is both necessary and sufficient for intracellular sequestration. - + 6517 Gene 74381 GLUT-4 - + 6517 Gene 74381 GLUT-4 - + 6517 Gene 74381 GLUT-4 - + 3630 Gene 173 insulin - + 6517 Gene 74381 GLUT-4 - + 6517 Gene 74381 GLUT-4 - + 6513 Gene 68520 GLUT-1 - + 6517 Gene 74381 GLUT-4 - + 6513 Gene 68520 GLUT-1 - + 6517 Gene 74381 GLUT-4 - + 6513 Gene 68520 GLUT-1 - + 6517 Gene 74381 GLUT-4 - + 6513 Gene 68520 GLUT-1 - + 6517 Gene 74381 GLUT-4 - + 6513 Gene 68520 GLUT-1 - + 6517 Gene 74381 GLUT-4 - + 3630 Gene 173 Insulin - + 11034 Species Sindbis virus - + 11034 Species Sindbis virus - + 3630 Gene 173 insulin - - MESH:D005947 - Chemical - - glucose - - - MESH:D005947 - Chemical - - glucose - - - MESH:D000641 - Chemical - - NH2 - - + CVCL:0213 CellLine CHO - + MESH:D005947 Chemical glucose - - MESH:D000596 - Chemical - - amino - - + CVCL:0213 CellLine CHO - - MESH:D000641 + + MESH:D005947 Chemical - - NH2 + + glucose - + MESH:D005947 Chemical @@ -74755,30 +75280,48 @@ abstract 174 Migration of polymorphonuclear leukocytes across epithelia is a hallmark of many inflammatory disease states. Neutrophils traverse epithelia by migrating through the paracellular space and crossing intercellular tight junctions. We have previously shown (Nash, S., J. Stafford, and J.L. Madara. 1987. J. Clin. Invest. 80:1104-1113), that leukocyte migration across T84 monolayers, a model human intestinal epithelium, results in enhanced tight junction permeability--an effect quantitated by the use of a simple, standard electrical assay of transepithelial resistance. Here we show that detailed time course studies of the transmigration-elicited decline in resistance has two components, one of which is unrelated to junctional permeability. The initial decrease in resistance, maximal 5-13 min after initiation of transmigration, occurs despite inhibition of transmigration by an antibody to the common beta subunit of neutrophil beta 2 integrins, and is paralleled by an increase in transepithelial short-circuit current. Chloride ion substitution and inhibitor studies indicate that the early- phase resistance decline is not attributable to an increase in tight junction permeability but is due to decreased resistance across epithelial cells resulting from chloride secretion. Since T84 cells are accepted models for studies of the regulation of Cl- and water secretion, our results suggest that neutrophil transmigration across mucosal surfaces (for example, respiratory and intestinal tracts) may initially activate flushing of the surface by salt and water. Equally important, these studies, by providing a concrete example of sequential transcellular and paracellular effects on transepithelial resistance, highlight the fact that this widely used assay cannot simply be viewed as a direct functional probe of tight junction permeability. - + 322842 Species L. Madara - + 9606 Species human - + MESH:D002712 Chemical chloride - + + MESH:D012492 + Chemical + + salt + + MESH:D002712 Chemical Chloride + + MESH:D014867 + Chemical + + water + + + MESH:D014867 + Chemical + + water + @@ -74806,101 +75349,89 @@ abstract 158 Peptidylglycine alpha-amidating monooxygenase (PAM: EC 1.14.17.3) is a bifunctional protein which catalyzes the COOH-terminal amidation of bioactive peptides; the NH2-terminal monooxygenase and mid-region lyase act in sequence to perform the peptide alpha-amidation reaction. Alternative splicing of the single PAM gene gives rise to mRNAs generating PAM proteins with and without a putative transmembrane domain, with and without a linker region between the two enzymes, and forms containing only the monooxygenase domain. The expression, endoproteolytic processing, storage, and secretion of this secretory granule-associated protein were examined after stable transfection of AtT-20 mouse pituitary cells with naturally occurring and truncated PAM proteins. The transfected proteins were examined using enzyme assays, subcellular fractionation, Western blotting, and immunocytochemistry. Western blots of crude membrane and soluble fractions of transfected cells demonstrated that all PAM proteins were endoproteolytically processed. When the linker region was present between the monooxygenase and lyase domains, monofunctional soluble enzymes were generated from bifunctional PAM proteins; without the linker region, bifunctional enzymes were generated. Soluble forms of PAM expressed in AtT-20 cells and soluble proteins generated through selective endoproteolysis of membrane-associated PAM were secreted in an active form into the medium; secretion of the transfected proteins and endogenous hormone were stimulated in parallel by secretagogues. PAM proteins were localized by immunocytochemistry in the perinuclear region near the Golgi apparatus and in secretory granules, with the greatest intensity of staining in the perinuclear region in cell lines expressing integral membrane forms of PAM. Monofunctional and bifunctional PAM proteins that were soluble or membrane-associated were all packaged into regulated secretory granules in AtT-20 cells. - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 10090 Species mouse - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - + 18484 Gene 37369 PAM - - MESH:C102887 - Chemical - - COOH - - - MESH:D000641 - Chemical - - NH2 - @@ -74973,163 +75504,169 @@ abstract 145 We have examined the role of feedback-regulation in the expression of the nonmuscle actin genes. C2 mouse myoblasts were transfected with the human beta- and gamma-actin genes. In gamma-actin transfectants we found that the total actin mRNA and protein pools remained unchanged. Increasing levels of human gamma-actin expression resulted in a progressive down-regulation of mouse beta- and gamma-actin mRNAs. Transfection of the beta-actin gene resulted in an increase in the total actin mRNA and protein pools and induced an increase in the levels of mouse beta-actin mRNA. In contrast, transfection of a beta- actin gene carrying a single-point mutation (beta sm) produced a feedback-regulatory response similar to that of the gamma-actin gene. Expression of a beta-actin gene encoding an unstable actin protein had no impact on the endogenous mouse actin genes. This suggests that the nature of the encoded actin protein determines the feedback-regulatory response of the mouse genes. The role of the actin cytoskeleton in mediating this feedback-regulation was evaluated by disruption of the actin network with Cytochalasin D. We found that treatment with Cytochalasin D abolished the down-regulation of mouse gamma-actin in both the gamma- and beta sm-actin transfectants. In contrast, a similar level of increase was observed for the mouse beta-actin mRNA in both control and transfected cells. These experiments suggest that the down- regulation of mouse gamma-actin mRNA is dependent on the organization of the actin cytoskeleton. In addition, the mechanism responsible for the down-regulation of beta-actin may be distinct from that governing gamma-actin. We conclude that actin feedback-regulation provides a biochemical assay for differences between the two nonmuscle actin genes. - + 728378 Gene 133745 beta- and gamma-actin - + 11465 Gene 74402 gamma-actin - + 11461;11465 Gene beta- and gamma-actin - + 11461 Gene 110648 beta-actin - + 11461 Gene 110648 beta-actin - + 11461 Gene 110648 beta- actin - + 11465 Gene 74402 gamma-actin - + 11461 Gene 110648 beta-actin - + 11465 Gene 74402 gamma-actin - + 11461 Gene 110648 beta-actin - + 11465 Gene 74402 gamma-actin - + 11461 Gene 110648 beta-actin - + 11465 Gene 74402 gamma-actin - + 10090 Species mouse - + 9606 Species human - + 9606 Species human - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 11465 Gene 74402 gamma-actin + + MESH:D015638 + Chemical + + Cytochalasin D + @@ -75151,60 +75688,48 @@ Hauft SM, Kim SH, Schmidt GH, Pease S, Rees S, Harris S, Roth KA, Hansbrough JR, Cohn SM, Ahnen DJ 0 Expression of SV-40 T antigen in the small intestinal epithelium of transgenic mice results in proliferative changes in the crypt and reentry of villus-associated enterocytes into the cell cycle but has no apparent effect on cellular differentiation programs and does not cause neoplastic transformation - + 10090 Species transgenic mice - - MESH:D002471 - Disease - - neoplastic transformation - ABSTRACT abstract 304 The mouse intestinal epithelium represents a unique mammalian system for examining the relationship between cell division, commitment, and differentiation. Proliferation and differentiation are rapid, perpetual, and spatially well-organized processes that occur along the crypt-to-villus axis and involve clearly defined cell lineages derived from a common multipotent stem cell located near the base of each crypt. Nucleotides -1178 to +28 of the rat intestinal fatty acid binding protein gene were used to establish three pedigrees of transgenic mice that expressed SV-40 large T antigen (TAg) in epithelial cells situated in the uppermost portion of small intestinal crypts and in already committed, differentiating enterocytes as they exited these crypts and migrated up the villus. T antigen production was associated with increases in crypt cell proliferation but had no apparent effect on commitment to differentiate along enterocytic, enteroendocrine, or Paneth cell lineages. Single- and multilabel- immunocytochemical studies plus RNA blot hybridization analyses suggested that the differentiation programs of these lineages were similar in transgenic mice and their normal littermates. This included enterocytes which, based on the pattern of [3H]thymidine and 5-bromo-2'- deoxyuridine labeling and proliferating nuclear antigen expression, had reentered the cell cycle during their migration up the villus. The state of cellular differentiation and/or TAg production appeared to affect the nature of the cell cycle; analysis of the ratio of S-phase to M-phase cells (collected by metaphase arrest with vincristine) and of the intensities of labeling of nuclei by [3H]thymidine indicated that the duration of S phase was longer in differentiating, villus- associated enterocytes than in the less well-differentiated crypt epithelial cell population and that there may be a block at the G2/M boundary. Sustained increases in crypt and villus epithelial cell proliferation over a 9-mo period were not associated with the development of gut neoplasms--suggesting that tumorigenesis in the intestine may require that the initiated cell have many of the properties of the gut stem cell including functional anchorage. - + 10090 Species mouse - + 9606 Species mammalian - + 10116 Species rat - + 10090 Species transgenic mice - + 10090 Species transgenic mice - - MESH:D014750 - Chemical - - vincristine - MESH:D001973 Chemical @@ -75212,17 +75737,41 @@ 5-bromo-2'- deoxyuridine - MESH:D013936 - Chemical - - [3H]thymidine + MESH:D009369 + Disease + + neoplasms - MESH:D013936 + MESH:D005227 + Chemical + + fatty acid + + + - Chemical [3H]thymidine + + MESH:D014750 + Chemical + + vincristine + + + MESH:D014316 + Chemical + + 3H + + + MESH:D013936 + Chemical + + thymidine + @@ -75482,137 +76031,125 @@ Forsayeth JR, Gu Y, Hall ZW 0 BiP forms stable complexes with unassembled subunits of the acetylcholine receptor in transfected COS cells and in C2 muscle cells - + 3309 Gene 3908 BiP - + CVCL:0529 CellLine C2 - + CVCL:0222 CellLine COS - - MESH:D000109 - Chemical - - acetylcholine - ABSTRACT abstract 131 We have investigated the role of the immunoglobulin-binding protein (BiP) in the folding and assembly of subunits of the acetylcholine receptor (AChR) in COS cells and in C2 muscle cells. Immunoprecipitation in COS cells showed that alpha, beta, and delta subunits are associated with BiP. In the case of the alpha subunit, which first folds to acquire toxin-binding activity and is then assembled with the other subunits to form the AChR, BiP was associated only with a form that is unassembled and does not bind alpha- bungarotoxin. Similar results were found in C2 cells. Although the alpha and beta subunits of the AChR are minor membrane proteins in C2 cells, they were prominent among the proteins immunoprecipitated by antibodies to BiP, suggesting that BiP could play a role in their maturation or folding. In pulse-chase experiments in C2 cells, however, labeled alpha subunit formed a stable complex with BiP that was first detected after most of the alpha subunit had acquired toxin-binding activity and whose amount continued to increase for several hours. These kinetics are not compatible with a role for the BiP complex in the folding or assembly pathway of the AChR, and suggest that BiP is associated with a misfolded form of the subunit that is slowly degraded. - + 3309 Gene 3908 immunoglobulin-binding protein - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + 3309 Gene 3908 BiP - + CVCL:0529 CellLine C2 - + CVCL:0529 CellLine C2 - + CVCL:0222 CellLine COS - + CVCL:0529 CellLine C2 - - MESH:D000109 - Chemical - - acetylcholine - - + CVCL:0529 CellLine C2 - + CVCL:0222 CellLine @@ -75651,23 +76188,41 @@ abstract 106 Polymerization of actin has been associated with development of polar shape in human neutrophils (PMN). To examine the relation of filamentous actin (F-actin) distribution to shape change in PMN, we developed a method using computerized video image analysis and fluorescence microscopy to quantify distribution of F-actin in single cells. PMN were labeled with fluorescent probe NBD-phallicidin to measure filamentous actin and Texas red to assess cell thickness. We show that Texas red fluorescence is a reasonable measure of cell thickness and that correction of the NBD-phallicidin image for cell thickness using the Texas red image permits assessment of focal F-actin content. Parameters were derived that quantify total F-actin content, movement of F-actin away from the center of the cell, asymmetry of F- actin distribution, and change from round to polar shape. The sequence of change in F-actin distribution and its relation to development of polar shape in PMN was determined using these parameters. After stimulation with chemotactic peptide at 25 degrees C, F-actin polymerized first at the rim of the PMN. This was followed by development of asymmetry of F-actin distribution and change to polar shape. The dominant pseudopod developed first in the region of lower F- actin concentration followed later by polymerization of actin in the end of the developed pseudopod. Asymmetric F-actin distribution was detected in round PMN before development of polar shape. Based upon these data, asymmetric distribution of F-actin is coincident with and probably precedes development of polar shape in PMN stimulated in suspension by chemotactic peptide. - + 9606 Species human - - MESH:C028877 + + - Chemical - - NBD-phallicidin + + phallicidin - - MESH:C028877 + + MESH:C034657 + Chemical + + Texas red + + + - + Chemical + + phallicidin + + + MESH:C034657 + Chemical + + Texas red + + + MESH:C034657 Chemical - - NBD-phallicidin + + Texas red @@ -75703,95 +76258,83 @@ abstract 95 The 47,000-D collagen-binding glycoprotein, heat shock protein 47 (HSP47), is a stress-inducible protein localized in the ER of collagen- secreting cells. The location and collagen-binding activity of this protein led to speculation that HSP47 might participate in collagen processing. Chemical crosslinking studies were used to test this hypothesis both before and after the perturbation of procollagen processing. The association of procollagen with HSP47 was demonstrated using cleavable bifunctional crosslinking reagents. HSP47 and procollagen were shown to be coprecipitated by the treatment of intact cells with anti-HSP47 or with anticollagen antibodies. Furthermore, several proteins residing in the ER were noted to be crosslinked to and coprecipitated with HSP47, suggesting that these ER-resident proteins may form a large complex in the ER. When cells were heat shocked, or when stable triple-helix formation was inhibited by treatment with alpha,alpha'-dipyridyl, coprecipitation of procollagen with HSP47 was increased. This increase was due to the inhibition of procollagen secretion and to the accumulation of procollagen in the ER. Pulse label and chase experiments revealed that coprecipitated procollagen was detectable as long as procollagen was present in the endoplasmic reticulum of alpha,alpha'-dipyridyl-treated cells. Under normal growth conditions, coprecipitated procollagen was observed to decrease after a chase period of 10-15 min, whereas total procollagen decreased only after 20-25 min. In addition, the intracellular association between HSP47 and procollagen was shown to be disrupted by a change in physiological pH, suggesting that the dissociation of procollagen from HSP47 is pH dependent. These findings support a specific role for HSP47 in the intracellular processing of procollagen, and provide evidence of a new category of "molecular chaperones" in terms of its substrate specificity and the dissociation mechanism. - + 871 Gene 20331 heat shock protein 47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - + 871 Gene 20331 HSP47 - - MESH:D015082 - Chemical - - alpha,alpha'-dipyridyl - - - MESH:D015082 - Chemical - - alpha,alpha'-dipyridyl - @@ -75928,15 +76471,15 @@ mouse - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ @@ -75945,114 +76488,124 @@ abstract 167 During early development, intracellular Ca2+ mobilization is not only essential for fertilization, but has also been implicated during other meiotic and mitotic events, such as germinal vesicle breakdown (GVBD) and nuclear envelope breakdown (NEBD). In this study, the roles of intracellular and extracellular Ca2+ were examined during meiotic maturation and reinitiation at parthenogenetic activation and during first mitosis in a single species using the same methodologies. Cumulus- free metaphase II mouse oocytes immediately resumed anaphase upon the induction of a large, transient Ca2+ elevation. This resumption of meiosis and associated events, such as cortical granule discharge, were not sensitive to extracellular Ca2+ removal, but were blocked by intracellular Ca2+ chelators. In contrast, meiosis I was dependent on external Ca2+; in its absence, the formation and function of the first meiotic spindle was delayed, the first polar body did not form and an interphase-like state was induced. GVBD was not dependent on external Ca2+ and showed no associated Ca2+ changes. NEBD at first mitosis in fertilized eggs, on the other hand, was frequently, but not always associated with a brief Ca2+ transient and was dependent on Ca2+ mobilization. We conclude that GVBD is Ca2+ independent, but that the dependence of NEBD on Ca2+ suggests regulation by more than one pathway. As cells develop from Ca(2+)-independent germinal vesicle oocytes to internal Ca(2+)-dependent pronuclear eggs, internal Ca2+ pools increase by approximately fourfold. - + 10090 Species mouse - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ - - MESH:D002118 - Chemical - - Ca2+ - - - MESH:C536875 - Disease - - meiosis I - - MESH:D002118 + MESH:D000069285 Chemical - - Ca(2+) + + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical Ca(2+) - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 - Chemical - - Ca2+ - - MESH:C536875 Disease - - meiosis + + meiosis I - - MESH:D002118 + + MESH:D000069285 Chemical - + Ca2+ + + Disease + + mitosis + - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical - - Ca2+ + + Ca(2+) - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 - Chemical - - Ca2+ + MESH:C536875 + Disease + + meiosis - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical Ca2+ - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical - + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + Ca2+ + + Disease + + mitosis + @@ -76093,7 +76646,7 @@ neurite initiation and neurite outgrowth - MESH:D010716 + - Chemical glycane-phosphatidylinositol @@ -76126,16 +76679,16 @@ 0 Regulation of alpha 2 beta 1-mediated fibroblast migration on type I collagen by shifts in the concentrations of extracellular Mg2+ and Ca2+ [published erratum appears in J Cell Biol 1992 Jul;118(1):219] - MESH:D002118 + - Chemical - - Ca2+ + + Mg2+ - MESH:D008274 + MESH:D000069285 Chemical - - Mg2+ + + Ca2+ @@ -76143,70 +76696,76 @@ abstract 204 Extracellular Ca2+ can reverse the Mg(2+)-dependent, alpha 2 beta 1- mediated adhesion of WI38 human fibroblasts to type I collagen substrates. Affinity chromatography data also demonstrate that Ca2+ can specifically elute the fibroblast alpha 2 beta 1 integrin bound to type I collagen-Sepharose in Mg2+. In modified Boyden chamber migration assays, Mg2+ alone supports the alpha 2 beta 1-mediated migration of fibroblasts on type I collagen substrates, while Ca2+ does not. However, a twofold enhancement in migration was observed when combinations of the two cations were used, with optimal migration observed when the Mg2+/Ca2+ ratio was higher than one. Inhibitory mAbs directed against various integrin subunits demonstrate that these observed cation effects appear to be mediated primarily by alpha 2 beta 1. These data, together with reports that under certain physiological conditions significant fluctuations in the concentrations of extracellular Ca2+ and Mg2+ can take place in vivo, suggest that the ratio between these two cations is involved in the up- and downregulation of integrin function, and thus, may influence cell migratory behavior. - + 9606 Species human - - MESH:D008274 - Chemical - - Mg2 - - MESH:D008274 + MESH:D000069285 Chemical - - Mg2+ + + Ca2+ - MESH:D008274 + - Chemical - + Mg2+ - MESH:D008274 + MESH:D000069285 Chemical - - Mg(2+) + + Ca2+ - MESH:D002118 + - Chemical - - Ca2+ + + Mg2+ - MESH:D002118 + - Chemical - - Ca2+ + + Mg(2+) - MESH:D008274 + MESH:D000069285 Chemical - - Mg2+ + + Ca2+ - MESH:D002118 + MESH:D000069285 Chemical - + Ca2+ - MESH:D002118 + - Chemical - - Ca2+ + + Mg2+ - MESH:D002118 + - Chemical - + + Mg2+ + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:D000069285 + Chemical + Ca2+ @@ -76230,145 +76789,187 @@ Troy CM, Greene LA, Shelanski ML 0 Neurite outgrowth in peripherin-depleted PC12 cells - + 24688 Gene 4559 peripherin + + CVCL:0481 + CellLine + + PC12 + ABSTRACT abstract 52 Peripherin is the major neuronal intermediate filament (IF) protein in PC12 cells and both its synthesis and amount increase during nerve growth factor (NGF) promoted neuronal differentiation. To address the question of the biological function of peripherin in neurite initiation we have used an antisense oligonucleotide complementary to the 5' region of peripherin mRNA to specifically inhibit its transcription. The oligonucleotide blocks both the synthesis of peripherin and its increase in response to NGF. Peripherin was found to be a stable protein with a cellular half-life of approximately 7 d. 6 wk of incubation with the oligonucleotide decreases peripherin to 11% of the level in naive control cells and to 3% of that in NGF-treated control cells. Despite the depletion, NGF elicits apparently normal neurite outgrowth from the oligonucleotide-treated cells. As evaluated by EM, there are few IFs in these cells, either in the cell bodies or neurites. There is no compensatory increase in NF-M, NF-L, or vimentin levels as a result of the inhibition of peripherin synthesis. These findings suggest that peripherin is not required for neurite formation, but is necessary for the formation of a cellular IF network which could be involved in process stability. They also demonstrate the utility of antisense oligonucleotides for the study of proteins with long half- lives. - + 24688 Gene 4559 Peripherin - + 24387 Gene 1554 intermediate filament (IF) - + 310738 Gene 1876 nerve growth factor - + 310738 Gene 1876 NGF - + 24688 Gene 4559 peripherin - + 24688 Gene 4559 peripherin - + 24688 Gene 4559 peripherin - + 310738 Gene 1876 NGF - + 24688 Gene 4559 Peripherin - + 24688 Gene 4559 peripherin - + 310738 Gene 1876 NGF - + 310738 Gene 1876 NGF - + 24588 Gene 137211 NF-M - + 83613 Gene 4487 NF-L - + 81818 Gene 2538 vimentin - + 24688 Gene 4559 peripherin - + 24688 Gene 4559 peripherin - + 24387 Gene 1554 IF + + CVCL:0481 + CellLine + + PC12 + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + @@ -76396,24 +76997,18 @@ abstract 122 Laminin self-assembles in vitro into a polymer by a reversible, entropy- driven and calcium-facilitated process dependent upon the participation of the short arm globular domains. We now find that this polymer is required for the structural integrity of the collagen-free basement membrane of cultured embryonal carcinoma cells (ECC) and for the supramolecular organization and anchorage of laminin in the collagen- rich basement membrane of the Engelbreth-Holm-Swarm tumor (EHS). First, low temperature and EDTA induced the dissolution of ECC basement membranes and released approximately 80% of total laminin from the EHS basement membrane. Second, laminin elastase fragments (E4 and E1') possessing the short arm globules of the B1, B2, and A chains selectively acted as competitive ligands that dissolved ECC basement membranes and displaced laminin from the EHS basement membrane into solution. The fraction of laminin released increased as a function of ligand concentration, approaching the level of the EDTA-reversible pool. The smaller (approximately 20%) residual pool of EHS laminin, in contrast, could only be effectively displaced by E1' and E4 if the collagenous network was first degraded with bacterial collagenase. The supramolecular architecture of freeze-etched and platinum/carbon replicated reconstituted laminin gel polymer, ECC, and collagenase- treated EHS basement membranes were compared and found to be similar, further supporting the biochemical data. We conclude that laminin forms a network independent of that of type IV collagen in basement membranes. Furthermore, in the EHS basement membrane four-fifths of laminin is anchored strictly through noncovalent bonds between laminin monomers while one-fifth is anchored through a combination of these bonds and laminin-collagen bridges. - + MESH:D002118 Chemical calcium - + MESH:D002244 Chemical carbon - - MESH:D009369 - Disease - - tumor - MESH:D010984 Chemical @@ -76423,21 +77018,33 @@ MESH:D004492 Chemical - + EDTA - MESH:D018236 - Disease - - embryonal carcinoma + MESH:D011108 + Chemical + + polymer + MESH:D009369 + Disease + + tumor + + MESH:D004492 Chemical - + EDTA + + MESH:D018236 + Disease + + embryonal carcinoma + @@ -76542,26 +77149,31 @@ Hamaguchi JR, Tobey RA, Pines J, Crissman HA, Hunter T, Bradbury EM 0 Requirement for p34cdc2 kinase is restricted to mitosis in the mammalian cdc2 mutant FT210 - + 983 Gene 68203 p34cdc2 - + 983 Gene 68203 cdc2 - + 9606 Species mammalian + + Disease + + mitosis + ABSTRACT @@ -76749,18 +77361,6 @@ cyclin A - - MESH:C102887 - Chemical - - COOH - - - MESH:C102887 - Chemical - - COOH - @@ -76864,7 +77464,7 @@ Semliki Forest virus - MESH:D014777 + MESH:D001102 Disease viral infections @@ -76890,54 +77490,24 @@ Klein F, Laroche T, Cardenas ME, Hofmann JF, Schweizer D, Gasser SM 0 Localization of RAP1 and topoisomerase II in nuclei and meiotic chromosomes of yeast - - MESH:C040523 - Chemical - - RAP1 - ABSTRACT abstract 85 Topoisomerase II (topoII) and RAP1 (Repressor Activator Protein 1) are two abundant nuclear proteins with proposed structural roles in the higher-order organization of chromosomes. Both proteins co-fractionate as components of nuclear scaffolds from vegetatively growing yeast cells, and both proteins are present as components of pachytene chromosome, co-fractionating with an insoluble subfraction of meiotic nuclei. Immunolocalization using antibodies specific for topoII shows staining of an axial core of the yeast meiotic chromosome, extending the length of the synaptonemal complex. RAP1, on the other hand, is located at the ends of the paired bivalent chromosomes, consistent with its ability to bind telomeric sequences in vitro. In interphase nuclei, again in contrast to anti-topoII, anti-RAP1 gives a distinctly punctate staining that is located primarily at the nuclear periphery. Approximately 16 brightly staining foci can be identified in a diploid nucleus stained with anti-RAP1 antibodies, suggesting that telomeres are grouped together, perhaps through interaction with the nuclear envelope. - + 4932 Species yeast - + 4932 Species yeast - - MESH:C040523 - Chemical - - RAP1 - - - MESH:C040523 - Chemical - - RAP1 - - - MESH:C040523 - Chemical - - RAP1 - - - MESH:C040523 - Chemical - - RAP1 - @@ -76983,54 +77553,93 @@ abstract 170 The spindle pole body (SPB) is the equivalent of the centrosome in fission yeast. In vivo it nucleates microtubules (MTs) during mitosis, but, unlike animal centrosomes, does not act as a microtubule organizing center (MTOC) during interphase. We have studied the MT- nucleating activity of SPBs in vitro and have found that SPBs in permeabilized cells retain in vivo characteristics. SPBs in cells permeabilized during mitosis can nucleate MTs, and are recognized by two antibodies: anti-gamma-tubulin and MPM-2 which recognizes phosphoepitopes. SPBs in cells permeabilized during interphase cannot nucleate MTs and are only recognized by anti-gamma-tubulin. Interphase SPBs which cannot nucleate can be converted to a nucleation competent state by incubation in cytostatic factor (CSF)-arrested Xenopus egg extracts. After incubation, they are recognized by MPM-2, and can nucleate MTs. The conversion does not occur in Xenopus interphase extract, but occurs in Xenopus interphase extract driven into mitosis by preincubation with exogenous cyclin B. The conversion is ATP dependent and inhibited by protein kinase inhibitors and alkaline phosphatase. Purified, active, cdc2 kinase/cyclin B complex in itself is not effective for activation of MT nucleation, although some interphase SPBs are now stained with MPM-2. These results suggest that the ability of SPBs in vitro to nucleate MTs after exposure to CSF- arrested extracts is activated through a downstream pathway which is regulated by cdc2 kinase. - + 4896 Species fission yeast - + 8355 Species Xenopus - + 8355 Species Xenopus - + 8355 Species Xenopus - - MESH:C476157 + + Disease + + mitosis + + + Disease + + mitosis + + + MESH:C042995 Chemical - - MPM-2 + + SPBs - - MESH:C476157 + + MESH:C042995 Chemical - - MPM-2 + + SPBs - + + MESH:C042995 + Chemical + + SPBs + + + Disease + + mitosis + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + MESH:D000255 Chemical ATP - - MESH:C476157 - Chemical - - MPM-2 - @@ -77064,27 +77673,34 @@ abstract 120 The protein product of the proto-oncogene c-src is a membrane- associated tyrosine kinase of unknown function. Identification of pp60c- src target membranes may elucidate the function of the c-src protein. The available evidence indicates that pp60c-src associates with distinct membranes within single cell types and has different distributions in different cell types. Our experiments demonstrate targeting of pp60c-src to an isolatable and biochemically identified membrane fraction in the neuroendocrine cell line PC12. The c-src protein was found to be specifically associated with synaptic vesicles since: (a) the pp60c-src immunofluorescent pattern overlapped with a synaptic vesicle marker, synaptophysin; (b) a significant proportion (44%) of the pp60c-src from PC12 but not fibroblast postnuclear supernatants was recovered in a small vesicle fraction; (c) an anti- synaptophysin cytoplasmic domain antibody immunodepleted all of the pp60c-src vesicles in this fraction, and (d) pp60c-src copurified during a 100-fold purification of PC12 synaptic vesicles. These results suggest a role for the c-src protein in the regulation of synaptic vesicle function. - + 6714 Gene 21120 proto-oncogene c-src - + 6714 Gene 21120 c-src - + 6714 Gene 21120 c-src + + 6855 + Gene + 2391 + + synaptophysin + 6714 Gene @@ -77093,35 +77709,25 @@ c-src - MESH:D016708 - Chemical + 6855 + Gene + 2391 synaptophysin - MESH:D016708 - Chemical - - synaptophysin - - CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - - Chemical - - tyrosine - - + CVCL:0481 CellLine @@ -77174,135 +77780,159 @@ abstract 100 We have used monolayers of control 3T3 cells and 3T3 cells expressing transfected human neural cell adhesion molecule (NCAM) or chick N- cadherin as a culture substrate for PC12 cells. NCAM and N-cadherin in the monolayer directly promote neurite outgrowth from PC12 cells via a G-protein-dependent activation of neuronal calcium channels. In the present study we show that ganglioside GM1 does not directly activate this pathway in PC12 cells. However, the presence of GM1 (12.5-100 micrograms/ml) in the co-culture was associated with a potentiation of NCAM and N-cadherin-dependent neurite outgrowth. Treatment of PC12 cells with GM1 (100 micrograms/ml) for 90 min led to trypsin-stable increases in both beta-cholera toxin binding to PC12 cells and an enhanced neurite outgrowth response to N-cadherin. The ganglioside response could be fully inhibited by treatment with pertussis toxin. These data are consistent with exogenous gangliosides enhancing neuritic growth by promoting cell adhesion molecule-induced calcium influx into neurons. - + 4684 Gene 40754 neural cell adhesion molecule - + 414745 Gene 20424 N- cadherin - + 24586 Gene 40754 NCAM - + 83501 Gene 20424 N-cadherin - + 24586 Gene 40754 NCAM - + 83501 Gene 20424 N-cadherin - + 83501 Gene 20424 N-cadherin - + 4684 Gene 40754 NCAM - + 9606 Species human - + 9031 Species chick - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + CVCL:0481 CellLine PC12 - + + MESH:D058225 + Disease + + neuritic + + CVCL:0594 CellLine 3T3 - + CVCL:0481 CellLine PC12 - + + MESH:D005677 + Chemical + + GM1 + + + MESH:D005677 + Chemical + + GM1 + + MESH:D002118 Chemical calcium - + CVCL:0594 CellLine 3T3 - + + MESH:D005732 + Chemical + + gangliosides + + CVCL:0481 CellLine PC12 - + MESH:D005732 Chemical ganglioside - - MESH:D002118 + + MESH:D005677 Chemical - - calcium + + GM1 - + MESH:D005732 Chemical @@ -77388,41 +78018,47 @@ abstract 85 Experiments are described supporting the proposition that the assembly of stress fibers in non-muscle cells and the assembly of myofibrils in cardiac cells share conserved mechanisms. Double staining with a battery of labeled antibodies against membrane-associated proteins, myofibrillar proteins, and stress fiber proteins reveals the following: (a) dissociated, cultured cardiac myocytes reconstitute intercalated discs consisting of adherens junctions (AJs) and desmosomes at sites of cell-cell contact and sub-sarcolemmal adhesion plaques (SAPs) at sites of cell-substrate contact; (b) each AJ or SAP associates proximally with a striated myofibril, and conversely every striated myofibril is capped at either end by an AJ or a SAP; (C) the invariant association between a given myofibril and its SAP is especially prominent at the earliest stages of myofibrillogenesis; nascent myofibrils are capped by oppositely oriented SAPs; (d) the insertion of nascent myofibrils into AJs or into SAPs invariably involves vinculin, alpha-actin, and sarcomeric alpha-actinin (s-alpha-actinin); (e) AJs are positive for A- CAM but negative for talin and integrin; SAPs lack A-CAM but are positive for talin and integrin; (f) in cardiac cells all alpha-actinin- containing structures invariably are positive for the sarcomeric isoform, alpha-actin and related sarcomeric proteins; they lack non-s- alpha-actinin, gamma-actin, and caldesmon; (g) in fibroblasts all alpha- actinin-containing structures are positive for the non-sarcomeric isoform, gamma-actin, and related non-sarcomeric proteins, including caldesmon; and (h) myocytes differ from all other types of adherent cultured cells in that they do not assemble authentic stress fibers; instead they assemble stress fiber-like structures of linearly aligned I-Z-I-like complexes consisting exclusively of sarcomeric proteins. - + 7414 Gene 7594 vinculin - + 87 Gene 55553 alpha-actinin - + 87 Gene 55553 alpha-actinin - + 87 Gene 55553 alpha-actinin - + 87 Gene 55553 alpha-actinin + + MESH:C580316 + Disease + + myofibrillar proteins + @@ -77504,96 +78140,78 @@ abstract 180 We have raised two monospecific antibodies against synthetic peptides derived from the membrane domain of the ER glycoprotein 3-hydroxy-3- methylglutaryl coenzyme A (HMG-CoA) reductase, the rate limiting enzyme in the cholesterol biosynthetic pathway. This domain, which was proposed to span the ER membrane seven times (Liscum, L., J. Finer- Moore, R. M. Stroud, K. L. Luskey, M. S. Brown, and J. L. Goldstein. 1985. J. Biol. Chem. 260:522-538), plays a critical role in the regulated degradation of the enzyme in the ER in response to sterols. The antibodies stain the ER of cells and immunoprecipitate HMG-CoA reductase and HMGal, a chimeric protein composed of the membrane domain of the reductase fused to Escherichia coli beta-galactosidase, the degradation of which is also accelerated by sterols. We show that the sequence Arg224 through Leu242 of HMG-CoA reductase (peptide G) faces the cytoplasm both in cultured cells and in rat liver, whereas the sequence Thr284 through Glu302 (peptide H) faces the lumen of the ER. This indicates that a sequence between peptide G and peptide H spans the membrane of the ER. Moreover, by epitope tagging with peptide H, we show that the loop segment connecting membrane spans 3 and 4 is sequestered in the lumen of the ER. These results demonstrate that the membrane domain of HMG-CoA reductase spans the ER eight times and are inconsistent with the seven membrane spans topological model. The approximate boundaries of the proposed additional transmembrane segment are between Lys248 and Asp276. Replacement of this 7th span in HMGal with the first transmembrane helix of bacteriorhodopsin abolishes the sterol-enhanced degradation of the protein, indicating its role in the regulated turnover of HMG-CoA reductase within the endoplasmic reticulum. - + 562 Species Escherichia coli - + 10116 Species rat - - MESH:C008047 - Chemical - - HMG-CoA - - - MESH:C026591 - Chemical - - Lys - - - MESH:C008047 + + - Chemical - - HMG-CoA + + Leu242 - + MESH:D013261 Chemical sterols - - MESH:C008047 + + - Chemical - - 3-hydroxy-3- methylglutaryl coenzyme A + + Thr284 - + MESH:D013261 Chemical sterol - + MESH:D013261 Chemical sterols - - MESH:C008047 - Chemical - - HMG-CoA - - - MESH:C008047 + + - Chemical - - HMG-CoA + + Arg224 - - MESH:C008047 + + - Chemical - - HMG-CoA + + Glu302 - - MESH:C505981 + + - Chemical Asp276 - + + - + Chemical + + Lys248 + + MESH:D002784 Chemical cholesterol - - MESH:C055175 - Chemical - - Thr - @@ -77615,42 +78233,18 @@ Pietrini G, Aggujaro D, Carrera P, Malyszko J, Vitale A, Borgese N 0 A single mRNA, transcribed from an alternative, erythroid-specific, promoter, codes for two non-myristylated forms of NADH-cytochrome b5 reductase - - MESH:D009243 - Chemical - - NADH - ABSTRACT abstract 147 Two forms of NADH-cytochrome b5 reductase are produced from one gene: a myristylated membrane-bound enzyme, expressed in all tissues, and a soluble, erythrocyte-specific, isoform. The two forms are identical in a large cytoplasmic domain (Mr approximately 30,000) and differ at the NH2-terminus, which, in the membrane form, is responsible for binding to the bilayer, and which contains the myristylation consensus sequence and an additional 14 uncharged amino acids. To investigate how the two differently targeted forms of the reductase are produced, we cloned a reductase transcript from reticulocytes, and studied its relationship to the previously cloned liver cDNA. The reticulocyte transcript differs from the liver transcript in the 5' non-coding portion and at the beginning of the coding portion, where the seven codons specifying the myristoylation consensus are replaced by a reticulocyte-specific sequence which codes for 13 non-charged amino acids. Analysis of genomic reductase clones indicated that the ubiquitous transcript is generated from an upstream "housekeeping" type promoter, while the reticulocyte transcript originates from a downstream, erythroid- specific, promoter. In vitro translation of the reticulocyte-specific mRNA generated two products: a minor one originating from the first AUG, and a major one starting from a downstream AUG, as indicated by mutational analysis. Both the AUGs used as initiation codons were in an unfavorable sequence context. The major, lower relative molecular mass product behaved as a soluble protein, while the NH2-terminally extended minor product interacted with microsomes in vitro. The generation of soluble reductase from a downstream AUG was confirmed in vivo, in Xenopus oocytes. Thus, differently localized products, with respect both to tissues and to subcellular compartments, are generated from the same gene by a combination of transcriptional and translational mechanisms. - + 8355 Species Xenopus - - MESH:D000641 - Chemical - - NH2 - - - MESH:D000641 - Chemical - - NH2 - - - MESH:D009243 - Chemical - - NADH - @@ -77684,79 +78278,85 @@ abstract 190 Ribophorins I and II are type I transmembrane glycoproteins of the ER that are segregated to the rough domains of this organelle. Both ribophorins appear to be part of the translocation apparatus for nascent polypeptides that is associated with membrane-bound ribosomes and participate in the formation of a proteinaceous network within the ER membrane that also includes other components of the translocation apparatus. The ribophorins are both highly stable proteins that lack O- linked sugars but each contains one high mannose N-linked oligosaccharide that remains endo H sensitive throughout their lifetimes. We have previously shown (Tsao, Y. S., N. E. Ivessa, M. Adesnik, D. D. Sabatini, and G. Kreibich. 1992. J. Cell Biol. 116:57- 67) that a COOH-terminally truncated variant of ribophorin I that contains only the first 332 amino acids of the luminal domain (RI332), when synthesized in permanent transformants of HeLa cells, undergoes a rapid degradation with biphasic kinetics in the ER itself and in a second, as yet unidentified nonlysosomal pre-Golgi compartment. We now show that in cells treated with brefeldin A (BFA) RI332 molecules undergo rapid O-glycosylation in a multistep process that involves the sequential addition of N-acetylgalactosamine, galactose, and terminal sialic acid residues. Addition of O-linked sugars affected all newly synthesized RI332 molecules and was completed soon after synthesis with a half time of about 10 min. In the same cells, intact ribophorins I and II also underwent O-linked glycosylation in the presence of BFA, but these molecules were modified only during a short time period immediately after their synthesis was completed, and the modification affected only a fraction of the newly synthesized polypeptides. More important, these molecules synthesized before the addition of BFA were not modified by O-glycosylation. The same is true for ribophorin I when overexpressed in HeLa cells although it is significantly less stable than the native polypeptide in control cells. We, therefore, conclude that soon after their synthesis, ribophorins lose their susceptibility to the relocated Golgi enzymes that effect the O-glycosylation, most likely as a consequence of a conformational change in the ribophorins that occurs during their maturation, although it cannot be excluded that rapid integration of these molecules into a supramolecular complex in the ER membrane leads to their inaccessibility to these enzymes. - + MESH:D000116 Chemical N-acetylgalactosamine - - MESH:D020126 - Chemical - - brefeldin A - - MESH:D002241 + - Chemical - - sugars + + RI332 - MESH:C102887 + MESH:D020126 Chemical - - COOH + + BFA - CVCL:0030 - CellLine - - HeLa + MESH:D020126 + Chemical + + brefeldin A - MESH:C524118 + MESH:D019158 Chemical - - BFA + + sialic acid - MESH:D002241 - Chemical - - sugars + CVCL:0030 + CellLine + + HeLa - MESH:D008358 + - Chemical - - mannose + + O-linked sugars + MESH:D020126 + Chemical + + BFA + + CVCL:0030 CellLine HeLa - - MESH:D012794 - Chemical - - sialic acid - - MESH:C524118 + MESH:D020126 Chemical - + BFA - MESH:C524118 + - Chemical - - BFA + + mannose N-linked oligosaccharide + - + Chemical + + O- linked sugars + + + - + Chemical + + RI332 + + MESH:D005690 Chemical @@ -77948,106 +78548,112 @@ abstract 106 Duchenne's muscular dystrophy (DMD) is caused by the absence or drastic decrease of the structural protein, dystrophin, and is characterized by sarcolemmal lesions in skeletal muscle due to the stress of contraction. Dystrophin has been localized to the sarcolemma, but its organization there is not known. We report immunofluorescence studies which show that dystrophin is concentrated, along with the major muscle isoform of beta-spectrin, in three distinct domains at the sarcolemma: in elements overlying both I bands and M lines, and in occasional strands running along the longitudinal axis of the myofiber. Vinculin, which has previously been found at the sarcolemma overlying the I bands and in longitudinal strands, was present in the same three structures as spectrin and dystrophin. Controls demonstrated that the labeling was intracellular. Comparison to labeling of the lipid bilayer and of the extracellular matrix showed that the labeling for spectrin and dystrophin is associated with the intact sarcolemma and is not a result of processing artifacts. Dystrophin is not required for this lattice- like organization, as similar domains containing spectrin but not dystrophin are present in muscle from the mdx mouse and from humans with Duchenne's muscular dystrophy. We discuss the possibility that dystrophin and spectrin, along with vinculin, may function to link the contractile apparatus to the sarcolemma of normal skeletal muscle. - + 1756 Gene 20856 dystrophin - + 1756 Gene 20856 dystrophin - + 7414 Gene 7594 Vinculin - + 1756 Gene 20856 dystrophin - + 1756 Gene 20856 dystrophin - + 13405 Gene 20856 Dystrophin - + 1756 Gene 20856 dystrophin - + 7414 Gene 7594 vinculin - + 1756 Gene 20856 Dystrophin - + 10090 Species mouse - + 9606 Species humans - + 1756 Gene 20856 dystrophin - + MESH:D020388 Disease Duchenne's muscular dystrophy - + MESH:D020388 Disease DMD - + MESH:D020388 Disease Duchenne's muscular dystrophy + + MESH:D008055 + Chemical + + lipid + @@ -78082,132 +78688,141 @@ abstract 91 To identify regulatory mechanisms potentially involved in formation of actomyosin structures in smooth muscle cells, the influence of F-actin on smooth muscle myosin assembly was examined. In physiologically relevant buffers, AMPPNP binding to myosin caused transition to the soluble 10S myosin conformation due to trapping of nucleotide at the active sites. The resulting 10S myosin-AMPPNP complex was highly stable and thick filament assembly was suppressed. However, upon addition to F- actin, myosin readily assembled to form thick filaments. Furthermore, myosin assembly caused rearrangement of actin filament networks into actomyosin fibers composed of coaligned F-actin and myosin thick filaments. Severin-induced fragmentation of actin in actomyosin fibers resulted in immediate disassembly of myosin thick filaments, demonstrating that actin filaments were indispensable for mediating myosin assembly in the presence of AMPPNP. Actomyosin fibers also formed after addition of F-actin to nonphosphorylated 10S myosin monomers containing the products of ATP hydrolysis trapped at the active site. The resulting fibers were rapidly disassembled after addition of millimolar MgATP and consequent transition of myosin to the soluble 10S state. However, reassembly of myosin filaments in the presence of MgATP and F-actin could be induced by phosphorylation of myosin P-light chains, causing regeneration of actomyosin fiber bundles. The results indicate that actomyosin fibers can be spontaneously formed by F-actin-mediated assembly of smooth muscle myosin. Moreover, induction of actomyosin fibers by myosin light chain phosphorylation in the presence of actin filament networks provides a plausible hypothesis for contractile fiber assembly in situ. - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + + MESH:D000266 Chemical - - Severin + + AMPPNP - - MESH:D009135 - Disease - - myosin thick filaments - - - MESH:D009135 - Disease - - myosin thick filaments - - + MESH:D000255 Chemical MgATP - + MESH:D000255 Chemical MgATP - + MESH:D000255 Chemical ATP + + MESH:D000266 + Chemical + + AMPPNP + @@ -78229,327 +78844,319 @@ Robinson LJ, Pang S, Harris DS, Heuser J, James DE 0 Translocation of the glucose transporter (GLUT4) to the cell surface in permeabilized 3T3-L1 adipocytes: effects of ATP insulin, and GTP gamma S and localization of GLUT4 to clathrin lattices - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 3630 Gene 173 insulin - - MESH:D005947 - Chemical - - glucose - - - MESH:D016244 - Chemical - - GTP gamma S - - + MESH:D000255 Chemical ATP + + MESH:D006160 + Chemical + + GTP + ABSTRACT abstract 192 Insulin stimulates the movement of two glucose transporter isoforms (GLUT1 and GLUT4) to the plasma membrane (PM) in adipocytes. To study this process we have prepared highly purified PM fragments by gently sonicating 3T3-L1 adipocytes grown on glass coverslips. Using confocal laser immunofluorescence microscopy we observed increased PM labeling for GLUT1 (2.3-fold) and GLUT4 (eightfold) after insulin treatment in intact cells. EM immunolabeling of PM fragments indicated that in the nonstimulated state GLUT4 was mainly localized to flat clathrin lattices. Whereas GLUT4 labeling of clathrin lattices was only slightly increased after insulin treatment, labeling of uncoated PM regions was markedly increased with insulin. These data suggest that GLUT4 recycles from the cell surface both in the presence and absence of insulin. In streptolysin-O permeabilized adipocytes, insulin, and GTP gamma S increased PM levels of GLUT4 to a similar extent as observed with insulin in intact cells. In the absence of an exogenous ATP source the magnitude of these effects was considerably reduced. Removal of ATP per se caused a significant increase in cell surface levels of GLUT4 suggesting that ATP may be required for intracellular sequestration of these transporters. When insulin and GTP gamma S were added together, in the presence of ATP, PM GLUT4 levels were similar to levels observed when either insulin or GTP gamma S was added individually. Addition of GTP gamma S was able to overcome this ATP dependence of insulin- stimulated GLUT4 movement. GTP gamma S had no effect on constitutive secretion of adipsin in permeabilized cells. In addition, there was no effect of insulin or GTP gamma S on GLUT4 movement to the PM in noninsulin sensitive streptolysin-O-permeabilized 3T3-L1 fibroblasts overexpressing GLUT4. We conclude that the insulin-stimulated movement of GLUT4 to the cell surface in adipocytes may require ATP early in the insulin signaling pathway and a GTP-binding protein(s) at a later step(s). We propose that the association of GLUT4 with clathrin lattices may be important in maintaining the exclusive intracellular location of this transporter in the absence of insulin. - + 6513 Gene 68520 GLUT1 - + 6517 Gene 74381 GLUT4 - + 6513 Gene 68520 GLUT1 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - - 3630 - Gene - 173 - - insulin - - + 3630 Gene 173 insulin - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 1675 Gene 20453 adipsin - + 3630 Gene 173 insulin or GTP gamma S - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - + 6517 Gene 74381 GLUT4 - - 3630 - Gene - 173 - - insulin - - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin - + 3630 Gene 173 Insulin - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin or GTP gamma S - + 3630 Gene 173 insulin - + 3630 Gene 173 insulin - - MESH:D016244 + + MESH:D006160 Chemical - - GTP gamma S + + GTP - + MESH:D016244 Chemical GTP gamma S - + MESH:D000255 Chemical ATP - + MESH:D000255 Chemical ATP - - MESH:D016244 - Chemical - - GTP gamma S - - + MESH:D000255 Chemical ATP - - MESH:D016244 - Chemical - - GTP gamma S + + MESH:D004832 + Disease + + absence of insulin - - MESH:D005947 + + MESH:D013455 Chemical - - glucose + + S - + MESH:D000255 Chemical ATP - + + MESH:D006160 + Chemical + + GTP + + MESH:D000255 Chemical ATP - + MESH:D000255 Chemical ATP + + MESH:D004832 + Disease + + absence of insulin + + + MESH:D006160 + Chemical + + GTP + @@ -78690,24 +79297,18 @@ abstract 114 The massive secretion of salt and water in cholera-induced diarrhea involves binding of cholera toxin (CT) to ganglioside GM1 in the apical membrane of intestinal epithelial cells, translocation of the enzymatically active A1-peptide across the membrane, and subsequent activation of adenylate cyclase located on the cytoplasmic surface of the basolateral membrane. Studies on nonpolarized cells show that CT is internalized by receptor-mediated endocytosis, and that the A1-subunit may remain membrane associated. To test the hypothesis that toxin action in polarized cells may involve intracellular movement of toxin- containing membranes, monolayers of the polarized intestinal epithelial cell line T84 were mounted in modified Ussing chambers and the response to CT was examined. Apical CT at 37 degrees C elicited a short circuit current (Isc: 48 +/- 2.1 microA/cm2; half-maximal effective dose, ED50 integral of 0.5 nM) after a lag of 33 +/- 2 min which bidirectional 22Na+ and 36Cl- flux studies showed to be due to electrogenic Cl- secretion. The time course of the CT-induced Isc response paralleled the time course of cAMP generation. The dose response to basolateral toxin at 37 degrees C was identical to that of apical CT but lag times (24 +/- 2 min) and initial rates were significantly less. At 20 degrees C, the Isc response to apical CT was more strongly inhibited (30-50%) than the response to basolateral CT, even though translocation occurred in both cases as evidenced by the formation of A1-peptide. A functional rhodamine-labeled CT-analogue applied apically or basolaterally at 20 degrees C was visualized only within endocytic vesicles close to apical or basolateral membranes, whereas movement into deeper apical structures was detected at 37 degrees C. At 15 degrees C, in contrast, reduction to the A1-peptide was completely inhibited and both apical and basolateral CT failed to stimulate Isc although Isc responses to 1 nM vasoactive intestinal peptide, 10 microM forskolin, and 3 mM 8Br- cAMP were intact. Re-warming above 32 degrees C restored CT-induced Isc. Preincubating monolayers for 30 min at 37 degrees C before cooling to 15 degrees C overcame the temperature block of basolateral CT but the response to apical toxin remained completely inhibited. These results identify a temperature-sensitive step essential to apical toxin action on polarized epithelial cells. We suggest that this event involves vesicular transport of toxin-containing membranes beyond the apical endosomal compartment. - - MESH:D000242 + + MESH:D012492 Chemical - - cAMP + + salt - + MESH:D005576 Chemical forskolin - - MESH:C027024 - Chemical - - rhodamine - MESH:D005732 Chemical @@ -78721,22 +79322,34 @@ diarrhea - MESH:D012211 + MESH:D014867 Chemical - - Re + + water - MESH:D000242 + MESH:D005677 Chemical - - cAMP + + GM1 - MESH:C068746 + MESH:D012235 Chemical - - 8Br + + rhodamine + + + MESH:D015124 + Chemical + + 8Br- cAMP + + + - + Chemical + + cAMP @@ -78914,84 +79527,61 @@ abstract 120 Wild-type and mutant chicken integrin beta 1 subunit (beta 1c) cDNAs were expressed in NIH 3T3 cells and assayed for localization in focal adhesions of cells plated on fibronectin substrates. Focal adhesion localization in stable transfected cells was assayed by indirect immunofluorescent staining with chicken-specific anti-beta 1c antibodies. Mutant beta 1c integrins containing internal deletions of 13 amino acids adjacent to the membrane, delta 759-771, and 20 centrally located amino acids, delta 771-790, localized in focal adhesions demonstrating that sequences required for direction to focal adhesion structures were not limited to one region of the cytoplasmic domain. Point mutations revealed three clusters of amino acids which contribute to localization in focal adhesions. These three clusters or signals are: cyto-1 (764-774), cyto-2 (785-788), and cyto-3 (797-800). The 11-residue cyto-1 signal is only found on integrin beta subunit sequences, except beta 4. Four residues within this region, D764, F768, F771, and E774, could not be altered without reducing focal adhesion staining intensities, and likely form a signal that occupies one side of an alpha helix. Mutations involving two cyto-1 residues, K770 and F771, also appeared to affect heterodimer affinity and specificity. Cyto-2 (785-788,), NPIY, is an NPXY signal that forms a tight turn motif. Cyto-2 provides a structural conformation, which when perturbed by proline removal or addition, inhibits integrin localization in focal adhesions. Cyto-3 (797-800), NPKY, resembles cyto-2, however, the nonconserved proline residue can be replaced without alteration of the localization phenotype. Cyto-3, therefore, constitutes a unique integrin signal, NXXY. Both serine and tyrosine residues at positions 790 and 788, respectively, which have been implicated in integrin phosphorylation/regulation, were conservatively replaced without detectable effect on focal adhesion localization. However, acidic replacements for these amino acids reduced focal adhesion staining intensities, suggesting that phosphorylation at these sites may negatively regulate integrin function. - + 14268 Gene 1533 fibronectin - + 9031 Species chicken - + 9031 Species chicken - + 16412 Gene 22999 integrin beta 1 - - MESH:C021597 - Chemical - - F771 - - - MESH:C047902 + + MESH:D011392 Chemical - - serine + + proline - - MESH:C021597 + + MESH:D011392 Chemical - - delta 759-771 + + proline - + CVCL:0594 CellLine NIH 3T3 - - MESH:C489032 - Chemical - - proline - - - MESH:C055434 - Chemical - - F768 - - + + MESH:D014443 Chemical tyrosine - - MESH:C021597 - Chemical - - F771 - - - MESH:C489032 + + MESH:D012694 Chemical - - proline + + serine @@ -79021,7 +79611,6 @@ KAR1 - OMIM:604588 Disease mitosis @@ -79032,90 +79621,82 @@ abstract 82 The yeast KAR1 gene is essential for mitotic growth and important for nuclear fusion. Mutations in KAR1 prevent duplication of the spindle pole body (SPB), and affect functions associated with both the nuclear and cytoplasmic microtubules. The localization of hybrid Kar1-lacZ proteins, described elsewhere (Vallen, E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press), suggest that the protein is associated with the SPB. In this paper, we report a deletion analysis demonstrating that the mitotic and karyogamy functions of KAR1 are separate and independent, residing in discrete functional domains. One region, here shown to be essential for mitosis, coincided with a part of the protein that is both necessary and sufficient to target Karl-lacZ hybrid proteins to the SPB (Vallen, E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press). Complementation testing demonstrated that deletions in this interval did not affect nuclear fusion. A second region, required only for karyogamy, was necessary for the localization of a Kar3-lacZ hybrid protein to the SPB. These data suggest a model for the roles of Kar1p and Kar3p, a kinesin-like protein, in nuclear fusion. Finally, a third region of KAR1 was found to be important for both mitosis and karyogamy. This domain included the hydrophobic carboxy terminus and is sufficient to target a lacZ-Kar1 hybrid protein to the nuclear envelope (Vallen E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press). Altogether, the essential mitotic regions of KAR1 comprised 20% of the coding sequence. We propose a model for Kar1p in which the protein is composed of several protein-binding domains tethered to the nuclear envelope via its hydrophobic tail. - + 855533 Gene KAR1 - + 855533 Gene KAR1 - + 855533 Gene Kar1 - + 855533 Gene KAR1 - + 855533 Gene KAR1 - + 855533 Gene KAR1 - + 855533 Gene Kar1 - + 4932 Species yeast - + 267260 Species D. Rose - + 267260 Species D. Rose - + 267260 Species D. Rose - - OMIM:604588 + Disease mitosis - - OMIM:604588 + Disease mitosis - - MESH:C015732 - Chemical - - carboxy - @@ -79163,151 +79744,145 @@ abstract 122 Our previous studies showed that in hepatic RER of young chickens, nascent apoAI is not associated with lipoprotein particles and only becomes part of these lipoprotein structures in the Golgi. In this study, we have used three different methodologies to determine the locations of apoAI and apoB in the RER and compared them to that of albumin. Immunoelectron microscopic examination of the RER cell fractions showed that both apoAI and apoB were associated only with the RER membrane whereas albumin was located both within the lumen and on the limiting membrane of the vesicles. To examine the possibility of membrane integration of nascent apoAI and apoB in the RER, we administered L-[3H]leucine to young chickens for 10 min, isolated RER, treated this cell fraction with buffers of varying pH, and measured the release of radioactive albumin, apoAI, and apoB. The majority of nascent apoAI (64%), nascent apoB (100%), and nascent albumin (97%) was released from RER vesicles at pH 11.2, suggesting that, like albumin, apolipoproteins are not integrated within the membrane. To determine if nascent apoproteins are exposed to the cytoplasmic surface, we administered L-[3H]leucine to young chickens and at various times isolated RER and Golgi cell fractions. Radioactive RER and Golgi cell fractions were treated with exogenous protease and the percent of nascent apoAI and apoB accessible to proteolysis was determined and compared to that of albumin. At 5, 10, and 20 min of labeling, 35-56% of nascent apoAI and 60-75% of apoB in RER were degraded, while albumin was refractive to this treatment. At all times both apolipoproteins and albumin present in Golgi cell fractions were protected from proteolysis. These biochemical and morphological findings indicate that apoAI and apoB are associated with the rough microsomal membrane and are partially exposed to the cytoplasmic surface at early stages of secretion. They may later enter the luminal side of the ER and, on entering the Golgi, form lipoprotein particles. - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396536 Gene 47900 apoAI - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 396535 Gene 328 apoB - + 9031 Species chickens - + 9031 Species chickens - + 9031 Species chickens - - MESH:C027819 - Chemical - - L-[3H]leucine - - - MESH:C027819 + + - Chemical L-[3H]leucine @@ -79345,55 +79920,49 @@ abstract 166 Membrane-cytoskeletons were prepared from Dictyostelium amebas, and networks of actin and myosin II filaments were visualized on the exposed cytoplasmic surfaces of the cell membranes by fluorescence staining (Yumura, S., and T. Kitanishi-Yumura. 1990. Cell Struct. Funct. 15:355-364). Addition of ATP caused contraction of the cytoskeleton with aggregation of part of actin into several foci within the network, but most of myosin II was released via the foci. However, in the presence of 10 mM MgCl2, which stabilized myosin II filaments, myosin II remained at the foci. Ultrastructural examination revealed that, after contraction, only traces of monomeric myosin II remained at the foci. By contrast, myosin II filaments remained in the foci in the presence of 10 mM MgCl2. These observations suggest that myosin II was released not in a filamentous form but in a monomeric form. Using [gamma 32P]ATP, we found that the heavy chains of myosin II released from membrane-cytoskeletons were phosphorylated, and this phosphorylation resulted in disassembly of myosin filaments. Using ITP (a substrate for myosin II ATPase) and/or ATP gamma S (a substrate for myosin II heavy-chain kinase [MHCK]), we demonstrated that phosphorylation of myosin heavy chains occurred at the foci within the actin network, a result that suggests that MHCK was localized at the foci. These results together indicate that, during contraction, the heavy chains of myosin II that have moved toward the foci within the actin network are phosphorylated by a specific MHCK, with the resultant disassembly of filaments which are finally released from membrane- cytoskeletons. This series of reactions could represent the mechanism for the relocation of myosin II from the cortical region to the endoplasm. - + 44689 Species Dictyostelium - + MESH:D015636 Chemical MgCl2 - - MESH:D013455 - Chemical - - S - - - MESH:D013455 - Chemical - - S - - - MESH:D007293 + + - Chemical ITP - + MESH:D015636 Chemical MgCl2 - + + MESH:C000615311 + Chemical + + 32P + + MESH:C022571 Chemical - - [gamma 32P]ATP + + ATP gamma S - + MESH:D000255 Chemical - + ATP - + MESH:D000255 Chemical @@ -79432,73 +80001,85 @@ abstract 118 The precise function of the yolk platelets of sea urchin embryos during early development is unknown. We have shown previously that the chemical composition of the yolk platelets remains unchanged in terms of phospholipid, triglyceride, hexose, sialic acid, RNA, and total protein content after fertilization and early development. However, the platelet is not entirely static because the major 160-kD yolk glycoprotein YP-160 undergoes limited, step-wise proteolytic cleavage during early development. Based on previous studies by us and others, it has been postulated that yolk platelets become acidified during development, leading to the activation of a cathepsin B-like yolk proteinase that is believed to be responsible for the degradation of the major yolk glycoprotein. To investigate this possibility, we studied the effect of addition of chloroquine, which prevents acidification of lysosomes. Consistent with the postulated requirement for acidification, it was found that chloroquine blocked YP-160 breakdown but had no effect on embryonic development. To directly test the possibility that acidification of the yolk platelets over the course of development temporally correlated with YP-160 proteolysis, we added 3-(2,4-dinitroanilo)-3-amino-N-methyldipropylamine (DAMP) to eggs or embryos. This compound localizes to acidic organelles and can be detected in these organelles by EM. The results of these studies revealed that yolk platelets did, in fact, become transiently acidified during development. This acidification occurred at the same time as yolk protein proteolysis, i.e., at 6 h after fertilization (64-cell stage) in Strongylocentrotus purpuratus and at 48 h after fertilization (late gastrula) in L. pictus. Furthermore, the pH value at the point of maximal acidification of the yolk platelets in vivo was equal to the pH optimum of the enzyme measured in vitro, indicating that this acidification is sufficient to activate the enzyme. For both S. purpuratus and Lytechinus pictus, the observed decrease in the pH was approximately 0.8 U, from 7.0 to 6.2. The trypsin inhibitor benzamidine was found to inhibit the yolk proteinase in vivo. By virtue of the fact that this inhibitor was reversible we established that the activity of the yolk proteinase is developmentally regulated even though the enzyme is present throughout the course of development. These findings indicate that acidification of yolk platelets is a developmentally regulated process that is a prerequisite to initiation of the catabolism of the major yolk glycoprotein. - + 7656 Species sea urchin - + 7668 Species Strongylocentrotus purpuratus - + 7653 Species L. pictus - + 7668 Species S. purpuratus - + 7653 Species Lytechinus pictus - - MESH:D012794 + + - Chemical - - sialic acid + + DAMP - - MESH:C033506 + + - Chemical 3-(2,4-dinitroanilo)-3-amino-N-methyldipropylamine - + MESH:D014280 Chemical triglyceride - + MESH:D002738 Chemical chloroquine - + MESH:C032157 Chemical benzamidine - + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D019158 + Chemical + + sialic acid + + MESH:D006601 Chemical hexose - + MESH:D002738 Chemical @@ -79525,12 +80106,7 @@ Pancake SJ, Holt GD, Mellouk S, Hoffman SL 0 Malaria sporozoites and circumsporozoite proteins bind specifically to sulfated glycoconjugates - - Chemical - - circumsporozoite - - + MESH:D008288 Disease @@ -79542,106 +80118,95 @@ abstract 96 Circumsporozoite (CS) proteins, which densely coat malaria (Plasmodia) sporozoites, contain an amino acid sequence that is homologous to segments in other proteins which bind specifically to sulfated glycoconjugates. The presence of this homology suggests that sporozoites and CS proteins may also bind sulfated glycoconjugates. To test this hypothesis, recombinant P. yoelii CS protein was examined for binding to sulfated glycoconjugate-Sepharoses. CS protein bound avidly to heparin-, fucoidan-, and dextran sulfate-Sepharose, but bound comparatively poorly to chondroitin sulfate A- or C-Sepharose. CS protein also bound with significantly lower affinity to a heparan sulfate biosynthesis-deficient mutant cell line compared with the wild- type line, consistent with the possibility that the protein also binds to sulfated glycoconjugates on the surfaces of cells. This possibility is consistent with the observation that CS protein binding to hepatocytes, cells invaded by sporozoites during the primary stage of malaria infection, was inhibited by fucoidan, pentosan polysulfate, and heparin. The effects of sulfated glycoconjugates on sporozoite infectivity were also determined. P. berghei sporozoites bound specifically to sulfatide (galactosyl[3-sulfate]beta 1-1ceramide), but not to comparable levels of cholesterol-3-sulfate, or several examples of neutral glycosphingolipids, gangliosides, or phospholipids. Sporozoite invasion into hepatocytes was inhibited by fucoidan, heparin, and dextran sulfate, paralleling the observed binding of CS protein to the corresponding Sepharose derivatives. These sulfated glycoconjugates blocked invasion by inhibiting an event occurring within 3 h of combining sporozoites and hepatocytes. Sporozoite infectivity in mice was significantly inhibited by dextran sulfate 500,000 and fucoidan. Taken together, these data indicate that CS proteins bind selectively to certain sulfated glycoconjugates, that sporozoite infectivity can be inhibited by such compounds, and that invasion of host hepatocytes by sporozoites may involve interactions with these types of compounds. - + 5861 Species P. yoelii - + 5821 Species P. berghei - + 10090 Species mice - + MESH:C007045 Chemical cholesterol-3-sulfate - + + MESH:D016264 + Chemical + + dextran sulfate + + + MESH:D012685 + Chemical + + Sepharose + + MESH:C007789 Chemical fucoidan - + MESH:D008288 Disease malaria infection - + MESH:C007789 Chemical fucoidan - + MESH:D006493 Chemical heparin - - MESH:D013431 - Chemical - - sulfate - - + + MESH:D012685 Chemical - - Circumsporozoite + + Sepharose - + MESH:D006028 Chemical glycosphingolipids - - MESH:D013431 - Chemical - - sulfate - - - MESH:C511581 - Chemical - - galactosyl[3-sulfate]beta 1-1ceramide - - - MESH:C061194 + + MESH:D006497 Chemical - - Sporozoite + + heparan sulfate - + MESH:D016264 Chemical dextran sulfate - - MESH:D013431 - Chemical - - sulfate - - MESH:C061194 + MESH:D016264 Chemical - - sporozoite + + dextran sulfate MESH:D005732 @@ -79656,54 +80221,90 @@ heparin + MESH:D010743 + Chemical + + phospholipids + + + - + Chemical + + galactosyl[3-sulfate]beta 1-1ceramide + + MESH:D006493 Chemical heparin - + MESH:D058456 Disease coat malaria - + + - + Chemical + + CS + + + - + Chemical + + CS + + MESH:C007789 Chemical fucoidan - + MESH:D010426 Chemical pentosan polysulfate - - MESH:C007789 + + - Chemical - - fucoidan + + CS - - MESH:D007239 - Disease - - Sporozoite infectivity + + - + Chemical + + CS - - MESH:C061194 + + MESH:D012685 Chemical - - sporozoite + + Sepharoses - + + MESH:D012685 + Chemical + + Sepharose + + + MESH:C007789 + Chemical + + fucoidan + + MESH:D002809 Chemical chondroitin sulfate A - + MESH:D013433 Chemical @@ -79802,38 +80403,26 @@ Drake SL, Klein DJ, Mickelson DJ, Oegema TR, Furcht LT, McCarthy JB 0 Cell surface phosphatidylinositol-anchored heparan sulfate proteoglycan initiates mouse melanoma cell adhesion to a fibronectin-derived, heparin-binding synthetic peptide - + 10090 Species mouse - + 14268 Gene 1533 fibronectin - + MESH:D008545 Disease melanoma - - MESH:D013431 - Chemical - - sulfate - - - MESH:D006493 - Chemical - - heparin - - + MESH:D010716 Chemical @@ -79845,385 +80434,367 @@ abstract 171 Cell surface heparan sulfate proteoglycan (HSPG) from metastatic mouse melanoma cells initiates cell adhesion to the synthetic peptide FN-C/H II, a heparin-binding peptide from the 33-kD A chain-derived fragment of fibronectin. Mouse melanoma cell adhesion to FN-C/H II was sensitive to soluble heparin and pretreatment of mouse melanoma cells with heparitinase. In contrast, cell adhesion to the fibronectin synthetic peptide CS1 is mediated through an alpha 4 beta 1 integrin and was resistant to heparin or heparitinase treatment. Mouse melanoma cell HSPG was metabolically labeled with [35S]sulfate and extracted with detergent. After HPLC-DEAE purification, 35S-HSPG eluted from a dissociative CL-4B column with a Kav approximately 0.45, while 35S- heparan sulfate (HS) chains eluted with a Kav approximately 0.62. The HSPG contained a major 63-kD core protein after heparitinase digestion. Polyclonal antibodies generated against HSPG purified from mouse melanoma cells grown in vivo also identified a 63-kD core protein. This HSPG is an integral plasma membrane component by virtue of its binding to Octyl Sepharose affinity columns and that anti-HSPG antibody staining exhibited a cell surface localization. The HSPG is anchored to the cell surface through phosphatidylinositol (PI) linkages, as evidenced in part by the ability of PI-specific phospholipase C to eliminate binding of the detergent-extracted HSPG to Octyl Sepharose. Furthermore, the mouse melanoma HSPG core protein could be metabolically labeled with 3H-ethanolamine. The involvement of mouse melanoma cell surface HSPG in cell adhesion to fibronectin was also demonstrated by the ability of anti-HSPG antibodies and anti-HSPG IgG Fab monomers to inhibit mouse melanoma cell adhesion to FN-C/H II. 35S- HSPG and 35S-HS bind to FN-C/H II affinity columns and require 0.25 M NaCl for elution. However, heparitinase-treated 125I-labeled HSPG failed to bind FN-C/H II, suggesting that HS, and not HSPG core protein, binds FN-C/H II. These data support the hypothesis that a phosphatidylinositol-anchored HSPG on mouse melanoma cells (MPIHP-63) initiates recognition to FN-C/H II, and implicate PI-associated signal transduction pathways in mediating melanoma cell adhesion to this defined ligand. - + 15529 Gene 2253 HSPG - + 14268 Gene 1533 fibronectin - + 14268 Gene 1533 fibronectin - + 70599 Gene 4912 CS1 - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 14268 Gene 1533 fibronectin - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 15529 Gene 2253 HSPG - + 10090 Species mouse - + 10090 Species Mouse - + 10090 Species mouse - + 10090 Species Mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + MESH:D008545 Disease melanoma - - MESH:D008545 - Disease - - melanoma HSPG core - - + MESH:D008545 Disease melanoma - + MESH:D008545 Disease melanoma - - MESH:C059676 + + MESH:C027176 Chemical - - Octyl + + Octyl Sepharose - - MESH:D006859 + + MESH:C000615320 Chemical - - H II + + 35S - - MESH:D004983 + + MESH:C027176 Chemical - - 3H-ethanolamine + + Octyl Sepharose - - MESH:D006859 + + MESH:D013431 Chemical - - H II + + sulfate - + MESH:D008545 Disease melanoma - - MESH:D013431 + + MESH:C000615320 Chemical - - sulfate + + 35S - + MESH:D012965 Chemical NaCl - - MESH:D006859 - Chemical - - H II - - + MESH:D010716 Chemical phosphatidylinositol - - MESH:D006493 + + MESH:C000615320 Chemical - - heparin + + 35S - - MESH:D006859 + + - Chemical - - H II + + CL-4B - - MESH:D006859 + + MESH:D006497 Chemical - - H II + + heparan sulfate - + MESH:D008545 Disease melanoma - - MESH:D006859 - Chemical - - H II - - - MESH:C031512 + + - Chemical - - [35S]sulfate + + 3H-ethanolamine - + MESH:D006493 Chemical heparin - - MESH:D013431 + + MESH:C000615320 Chemical - - sulfate + + 35S - + + MESH:D006497 + Chemical + + HS + + + MESH:C007369 + Chemical + + DEAE + + MESH:D008545 Disease melanoma - - MESH:D006859 - Chemical - - H II + + MESH:D008545 + Disease + + melanoma HSPG - + MESH:D010716 Chemical phosphatidylinositol - + MESH:D008545 Disease melanoma - + MESH:D008545 Disease melanoma - - MESH:C059676 - Chemical - - Octyl - - - MESH:C495909 + + MESH:C000615320 Chemical - - MPIHP-63 + + 35S - + MESH:D008545 Disease melanoma - - MESH:D006493 - Chemical - - heparin - - - MESH:C492712 + + MESH:D006497 Chemical - - 125I + + HS @@ -80252,53 +80823,47 @@ abstract 131 The 100-110-kD proteins (alpha-, beta-, beta'-, and gamma-adaptins) of clathrin-coated vesicles and the 110-kD protein (beta-COP) of the nonclathrin-coated vesicles that mediate constitutive transport through the Golgi have homologous protein sequences. To determine whether homologous processes are involved in assembly of the two types of coated vesicles, the membrane binding properties of their coat proteins were compared. After treatment of MDBK cells with the fungal metabolite Brefeldin A (BFA), beta-COP was redistributed to the cytoplasm within 15 s, gamma-adaptin and clathrin in the trans-Golgi network (TGN) dispersed within 30 s, but the alpha-adaptin and clathrin present on coated pits and vesicles derived from the plasma membrane remained membrane associated even after a 15-min exposure to BFA. In PtK1 cells and MDCK cells, BFA did not affect beta-COP binding or Golgi morphology but still induced redistribution of gamma-adaptin and clathrin from TGN membranes to the cytoplasm. Thus BFA affects the binding of coat proteins to membranes in the Golgi region (Golgi apparatus and TGN) but not plasma membranes. However, the Golgi binding interactions of beta- COP and gamma-adaptin are distinct and differentially sensitive to BFA. BFA treatment did not release gamma-adaptin or clathrin from purified clathrin-coated vesicles, suggesting that their distribution to the cytoplasm after BFA treatment of cells was due to interference with their rebinding to TGN membranes after a normal cycle of disassembly. This was confirmed using an in vitro assay in which gamma-adaptin binding to TGN membranes was blocked by BFA and enhanced by GTP gamma S, similar to the binding of beta-COP to Golgi membranes. These results suggest the involvement of GTP-dependent proteins in the association of the 100-kD coat proteins with membranes in the Golgi region of the cell. - + 281706 Gene 3499 beta-COP - + 281706 Gene 3499 beta-COP - + 281706 Gene 3499 beta-COP - + 281706 Gene 3499 beta- COP - + 281706 Gene 3499 beta-COP - + CVCL:0422 CellLine MDCK - - MESH:D016244 - Chemical - - GTP gamma S - MESH:D020126 Chemical @@ -80306,71 +80871,83 @@ Brefeldin A - MESH:C524118 + MESH:D006160 Chemical - - BFA + + GTP - MESH:C524118 + MESH:D020126 Chemical - + BFA - MESH:C020361 + MESH:D020126 Chemical - - gamma-adaptins + + BFA - MESH:C524118 + MESH:D020126 Chemical - + BFA - MESH:C524118 + MESH:D006160 Chemical - - BFA + + GTP - MESH:C524118 + MESH:D020126 Chemical - + BFA - MESH:C524118 + MESH:D013455 Chemical - - BFA + + S - MESH:D058456 + MESH:C536528 Disease - - coated pits + + pits - CVCL:0421 - CellLine - - MDBK + MESH:D020126 + Chemical + + BFA - MESH:C524118 + MESH:D020126 Chemical - + BFA - MESH:C524118 + MESH:D020126 Chemical BFA + + CVCL:0421 + CellLine + + MDBK + + + MESH:D020126 + Chemical + + BFA + @@ -80576,219 +81153,213 @@ Hennemann H, Suchyna T, Lichtenberg-Fraté H, Jungbluth S, Dahl E, Schwarz J, Nicholson BJ, Willecke K 0 Molecular cloning and functional expression of mouse connexin40, a second gap junction gene preferentially expressed in lung - + 14613 Gene 3856 connexin40 - + 10090 Species mouse - - MESH:D008171 - Disease - - lung - ABSTRACT abstract 125 From a mouse genomic library, a clone has been isolated that codes for a connexin-homologous sequence of 358 amino acids. Because of its theoretical molecular mass of 40.418 kD it is named connexin40 (Cx40). Based on both protein and nucleotide sequence, mouse Cx40 is more closely related to mouse Cx43 (alpha subgroup of connexins) than to mouse Cx32 (beta subgroup). The highest overall homology detected, however, was to chick Cx42 (67% amino acid and 86% nucleotide identity), raising the possibility that Cx40 may be the mouse analogue. The coding region of Cx40 is uninterrupted by introns and is detected as a single copy gene in the mouse genome. High stringency hybridization of Northern blots with the coding sequence of Cx40 identified a single transcript of 3.5 kb that is at least 16-fold more abundant in lung-similar to mouse Cx37-than in other adult tissues (kidney, heart, and skin). In embryonic kidney, skin, and liver the level of the Cx40 transcript is two- to fourfold higher than in the corresponding adult tissues. Microinjection of Cx40 cRNA into Xenopus oocytes induced functional cell-to-cell channels between pairs. These channels show a symmetrical and markedly cooperative closure in response to transjunctional voltage (Boltzmann parameters of Vo = +/- 35 mV; A = 0.32) which is also fast relative to other connexin channels recorded similarly (tau = 580 ms at Vj of +/- 50 mV). Although Cx40- expressing oocytes did not couple efficiently with oocytes expressing endogenous connexins, they did couple well to Cx37-expressing oocytes. The heterotypic channels which formed had voltage-gating properties modified from those of the original homotypic forms. Transfection of mouse Cx40 DNA, under control of the SV-40 early promoter, into coupling-deficient human HeLa or SK-Hep-1 cells resulted in expression of the expected transcript and restoration of fluorescent dye transfer in transfected clones. - + 14613 Gene 3856 connexin40 - + 14613 Gene 3856 Cx40 - + 14613 Gene 3856 Cx40 - + 14611 Gene 9670 Cx43 - + 14618 Gene 137 Cx32 - + 396502 Gene 3856 Cx42 - + 14613 Gene 3856 Cx40 - + 14613 Gene 3856 Cx40 - + 14613 Gene 3856 Cx40 - + 14612 Gene 1556 Cx37 - + 14613 Gene 3856 Cx40 - + 108707841 Gene Cx40 - + 14613 Gene 3856 Cx40 - + 14612 Gene 1556 Cx37 - + 14613 Gene 3856 Cx40 - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 9031 Species chick - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mouse - + + 9606 + Species + + human + + 8355 Species Xenopus - + MESH:D007674 Disease embryonic kidney - - MESH:D015658 - Disease - - coupling-deficient human HeLa - - - MESH:D008171 - Disease - - lung - - + CVCL:0525 CellLine SK-Hep-1 + + CVCL:0030 + CellLine + + HeLa + @@ -80810,145 +81381,121 @@ Navone F, Niclas J, Hom-Booher N, Sparks L, Bernstein HD, McCaffrey G, Vale RD 0 Cloning and expression of a human kinesin heavy chain gene: interaction of the COOH-terminal domain with cytoplasmic microtubules in transfected CV-1 cells - + 3799 Gene 55829 kinesin heavy chain - + 9606 Species human - + CVCL:0229 CellLine CV-1 - - MESH:C102887 - Chemical - - COOH - ABSTRACT abstract 156 To understand the interactions between the microtubule-based motor protein kinesin and intracellular components, we have expressed the kinesin heavy chain and its different domains in CV-1 monkey kidney epithelial cells and examined their distributions by immunofluorescence microscopy. For this study, we cloned and sequenced cDNAs encoding a kinesin heavy chain from a human placental library. The human kinesin heavy chain exhibits a high level of sequence identity to the previously cloned invertebrate kinesin heavy chains; homologies between the COOH-terminal domain of human and invertebrate kinesins and the nonmotor domain of the Aspergillus kinesin-like protein bimC were also found. The gene encoding the human kinesin heavy chain also contains a small upstream open reading frame in a G-C rich 5' untranslated region, features that are associated with translational regulation in certain mRNAs. After transient expression in CV-1 cells, the kinesin heavy chain showed both a diffuse distribution and a filamentous staining pattern that coaligned with microtubules but not vimentin intermediate filaments. Altering the number and distribution of microtubules with taxol or nocodazole produced corresponding changes in the localization of the expressed kinesin heavy chain. The expressed NH2-terminal motor and the COOH-terminal tail domains, but not the alpha-helical coiled coil rod domain, also colocalized with microtubules. The finding that both the kinesin motor and tail domains can interact with cytoplasmic microtubules raises the possibility that kinesin could crossbridge and induce sliding between microtubules under certain circumstances. - + 3799 Gene 55829 kinesin heavy chain - + 3799 Gene 55829 kinesin heavy chain - + 9606 Species human - + 9606 Species human - + 9606 Species human - + 9606 Species human - + 3799 Gene 55829 kinesin heavy chain - + 3799 Gene 55829 kinesin heavy chain - + 3799 Gene 55829 kinesin heavy chain - + 3799 Gene 55829 kinesin heavy chain - + CVCL:0229 CellLine CV-1 - + MESH:D015739 Chemical nocodazole - + CVCL:0229 CellLine CV-1 - - MESH:C102887 - Chemical - - COOH - - + MESH:D017239 Chemical taxol - - MESH:C102887 - Chemical - - COOH - - - MESH:D000641 - Chemical - - NH2 - @@ -80982,41 +81529,47 @@ abstract 146 A cDNA encoding a unique hyaluronan receptor has been molecularly cloned from a lambda GT11 3T3 cDNA expression library. Immunoblot analyses of cell lysates, using antibodies to peptides encoded in the cDNA, specifically react with a 58-kD protein. This protein is regulated by the mutant H-ras gene in cells containing a metallothionein promoter H-ras hybrid gene. Further, antibodies to peptide sequences encoded in the cDNA block the increase in locomotion resulting from induction of the mutant H-ras gene in this cell line. In a transblot assay, the bacterially expressed protein binds to biotinylated hyaluronan. Antibodies to peptides encoded in the cDNA react in immunoblot assays with the 58- and 52-kD proteins of a novel hyaluronan receptor complex previously implicated in cell locomotion. Furthermore, antibodies specific to the 58- and 52-kD proteins, which block ras-induced locomotion, also cross-react with the expressed, encoded protein. The gene product described here appears to be a new type of hyaluronan receptor that is involved in cell locomotion. It is named RHAMM, an acronym for receptor for hyaluronan-mediated motility. - + 3265 Gene 55890 H-ras - + 3265 Gene 55890 H-ras - + 3161 Gene 8271 RHAMM - + 3161 Gene 8271 receptor for hyaluronan-mediated motility - + 3265 Gene 55890 H-ras + + MESH:D006820 + Chemical + + hyaluronan + @@ -81038,8 +81591,14 @@ Doberstein SK, Pollard TD 0 Localization and specificity of the phospholipid and actin binding sites on the tail of Acanthamoeba myosin IC - - MESH:D015823 + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D000562 Disease Acanthamoeba myosin IC @@ -81050,90 +81609,120 @@ abstract 111 We used bacterially expressed beta-galactosidase fusion proteins to localize the phospholipid binding domain of Acanthamoeba myosin IC to the region between amino acids 701 and 888 in the NH2-terminal half of the tail. Using a novel immobilized ligand lipid binding assay, we determined that myosin I can bind to several different acidic phospholipids, and that binding requires a minimum of 5 mol% acidic phospholipid in a neutral lipid background. The presence of di- and triglycerides and sterols in the lipid bilayer do not contribute to the affinity of myosin I for membranes. We confirm that the ATP-insensitive actin binding site is contained in the COOH-terminal 30 kD of the tail as previously shown for Acanthamoeba myosin IA. We conclude that the association of the myosin IC tail with acidic phospholipid head groups supplies much of the energy for binding myosin I to biological membranes, but probably not specificity for targeting myosin I isoforms to different cellular locations. - + 2720 Gene 47922 beta-galactosidase - + 4641 Gene 32046 myosin IC - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - - MESH:D014280 - Chemical - - triglycerides - - - MESH:C102887 + + MESH:D008055 Chemical - - COOH + + lipid - + MESH:D000255 Chemical ATP - + MESH:D013261 Chemical sterols - - MESH:D015823 + + MESH:D008055 + Chemical + + lipid + + + MESH:D000562 Disease Acanthamoeba myosin IA - - MESH:D000641 + + MESH:D010743 Chemical - - NH2 + + phospholipid - - MESH:D015823 + + MESH:D008055 + Chemical + + lipid + + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D010743 + Chemical + + phospholipids + + + MESH:D000562 Disease Acanthamoeba myosin IC + + MESH:D010743 + Chemical + + phospholipid + + + - + Chemical + + di- and triglycerides + @@ -81155,7 +81744,13 @@ Dentler WL, Adams C 0 Flagellar microtubule dynamics in Chlamydomonas: cytochalasin D induces periods of microtubule shortening and elongation; and colchicine induces disassembly of the distal, but not proximal, half of the flagellum - + + MESH:D015638 + Chemical + + cytochalasin D + + MESH:D003078 Chemical @@ -81167,42 +81762,152 @@ abstract 212 To study the mechanisms responsible for the regulation of flagellar length, we examined the effects of colchicine and Cytochalasin D (CD) on the growth and maintenance of Chlamydomonas flagella on motile wild type cells as well as on pf 18 cells, whose flagella lack the central microtubules and are immobile. CD had no effect on the regeneration of flagella after deflagellation but it induced fully assembled flagella to shorten at an average rate of 0.03 microns-min. Cells remained fully motile in CD and even stubby flagella continued to move, indicating that flagellar shortening did not selectively disrupt machinery necessary for motility. To observe the effects of the drug on individual cells, pf 18 cells were treated with CD and flagella on cells were monitored by direct observation over a 5-hour period. Flagella on control pf 18 cells maintained their initial lengths throughout the experiment but flagella on CD-treated cells exhibited periods of elongation, shortening, and regrowth suggestive of the dynamic behavior of cytoplasmic microtubules observed in vitro and in vitro. Cells behaved individually, with no two cells exhibiting the same flagellar behavior at any given time although both flagella on any single cell behaved identically. The rate of drug-induced flagellar shortening and elongation in pf 18 cells varied from 0.08 to 0.17 microns-min-1, with each event occurring over 10-60-min periods. Addition of colchicine to wild type and pf 18 cells induced flagella to shorten at an average rate of 0.06 microns-min-1 until the flagella reached an average of 73% of their initial length, after which they exhibited no further shortening or elongation. Cells treated with colchicine and CD exhibited nearly complete flagellar resorption, with little variation in flagellar length among cells. The effects of these drugs were reversible and flagella grew to normal stable lengths after drug removal. Taken together, these results show that the distal half to one-third of the Chlamydomonas flagellum is relatively unstable in the presence of colchicine but that the proximal half to two-thirds of the flagellum is stable to this drug. In contrast to colchicine, CD can induce nearly complete flagellar microtubule disassembly as well as flagellar assembly. Flagellar microtubules must, therefore, be inherently unstable, and flagellar length is stabilized by factors that are sensitive, either directly or indirectly, to the effects of CD. - + + Disease + + flagella + + MESH:D003078 Chemical colchicine - - MESH:D003078 + + Disease + + stubby flagella + + + MESH:D015638 Chemical - - colchicine + + CD - + + MESH:D001259 + Disease + + flagella lack the central microtubules + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + MESH:D003078 Chemical colchicine - - MESH:D001259 + Disease - - flagella lack + + flagella - + + MESH:D015638 + Chemical + + Cytochalasin D + + + MESH:D015638 + Chemical + + CD + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + MESH:D003078 Chemical colchicine - + MESH:D003078 Chemical colchicine + + MESH:D015638 + Chemical + + CD + + + MESH:C535850 + Disease + + flagellar shortening + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + + MESH:D003078 + Chemical + + colchicine + + + MESH:C535850 + Disease + + flagellar shortening + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + @@ -81242,50 +81947,14 @@ abstract 92 Dipeptidyl peptidase IV (DPPIV) is mainly vectorially targeted to the apical surface in MDCK cells. BFA was found to abolish the apical targeting of DPPIV. This BFA effect could be achieved under conditions where the ER to Golgi transport and the total surface expression of DPPIV were essentially unaffected. BFA executed its effect during the transport from the trans-Golgi network (TGN) to the surface. The inhibition of apical targeting resulted in enhanced mis-targeting to the basolateral surface. The mistargeted DPPIV was transcytosed back to the apical domain only after BFA withdrawal. In contrast, the basolateral targeting of uvomorulin was unaffected by BFA. These results established that the apical targeting of DPPIV was selectively abolished by BFA. - + 442858 Gene 20917 uvomorulin - - MESH:C524118 - Chemical - - BFA - - - MESH:C524118 - Chemical - - BFA - - - MESH:C524118 - Chemical - - BFA - - - MESH:C524118 - Chemical - - BFA - - - MESH:C524118 - Chemical - - BFA - - - MESH:C524118 - Chemical - - BFA - - + CVCL:0422 CellLine @@ -81363,89 +82032,107 @@ Metsäranta M, Garofalo S, Decker G, Rintala M, de Crombrugghe B, Vuorio E 0 Chondrodysplasia in transgenic mice harboring a 15-amino acid deletion in the triple helical domain of pro alpha 1(II) collagen chain - + 10090 Species transgenic mice - + MESH:D010009 Disease Chondrodysplasia - - MESH:D000596 - Chemical - - 15-amino acid - ABSTRACT abstract 134 We have generated transgenic mice by microinjection of a 39-kb mouse pro alpha 1(II) collagen gene construct containing a deletion of exon 7 and intron 7. This mutation was expected to disturb the assembly and processing of the homotrimeric type II collagen molecule in cartilage. Expression of transgene mRNA at levels equivalent or higher than the endogenous mRNA in the offspring of two founder animals resulted in a severe chondrodysplastic phenotype with short limbs, hypoplastic thorax, abnormal craniofacial development, and other skeletal deformities. The affected pups died at birth due to respiratory distress. Light microscopy of epiphyseal growth plates of transgenic pups demonstrated a marked reduction in cartilaginous extracellular matrix and disruption of the normal organization of the growth plate. The zone of proliferating chondrocytes was greatly reduced whereas the zone of hypertrophic chondrocytes was markedly increased extending deep into the diaphysis suggestive of a defect in endochondral ossification. Electron microscopic examination revealed chondrocytes with extended RER, a very severe reduction in the amount of cartilage collagen fibrils, and abnormalities in their structure. We postulate that the deletion in the alpha 1(II) collagen acts as a dominant negative mutation disrupting the assembly and secretion of type II collagen molecules. The consequences of the mutation include interference with normal endochondral ossification. These mice constitute a valuable model to study the mechanisms underlying human chondrodysplasias and normal bone formation. - + 10090 Species transgenic mice - + 10090 Species mouse - + 10090 Species mice - + 9606 Species human - + 10090 Species transgenic - - MESH:D009139 - Disease - - skeletal deformities - - - MESH:D012128 - Disease - - respiratory distress - + MESH:D000741 Disease - - chondrodysplastic + + hypoplastic + MESH:D019465 + Disease + + abnormal craniofacial development + + MESH:D010009 Disease chondrodysplasias - + MESH:D006984 Disease hypertrophic + + MESH:C535964 + Disease + + cartilage collagen fibrils + + + MESH:D009139 + Disease + + skeletal deformities + + + MESH:D015831 + Disease + + cartilaginous + + + MESH:D002357 + Disease + + cartilage + + + Disease + + chondrodysplastic + @@ -81479,47 +82166,59 @@ abstract 67 Export of mRNA from the nucleus to the cytoplasm was studied in mature Xenopus laevis oocytes. In vitro transcribed, capped 32P-labeled mRNA was microinjected into nuclei, and its appearance in the cytoplasm measured by counting radioactivity or by RNA extraction and gel electrophoresis. Both for a 5.0-kb transferrin receptor mRNA and a 2.0- kb 4F2 antigen heavy chain mRNA we found saturable transport with an apparent Km of 3.6 x 10(8) molecules per oocyte nucleus. Under non- saturating conditions the half-time for mRNA export from the nucleus was approximately 2 min at 20 degrees C. At higher concentrations of injected mRNA this half-time was prolonged, and the maximal transport rate was reached at approximately 1.6 x 10(8) molecules/min. mRNA transport showed properties of an energy-dependent mechanism, since it was inhibited at 4 degrees C or by ATP depletion. Co-injection of the cap dinucleotide m7GpppG blocked the export effectively, suggesting a role for the cap in this process. The export was also inhibited by the pre-injection of wheat germ agglutinin. The effect of the lectin was specific and abolished by co-injection of N-acetylglucosamine. Finally, we found significant competitive inhibition in mRNA export by the presence of tRNA. Our results suggest that mRNA transport is a facilitated process which may share common steps with tRNA transport. Preliminary gel retardation experiments show that injected mRNA associates with endogenous nuclear proteins and suggest an exchange of some of the bound components during the transport to the cytoplasm. - + 379502 Gene transferrin - + 379133 Gene 4F2 - + 8355 Species Xenopus laevis - + 4565 Species wheat - + MESH:D008607 Disease retardation - + + MESH:D000117 + Chemical + + N-acetylglucosamine + + MESH:D000255 Chemical ATP - - MESH:C014088 + + MESH:C000615311 Chemical - - N-acetylglucosamine + + 32P + + + - + Chemical + + dinucleotide m7GpppG @@ -81562,70 +82261,64 @@ abstract 114 The neural cell adhesion molecule (N-CAM) engages in diverse functional roles in neural cell interactions. Its extracellular part consists of five Ig-like domains and two fibronectin type III homologous (type III) repeats. To investigate the functional properties of the different structural domains of the molecule in cell interactions and signal transduction to the cell interior, we have synthesized, in a bacterial expression system, the individual domains and tandem sets of individual domains as protein fragments. These protein fragments were tested for their capacity to influence adhesion and spreading of neuronal cell bodies, promote neurite outgrowth, and influence cellular migration patterns from cerebellar microexplants in vitro. Ig-like domains I and II and the combined type III repeats I-II were most efficient for adhesion of neuronal cell bodies, when coated as substrates. Neurite outgrowth was best on the substrate-coated combined type III repeats I- II, followed by the combined Ig-like domains I-V and Ig-like domain I. Spreading of neuronal cell bodies was best on substrate-coated combined type III repeats I-II, followed by Ig-like domain I and the combined Ig- like domains I-V. The cellular migration pattern from cerebellar microexplant cultures plated on a mixture of laminin and poly-L-lysine was modified by Ig-like domains I, III, and IV, while Ig-like domains II and V and the combined type III repeats I-II did not show significant modifications, when added as soluble fragments. Outgrowth of astrocytic processes from the explant core was influenced only by Ig- like domain I. Metabolism of inositol phosphates was strongly increased by Ig-like domain I and less by the Ig-like domains II, III, IV, and V, and not influenced by the combined type III repeats I-II. Intracellular concentrations of Ca2+ and pH values were increased only by the Ig-like domains I and II. Intracellular levels of cAMP and GMP were not influenced by any protein fragment. These experiments indicate that different domains of N-CAM subserve different functional roles in cell recognition and signal transduction, and are functionally competent without nervous system-derived carbohydrate structures. - + 4684 Gene 40754 neural cell adhesion molecule - + 4684 Gene 40754 N-CAM - + 2335 Gene 1533 fibronectin - + 4684 Gene 40754 N-CAM - - MESH:D000242 - Chemical - - cAMP - - - MESH:D007295 - Chemical - - inositol phosphates - - - MESH:D002118 + + MESH:D000069285 Chemical Ca2+ - - MESH:D011107 - Chemical - - poly-L-lysine - - + MESH:D002241 Chemical carbohydrate - - MESH:C068152 + + MESH:C066524 Chemical GMP + + MESH:D007295 + Chemical + + inositol phosphates + + + - + Chemical + + cAMP + @@ -81647,29 +82340,30 @@ Goday C, González-García JM, Esteban MR, Giovinazzo G, Pimpinelli S 0 Kinetochores and chromatin diminution in early embryos of Parascaris univalens - - Disease - - Parascaris univalens - ABSTRACT abstract 79 In Parascaris the mitotic chromosomes of gonial germline cells are holocentric and possess a continuous kinetochore along their entire length. By contrast, in meiotic cells, the centromeric activity is restricted to the heterochromatic tips where direct insertion of spindle microtubules into chromatin without any kinetochore plate is seen. In the presomatic cells of early embryos, which undergo heterochromatin elimination, only euchromatin shows kinetic activity. After developing a technique to separate the very resistant egg shell from the embryos, we studied the cell divisions during early embryogenesis by immunochemical and EM approaches. The results reported here show that in presomatic cells microtubules bind only the euchromatin where a continuous kinetochore plate is present. We also report observations suggesting that the binding of the long kinetochores to the mitotic spindle initiates to a limited number of sites and extends along the entire length, during chromosome condensation. The existence of different centromere stages in different cell types, rends Parascaris chromosomes a very good model to study centromere organization. - + 6257 Species Parascaris - + 6257 Species Parascaris + + MESH:D060725 + Disease + + heterochromatic tips + @@ -81697,26 +82391,32 @@ abstract 141 mAb DSS-8 binds to a 164-kD developmental stage-specific cell surface antigen in the nervous system of the cockroach, Periplaneta americana. The antigen is localized to different subsets of cells at various stages of development. The spatial and temporal distributions of DSS-8 binding were determined and are consistent with this antigen playing multiple roles in the development of the nervous system. Direct identification of some of these functions was made by perturbation experiments in which pioneer axon growth occurs in embryos that are cultured in vitro in the presence of mAb DSS-8 or its Fab fragment. Under these conditions the pioneer axons of the median fiber tract grow but follow altered pathways. In a smaller percentage of the ganglia, the immunoreagents additionally produce defasciculation of a subset of DSS-8 labeled axons. Therefore, direct roles for the DSS-8 antigen in both the guidance of pioneer axons and selective fasciculation have been demonstrated. - + 2187 Gene 51880 Fab - + 6978 Species Periplaneta americana - + + - + Chemical + + DSS-8 + + MESH:D005207 Disease fasciculation - + Disease defasciculation @@ -81748,96 +82448,101 @@ abstract 70 We identified two new Saccharomyces cerevisiae kinesin-related genes, KIP1 and KIP2, using polymerase chain reaction primers corresponding to highly conserved regions of the kinesin motor domain. Both KIP proteins are expressed in vivo, but deletion mutations conferred no phenotype. Moreover, kip1 kip2 double mutants and a triple mutant with kinesin- related kar3 had no synthetic phenotype. Using a genetic screen for mutations that make KIP1 essential, we identified another gene, KSL2, which proved to be another kinesin-related gene, CIN8. KIP1 and CIN8 are functionally redundant: double mutants arrested in mitosis whereas the single mutants did not. The microtubule organizing centers of arrested cells were duplicated but unseparated, indicating that KIP1 or CIN8 is required for mitotic spindle assembly. Consistent with this role, KIP1 protein was found to colocalize with the mitotic spindle. - + 852216 Gene 136830 KIP1 - + 855948 Gene 137176 KIP2 - + 852216 Gene 136830 kip1 - + 855948 Gene 137176 kip2 - + 852216 Gene 136830 KIP1 - + 856648 Gene 136907 KSL2 - + 856648 Gene 136907 CIN8 - + 852216 Gene 136830 KIP1 - + 856648 Gene 136907 CIN8 - + 852216 Gene 136830 KIP1 - + 856648 Gene 136907 CIN8 - + 852216 Gene 136830 KIP1 - + 4932 Species Saccharomyces cerevisiae + + Disease + + mitosis + @@ -81940,43 +82645,37 @@ abstract 106 alpha-Lytic protease is a bacterial serine protease of the trypsin family that is synthesized as a 39-kD preproenzyme (Silen, J. L., C. N. McGrath, K. R. Smith, and D. A. Agard. 1988. Gene (Amst.). 69: 237- 244). The 198-amino acid mature protease is secreted into the culture medium by the native host, Lysobacter enzymogenes (Whitaker, D. R. 1970. Methods Enzymol. 19:599-613). Expression experiments in Escherichia coli revealed that the 166-amino acid pro region is transiently required either in cis (Silen, J. L., D. Frank, A. Fujishige, R. Bone, and D. A. Agard. 1989. J. Bacteriol. 171:1320-1325) or in trans (Silen, J. L., and D. A. Agard. 1989. Nature (Lond.). 341:462-464) for the proper folding and extracellular accumulation of the enzyme. The maturation process is temperature sensitive in E. coli; unprocessed precursor accumulates in the cells at temperatures above 30 degrees C (Silen, J. L., D. Frank, A. Fujishige, R. Bone, and D. A. Agard. 1989. J. Bacteriol. 171:1320-1325). Here we show that full- length precursor produced at nonpermissive temperatures is tightly associated with the E. coli outer membrane. The active site mutant Ser 195----Ala (SA195), which is incapable of self-processing, also accumulates as a precursor in the outer membrane, even when expressed at permissive temperatures. When the protease domain is expressed in the absence of the pro region, the misfolded, inactive protease also cofractionates with the outer membrane. However, when the folding requirement for either wild-type or mutant protease domains is provided by expressing the pro region in trans, both are efficiently secreted into the extracellular medium. Attempts to separate folding and secretion functions by extensive deletion mutagenesis within the pro region were unsuccessful. Taken together, these results suggest that only properly folded and processed forms of alpha-lytic protease are efficiently transported to the medium. - + 69 Species Lysobacter enzymogenes - + 562 Species Escherichia coli - + 562 Species E. coli - + 562 Species E. coli - - MESH:C478787 - Chemical - - 166-amino acid - - - MESH:C047902 + + - Chemical - - serine + + SA195 - + p.S195A ProteinMutation @@ -82016,63 +82715,52 @@ abstract 109 High efficiency gene transfer (greater than 90%) in chicken dorsal root ganglion neurons has been obtained by DNA calcium phosphate co- precipitation, hence providing an important tool to study control of gene expression in primary neurons. Transfection with c-fos promoter sequences linked to the chloramphenicol acetyltransferase reporter gene showed that the serum responsive element functions as a strong transcriptional enhancer. Transcription from this element is developmentally regulated, and mediates the genetic response to nerve growth factor (NGF) in developing avian sensory neurons. Furthermore, NGF exerts a negative effect on transcription from the cyclic AMP responsive element, thereby supporting the involvement of tyrosine kinase activation by NGF in primary sensory neurons. - + 396466 Gene 1876 nerve growth factor - + 396466 Gene 1876 NGF - + 396466 Gene 1876 NGF - + 396466 Gene 1876 NGF - + 9031 Species chicken - - Chemical - - tyrosine - - + MESH:D000242 Chemical cyclic AMP - + MESH:C020243 Chemical calcium phosphate - - MESH:D002701 - Chemical - - chloramphenicol - @@ -82100,32 +82788,20 @@ abstract 139 After Listeria is phagocytosed by a macrophage, it dissolves the phagosomal membrane and enters the cytoplasm. The Listeria then nucleates actin filaments from its surface. These actin filaments rearrange to form a tail with which the Listeria moves to the macrophage surface as a prelude to spreading. Since individual actin filaments appear to remain in their same positions in the tail in vitro after extraction with detergent, the component filaments must be cross- bridged together. From careful examination of the distribution of actin filaments attached to the surface of Listeria and in the tail, and the fact that during and immediately after division filaments are not nucleated from the new wall formed during septation, we show how a cloud of actin filaments becomes rearranged into a tail simply by the mechanics of growth. From lineage studies we can relate the length of the tail to the age of the surface of Listeria and make predictions as to the ratio of Listeria with varying tail lengths at a particular time after the initial infection. Since we know that division occurs about every 50 min, after 4 h we would predict that if we started with one Listeria in a macrophage, 16 bacteria would be found, two with long tails, two with medium tails, four with tiny tails, and eight with no tails or a ratio of 1:1:2:4. We measured the lengths of the tails on Listeria 4 h after infection in serial sections and confirmed this prediction. By decorating the actin filaments that make up the tail of Listeria with subfragment 1 of myosin we find (a) that the filaments are indeed short (maximally 0.3 microns in length); (b) that the filament length is approximately the same at the tip and the base of the tail; and (c) that the polarity of these filaments is inappropriate for myosin to be responsible or to facilitate movement through the cytoplasm, but the polarity insures that the bacterium will be located at the tip of a pseudopod, a location that is essential for spreading to an adjacent cell. Putting all this information together we can begin to unravel the problem of how the Listeria forms the cytoskeleton and what is the biological purpose of this tail. Two functions are apparent: movement and pseudopod formation. - + 79784 Gene 23480 myosin - + 79784 Gene 23480 myosin - - MESH:D007239 - Disease - - infection - - - MESH:D007239 - Disease - - infection - @@ -82171,165 +82847,171 @@ abstract 61 Parkinson's disease (PD) is not only characterized by motor disturbances but also, by cognitive, sensory, psychiatric and autonomic dysfunction. It has been proposed that some of these symptoms might be related to the widespread pathology of alpha-synuclein (alpha-syn) aggregation in different nuclei of the central and peripheral nervous system. However, the pathogenic formation of alpha-syn aggregates in different brain areas of PD patients is poorly understood. Most experimental models of PD are valuable to assess specific aspects of its pathogenesis, such as toxin-induced dopaminergic neurodegeneration. However, new models are required that reflect the widespread and progressive formation of alpha-syn aggregates in different brain areas. Such alpha-syn aggregation is induced in only a few animal models, for example perikaryon inclusions are found in rats administered rotenone, aggregates with a neuritic morphology develop in mice overexpressing either mutated or wild-type alpha-syn, and in Smad3 deficient mice, aggregates form extensively in the perikaryon and neurites of specific brain nuclei. In this review we focus on alpha-syn aggregation in the human disorder, its genetics and the availability of experimental models. Indeed, evidences show that dopamine (DA) metabolism may be related to alpha-syn and its conformational plasticity, suggesting an interesting link between the two pathological hallmarks of PD: dopaminergic neurodegeneration and Lewy body (LB) formation. - + 20617 Gene 293 alpha-synuclein - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 29219 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 6622 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 9606 Species patients - + 10116 Species rats - + 10090 Species mice - + 10090 Species mice - + 9606 Species human - + MESH:D001523 Disease psychiatric - + MESH:D010300 Disease PD - + + MESH:D001342 + Disease + + autonomic dysfunction + + MESH:D010300 Disease PD - + + MESH:D004298 + Chemical + + DA + + MESH:D010300 Disease Parkinson's disease - + MESH:D010300 Disease PD - + MESH:D004298 Chemical dopamine - + MESH:D012402 Chemical rotenone - + MESH:D019636 Disease - + neurodegeneration - + MESH:D019636 Disease - - dopaminergic neurodegeneration + + neurodegeneration - + MESH:D010300 Disease PD - - MESH:D012735 - Disease - - autonomic dysfunction - INTRO @@ -82342,61 +83024,61 @@ paragraph 1573 While the first description of Parkinson's disease (PD) may date back to ancient Indian and Chinese texts from 1000 BC, the first clear medical description of this disorder was presented by James Parkinson in 1817. Some years later, in the mid-1800s, Jean-Martin Charcot separated PD from multiple sclerosis and other disorders that are also characterized by tremor, and in 1895 Brissaud formulated the hypothesis that the substantia nigra (SN) is the main brain nucleus pathologically affected in PD. Subsequently, it was Friedrich Lewy who first described the protein aggregates that form in different areas of the brain of PD patients, including the dorsal vagal nucleus, locus coeruleus and globus pallidus. Not long after, Tretiakoff validated Brissaud's hypothesis in 1919 and, by examining post-mortem tissue, he described the protein aggregates in the SN and called them Lewy bodies. Despite this long history, even today the etiology of idiopathic PD remains unknown and given the diversity of the molecular mechanisms proposed, it has been suggested that multiple factors may cause the disease. - + 9606 Species patients - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010302 Disease Parkinson - + MESH:D010300 Disease PD - + + MESH:D009103 + Disease + + multiple sclerosis + + MESH:D010300 Disease PD - - MESH:D012598 - Disease - - sclerosis - - + MESH:D010300 Disease PD - + MESH:D010300 Disease Parkinson's disease - + MESH:D014202 Disease @@ -82408,86 +83090,86 @@ paragraph 2683 PD is the second most common neurodegenerative disorder that affects the human brain. It is primarily characterized by motor symptoms like akinesia, rigidity, resting tremor and postural instability, manifestations that are mainly derived from the progressive degeneration of dopaminergic neurons in the SN pars compacta. Non-motor symptoms also develop that are associated with cognitive deficits (ranging from memory impairment to dementia), emotional changes (depression, apathy and anxiety), sleep perturbations, autonomic dysfunction (bladder disturbances, orthostatic hypotension, sweating), sensory symptoms (pain, visual impairment, olfactory deficit, paresthesia, ageusia) and gastrointestinal symptoms (constipation, dribbling of saliva:. - + 9606 Species human - + MESH:D014202 Disease tremor - + MESH:D010300 Disease PD - + MESH:D019636 Disease neurodegenerative disorder - + MESH:D004409 Disease akinesia - + MESH:D009127 Disease rigidity - - MESH:D009410 - Disease - - degeneration of dopaminergic neurons - INTRO paragraph 3432 Although the primary motor symptoms are shared by patients, both the full presentation of the disorder and the response to treatment are quite heterogeneous. It must be borne in mind that non-dopaminergic neuronal loss is also detected in some areas of the brain, for example, that of monoaminergic cells in the locus coeruleus and raphe nuclei, cholinergic cells in the nucleus basalis of Meynert and in the pedunculopontine tegmental nucleus, as well as the loss of hypocretin cells in the hypothalamus. Indeed, the other pathological changes observed are widespread, with the appearance of LB inclusions in different areas of the brain (mesostriatal system, cortex, thalamus, hypothalamus, olfactory bulb or brainstem), or alterations in the autonomic system (the spinal cord, sympathetic ganglia and myenteric plexus in the gastrointestinal tract). The widespread nature of this pathology is indicative that the disorder is not just a motor alteration but rather, a sensory, cognitive, psychiatric and autonomic disorder. - + 15171 Gene 1166 hypocretin - + 9606 Species patients - + MESH:D007029 Disease hypothalamus - + + MESH:D004067 + Disease + + gastrointestinal tract + + MESH:D009410 Disease neuronal loss - + MESH:D001523 Disease psychiatric and autonomic disorder - + MESH:D007029 Disease @@ -82499,7 +83181,7 @@ title_1 4458 Lewy bodies in PD - + MESH:D010300 Disease @@ -82511,48 +83193,24 @@ paragraph 4476 A common neuropathological feature of some neurodegenerative diseases is the presence of proteinaceous inclusion bodies caused by misfolded and intracellular aggregation of proteins in many brain regions. These abnormal protein deposits may provoke LB pathologies that involve the deposition of LBs in cell bodies, or the formation of Lewy neurites (LNs) and Papp-Lantos inclusions. While the presence of LBs is a histological hallmark of PD, they are also associated with disorders such as dementia with LBs, multiple system atrophy, Alzheimer's disease, Down's syndrome, neurodegeneration with brain iron accumulation type I (Hallervorden-Spatz disease), progressive autonomic failure, rapid eye movement sleep disorder, parkinsonism-dementia complex of Guam, Gaucher's disease or Pick's disease. - + MESH:D020774 Disease Pick's disease - - MESH:D000544 - Disease - - Alzheimer's disease - - - MESH:D015835 + + MESH:D010300 Disease - - eye movement sleep disorder + + 's syndrome - + MESH:D019636 Disease neurodegenerative diseases - - MESH:D019578 - Disease - - multiple system atrophy - - - MESH:D012791 - Disease - - progressive autonomic failure - - - MESH:D010300 - Disease - - PD - MESH:D006211 Disease @@ -82560,35 +83218,71 @@ Hallervorden-Spatz disease - MESH:C537240 + MESH:D020961 Disease - - parkinsonism-dementia + + Lewy neurites - MESH:D003704 + MESH:D006211 Disease - - dementia + + brain iron accumulation type I - MESH:D007501 - Chemical - - brain iron + MESH:D001284 + Disease + + atrophy + MESH:D003704 + Disease + + dementia + + MESH:D019636 Disease neurodegeneration - + MESH:D005776 Disease Gaucher's disease + + MESH:D000544 + Disease + + Alzheimer's disease + + + MESH:D010300 + Disease + + PD + + + MESH:D020187 + Disease + + rapid eye movement sleep disorder + + + MESH:C537240 + Disease + + parkinsonism-dementia + + + MESH:D012791 + Disease + + autonomic failure + INTRO @@ -82601,44 +83295,44 @@ paragraph 5297 LBs are morphologically heterogeneous, with classic LB arising in the brainstem as cytoplasmic inclusions of 8-30 mum in diameter, with a dense eosinophilic core and a narrow pale stained rim. On haematoxylin/eosin staining, classic LBs are observed as a spherical body with a dense core surrounded by a halo, whereas the cortical LBs present in layers V-VI of the temporal, insular and cingulate cortex have no obvious halo. A third type of LBs are known as pale bodies, that are rounded, pale, eosinophilic granules which lack the eosinophilic core of classic LBs, and that are only weakly and diffusely stained with eosin. These pale bodies are thought to be precursor LBs. In addition, dystrophic LNs are present in axonal processes with both thread-like and spheroid forms. - + MESH:D020388 Disease dystrophic LNs - + MESH:D004801 Chemical eosin - + + MESH:D006416 + Chemical + + haematoxylin + + MESH:D004801 Chemical eosin - - MESH:C018328 - Chemical - - haematoxylin - INTRO paragraph 6079 In dementia with LB disease, inclusions seem to be morphologically homogeneous in the neocortical and paralimbic regions of the brain. In the SN, the LBs immunolabeled with alpha-syn and ubiquitin fall within the spectrum from diffuse to pale bodies, and to classic LBs. Indeed, this morphological diversity in the SN may represent different stages in the formation of LBs. - + 20617 Gene 293 alpha-syn - + MESH:D003704 Disease @@ -82650,7 +83344,7 @@ paragraph 6453 Electron microscopy has shown that the pale rim and dense core of classic LBs correspond to zones of radially-oriented straight filaments, and zones of circular profiles, respectively. Cortical LBs and LNs also contain filaments. Three-dimensional reconstruction from serial confocal images reflects that the LB core, immunolabeled with alpha-syn and ubiquitin, has a concentric layered structure, with neurofilament encircling these inner layers. Indeed, a frequent continuity between LBs and LNs has been detected, indicating that LNs may evolve into LBs. - + 20617 Gene 293 @@ -82669,84 +83363,84 @@ paragraph 7026 LBs are thought to be mainly composed of alpha-syn and the morphological characterization of LBs is mostly based on immunochemistry for alpha-syn, ubiquitin and neurofilament. Furthermore, the alpha-syn in LBs undergoes post-translational modifications, such as phosphorylation, ubiquitination and oxidative nitration. However, LBs also contain many different proteins, such as the Leucine-rich repeat kinase 2 (LRRK2:, histone deacetylase 6 (HDAC6: and charged multivesicular body protein 2B (CHMP2B:, all of which are found in the core. Indeed, LRRK2 is associated with the endoplasmic reticulum of dopaminergic neurons, HDAC6 is considered a sensor of proteasomal inhibition that plays a central role in autophagy, and CHMP2B is a component of the endosomal sorting complex involved in protein degradation. The identification of proteins present in LBs may offer molecular clues to the processes that may participate in LBs formation. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 Leucine-rich repeat kinase 2 - + 66725 Gene 18982 LRRK2 - + 15185 Gene 31353 histone deacetylase 6 - + 15185 Gene 31353 HDAC6 - + 68942 Gene 8534 charged multivesicular body protein 2B - + 68942 Gene 8534 CHMP2B - + 66725 Gene 18982 LRRK2 - + 15185 Gene 31353 HDAC6 - + 68942 Gene 8534 @@ -82759,26 +83453,26 @@ paragraph 7964 Nevertheless, the comprehensive molecular composition of LBs is still unclear, although new proteomics approaches based on mass spectrometry provide an interesting approach to discover proteins not yet described in aggregates. However, the success of such approaches will depend on the techniques used to isolate LBs. One study used cortical LB-enriched fractions derived from the LB variant of Alzheimer's disease to identify 40 LB proteins involved in phosphorylation, ubiquitination, oxidative stress or protein trafficking. Laser capture microdissection of cortical LBs from neurons located in the temporal cortex of dementia with LB disease patients, allowed 296 proteins to be detected, including the chaperone HSC71, confirming the presence of chaperone molecules in LBs. - + 3312 Gene 68524 HSC71 - + 9606 Species patients - + MESH:D000544 Disease Alzheimer's disease - + MESH:D003704 Disease @@ -82796,6 +83490,11 @@ paragraph 8778 Stage 1: At the earliest stage, only the dorsal motor nucleus of the vagus nerve (in the lower medulla oblongata) and the anterior olfactory nucleus are affected, suggesting that the brain pathology originates in these structures. + + Disease + + oblongata + INTRO @@ -82814,7 +83513,7 @@ paragraph 9351 Stage 4: Cortical LBs appear in the temporal mesocortex, in the allocortical CA2 of the hippocampus and in thalamic structures. - + 12349 Gene 37256 @@ -82827,14 +83526,14 @@ paragraph 9479 Stage 5: The anterior cingulate, insular and subgenual mesocortex is affected, and the neocortex appears to be affected for the first time, with LBs in high order sensory association structures and in the prefrontal neocortex. The hippocampus is clearly affected in the CA1, CA3 and entorhinal cortex. - + 12346 Gene 20414 CA1 - + 12350 Gene 31298 @@ -82853,13 +83552,13 @@ paragraph 9901 Studies of the topographic distribution of LBs during the course of sporadic PD has enabled a classification of the stages of disease progression to be drawn up. Neurons susceptible to contain LBs have long, thin, unmyelinated or poorly myelinated axons and the parkinsonian brains analyzed can nearly all be categorized into one of six different stages, based on the location of the inclusion bodies: - - MESH:D010300 + + MESH:D010302 Disease - - parkinsonian brains + + parkinsonian - + MESH:D010300 Disease @@ -82873,7 +83572,7 @@ table_caption 10303 Overview of LB-like formation in rodent models. Brain areas with the presence of DA receptors and LB-like and/or LN-like aggregates may suggest an interaction between the dopaminergic system and alpha-syn (see text) - + 20617 Gene 293 @@ -82888,102 +83587,60 @@ Tab1.xml 10521 Brain Nucleus Braak's stage PrP-A53T-alpha-syn mice Rotenone in rats Smad3 null mice DA system X-XII 1 +++ LB- LN Yes OB 1 ++ LB-LN Yes sp5 2 ++ LN ++ LN Yes ll 2 ++ LN Yes Sc 2 ++++ LB-LN ++ LB Yes RF/LC 2 +++ LB- LN ++ LN Yes RN 2, 3 +++ LB-LN Yes VTA 3 + LB- LN ++ LN Yes CPu 3 ++ LB- LN +++++ LB- LN ++++ LN Yes SN 3 + LB- LN +++++ LB- LN +++++ LN Yes Hp 3,4 ++++ LB-LN Yes Th 4 +++ LB-LN Yes M1-M2 5,6 ++++ LB-LN Yes Cg 5,6 +++ LN Yes cc-ic-cp -- +++ LN Pn -- ++ LB Yes CB -- ++++ LB-LN ++ LN Yes Pir -- +++ LN Yes IC -- +++ LB-LN SC -- +++ LB-LN Yes - + 25631 Gene 55937 Smad3 - + 110027 Gene CPu 3 - + 185869 Species T-alpha - + 10090 Species mice - + 10116 Species rats - + 10090 Species mice - + 16473 Gene syn - + MESH:D012402 Chemical Rotenone - - MESH:C045084 - Chemical - - LN ++++ - - MESH:D012538 - Chemical - - Sc 2 +++ - - - MESH:D013910 - Chemical - - Th 4 - - - MESH:C045084 - Chemical - - LN ++++ - - - MESH:D009765 - Disease - - OB - - - MESH:C045084 - Chemical - - LN Pn - - - MESH:C045084 - Chemical - - LN ++++ - - - MESH:C045084 + MESH:C028505 Chemical - - LN +++ + + Cg @@ -82991,50 +83648,50 @@ paragraph 11154 This model reflects the pathological progression in PD patients (Braak PD stage 4-6), with a caudo-rostral gradient in LB deposition from the lower brainstem to the neocortex, and with both dopaminergic and non-dopaminergic areas being affected (Table 1). Some patients do not have clinical PD symptoms, but display alpha-syn deposition at autopsy in areas according to Braak PD stage 1-3. They are referred to as showing incidental LB disease (iLBD), considered a prodromal state of PD. - + 6622 Gene 293 alpha-syn - + 9606 Species patients - + 9606 Species patients - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010300 Disease - - Braak PD + + PD - + MESH:D010300 Disease - - Braak PD + + PD - + MESH:D010300 Disease @@ -83046,21 +83703,21 @@ paragraph 11644 Besides neuronal alpha-syn deposition in LBs and LNs, the presence of inclusions in astrocytes has been detected by silver staining. Glial inclusions are widespread in PD, both in areas with neuronal loss and gliosis (substantia nigra, locus coeruleus and dorsal vagal nucleus), and in areas with no clear neurodegeneration or gliosis (cerebral cortex, cerebral white matter, striatum, globus pallidus, thalamus, cerebellum and spinal cord:. Indeed, alpha-syn-immunoreactive (-ir) astrocytes adopt a topographical distribution that closely parallels that of the cortical LBs. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + MESH:D012834 Chemical @@ -83072,13 +83729,13 @@ paragraph 12220 However, we should consider that this disorder has a very heterogeneous clinical manifestation and as yet, there is no clear significance attributed to the presence of LBs and LNs, as direct determinants of the clinical manifestations and symptoms evident in patients with PD. - + 9606 Species patients - + MESH:D010300 Disease @@ -83096,88 +83753,88 @@ paragraph 12509 It is considered that most patients with PD develop the idiopathic form of the disease as opposed to that which is genetically inherited. Although, 10-30% of PD patients report a first-degree relative with the disorder, this does not necessarily reflect genetic inheritance as it may correspond with exposure to a common environmental factor. However, it is increasingly clear that genetic factors contribute to the pathogenesis of PD and it is currently considered a multifactorial complex disorder, caused by interactions between genetic and environmental risk factors. Alternatively, only 10% of cases develop the rarer form of PD that follows Mendelian inheritance, representing 2% of late-onset and 50% of early-onset familial PD. Studies into these familial forms have identified several causative genes related to mitochondrial or lysosomal dysfunction, protein aggregation, the proteasome system and kinase signaling. - + 9606 Species patients - + 9606 Species patients - - MESH:C537708 - Disease - - multifactorial complex disorder - - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - - MESH:D058495 - Disease - - protein aggregation - - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + + MESH:D066263 + Disease + + protein aggregation + + MESH:D010300 Disease familial PD + + MESH:D016464 + Disease + + lysosomal dysfunction + INTRO paragraph 13435 LB deposition is associated with specific mutations in some of these already known genes, including alpha-syn (SNCA: and LRRK2. Moreover, the distinction between the familial and idiopathic form of PD is currently not clear in all cases, as some evidence suggests that SNCA or LRKK2 mutations also participate in the sporadic disease. - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 20617 Gene 293 SNCA - + 20617 Gene 293 @@ -83196,256 +83853,268 @@ paragraph 13775 Several dominant mutations have been described in the gene encoding alpha-syn, SNCA, with varying penetrance. In 1997, the A53T missense mutation was reported in a large Italian family associated with familial PD. Subsequently, two additional missense mutations in SNCA were also seen to be associated to familial PD, A30P and E46K, although these point mutations are extremely rare. Duplicate and triplicate loci of SNCA of different sizes (from 0.4 to 4.5 Mb) have been seen to give rise to PD, which has been related to the overexpression of the wild-type protein, influencing the age of onset and severity of the disorder. Indeed, SNCA duplications have also been reported in apparently sporadic PD patients and interestingly, patients carrying these mutations present a broad clinical phenotype, even within a given family, suggesting an influence of genetic modifiers. In addition, a dinucleotide repeat polymorphism (Rep1) has been described in the promoter region, 10 Kb upstream of the start codon, which induces SNCA overexpression and that may account for increased risk in 3% of non-familiar PD. Moreover, specific haplotypes, primarily in the 3' UTR region, may also be associated to sporadic PD. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 SNCA - + 20617 Gene 293 SNCA - + 20617 Gene 293 SNCA - + 6622 Gene 293 SNCA - + 104354 Gene Rep1 - + 9606 Species patients - + 9606 Species patients - + 20617 Gene 293 SNCA - + MESH:D010300 Disease PD - + + MESH:D015226 + Chemical + + dinucleotide + + p.A30P;RS#:104893878 ProteinMutation A30P - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + p.E46K;RS#:104893875 ProteinMutation E46K - + + MESH:D010300 + Disease + + PD + + c.53A>T;RS#:104893877 DNAMutation A53T - - MESH:D010300 - Disease - - familial PD - INTRO paragraph 14985 Although SNCA mutations are very rare, their identification led to the association of alpha-syn with LBs. This protein is predominantly localized in presynaptic nerve terminals and missense mutations may reduce its affinity for lipids, enhancing its propensity to adopt a beta-sheet conformation and promoting self-assemble into oligomers and fibril formation. In this sense, the A53T mutation of SNCA is associated with the predominantly neuritic aggregation of alpha-syn in the human brain and in a mouse model, as well as with severe motor impairments. Alternatively, SNCA overexpression may decrease the density of dopaminergic vesicles and of synaptic contacts. Neuropathological diagnosis of PD requires both dopaminergic neurodegeneration in the SNpc and the presence of LBs, and as described previously, the severity of the clinical manifestation has been correlated with the broad distribution of LBs, which may be related to the number of alleles. Both triplication and point mutations in SNCA are associated with the formation of cortical and subcortical LBs, and a clinical diagnosis of PD with dementia. - + 20617 Gene 293 SNCA - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 SNCA - + 6622 Gene 293 alpha-syn - + 20617 Gene 293 SNCA - + 9606 Species human - + 10090 Species mouse - + 20617 Gene 293 SNCA - + c.53A>T;RS#:104893877 DNAMutation A53T - + MESH:D010300 Disease PD - - MESH:D008569 - Disease - - impairments - - - MESH:D003704 - Disease - - dementia - - + MESH:D010300 Disease PD - + MESH:D019636 Disease neurodegeneration - + + MESH:D008055 + Chemical + + lipids + + MESH:D058225 Disease neuritic aggregation + + MESH:D003704 + Disease + + dementia + + + MESH:D009422 + Disease + + impairments + INTRO paragraph 16102 The SNCA gene and that encoding the microtubule-associated protein tau (MAPT) have consistently been associated in different populations of sporadic PD, mainly those of European origin. However, such an associated was not identified in Japanese population, suggesting that population-specific differences may exist in the genetics of PD. - + 20617 Gene 293 SNCA - + 17762 Gene 74962 microtubule-associated protein tau - + 17762 Gene 74962 MAPT - + MESH:D010300 Disease PD - + MESH:D010300 Disease @@ -83457,14 +84126,14 @@ title_3 16440 Leucine-rich repeat kinase 2 (LRRK2) - + 66725 Gene 18982 Leucine-rich repeat kinase 2 - + 66725 Gene 18982 @@ -83477,117 +84146,117 @@ paragraph 16477 Point mutations in the LRRK2 gene are frequently associated to PD, and they are found in both late onset familial and sporadic PD. LRRK2 mutations have the highest prevalence rate in PD patients discovered to date, having been found in 10% of cases with autosomal dominant familial PD, in 3.6% of sporadic PD cases and even in 1.8% of healthy controls. The presence of mutations in healthy controls may suggest a reduced and incomplete age-dependent penetrance. However, while over 80 missense variants have been identified, only seven mutations are considered pathogenic (N1437H, R1441G/C/H, Y1699C, G2019S and I2020T), most of them lying in the C-terminal half of the protein, while two others are considered as risk factors (G2385R and R1628P:. Of these, G2019S is the most frequent mutation in the Caucasian population, which explains 1% of PD cases. The prevalence of this mutation is strongly influenced by ethnicity and as such, it is more frequent in patients of Southern European or North African origin, and in Ashkenazi Jews (18-40% PD cases), yet it is very rare in Asians or Northern Europeans. The risk of PD for a person who inherits this mutation is 28% at age 59 years, 51% at 69 years and 74% at 79 years. The G2019S mutation gives rise to a uniform clinical phenotype that resembles sporadic PD, both in homozygous and heterozygous carriers. - + 66725 Gene 18982 LRRK2 - + 120892 Gene 18982 LRRK2 - + 9606 Species patients - + 9606 Species patients - + 9606 Species person - + p.N1437H;RS#:74163686 ProteinMutation N1437H - + p.R1628P;RS#:33949390 ProteinMutation R1628P - + p.I2020T;RS#:35870237 ProteinMutation I2020T - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:D010300 Disease PD - + p.Y1699C;RS#:35801418 ProteinMutation Y1699C - + p.G2385R;RS#:34778348 ProteinMutation G2385R - + MESH:D010300 Disease PD - - p.R1441G,C,H;RS#:33939927;34995376 - ProteinMutation - - R1441G/C/H - - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:D010300 Disease autosomal dominant familial PD - + + p.R1441G,C,H;RS#:33939927 + ProteinMutation + + R1441G/C/H + + p.G2019S;RS#:34637584 ProteinMutation @@ -83599,105 +84268,105 @@ paragraph 17841 LRRK2 is a gene with 51 exons and it encodes a large protein with two different enzymatic activities in the same molecule: that of a kinase and a GTPase. LRRK2 also has multiple protein interaction domains suggesting that it may serve as a scaffold for the assembly of other proteins. The G2019S mutation affects the kinase domain of the protein, with other common mutations lying in the GTPase domain or in the protein interaction domains. The function of LRRK2 remains largely elusive but G2019S mutations enhance its kinase activity, which may mediate neural toxicity. LRRK2 is widely expressed in the healthy adult brain, mainly in the endoplasmic reticulum. Interestingly, in sporadic PD patients LRRK2 is found in the core of LBs in the SN and locus coeruleus, suggesting it may contribute to LB formation. Indeed, some PD patients with LRRK2 mutations have LB pathology but this is not always the case, even within a family. How LRRK2 might participate in LB formation remains unknown, although dysfunctional autophagy has been proposed, as well as altered solubility and aggregation of alpha-syn. - + 66725 Gene 18982 LRRK2 - + 66725 Gene 18982 LRRK2 - + 66725 Gene 18982 LRRK2 - + 66725 Gene 18982 LRRK2 - + 120892 Gene 18982 LRRK2 - + 120892 Gene 18982 LRRK2 - + 66725 Gene 18982 LRRK2 - + 20617 Gene 293 alpha-syn - + 9606 Species patients - + 9606 Species patients - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:C564093 Disease dysfunctional autophagy - + MESH:D010300 Disease PD - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:D010300 Disease PD - + MESH:D064420 Disease @@ -83715,57 +84384,57 @@ paragraph 18966 Homozygous or compound heterozygous mutations in the recessive genes Parkin, PINK and DJ-1 are associated to relatively rare forms of familial PD, and these result in early onset PD and nigral dopaminergic neuronal loss. Parkin mutations account for around 50% of familial juvenile and early onset PD, reaching 80% in patients with onset before the age of 20 and decreasing with increasing age at onset, becoming very rare when onset occurs after the age of 50. At least 170 mutations have been described in Parkin, including point mutations, exon rearrangements, "indels" and duplications. Homozygous mutations with loss-of-function predominantly induce neuronal loss in the SN and locus coeruleus, in the absence of LBs. However, compound heterozygous mutations with LB pathology have been described in a minority of patients. This variability may indicate some mutation-specific effect, as it has been described loss-of-function, inactivating and activating mutations. Parkin is a cytosolic E3 ubiquitin ligase that transfers ubiquitin to specific protein substrates for proteasomal and autophagic degradation. Mutations may impair Parkin activity, which might reduce the clearance and aggregation of proteins. As such, these mutations could participate in decreased alpha-synuclein clearance or alternatively, Parkin mutations might decrease its solubility and lead to aggregate formation. - + 57320 Gene 38295 DJ-1 - + 20617 Gene 293 alpha-synuclein - + 9606 Species patients - + 9606 Species patients - + MESH:D009410 Disease neuronal loss - + MESH:D010300 Disease PD - + MESH:D009410 Disease neuronal loss - + MESH:D010300 Disease familial PD - + MESH:D010300 Disease @@ -83777,14 +84446,14 @@ paragraph 20361 Other recessive mutations are less common, such as PINK-1 and DJ-1, producing clinical manifestations broadly similar to Parkin mutations and in the absence of LBs. - + 68943 Gene 32672 PINK-1 - + 57320 Gene 38295 @@ -83797,7 +84466,7 @@ title_3 20526 GBA mutations - + 14466 Gene 68040 @@ -83810,108 +84479,108 @@ paragraph 20540 Glucocerebrosidase (GBA) mutations were first described in Gaucher's disease, a recessive lysosomal storage disorder that may develop into parkinsonism, and PD patients may have an increased frequency of GBA mutations. Indeed, carriers of GBA mutations exhibit clinical features related to early-onset, levodopa-responsive PD, experiencing hallucinations and symptoms of cognitive decline or dementia, with the abundant presence of LBs in the neocortex. GBA mutations in PD patients confer increased susceptibility to an earlier disease onset, to have affected relatives and to develop atypical clinical manifestations. - + 2629 Gene 68040 Glucocerebrosidase - + 2629 Gene 68040 GBA - + 2629 Gene 68040 GBA - + 14466 Gene 68040 GBA - + 2629 Gene 68040 GBA - + 9606 Species patients - + 9606 Species patients - + MESH:D003072 Disease cognitive decline - + MESH:D016464 Disease recessive lysosomal storage disorder - + MESH:D006212 Disease hallucinations - + MESH:D007980 Chemical levodopa - + MESH:D010300 Disease PD - + MESH:D005776 Disease Gaucher's disease - + MESH:D003704 Disease dementia - + MESH:D010300 Disease PD - + MESH:D010302 Disease parkinsonism - + MESH:D010300 Disease @@ -83923,7 +84592,7 @@ title_1 21162 Animal models of alpha-syn aggregation - + 20617 Gene 293 @@ -83936,66 +84605,61 @@ paragraph 21201 Animal models are used in order to study the neurobiological basis of human disorders and to develop new treatments. When examining the validity of an animal model we can consider face validity (similarity between neurological and behavioral phenotypes seen in an animal model and the human patient), construct validity (consistent with a theoretical rational, such that the molecular and cellular changes that result from genetic manipulations should be the same as those that occur in humans), and predictive validity (i.e. it should respond to pharmacological treatment as in humans). Face validity is a major criterion for model evaluation. One can argue that modeling of LB deposition and dopaminergic neurodegeneration may correlate to the human pathology. However, the behavioral phenotypes differ considerably between animals such as mice and humans. Thus, face validity may prove to be an unrealistic criterion for some symptoms of the disease such as motor and cognitive deficits or emotional changes. Although some researchers advocate the primacy of one of these approaches, in practice, the validity of a model should consider all three sources of evidence. - + 9606 Species human - + 9606 Species human - + 9606 Species patient - + 9606 Species humans - + 9606 Species humans - + 9606 Species human - + 10090 Species mice - + 9606 Species humans - + MESH:D003072 Disease cognitive deficits - - Chemical - - correlate - - + MESH:D019636 Disease @@ -84007,38 +84671,38 @@ paragraph 22372 Considering the histopathological hallmarks of PD, models of the disorder should address the progressive loss of dopaminergic neurons in the SNpc, reduced striatal dopamine content and altered catabolism, and it should be age dependent and with progressive alteration of the animal's movement. Furthermore, the Braak staging scheme has strongly influenced what we think would be a good animal model for PD. Indeed, the presence of LBs and LNs, the post-translational modifications observed in the human disorder, and the composition and histological localization of LBs are all central features that an animal model of PD must address. In this review we will focus on those models related to alpha-syn aggregation and LB-like formation. - + 20617 Gene 293 alpha-syn - + 9606 Species human - + MESH:D004298 Chemical dopamine - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010300 Disease @@ -84050,226 +84714,231 @@ paragraph 23109 Classical neurotoxin-induced rodent models of PD have been studied in depth and they involve treatment with 6-hydroxydopamine (oxidative stress), MPTP, rotenone and paraquat (mitochondrial complex I inhibition), PSI and epoxomicin (proteasomal inhibition), and lipopolysaccharide (glial activation). Some of these models address the toxic insult hypothesis, whereby pesticide and herbicide use can increase the risk of PD. Most of these chemical models induce degeneration of the dopaminergic neurons in the rodent and primate SN, inducing a motor syndrome that can be modulated by anti-parkinsonian medication. Indeed, the MPTP-treated primate is still the animal model used to test drugs during their selection for clinical trials in humans. These models have been useful to propose pathogenic events that occur in the disease and each mechanism (oxidative stress, mitochondrial complex I inhibition, etc.) is thought to contribute to the pathogenesis of the disorder. While all these models usually display robust nigro-striatal degeneration, the formation of LB inclusions is not a common feature. In this sense, MPTP and 6-OHDA neurotoxic models have face validity related to motor features, although construct (also known as aetiological) validity are limited. Indeed, the poor predictive validity of these models seems to be related to the high failure rate of new treatments in clinical trials. - + 9606 Species humans - + MESH:D015632 Chemical MPTP - + MESH:D012402 Chemical rotenone - - MESH:D010300 - Disease - - parkinsonian - - - MESH:C078846 - Chemical - - epoxomicin - - + MESH:D016627 Chemical 6-OHDA - + MESH:D015632 Chemical MPTP - + + MESH:D001259 + Disease + + motor syndrome + + MESH:D010300 Disease PD - + MESH:D015632 Chemical MPTP - + + MESH:D008070 + Chemical + + lipopolysaccharide + + MESH:D016627 Chemical 6-hydroxydopamine - + MESH:D010300 Disease PD - + MESH:D020258 Disease neurotoxic - + + Disease + + PSI + + MESH:D010269 Chemical paraquat + + MESH:D010302 + Disease + + parkinsonian + INTRO paragraph 24512 The discovery of mutations in patients has led to the generation of different genetic models, which may prove to be a more realistic approach to study PD. Many different species and cell models are useful for genetic manipulation, including mice, Drosophila melanogaster and Caenorhabditis elegans. In this sense, several transgenic models have been described that carry SNCA or LRRK2 mutations, and while many of these develop inclusions they fail to display robust neurodegeneration. Recessive models using knockout mice of PINK-1, Parkin or DJ-1 similarly fail to exhibit a nigro-striatal pathology. Animal models carrying mutations in GBA have recently been described in the context of PD, while other animal models focus on altering the intracellular signaling of neurotrophic factors like Smad3 or on the mechanisms of aggregation of alpha-syn. It should be noted that most models do not capture the main hallmarks of PD in the same animal and hence, most models are only suitable to address one particular issue. However, interesting approximations for LB-like formation have been obtained with rotenone treatment, alpha-syn transgenesis with either wild-type or A53T mutation and in Smad3 deficient mice. Thus, the current models do appear to be useful to asses not only motor symptoms but also aetiological and predictive validity of neuroprotective molecules that could halt the pathological progression of the disease. - + 20617 Gene 293 SNCA - + 66725 Gene 18982 LRRK2 - + 68943 Gene 32672 PINK-1 - + 57320 Gene 38295 DJ-1 - + 14466 Gene 68040 GBA - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 9606 Species patients - + 10090 Species mice - + 6239 Species Caenorhabditis elegans - + 10090 Species mice - + 10090 Species mice - + 10090 Species transgenic - + MESH:D019636 Disease neurodegeneration - - MESH:D009133 - Disease - - neurotrophic - - + c.53A>T;RS#:104893877 DNAMutation A53T - - MESH:D010300 - Disease - - PD - - + MESH:D012402 Chemical rotenone - + MESH:D010300 Disease PD - + + MESH:D010300 + Disease + + PD + + MESH:D010300 Disease @@ -84281,7 +84950,7 @@ title_2 25942 Non-human primates - + 9606 Species @@ -84293,98 +84962,86 @@ paragraph 25961 MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin used in non-human primates as the most relevant animal model of PD, given its ability to induce persistent parkinsonism in humans and on the induction of selective destruction of midbrain dopaminergic neurons. Cardinal motor symptoms of PD are reproduced in this model, as well as other non-motor symptoms such as constipation, salivation, sleep disturbance and cognitive deficits. The MPTP model has limitations as the rapid toxicity results in the acute onset of neurodegeneration and neurological symptoms. Although increased alpha-syn immunoreactivity (-ir) within surviving neurons is detected, MPTP-affected dopaminergic neurons do not develop LBs or LNs. - + 20617 Gene 293 alpha-syn - + 9606 Species human - + 9606 Species humans - + MESH:D010300 Disease PD - - MESH:D003072 - Disease - - cognitive deficits - - + MESH:D015632 Chemical MPTP - + + MESH:D020271 + Disease + + neurodegeneration and neurological symptoms + + MESH:D064420 Disease toxicity - + MESH:D010300 Disease PD - + MESH:D003248 Disease constipation - + + MESH:D003072 + Disease + + sleep disturbance and cognitive deficits + + MESH:D015632 Chemical 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine - - MESH:D019636 - Disease - - neurodegeneration - - + MESH:D015632 Chemical MPTP - + MESH:D010302 Disease parkinsonism - - MESH:D009422 - Disease - - neurological symptoms - - - MESH:D012893 - Disease - - sleep disturbance - - + MESH:D015632 Chemical @@ -84402,7 +85059,7 @@ title_3 26699 MPTP in mice - + MESH:D015632 Chemical @@ -84414,89 +85071,107 @@ paragraph 26712 In mice, the MPTP model must be induced in specific strains for it to be a consistent model, such as C57 and Swiss Webster mice. The loss of dopaminergic neurons depends on the administration regime (from acute to chronic), ranging from 60% to 90%. Acute or sub-chronic administration does not lead to the formation of inclusions and while alpha-syn inclusions have been detected in the study of some chronic models, this is not always replicated. In this sense, alpha-syn-ir was detected in dopaminergic neurons when chronic MPTP treatment was coupled with probenecid administration to block the rapid clearance of MPTP and its derivates. On examination by electron microscopy, the neurons that accumulate alpha-syn appeared to contain lipid droplets or secondary lysosomes covered by proteins. Although this is quite a distinct morphology to the straight filaments detected in human LBs, it was suggested that lipofuscins may be important for the development of LBs. When MPTP was administered continuously using an osmotic minipump, ubiquitin-ir and alpha-syn-ir nigral inclusions formed, although their co-localization in the same cell or aggregate was not studied. Ultrastructurally, these inclusions appeared as concentric membranes containing alpha-syn that was transformed into fibrillar morphology. However, other studies failed to find such inclusions following acute, semi-chronic or chronic MPTP treatment with probenecid or using osmotic minipumps. Thus, further work is required before this model can be used to study aggregate formation. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 10090 Species mice - + 10090 Species mice - + 9606 Species human - + + MESH:D011339 + Chemical + + probenecid + + MESH:D015632 Chemical MPTP - + MESH:D015632 Chemical MPTP - + MESH:D015632 Chemical MPTP - + + MESH:D008055 + Chemical + + lipid + + MESH:D008062 Chemical lipofuscins - + MESH:D015632 Chemical MPTP + + MESH:D011339 + Chemical + + probenecid + MESH:D015632 Chemical @@ -84521,144 +85196,144 @@ paragraph 28282 Rotenone is an insecticide that accumulates in the mitochondria, inhibiting complex I and promoting oxidative stress, as well as inhibiting the ubiquitin-proteasome system in vitro. While this model is one of the most promising models for PD studies, it has several drawbacks since rotenone is not a selective dopaminergic neurotoxin, it produces high rates of mortality (~30%) and there is also strong variability in neurodegeneration (only observed in 50% of rats). Interestingly, peripheral treatment with rotenone induces the formation of alpha-syn inclusions, with a dense core and fibrillar surrounding that resembles those observed in PD. The aggregation of alpha-syn may be enhanced by neonatal lipopolysaccharide (LPS) treatment, suggesting some cooperation between perinatal inflammatory processes and exposure to this pesticide. In vitro studies have shown that rotenone induces a conformational change in alpha-syn and that it accelerates the rate of fibril formation. Indeed, like the human pathology alpha-syn deposits are observed in the myenteric plexus. In order to avoid peripheral rotenone toxicity and hence, the high mortality rate in rats, intracerebral administration of this toxin has been used, showing significant nigro-striatal neurodegeneration but not the appearance of alpha-syn inclusions. Indeed, variability in the results obtained following peripheral administration (subcutaneous versus intravenous infusion) has limited the use of this model as a reliable tool to study PD. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 alpha-syn - + 29219 Gene 293 alpha-syn - + 21898 Gene 41317 LPS - + 10116 Species rats - + 9606 Species human - + 10116 Species rats - + MESH:D019636 Disease neurodegeneration - + MESH:D012402 Chemical rotenone - + MESH:D010300 Disease PD - + + MESH:D008070 + Chemical + + lipopolysaccharide + + MESH:D012402 Chemical rotenone - + MESH:D012402 Chemical rotenone - + MESH:D010300 Disease PD - + MESH:D064420 Disease toxicity - + MESH:D012402 Chemical Rotenone - + MESH:D012402 Chemical rotenone - - MESH:D010300 - Disease - - PD - - + MESH:D019636 Disease neurodegeneration + + MESH:D010300 + Disease + + PD + INTRO paragraph 29792 Combined administration of other herbicides and fungicides, such as paraquat and Maneb also produces a high mortality rate and modest nigro-striatal neurodegeneration without LB deposition. - - MESH:D010269 - Chemical - - paraquat - MESH:D019636 Disease @@ -84731,22 +85406,21 @@ mice + Disease + + PSI + + MESH:D064420 Disease toxicity - + MESH:D010300 Disease - - familial PD - - - MESH:C078846 - Chemical - - epoxomicin + + PD MESH:D010300 @@ -84772,155 +85446,149 @@ paragraph 31603 A loss of mitochondrial complex I is observed in sporadic PD that may lead to selective dopaminergic neurodegeneration due to oxidative stress. Midbrain dopaminergic neurons of PD patients and elderly humans carry high levels of somatic mtDNA mutations. Indeed, neurotoxins like MPTP and rotenone inhibit this mitochondrial complex. Transgenic mice overexpressing wild-type alpha-syn induces mitochondrial fragmentation, which may predispose to neural degeneration, and transgenic mice overexpressing human A53T alpha-syn mutation also display mitochondrial abnormalities that may explain some aspects of the aggregated alpha-syn toxicity. Disruption of the respiratory chain has been modeled in mice by deleting the gene encoding the mitochondrial transcription factor A (TFAM) in dopaminergic neurons, thereby inhibiting the transcription of mtDNA genes. Although these mice have an interesting phenotype, with impaired motor function and neurodegeneration, alpha-syn is not needed to express this phenotype. Indeed, the inclusions found in dopaminergic neurons have no alpha-syn but rather, abnormal mitochondrial membranes. - + 20617 Gene 293 alpha-syn - + 21780 Gene 31139 TFAM - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 9606 Species patients - + 9606 Species humans - + 10090 Species Transgenic mice - + 10090 Species transgenic mice - + 9606 Species human - + 10090 Species mice - + 10090 Species mice - + 185869 Species T alpha - + 16473 Gene syn - + 16473 Gene syn - - MESH:D009410 - Disease - - neural degeneration - - + MESH:D019636 Disease neurodegeneration - + MESH:D064420 Disease toxicity - + + MESH:C565640 + Disease + + neural degeneration + + MESH:D010300 Disease PD - + MESH:D019636 Disease neurodegeneration - + MESH:D015632 Chemical MPTP - + MESH:D010300 Disease PD - + MESH:D012402 Chemical rotenone - - MESH:D028361 - Disease - - mitochondrial abnormalities - INTRO title_3 32731 alpha-syn transgenesis - + 20617 Gene 293 @@ -84933,218 +85601,200 @@ paragraph 32754 Different transgenic models have been developed that overexpress wild type, A53T, A30P and truncated alpha-syn. However, despite initial results with mutated alpha-syn in rodents, they have failed to translate into truly effective transgenic models of PD. Attempts to overexpress or to knock-out alpha-syn in rodents have produced a variety of pathological abnormalities, including aggregate formation. However, no clear loss of dopaminergic neurons is observed. On the other hand, viral transduction induces rapid neurodegeneration but it is limited to the region targeted. The use of cell-type promoters, such as tyrosine hydroxylase, is a limitation to transgenic overexpression that prevents the induction of the broad brain pathology observed in humans. However, this localized overexpression of alpha-syn (or truncated forms) may be useful to address specific features of PD. A variety of transgenic models using different promoters that drive broad expression have induced the graded appearance of alpha-syn aggregations with no fibrillar morphology. One particularly interesting model is the transgenic A53T alpha-syn under the control of the mouse prion promoter, which overexpresses the mutant protein in neurons. These mice develop progressive motor failure at 8 months of age leading to paralysis and death. Inclusions have been detected in different brain areas like the SN, raphe, pons, pontine reticular nuclei, locus coeruleus and deep cerebellar nuclei. The biochemical characterization of these inclusions showed SDS-insoluble alpha-syn aggregation into dimers, trimers and multimers, and these LB-like inclusions have a filamentous structure like human LBs. However, as indicated, a major drawback of this model is that no dopaminergic neurodegeneration is detected in the SN. Conversely, there is neurodegeneration in areas not affected in PD, such as among motor neurons. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 21823 Gene 307 tyrosine hydroxylase - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 alpha-syn - + 9606 Species humans - + 10090 Species mouse - + 36469 Species prion - + 10090 Species mice - + 9606 Species human - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 10090 Species transgenic - + 185869 Species T alpha - + 16473 Gene syn - + p.A30P;RS#:104893878 ProteinMutation A30P - + MESH:D019636 Disease neurodegeneration - - MESH:C579880 - Disease - - filamentous - - + MESH:D010300 Disease PD - + c.53A>T;RS#:104893877 DNAMutation A53T - + MESH:D019636 Disease neurodegeneration - - MESH:D017093 + + MESH:D001259 Disease motor failure - - MESH:C032259 - Chemical - - SDS - - + MESH:D010300 Disease PD - - MESH:D003643 - Disease - - death - - - MESH:D013568 - Disease - - pathological abnormalities + + MESH:D012967 + Chemical + + SDS - + MESH:D010243 Disease paralysis - - MESH:D019636 - Disease - - neurodegeneration - - + MESH:D010300 Disease PD + + MESH:D019636 + Disease + + neurodegeneration + INTRO title_3 34647 LRRK2 transgenesis - + 66725 Gene 18982 @@ -85157,117 +85807,123 @@ paragraph 34666 LRRK2 null mice have no overt dopaminergic deficit or clear pathology in the brain, although age-dependent renal atrophy is observed that is associated with the aggregation of alpha-syn in renal tubules of the aged kidney. The R1441C mutation in LRRK2 generated by a knock-in strategy in mice does not induce dopaminergic neurodegeneration but rather, DA neurotransmission and D2 receptor dysfunction. However, no obvious accumulation of alpha-syn is detected. Mice overexpressing the G2019S mutation in LRRK2 do not reproduce any obvious gross neuropathological phenotypes nor is alpha-syn aggregation observed, although this LRRK2 mutation may accelerate the pathogenic phenotype of A53T alpha-syn mice. - + 66725 Gene 18982 LRRK2 - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 13489 Gene 22561 D2 receptor - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 10090 Species mice - + 10090 Species mice - + 10090 Species Mice - + 10090 Species mice - + 185869 Species T alpha - + 16473 Gene syn - + + MESH:D009461 + Disease + + deficit + + p.R1441C;RS#:33939927 ProteinMutation R1441C - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:D007674 Disease renal atrophy - + MESH:D019636 Disease @@ -85285,34 +85941,34 @@ paragraph 35398 Several studies have suggested a role for autophagy in PD. The autophagy-lysosome system is a catabolic pathway involved in protein and organelle degradation. Several types of autophagy have been described, including: microautophagy; macroautophagy, engulfing large structures; and chaperone-mediated autophagy that degrades only soluble proteins in a selective manner. Cellular homeostasis of autophagy is crucial to maintain the balance between healthy and unhealthy cells. In PD and related LB diseases an accumulation of autophagosomes has been described, coupled with a reduction of lysosomal markers in nigral dopaminergic neurons, suggesting a defect in lysosome-mediated clearance of alpha-syn aggregates. Indeed, alpha-syn is physiologically degraded by both the ubiquitin-proteasome and the autophagy-lysosome system (even by the chaperone-mediated mechanism), although the mutant forms of alpha-syn appear to inhibit their own degradation. Hence, these pathways would appear to participate in LB formation. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + MESH:D010300 Disease PD - + MESH:D010300 Disease @@ -85324,107 +85980,107 @@ paragraph 36416 In mice, deletion of the gene essential for macroautophagy (Atg7) in dopaminergic neurons induces progressive moderate dopaminergic loss in the SN and striatal DA depletion at an age of 9 months, although most dopaminergic neurons are resistant to the long-term stress induced by impairing autophagy. These mice have ubiquitinated-SQSTM1 inclusions in dopaminergic neurons but no abnormal aggregation of alpha-syn. In PD patients, the translocation to the nucleus of the TFEB transcription factor (a regulator of autophagy), is dampened in midbrain dopaminergic neurons and it co-localizes with alpha-syn in LBs. Using adeno-associated viral vectors to model alpha-syn transgenesis in rats, under the control of the synapsin-1 promoter and the WPRE enhancer, stimulation of TFEB and Beclin (an activator of autophagy) was shown to rescue nigral DA neurons from alpha-syn toxicity. These results identify interesting new mechanisms, however no clear alpha-syn aggregates have yet been formed by manipulating the autophagy-lysosome system. - + 74244 Gene 4662 Atg7 - + 18412 Gene 31202 SQSTM1 - + 20617 Gene 293 alpha-syn - + 7942 Gene 5182 TFEB - + 6622 Gene 293 alpha-syn - + 29219 Gene 293 alpha-syn - + 24949 Gene 48483 synapsin-1 - + 316214 Gene 5182 TFEB - + 29219 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 10090 Species mice - + 10090 Species mice - + 9606 Species patients - + 10116 Species rats - + MESH:D010300 Disease PD - + MESH:D064420 Disease @@ -85436,7 +86092,7 @@ title_3 37454 Neurotrophic factors: Smad3 deficiency - + 17127 Gene 55937 @@ -85449,107 +86105,107 @@ paragraph 37493 GDNF and its close relative Neurturin provide functional rescue of nigro-striatal dopaminergic neurons after 6-OHDA or MPTP treatment. These results led to clinical trials into the use of these molecules in patients with PD, although the use of exogenous GDNF in clinical trials produced inconclusive results. The neuroprotective effect of this neurotrophic factor may not be translated to the clinic as GDNF fails to protect against alpha-syn-induced toxicity, probably due to the blockade of GDNF signaling by alpha-syn. Indeed, transgenesis of alpha-syn induced by viral overexpression in the nigro-striatum, induces dystrophic terminals in the striatum containing alpha-syn aggregates, which are not modified by exogenous administration of GDNF. - + 14573 Gene 433 GDNF - + 2668 Gene 433 GDNF - + 14573 Gene 433 neurotrophic factor - + 14573 Gene 433 GDNF - + 20617 Gene 293 alpha-syn - + 14573 Gene 433 GDNF - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 14573 Gene 433 GDNF - + 9606 Species patients - + MESH:D015632 Chemical MPTP - + MESH:D064420 Disease toxicity - + MESH:D020388 Disease dystrophic - + MESH:D016627 Chemical 6-OHDA - + MESH:D010300 Disease @@ -85561,404 +86217,404 @@ paragraph 38243 Recently, intracellular TGF-beta1 signaling has been implicated in different pathological events related to PD in a mouse model, including LB-like formation. In humans, TGF-beta1 is up-regulated in the striatum and in the ventricular cerebrospinal fluid of patients with PD. Moreover, active TGF-beta1 overexpression in the nigro-striatal system of MPTP-treated mice using adenoviral vectors produces poorer survival of dopaminergic neurons and higher levels of striatal DA depletion. However, since the effects of TGF-beta1 are dose- and context-dependent, its overexpression may introduce a bias in studies on animal models. Smad3 deficiency, a molecule involved in the intracellular TGF-beta1 signaling cascade, provides us with a new and interesting model of PD, as it promotes selective postnatal neurodegeneration of dopaminergic midbrain neurons, strong MAO-mediated catabolism of DA in the striatum and oxidative stress, as well as dampening the trophic and astrocytic support to dopaminergic neurons. Interestingly, Smad3 deficiency induces the formation of alpha-syn inclusions in selected brain areas, which accumulate with age in a progressive and gene dosage dependent manner. These alpha-syn inclusions appear in both the perikaryon (SN and paralemniscal nucleus) and in neurites (in the motor and cingulate cortices, striatum, corpus callosum and spinal cord). Indeed, these alpha-syn deposits are phosphorylated at Ser129 and ubiquitinated, and they form a core/halo distribution that resembles the deposits observed in human LBs. In other brain areas alpha-syn expression is associated with an irregular morphology, increased alpha-syn-ir staining and neurite thickness (pontine nuclei, cerebellar white matter, cerebral peduncle, diencephalic nuclei and internal capsule). Moreover, alpha-syn inclusions are also detected in glial cells in the cerebellum and spinal cord. Biochemical analyses show the presence of detergent-insoluble dimers, trimers and oligomers of alpha-syn in the ventral midbrain, motor cortex and spinal cord. - + 21803 Gene 540 TGF-beta1 - + 7040 Gene 540 TGF-beta1 - + 21803 Gene 540 TGF-beta1 - + 21803 Gene 540 TGF-beta1 - + 17127 Gene 55937 Smad3 - + 21803 Gene 540 TGF-beta1 - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 10090 Species mouse - + 9606 Species humans - + 9606 Species patients - + 10090 Species mice - + 9606 Species human - + + - + Chemical + + Ser129 + + MESH:D015632 Chemical MPTP - - MESH:D014693 - Disease - - ventricular cerebrospinal - - - MESH:C530429 + + MESH:D004298 Chemical - - Ser + + DA - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - - MESH:D007027 - Disease - - diencephalic nuclei - - + MESH:D019636 Disease neurodegeneration + + MESH:D007027 + Disease + + diencephalic + INTRO paragraph 40293 There is currently increasing attention being paid to the non-motor symptoms of PD, such as cognitive impairment and behavioral disorders. The hippocampus is implicated in physiological learning and memory, as well as in the cognitive dysfunction seen in some PD patients where there is an interaction with the dopaminergic system. The hippocampus of PD patients is also affected by the presence of LBs and in mouse models, alpha-syn modulates adult neurogenesis in the dentate gyrus. Indeed, the accumulation of extracellular alpha-syn oligomers impairs hippocampal LTP and it enhances basal synaptic transmission. Both triplication and point mutations of SNCA are associated with cortical and subcortical LB accumulation, and such patients are clinically diagnosed as PD with dementia. These data suggest a role for the alpha-syn aggregates in the cognitive impairment observed in demented PD patients. Similarly, Smad3 deficient mice have alpha-syn inclusions in the neuronal layers of the hippocampus. Smad3 is strongly expressed in hippocampal neurons and Smad3 deficiency induces a strong decrease in adult neurogenesis in the dentate gyrus, inducing the apoptosis of early stage and highly proliferative intermediate precursor cells (IPCs). Indeed, Smad3 deficiency abolishes the induction of LTP in the dentate gyrus but not in the CA1, highlighting the specificity of this effect. Both neurogenesis and LTP induction in the adult hippocampus are two aspects of hippocampal brain plasticity related to learning and memory that decline with age, and as a result of neurological disorders. Smad3 deficiency sheds light on a new interesting pathological mechanism and provides a new model of PD to be explored in which dopaminergic dysfunction, widespread alpha-syn inclusions and cognitive impairment co-exist. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 6622 Gene 293 SNCA - + 6622 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 17127 Gene 55937 Smad3 - + 17127 Gene 55937 Smad3 - + 12346 Gene 20414 CA1 - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 109912 Gene LTP - + 9606 Species patients - + 9606 Species patients - + 10090 Species mouse - + 9606 Species patients - + 9606 Species patients - + 10090 Species mice - + 109912 Gene LTP - + 109912 Gene LTP - + MESH:D003072 Disease cognitive impairment - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - + MESH:D003072 Disease cognitive impairment - + MESH:D003704 Disease dementia - + MESH:D003072 Disease cognitive dysfunction - + MESH:D010300 Disease PD - - MESH:D002653 - Disease - - behavioral disorders - - + MESH:D003072 Disease cognitive impairment - + + MESH:D010300 + Disease + + PD + + + MESH:D001523 + Disease + + behavioral disorders + + MESH:D009422 Disease neurological disorders - + MESH:D010300 Disease PD - + MESH:D010300 Disease PD - - MESH:D010300 - Disease - - demented PD - INTRO @@ -85971,109 +86627,121 @@ paragraph 42130 Several invertebrate models have been developed that recapitulate key features of human PD. Drosophila models of alpha-syn overexpression show loss of dopaminergic neurons, locomotor dysfunction and formation of alpha-syn inclusions. Indeed, these inclusions are observed as a core with peripherally radiating filaments. Despite the simplicity of the nematode model Caenorhabditis elegans, the presence of eight dopaminergic neurons has allowed their evaluation following parkinsonian toxic and genetic insult. Overexpression of human wild-type or mutant alpha-syn on C. elegans induces dopaminergic neurodegeneration and the formation of alpha-syn aggregates with fibrillar morphology. It is interesting to note that both Drosophila and C. elegans do not express alpha-syn and thus, other genes like LRRK2 cannot be studied in the context of alpha-syn inclusion formation. These models also lack the complexity of vertebrates and hence, they are not perfect models of PD but rather, they may be useful for comprehensive genetic analysis and drug screening. - + 6622 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 9606 Species human - + 6239 Species Caenorhabditis elegans - + 9606 Species human - + 6239 Species C. elegans - + 6239 Species C. elegans - + + 7227 + Species + + Drosophila + + + 7227 + Species + + Drosophila + + 16473 Gene syn - + 16473 Gene syn - + 16473 Gene syn - + 16473 Gene syn - + 16473 Gene syn - + + MESH:D010302 + Disease + + parkinsonian + + MESH:D010300 Disease PD - - MESH:D019636 - Disease - - neurodegeneration - - + MESH:D010300 Disease - - parkinsonian toxic + + PD - + MESH:D009069 Disease locomotor dysfunction - - MESH:D010300 + + MESH:D019636 Disease - - PD + + neurodegeneration @@ -86081,7 +86749,7 @@ title_1 43188 Dopamine metabolism and alpha-syn - + MESH:D004298 Chemical @@ -86128,16 +86796,28 @@ toxicity - MESH:D007980 + MESH:D004298 Chemical - - L-Dopa + + DA - MESH:D065105 + MESH:D004298 + Chemical + + DA + + + MESH:D004298 Chemical - - aromatic amino acid + + DA + + + MESH:D007980 + Chemical + + L-Dopa @@ -86145,211 +86825,236 @@ paragraph 43583 The Smad3 deficient mouse model described earlier also associates alpha-syn and DA metabolism, whereby Smad3 deficiency drives alpha-syn overexpression and aggregation, as well as the deregulation of DA turnover by inducing MAO-dependent DA catabolism, provoking a loss of dopaminergic neurons. Other mouse models, such as alpha-syn, parkin, DJ-1 or PINK1 null mice, also display deregulated striatal DA metabolism, release or re-uptake, although without dopaminergic neuronal loss. Increased extracellular DA, decreased DAT, increased GSH levels and differences in the catabolism of DA have all been described in Parkin deficient mice. However, neither the number of dopaminergic neurons nor the striatal DA levels and turnover are altered in DJ-1 or PINK knock-out mice. Nevertheless, the evoked DA overflow in these mice diminishes due to increased DA uptake or to the decreased quantal release of DA, respectively. The alpha-syn knock-out mice have less striatal DA, enhanced activity-dependent DA release and smaller reserve pools of synaptic vesicles, which may be due to a uninhibited DA vesicular release. All these mutant mice illustrate that impaired presynaptic DA metabolism, release or re-uptake may be common to PD, although the presence of alpha-syn aggregates and DA neuronal loss is only detected in Smad3 deficient mice. Indeed, 3,4-dihydroxyphenylacetaldehyde (DOPAL), a MAO intermediate metabolite of DA, can induce toxic aggregation of alpha-synuclein. These data suggest that DA metabolism and alpha-syn interactions may underlie the susceptibility of SN neurodegeneration in PD. - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 57320 Gene 38295 DJ-1 - + 68943 Gene 32672 PINK1 - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-synuclein - + 20617 Gene 293 alpha-syn - + 13162 Gene 55547 DAT - + 10090 Species mouse - + 10090 Species mouse - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 10090 Species mice - + 57320 Gene 38295 DJ-1 - + MESH:D009410 Disease neuronal loss - + + MESH:D004298 + Chemical + + DA + + Disease - - Parkin deficient + + SN neurodegeneration - + MESH:D010300 Disease PD - - MESH:D006402 - Disease - - toxic aggregation + + MESH:D004298 + Chemical + + DA - + + MESH:D004298 Chemical - - DOPAL + + DA - + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + MESH:D010300 Disease PD - - MESH:D005978 + + MESH:C007430 Chemical - - GSH + + DOPAL - - MESH:D019636 - Disease - - SN neurodegeneration + + MESH:D004298 + Chemical + + DA - + MESH:D009410 Disease neuronal loss - + + - + Chemical + + GSH + + MESH:C007430 Chemical @@ -86361,133 +87066,145 @@ paragraph 45185 It was recently suggested that the spread of the pathological elements in PD, as described in the Braak stages, can occur by neuron-to-neuron transmission of aggregates to healthy cells. In this hypothesis, preformed fibrils of alpha-syn enter neurons, probably by endocytosis, to recruit soluble endogenous alpha-syn into insoluble LB- and LN-like aggregates. Interestingly, peripheral inoculation of alpha-syn fibrils by intramuscular injections can propagate the pathogenic protein to brain nuclei. However, another study showed that alpha-syn fibrils injected into transgenic mice overexpressing alpha-syn mutations promotes the widespread formation of alpha-syn inclusions in the brain of A53T but not E46K mutant mice, nor in non-transgenic mice. The authors suggest inespecificity with neurofilament of the antibodies used and that only A53T mutant mice have the capacity to induce alpha-syn aggregation upon exogenous administration of alpha-syn. This aggregate transmission may be related to the existence of extracellular forms of alpha-syn that are released from neurons and glia by exocytosis. Indeed, extracellular alpha-syn may induce DA release in the striatum, establishing a new link between alpha-syn and DA metabolism. Furthermore, alpha-syn function may be related to the reorganization of plasma membrane microdomains. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 10090 Species transgenic mice - + 10090 Species mice - + 10090 Species transgenic mice - + 10090 Species mice - + c.53A>T;RS#:104893877 DNAMutation A53T - + c.53A>T;RS#:104893877 DNAMutation A53T - + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + MESH:D010300 Disease PD - + p.E46K;RS#:104893875 ProteinMutation @@ -86505,55 +87222,61 @@ paragraph 46564 Advances in cell-free systems and model cell systems have shed light on the process of alpha-syn aggregation. The role proposed for alpha-syn is in the modulation of neurotransmitter release in the presynaptic nerve terminal, as well as influencing DA neurotransmitter biosynthesis, vesicle trafficking and exocytosis. In the cytoplasm and/or vesicle lumen, alpha-syn is present as an intrinsically disordered protein (IDP: i.e. a monomer that lacks a well-organized secondary structure), yet when bound to membranes it adopts several conformations, such as an extended alpha-helix or a broken-helix. Indeed, despite the overwhelming evidence that alpha-syn is a disordered monomer in solution, two recent reports suggest that the native protein exists as a helical tetramer under physiological conditions, with reduced aggregation tendencies, and that the dissociation of the tetramer into monomeric subunits promotes toxic aggregation. Indeed, when monomeric alpha-syn binds to membranes, it changes its conformation to a partially helical form. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 15926 Gene 21195 IDP - - MESH:D006402 + + MESH:D030342 Disease - - toxic aggregation + + disordered - + + MESH:D030342 + Disease + + disordered + + MESH:C536965 Disease @@ -86565,14 +87288,14 @@ paragraph 47612 Many IDPs are known to interact with a large number of proteins, serving as a nodes or hubs, in a way that IDPs undergo a disorder-to-order transition upon interaction with specific partners. More than 50 proteins have been reported to interact with alpha-syn, although it is unknown the proportion unfolded alpha-syn or that which has adopted a secondary structure as a consequence of binding within the cell. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 @@ -86585,37 +87308,31 @@ paragraph 48023 The alpha-syn peptide has 140 amino acids, with 3 distinct regions: a N-terminal (1-60 residues) that contain four imperfect repeats of KTKEGV motifs; a NAC region (61-95 residues), with 3 additional KTKEGV repeats, and the hydrophobic and amyloidogenic NAC region; a C-terminus (96-140 residues) that is enriched in acidic and proline residues, and that facilitates interactions with different proteins. - + 20617 Gene 293 alpha-syn - - MESH:D009584 + + - Chemical - - N + + NAC - - MESH:C489032 + + MESH:D011392 Chemical proline - - MESH:C086501 + + - Chemical NAC - - MESH:C086501 - Chemical - - NAC - FIG @@ -86624,127 +87341,139 @@ fig_caption 48431 Models of alpha-syn aggregation and LBs formation. A alpha-Syn is present in the vesicle lumen and in the cytoplasma as an intrinsically disordered protein. alpha-Syn bound to membranes has distinct conformation such as an extended alpha-helix or a broken-helix. In the pathological context, disordered monomers may lead to oligomerization and fibril formation, following a nucleation-dependent process, in which monomers are added to existing aggregates. B Rotenone administration in rats, A53T alpha-syn transgenesis in mice and Smad3 deficient mice are interesting models to study LB formation. While A53T alpha-syn transgenesis and Smad3 deficiency can modulate DA metabolism, rotenone and Smad3 deficiency induce oxidative stress, mechanisms that may participate in LBs formation. Indeed, proteasome and autophagy inhibitors may impair degradation of alpha-syn. LRRK2 mutations may participate in LB formation by altering autophagy and alpha-syn solubility. - + 20617 Gene 293 alpha-syn - + 16473 Gene Syn - + 16473 Gene Syn - + 17127 Gene 55937 Smad3 - + 17127 Gene 55937 Smad3 - + 17127 Gene 55937 Smad3 - + 20617 Gene 293 alpha-syn - + 66725 Gene 18982 LRRK2 - + 20617 Gene 293 alpha-syn - + 14960 Gene 123820 A alpha - + 10116 Species rats - + 10090 Species mice - + 10090 Species mice - + 185869 Species T alpha - + 185869 Species T alpha - + 16473 Gene syn - + 16473 Gene syn - + + MESH:D030342 + Disease + + disordered + + MESH:D012402 Chemical rotenone - + MESH:D012402 Chemical - - B Rotenone + + Rotenone + + + MESH:D030342 + Disease + + disordered @@ -86752,22 +87481,28 @@ paragraph 49395 The major constituent of LBs is a fibrillar form of alpha-syn that adopts a beta-sheet structure and hence, the disordered monomer or helical tetramer is transformed into highly organized fibrils in the course of this pathology. In vitro studies suggest a nucleation-dependent process due to a conformational transition to anti-parallel beta-sheet structures, including a committed step of alpha-syn dimer formation. Nucleation follows a sigmoidal growth profile, with an initial lag phase where the protein changes to a partially folded intermediate to form nuclei with oligomers (Figure 1). This conformational change exposes the NAC domain that can participate in hydrophobic interactions that may initiate the aggregation process. During the growth phase, the nucleus adds monomers to form larger oligomers, polarized protofibrils and finally, fibrils. In the last steady-state phase both the fibrils and monomers appear to be in equilibrium, this model therefore reflecting the addition of monomers to existing aggregates. Recently, a key conformational intermediate was characterized after oligomer formation, with the appearance of stable and compact oligomers that are more damaging to cells (Figure 1). Indeed, it seems that the assembly process can be reversed and that fibrils may disaggregate to form these stable, cytotoxic oligomers. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - - MESH:C086501 + + MESH:D030342 + Disease + + disordered + + + - Chemical NAC @@ -86778,56 +87513,56 @@ paragraph 50743 The pathogenic aggregation of alpha-syn can be modulated by endogenous and exogenous factor, such as metals and pesticides, genetic mutations in SNCA, post-translational modifications and protein-protein interactions. It is not clear how alpha-syn bound to membranes, such as synaptic vesicles or presynaptic plasma membrane, can aggregate. One model proposes that the broken-helix membrane bound state of alpha-syn releases its C-terminal region, converting the protein into a partially helical membrane bound state, which may lead to oligomerization and fibril formation. Alternatively, alpha-helical forms of alpha-syn bound to membranes may inhibit fibril formation, and membrane-bound alpha-syn monomers may be protected from aggregation. Indeed, oligomers have a high propensity to bind to membranes, which may promote permeabilization and disrupt cellular homeostasis. Furthermore, alpha-syn localizes to the nerve terminal, where modest increments in alpha-syn (such as those produced by gene duplication) inhibit neurotransmitter release by reducing synaptic vesicle density and by altering vesicle reclustering after endocytosis. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 SNCA - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 @@ -86840,45 +87575,45 @@ paragraph 51883 By contrast, oligomers might be kinetically detained by interactions with small molecules, inducing secondary structures such as annular pores or spheres. The point mutations detected in the SNCA gene of PD patients are also endogenous factors that could accelerate alpha-syn aggregation in vitro, with A53T and G46L forming oligomers and fibrils, and A30P forming oligomers but not fibrils. - + 6622 Gene 293 SNCA - + 6622 Gene 293 alpha-syn - + 9606 Species patients - + p.G46L ProteinMutation G46L - + MESH:D010300 Disease PD - + p.A30P;RS#:104893878 ProteinMutation A30P - + c.53A>T;RS#:104893877 DNAMutation @@ -86896,107 +87631,119 @@ paragraph 52320 Several post-translational modifications of alpha-syn may occur, such as phosphorylation, truncation, ubiquitination, nitration, sumoylation and enzymatic cross-linking. In LBs, phosphorylation at serine 129 is a common alpha-syn modification, although its role is unclear if we consider that overexpression of the phosphorylated serine 129 isoform in animal models does not produce toxicity. The majority of alpha-syn is mono- to tri-ubiquinated, and while poly-ubiquitination serves as a signal for alpha-syn degradation by the proteasome, it does not seem to be required for alpha-syn fibrillation and LB formation. Nevertheless, an interplay between phosphorylation and ubiquitination may render the protein more susceptible to aggregation. It is estimated that 85% of all human proteins undergo Nalpha-acetylation due to the activity of Nalpha-acetyltransferases, probably influencing the subcellular localization of proteins, their rate of synthesis and protein-protein interactions. Tetramers of alpha-syn seem to be Nalpha-acetylated, as is probably are the aggregated alpha-syn isolated from PD deposits. However, Nalpha-acetylation does not seem to alter protein aggregation but more likely, lipid binding. Small amounts of various C-terminal truncated forms of alpha-syn have been detected in LBs, which exhibit greater fibrillation capacity. Furthermore, four alternatively spliced forms add another level of complexity. Compared to the canonical alpha-syn, the three alternative isoforms aggregate significantly less, forming shorter fibrils that are arranged in parallel or with anular structures. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 9606 Species human - + + MESH:D014693 + Disease + + fibrillation + + MESH:D064420 Disease toxicity - + MESH:D010300 Disease PD deposits - - MESH:C047902 + + MESH:D012694 Chemical serine - - MESH:C017937 + + MESH:D014693 + Disease + + fibrillation + + + MESH:D008055 Chemical - - poly + + lipid - - MESH:C047902 + + MESH:D012694 Chemical serine - - MESH:D058495 + + MESH:D066263 Disease protein aggregation @@ -87007,53 +87754,53 @@ paragraph 53932 Despite the huge amount of research into the properties of alpha-syn aggregation, there is still no coherent picture on the structure, dynamics, and the physiological and pathological roles of alpha-syn. New animal models for PD need to explore LB formation in the context of neurodegeneration and DA metabolism, such as the rotenone, alpha-syn transgenesis and Smad3 deficient mice, in order to clearly understand the pathological mechanism of LB formation as well as to attain effective therapies for this disease. - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 20617 Gene 293 alpha-syn - + 17127 Gene 55937 Smad3 - + 10090 Species mice - + MESH:D010300 Disease PD - + MESH:D012402 Chemical rotenone - + MESH:D019636 Disease @@ -87077,7 +87824,7 @@ paragraph 54466 Dopamine - + MESH:D004298 Chemical @@ -87095,6 +87842,12 @@ paragraph 54479 Intrinsically disordered protein + + MESH:D030342 + Disease + + disordered + ABBR @@ -87125,7 +87878,7 @@ paragraph 54541 PD - + MESH:D010300 Disease @@ -87137,7 +87890,7 @@ paragraph 54544 Parkinson's disease - + MESH:D010300 Disease @@ -87155,11 +87908,6 @@ paragraph 54569 Substantia nigra - - Disease - - Substantia nigra - ABBR @@ -87202,7 +87950,7 @@ ref 54702 The history of Parkinson's disease: early clinical descriptions and neurological therapies - + MESH:D010300 Disease @@ -87220,7 +87968,7 @@ 933 54795 Paralysis agitans - + MESH:D010300 Disease @@ -87240,17 +87988,12 @@ 319 54813 Substantia nigra and Parkinson's disease: a brief history of their long and intimate relationship - + MESH:D010300 Disease Parkinson's disease - - Disease - - Substantia nigra - surname:Ide;given-names:SE @@ -87283,14 +88026,14 @@ surname:Dehejia;given-names:A 54913 Mutation in the alpha-synuclein gene identified in families with Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -87309,13 +88052,13 @@ 250 55000 Multiple hit hypotheses for dopamine neuron loss in Parkinson's disease - + MESH:D004298 Chemical dopamine - + MESH:D010300 Disease @@ -87335,7 +88078,7 @@ 474 55074 Non-motor symptoms of Parkinson's disease: dopaminergic pathophysiology and treatment - + MESH:D010300 Disease @@ -87356,7 +88099,7 @@ 145 55162 The heterogeneity of idiopathic Parkinson's disease - + MESH:D010300 Disease @@ -87378,23 +88121,17 @@ 341 55216 Neuronal loss is greater in the locus coeruleus than nucleus basalis and substantia nigra in Alzheimer and Parkinson diseases - - MESH:D000544 - Disease - - Alzheimer - - + MESH:D009410 Disease Neuronal loss - - MESH:D010300 + + MESH:D000544 Disease - - Parkinson diseases + + Alzheimer and Parkinson diseases @@ -87417,13 +88154,13 @@ surname:Kalbe;given-names:E 55342 Dementia in Parkinson disease: functional imaging of cholinergic and dopaminergic pathways - + MESH:D010300 Disease Parkinson disease - + MESH:D003704 Disease @@ -87446,7 +88183,7 @@ surname:Roytta;given-names:M 55433 Loss of cholinergic neurons in the pedunculopontine nucleus in Parkinson's disease is related to disability of the patients - + MESH:D010300 Disease @@ -87467,14 +88204,14 @@ 1595 55559 Hypocretin (orexin) cell loss in Parkinson's disease - + 15171 Gene 1166 Hypocretin - + MESH:D010300 Disease @@ -87506,11 +88243,11 @@ 272 55626 Neuropathology of Parkinson's disease - + MESH:D010300 Disease - - Parkinson's disease + + Neuropathology of Parkinson's disease @@ -87529,14 +88266,14 @@ surname:Hyman;given-names:BT 55666 alpha-Synuclein immunoreactivity in dementia with Lewy bodies: morphological staging and comparison with ubiquitin immunostaining - + 20617 Gene 293 alpha-Synuclein - + MESH:D003704 Disease @@ -87556,11 +88293,11 @@ S12 55796 Neuropathological spectrum of synucleinopathies - - MESH:D002659 + + MESH:D000080874 Disease - - Neuropathological spectrum of synucleinopathies + + synucleinopathies @@ -87582,18 +88319,24 @@ surname:Kikuchi;given-names:E 55844 Heterogeneity of nigral and cortical Lewy bodies differentiated by amplified triple-labeling for alpha-synuclein, ubiquitin, and thiazin red - + 20617 Gene 293 alpha-synuclein - + MESH:C419142 Chemical - - thiazin + + thiazin red + + + MESH:D020961 + Disease + + cortical Lewy bodies @@ -87613,7 +88356,7 @@ surname:Mizusawa;given-names:H 55985 Pale neurites, premature alpha-synuclein aggregates with centripetal extension from axon collaterals - + 20617 Gene 293 @@ -87634,13 +88377,13 @@ 684 56086 Role of oxidative damage in protein aggregation associated with Parkinson's disease and related disorders - - MESH:D058495 + + MESH:D066263 Disease protein aggregation - + MESH:D010300 Disease @@ -87680,33 +88423,27 @@ surname:Izumiyama;given-names:Y 56233 Immunoelectron-microscopic demonstration of NACP/alpha-synuclein-epitopes on the filamentous component of Lewy bodies in Parkinson's disease and in dementia with Lewy bodies - + 20617 Gene 293 NACP - + 20617 Gene 293 alpha-synuclein - + MESH:D003704 Disease dementia - - MESH:C579880 - Disease - - filamentous - - + MESH:D010300 Disease @@ -87763,7 +88500,7 @@ surname:Jakes;given-names:R 56603 Alpha-synuclein in Lewy bodies - + 20617 Gene 293 @@ -87784,14 +88521,14 @@ 524 56634 alpha-Synuclein posttranslational modification and alternative splicing as a trigger for neurodegeneration - + 20617 Gene 293 alpha-Synuclein - + MESH:D019636 Disease @@ -87817,14 +88554,14 @@ surname:Rovelli;given-names:G 56741 Leucine-rich repeat kinase 2 is associated with the endoplasmic reticulum in dopaminergic neurons and accumulates in the core of Lewy bodies in Parkinson disease - + 66725 Gene 18982 Leucine-rich repeat kinase 2 - + MESH:D010300 Disease @@ -87848,7 +88585,7 @@ surname:Takahashi;given-names:H 56903 Accumulation of histone deacetylase 6, an aggresome-related protein, is specific to Lewy bodies and glial cytoplasmic inclusions - + 15185 Gene 31353 @@ -87873,18 +88610,18 @@ surname:Takahashi;given-names:H 57032 Endosomal sorting related protein CHMP2B is localized in Lewy bodies and glial cytoplasmic inclusions in alpha-synucleinopathy - + 68942 Gene 8534 CHMP2B - - MESH:D005935 + + MESH:D000080874 Disease - - alpha-synucleinopathy + + synucleinopathy @@ -87943,7 +88680,7 @@ 23 57297 Poor and protracted myelination as a contributory factor to neurodegenerative disorders - + MESH:D019636 Disease @@ -87975,14 +88712,14 @@ surname:Berendse;given-names:HW 57386 Patterns of alpha-synuclein pathology in incidental cases and clinical subtypes of Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88004,17 +88741,17 @@ 372 57489 Widespread occurrence of argyrophilic glial inclusions in Parkinson's disease - + MESH:D010300 Disease Parkinson's disease - - MESH:D004194 + + MESH:C537394 Disease - - argyrophilic glial inclusions + + argyrophilic glial @@ -88031,14 +88768,14 @@ 241 57569 Development of alpha-synuclein immunoreactive astrocytes in the forebrain parallels stages of intraneuronal pathology in sporadic Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88062,7 +88799,7 @@ surname:Revesz;given-names:T 57721 A clinico-pathological study of subtypes in Parkinson's disease - + MESH:D010300 Disease @@ -88083,7 +88820,7 @@ 491 57787 A critical evaluation of the Braak staging scheme for Parkinson's disease - + MESH:D010300 Disease @@ -88105,13 +88842,13 @@ 415 57863 The progression of pathology in longitudinally followed patients with Parkinson's disease - + 9606 Species patients - + MESH:D010300 Disease @@ -88137,13 +88874,13 @@ surname:Ahlskog;given-names:JE 57955 Familial aggregation of Parkinson's disease: The Mayo Clinic family study - + 162683 Species Mayo - + MESH:D010300 Disease @@ -88170,7 +88907,7 @@ surname:Gugmundsson;given-names:G 58031 Familial aggregation of Parkinson's disease in Iceland - + MESH:D010300 Disease @@ -88189,7 +88926,7 @@ 318 58088 Genetics of Parkinson disease: paradigm shifts and future prospects - + MESH:D010300 Disease @@ -88211,11 +88948,11 @@ 271 58156 Review: Familial Parkinson's disease-genetics, clinical phenotype and neuropathology in relation to the common sporadic form of the disease - + MESH:D010300 Disease - - Parkinson's disease + + Familial Parkinson's disease @@ -88232,7 +88969,7 @@ 1218 58300 What genetics tells us about the causes and mechanisms of Parkinson's disease - + MESH:D010300 Disease @@ -88271,14 +89008,14 @@ surname:Hague;given-names:S 58380 alpha-Synuclein locus triplication causes Parkinson's disease - + 20617 Gene 293 alpha-Synuclein - + MESH:D010300 Disease @@ -88317,11 +89054,11 @@ surname:Simon;given-names:J 58444 Cloning of the gene containing mutations that cause PARK8-linked Parkinson's disease - + MESH:D010300 Disease - - Parkinson's disease + + PARK8-linked Parkinson's disease @@ -88357,15 +89094,15 @@ surname:Farrer;given-names:M 58531 Mutations in LRRK2 cause autosomal-dominant parkinsonism with pleomorphic pathology - + 66725 Gene 18982 LRRK2 - - MESH:D010300 + + MESH:C537176 Disease autosomal-dominant parkinsonism @@ -88392,14 +89129,14 @@ surname:Graeber;given-names:M 58615 Ala30Pro mutation in the gene encoding alpha-synuclein in Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88432,25 +89169,25 @@ surname:Ros;given-names:R 58695 The new mutation, E46K, of alpha-synuclein causes Parkinson and Lewy body dementia - + 20617 Gene 293 alpha-synuclein - - MESH:D010300 - Disease - - Parkinson and Lewy body dementia - - + p.E46K;RS#:104893875 ProteinMutation E46K + + MESH:D020961 + Disease + + Parkinson and Lewy body dementia + surname:Thomson;given-names:AJ @@ -88477,14 +89214,14 @@ surname:Burdon;given-names:T 58778 Parkinson's disease induced pluripotent stem cells with triplication of the alpha-synuclein locus - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88520,14 +89257,14 @@ surname:Hulihan;given-names:MM 58878 Genomic investigation of alpha-synuclein multiplication and parkinsonism - + 20617 Gene 293 alpha-synuclein - + MESH:D010302 Disease @@ -88557,14 +89294,14 @@ surname:Lee;given-names:DS 58951 Alpha-Synuclein gene duplication is present in sporadic Parkinson disease - + 20617 Gene 293 Alpha-Synuclein - + MESH:D010300 Disease @@ -88596,13 +89333,13 @@ surname:Muller;given-names:T 59025 Familial parkinsonism with synuclein pathology: clinical and PET studies of A30P mutation carriers - + p.A30P;RS#:104893878 ProteinMutation A30P - + MESH:D010302 Disease @@ -88628,30 +89365,30 @@ surname:Katechalidou;given-names:L 59124 Clinical, neuropathological and genotypic variability in SNCA A53T familial Parkinson's disease. Variability in familial Parkinson's disease - + 20617 Gene 293 SNCA - + c.53A>T;RS#:104893877 DNAMutation A53T - + MESH:D010300 Disease familial Parkinson's disease - + MESH:D010300 Disease - - Parkinson's disease + + familial Parkinson's disease @@ -88693,14 +89430,14 @@ surname:Ioannidis;given-names:JP 59269 Collaborative analysis of alpha-synuclein gene promoter variability and Parkinson disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88721,27 +89458,27 @@ 116 59359 Australian data and meta-analysis lend support for alpha-synuclein (NACP-Rep1) as a risk factor for Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + 104354 Gene Rep1 - + 20617 Gene 293 NACP - + MESH:D010300 Disease @@ -88769,14 +89506,14 @@ surname:Lesnick;given-names:TG 59481 alpha-Synuclein gene haplotypes are associated with Parkinson's disease - + 20617 Gene 293 alpha-Synuclein - + MESH:D010300 Disease @@ -88808,14 +89545,14 @@ surname:Tan;given-names:C 59555 Alpha-synuclein haplotypes implicated in risk of Parkinson's disease - + 20617 Gene 293 Alpha-synuclein - + MESH:D010300 Disease @@ -88858,14 +89595,14 @@ surname:Hu;given-names:J 59658 Mapping the subcellular distribution of alpha-synuclein in neurons using genetically encoded probes for correlated light and electron microscopy: implications for Parkinson's disease pathogenesis - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88887,14 +89624,14 @@ 724 59856 Familial Parkinson disease mutations influence alpha-synuclein assembly - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -88917,15 +89654,21 @@ surname:Nicholson;given-names:GA 59928 Clinical and pathological features of a Parkinsonian syndrome in a family with an Ala53Thr alpha-synuclein mutation - + 20617 Gene 293 alpha-synuclein - - MESH:D010300 + + p.A53T;RS#:104893877 + ProteinMutation + + Ala53Thr + + + MESH:D010302 Disease Parkinsonian syndrome @@ -88950,26 +89693,20 @@ surname:Shannon;given-names:KM 60044 Abundant neuritic inclusions and microvacuolar changes in a case of diffuse Lewy body disease with the A53T mutation in the alpha-synuclein gene - + 20617 Gene 293 alpha-synuclein - - MESH:D058225 - Disease - - Abundant neuritic inclusions - - + c.53A>T;RS#:104893877 DNAMutation A53T - + MESH:D020961 Disease @@ -88993,36 +89730,36 @@ surname:Trojanowski;given-names:JQ 60189 Neuronal alpha-synucleinopathy with severe movement disorder in mice expressing A53T human alpha-synuclein - + 10090 Species mice - + 9606 Species human - + MESH:D009069 Disease movement disorder - + + MESH:D000080874 + Disease + + synucleinopathy + + c.53A>T;RS#:104893877 DNAMutation A53T - - MESH:D009410 - Disease - - Neuronal alpha-synucleinopathy - surname:Mohanna;given-names:S @@ -89047,24 +89784,19 @@ surname:Ardah;given-names:MT 60296 Nigrostriatal overabundance of alpha-synuclein leads to decreased vesicle density and deficits in dopamine release that correlate with reduced motor activity - + 20617 Gene 293 alpha-synuclein - + MESH:D004298 Chemical dopamine - - Chemical - - correlate - surname:Maraganore;given-names:D @@ -89085,25 +89817,25 @@ surname:Muenter;given-names:M 60454 Distinctive neuropathology revealed by alpha-synuclein antibodies in hereditary parkinsonism and dementia linked to chromosome 4p - + 20617 Gene 293 alpha-synuclein - - MESH:D009386 - Disease - - hereditary parkinsonism - - + MESH:D003704 Disease dementia + + MESH:D010302 + Disease + + hereditary parkinsonism + surname:Burt;given-names:A @@ -89134,21 +89866,21 @@ surname:Powell;given-names:EH 60584 Genome-wide association study confirms SNPs in SNCA and the MAPT region as common risk factors for Parkinson disease - + 17762 Gene 74962 MAPT - + 20617 Gene 293 SNCA - + MESH:D010300 Disease @@ -89196,7 +89928,7 @@ surname:Gibbs;given-names:JR 60701 Genome-wide association study reveals genetic risk underlying Parkinson's disease - + MESH:D010300 Disease @@ -89236,7 +89968,7 @@ surname:Ito;given-names:C 60785 Genome-wide association study identifies common variants at four loci as genetic risk factors for Parkinson's disease - + MESH:D010300 Disease @@ -89259,14 +89991,14 @@ surname:Singleton;given-names:AB 60905 Comprehensive analysis of LRRK2 in publicly available Parkinson's disease cases and neurologically normal controls - + 66725 Gene 18982 LRRK2 - + MESH:D010300 Disease @@ -89287,13 +90019,13 @@ 393 61022 Genetics of Parkinson disease and essential tremor - + MESH:D014202 Disease tremor - + MESH:D010300 Disease @@ -89312,14 +90044,14 @@ 1160 61073 LRRK2 gene variation and its contribution to Parkinson disease - + 66725 Gene 18982 LRRK2 - + MESH:D010300 Disease @@ -89362,14 +90094,14 @@ surname:Durr;given-names:A 61136 Phenotype, genotype, and worldwide genetic penetrance of LRRK2-associated Parkinson's disease: a case-control study - + 66725 Gene 18982 LRRK2 - + MESH:D010300 Disease @@ -89393,14 +90125,14 @@ surname:Dawson;given-names:TM 61256 Kinase activity of mutant LRRK2 mediates neuronal toxicity - + 66725 Gene 18982 LRRK2 - + MESH:D064420 Disease @@ -89429,14 +90161,14 @@ surname:Papapetropoulos;given-names:S 61315 Lrrk2 and Lewy body disease - + 66725 Gene 18982 Lrrk2 - + MESH:D020961 Disease @@ -89462,21 +90194,21 @@ surname:Poulose;given-names:SM 61343 Disrupted autophagy leads to dopaminergic axon and dendrite degeneration and promotes presynaptic accumulation of alpha-synuclein and LRRK2 in the brain - + 20617 Gene 293 alpha-synuclein - + 66725 Gene 18982 LRRK2 - + MESH:D007635 Disease @@ -89502,31 +90234,18 @@ surname:Lees;given-names:A 61496 Divergent alpha-synuclein solubility and aggregation properties in G2019S LRRK2 Parkinson's disease brains with Lewy Body pathology compared to idiopathic cases - + 20617 Gene 293 alpha-synuclein - - 66725 - Gene - 18982 - - LRRK2 - - + MESH:D010300 Disease - - Parkinson's disease - - - p.G2019S;RS#:34637584 - ProteinMutation - - G2019S + + G2019S LRRK2 Parkinson's disease @@ -89552,11 +90271,11 @@ surname:Eskelson;given-names:C 61659 Lewy body Parkinson's disease in a large pedigree with 77 Parkin mutation carriers - + MESH:D010300 Disease - - Parkinson's disease + + Lewy body Parkinson's disease @@ -89580,7 +90299,7 @@ surname:Manczyk;given-names:N 61744 A molecular explanation for the recessive nature of parkin-linked Parkinson's disease - + MESH:D010300 Disease @@ -89601,7 +90320,7 @@ ref 61832 Ubiquitination increases parkin activity to promote autophagic alpha-synuclein clearance - + 20617 Gene 293 @@ -89629,14 +90348,14 @@ surname:Crews;given-names:L 61921 alpha-Synuclein aggregates interfere with Parkin solubility and distribution: role in the pathogenesis of Parkinson disease - + 20617 Gene 293 alpha-Synuclein - + MESH:D010300 Disease @@ -89679,14 +90398,14 @@ surname:Gwinn-Hardy;given-names:K 62045 Analysis of the PINK1 gene in a large cohort of cases with Parkinson disease - + 68943 Gene 32672 PINK1 - + MESH:D010300 Disease @@ -89720,18 +90439,18 @@ surname:Tarantino;given-names:P 62122 DJ-1 mutations and parkinsonism-dementia-amyotrophic lateral sclerosis complex - + 57320 Gene 38295 DJ-1 - + OMIM:105500 Disease - - parkinsonism-dementia-amyotrophic lateral sclerosis complex + + parkinsonism-dementia-amyotrophic lateral sclerosis @@ -89748,14 +90467,14 @@ 493 62201 Exploring the link between glucocerebrosidase mutations and parkinsonism - + 14466 Gene 68040 glucocerebrosidase - + MESH:D010302 Disease @@ -89790,14 +90509,14 @@ surname:Parkkinen;given-names:L 62274 Glucocerebrosidase mutations in clinical and pathologically proven Parkinson's disease - + 14466 Gene 68040 Glucocerebrosidase - + MESH:D010300 Disease @@ -89845,14 +90564,14 @@ surname:Annesi;given-names:G 62363 Multicenter analysis of glucocerebrosidase mutations in Parkinson's disease - + 14466 Gene 68040 glucocerebrosidase - + MESH:D010300 Disease @@ -89872,19 +90591,19 @@ 366 62441 How many ways an mouse behavioral experiments go wrong? Confounding variables in mouse models of neurodegenerative diseases and how to control them - + 10090 Species mouse - + 10090 Species mouse - + MESH:D019636 Disease @@ -89904,7 +90623,7 @@ ref 62589 Evaluation of animal models of neurobehavioral disorders - + MESH:D019954 Disease @@ -89924,8 +90643,8 @@ 188 62646 The validity of animal models of predisposition to depression - - MESH:D003866 + + MESH:D000275 Disease depression @@ -89944,7 +90663,7 @@ 1391 62708 Animal models of Parkinson's disease: a source of novel treatments and clues to the cause of the disease - + MESH:D010300 Disease @@ -89964,19 +90683,19 @@ 312 62815 Animal models of Parkinson's disease and L-dopa induced dyskinesia: how close are we to the clinic? - + MESH:D007980 Chemical L-dopa - + MESH:D010300 Disease Parkinson's disease - + MESH:D004409 Disease @@ -89996,7 +90715,7 @@ 241 62917 Why hasn't neuroprotection worked in Parkinson's disease? - + MESH:D010300 Disease @@ -90018,19 +90737,19 @@ 3247 62977 Augmentation of phenotype in a transgenic Parkinson mouse heterozygous for a Gaucher mutation - + 10090 Species mouse - + 10090 Species transgenic - + MESH:D010302 Disease @@ -90060,14 +90779,20 @@ surname:Akbarian;given-names:S 63071 Neuroinflammation and alpha-synuclein accumulation in response to glucocerebrosidase deficiency are accompanied by synaptic dysfunction - + 20617 Gene 293 alpha-synuclein - + + MESH:C536122 + Disease + + synaptic dysfunction + + MESH:D005776 Disease @@ -90099,21 +90824,21 @@ surname:Altmeppen;given-names:H 63207 LIMP-2 expression is critical for beta-glucocerebrosidase activity and alpha-synuclein clearance - + 12492 Gene 48353 LIMP-2 - + 14466 Gene 68040 beta-glucocerebrosidase - + 20617 Gene 293 @@ -90133,13 +90858,13 @@ 137 63304 The contribution of the MPTP-treated primate model to the development of new treatment strategies for Parkinson's disease - + MESH:D015632 Chemical MPTP - + MESH:D010300 Disease @@ -90159,7 +90884,7 @@ S29 63428 Functional models of Parkinson's disease: a valuable tool in the development of novel therapies - + MESH:D010300 Disease @@ -90184,13 +90909,13 @@ surname:Kilpatrick;given-names:G 63526 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced parkinsonism in the common marmoset - + MESH:D010302 Disease parkinsonism - + MESH:D015632 Chemical @@ -90210,7 +90935,7 @@ 606 63615 Animal models of the non-motor features of Parkinson's disease - + MESH:D010300 Disease @@ -90237,13 +90962,7 @@ surname:Ros-Bernal;given-names:F 63680 No Lewy pathology in monkeys with over 10 years of severe MPTP Parkinsonism - - MESH:D015632 - Chemical - - MPTP - - + MESH:D010302 Disease @@ -90263,25 +90982,25 @@ 354 63756 Neurotoxic effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and methamphetamine in several strains of mice - + MESH:D015632 Chemical 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine - + MESH:D008694 Chemical methamphetamine - + MESH:D020258 Disease Neurotoxic - + MESH:D015632 Chemical @@ -90305,20 +91024,20 @@ surname:Callison;given-names:RC;suffix:Jr 63877 Lysosomal malfunction accompanies alpha-synuclein aggregation in a progressive mouse model of Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + 10090 Species mouse - + MESH:D010300 Disease @@ -90341,24 +91060,18 @@ S115 63993 Modeling PD pathogenesis in mice: advantages of a chronic MPTP protocol - + 10090 Species mice - + MESH:D010300 Disease PD - - MESH:D015632 - Chemical - - MPTP - surname:Gesi;given-names:M @@ -90386,13 +91099,13 @@ surname:Ruffoli;given-names:R 64065 Parkinson-like syndrome induced by continuous MPTP infusion: convergent roles of the ubiquitin-proteasome system and alpha-synuclein - - MESH:D020734 + + MESH:D010302 Disease Parkinson-like syndrome - + MESH:D015632 Chemical @@ -90419,18 +91132,24 @@ surname:Marien;given-names:M 64198 Modelling Parkinson-like neurodegeneration via osmotic minipump delivery of MPTP and probenecid - + MESH:D015632 Chemical MPTP - + MESH:D020721 Disease Modelling Parkinson-like neurodegeneration + + MESH:D011339 + Chemical + + probenecid + surname:Levesque;given-names:D @@ -90449,26 +91168,20 @@ surname:Rouillard;given-names:C 64294 Differences between subacute and chronic MPTP mice models: investigation of dopaminergic neuronal degeneration and alpha-synuclein inclusions - + 20617 Gene 293 alpha-synuclein - + 10090 Species mice - - MESH:D015632 - Chemical - - MPTP - - + MESH:D009410 Disease @@ -90491,19 +91204,19 @@ surname:Dawson;given-names:TM 64436 Absence of inclusion body formation in the MPTP mouse model of Parkinson's disease - + 10090 Species mouse - + MESH:D015632 Chemical MPTP - + MESH:D010300 Disease @@ -90531,19 +91244,19 @@ surname:Richardson;given-names:JR 64521 Mechanism of toxicity in rotenone models of Parkinson's disease - + MESH:D012402 Chemical rotenone - + MESH:D064420 Disease toxicity - + MESH:D010300 Disease @@ -90565,7 +91278,7 @@ 205 64587 Inhibitory effects of pesticides on proteasome activity: implication in Parkinson's disease - + MESH:D010300 Disease @@ -90589,7 +91302,7 @@ surname:Panov;given-names:AV 64681 Chronic systemic pesticide exposure reproduces features of Parkinson's disease - + MESH:D010300 Disease @@ -90611,14 +91324,14 @@ 16 64762 Subcutaneous rotenone exposure causes highly selective dopaminergic degeneration and alpha-synuclein aggregation - + 20617 Gene 293 alpha-synuclein - + MESH:D012402 Chemical @@ -90642,25 +91355,25 @@ surname:Bhatt;given-names:AJ 64875 Neonatal exposure to lipopolysaccharide enhances accumulation of alpha-synuclein aggregation and dopamine transporter protein expression in the substantia nigra in responses to rotenone challenge in later life - + 20617 Gene 293 alpha-synuclein - - MESH:D004298 - Chemical - - dopamine - - + MESH:D012402 Chemical rotenone + + MESH:D008070 + Chemical + + lipopolysaccharide + surname:Uversky;given-names:VN @@ -90677,14 +91390,14 @@ 732 65085 Biophysical Characterization of alpha-Synuclein and Rotenone Interaction - + 20617 Gene 293 alpha-Synuclein - + MESH:D012402 Chemical @@ -90705,14 +91418,14 @@ 108 65158 Pesticides directly accelerate the rate of alpha-synuclein fibril formation: a possible factor in Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -90734,13 +91447,13 @@ 102 65278 Chronic rotenone exposure reproduces Parkinson's disease gastrointestinal neuropathology - + MESH:D010300 Disease Parkinson's disease gastrointestinal neuropathology - + MESH:D012402 Chemical @@ -90763,13 +91476,13 @@ surname:Dowd;given-names:E 65369 Characterisation of a novel model of Parkinson's disease by intra-striatal infusion of the pesticide rotenone - + MESH:D010300 Disease Parkinson's disease - + MESH:D012402 Chemical @@ -90790,7 +91503,7 @@ 483 65481 Environmental toxins and Parkinson's disease: what have we learned from pesticide-induced animal models? - + MESH:D010300 Disease @@ -90815,13 +91528,13 @@ surname:Gould;given-names:P 65588 Rotenone induces non-specific central nervous system and systemic toxicity - + MESH:D012402 Chemical Rotenone - + MESH:D064420 Disease @@ -90857,7 +91570,7 @@ 162 65722 Systemic exposure to proteasome inhibitors causes a progressive model of Parkinson's disease - + MESH:D010300 Disease @@ -90893,19 +91606,19 @@ surname:Powe;given-names:DG 65817 Depletion of 26S proteasomes in mouse brain neurons causes neurodegeneration and Lewy-like inclusions resembling human pale bodies - + 10090 Species mouse - + 9606 Species human - + MESH:D019636 Disease @@ -90929,13 +91642,13 @@ surname:Mayer;given-names:RJ 65948 Pale body-like inclusion formation and neurodegeneration following depletion of 26S proteasomes in mouse brain neurones are independent of alpha-synuclein - + 10090 Species mouse - + MESH:D019636 Disease @@ -90957,13 +91670,13 @@ 663 66103 Mitochondrial dysfunction in the limelight of Parkinson's disease pathogenesis - + MESH:D010300 Disease Parkinson's disease - + MESH:D028361 Disease @@ -90993,7 +91706,7 @@ surname:Reeve;given-names:AK 66184 High levels of mitochondrial DNA deletions in substantia nigra neurons in aging and Parkinson disease - + MESH:D010300 Disease @@ -91013,24 +91726,18 @@ 33 66286 Neurotoxic in vivo models of Parkinson's disease recent advances - + MESH:D010300 Disease Parkinson's disease - + MESH:D020258 Disease Neurotoxic - - MESH:D020178 - Disease - - advances - surname:Skibinski;given-names:G @@ -91059,11 +91766,11 @@ surname:Levy;given-names:JM 66353 Direct membrane association drives mitochondrial fission by the Parkinson disease-associated protein alpha-synuclein - - MESH:D010302 + + MESH:D010300 Disease - - Parkinson + + Parkinson disease @@ -91080,7 +91787,7 @@ 661 66470 Genetic animal models of Parkinson's disease - + MESH:D010300 Disease @@ -91113,19 +91820,25 @@ surname:Hofstetter;given-names:C 66517 Progressive parkinsonism in mice with respiratory-chain-deficient dopamine neurons - + 10090 Species mice - + + MESH:D028361 + Disease + + respiratory-chain-deficient + + MESH:D010302 Disease - - Progressive parkinsonism + + parkinsonism - + MESH:D004298 Chemical @@ -91144,14 +91857,14 @@ 27 66600 In vivo alpha-synuclein overexpression in rodents: a useful model of Parkinson's disease? - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -91186,20 +91899,20 @@ surname:Yu;given-names:J 66692 Conditional expression of Parkinson's disease-related mutant alpha-synuclein in the midbrain dopaminergic neurons causes progressive neurodegeneration and degradation of transcription factor nuclear receptor related 1 - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease Parkinson's disease - + MESH:D019636 Disease @@ -91219,7 +91932,7 @@ 1118 66912 Modelling of Parkinson's disease in mice - + MESH:D010300 Disease @@ -91247,26 +91960,26 @@ surname:Gannon;given-names:P 66955 Distinct region-specific alpha-synuclein oligomers in A53T transgenic mice: implications for neurodegeneration - + 20617 Gene 293 alpha-synuclein - + 10090 Species transgenic mice - + c.53A>T;RS#:104893877 DNAMutation A53T - + MESH:D019636 Disease @@ -91301,32 +92014,32 @@ surname:Soto;given-names:O 67066 Motor dysfunction and gliosis with preserved dopaminergic markers in human alpha-synuclein A30P transgenic mice - + 6622 Gene 293 alpha-synuclein - + 9606 Species human - + p.A30P;RS#:104893878 ProteinMutation A30P - - MESH:D004194 + + MESH:D000068079 Disease Motor dysfunction - + MESH:D005911 Disease @@ -91352,37 +92065,31 @@ surname:Copeland;given-names:NG 67178 Parkinson's disease alpha-synuclein transgenic mice develop neuronal mitochondrial degeneration and cell death - + 20617 Gene 293 alpha-synuclein - + 10090 Species transgenic mice - + MESH:D010300 Disease Parkinson's disease - + MESH:D009410 Disease neuronal mitochondrial degeneration - - MESH:D003643 - Disease - - death - surname:Li;given-names:X @@ -91410,32 +92117,32 @@ surname:Lin;given-names:BM 67291 Unexpected lack of hypersensitivity in LRRK2 knock-out mice to MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) - + 66725 Gene 18982 LRRK2 - + 10090 Species mice - + MESH:D004342 Disease - - hypersensitivity + + hypersensitivity in - + MESH:D015632 Chemical 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine - + MESH:D015632 Chemical @@ -91460,26 +92167,20 @@ surname:Kopan;given-names:R 67406 Loss of leucine-rich repeat kinase 2 causes impairment of protein degradation pathways, accumulation of alpha-synuclein, and apoptotic cell death in aged mice - + 66725 Gene 18982 leucine-rich repeat kinase 2 - + 20617 Gene 293 alpha-synuclein - - MESH:D003643 - Disease - - death - surname:Karouani;given-names:M @@ -91499,14 +92200,14 @@ surname:Yamaguchi;given-names:H 67565 R1441C mutation in LRRK2 impairs dopaminergic neurotransmission in mice - + 66725 Gene 18982 LRRK2 - + p.R1441C;RS#:33939927 ProteinMutation @@ -91538,29 +92239,24 @@ surname:Stemmelen;given-names:C 67637 High LRRK2 levels fail to induce or exacerbate neuronal alpha-synucleinopathy in mouse brain - + 66725 Gene 18982 LRRK2 - + 10090 Species mouse - - MESH:D005935 + + MESH:D000080874 Disease - - alpha-synucleinopathy - - - Chemical - - exacerbate + + synucleinopathy @@ -91592,14 +92288,14 @@ surname:Shim;given-names:H 67730 Leucine-rich repeat kinase 2 regulates the progression of neuropathology induced by Parkinson's-disease-related mutant alpha-synuclein - + 66725 Gene 18982 Leucine-rich repeat kinase 2 - + MESH:D010300 Disease @@ -91639,31 +92335,25 @@ surname:Kim;given-names:J 67867 Dopaminergic neuronal loss, reduced neurite complexity and autophagic abnormalities in transgenic mice expressing G2019S mutant LRRK2 - + 10090 Species transgenic mice - + p.G2019S;RS#:34637584 ProteinMutation G2019S - + MESH:D009410 Disease neuronal loss - - MESH:C536522 - Disease - - autophagic abnormalities - - + MESH:D058225 Disease @@ -91685,6 +92375,12 @@ 1075 68001 Autophagy fights disease through cellular self-digestion + + MESH:C564093 + Disease + + Autophagy fights disease + surname:Olanow;given-names:CW @@ -91702,14 +92398,14 @@ surname:Kordower;given-names:JH 68058 Alterations in lysosomal and proteasomal markers in Parkinson's disease: relationship to alpha-synuclein inclusions - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -91734,7 +92430,7 @@ surname:Recasens;given-names:A 68176 Pathogenic lysosomal depletion in Parkinson's disease - + MESH:D010300 Disease @@ -91757,7 +92453,7 @@ surname:Sulzer;given-names:D 68232 Impaired degradation of mutant alpha-synuclein by chaperone-mediated autophagy - + 20617 Gene 293 @@ -91802,27 +92498,27 @@ surname:Jakobsson;given-names:J 68444 TFEB-mediated autophagy rescues midbrain dopamine neurons from alpha-synuclein toxicity - + 21425 Gene 5182 TFEB - + 20617 Gene 293 alpha-synuclein - + MESH:D064420 Disease toxicity - + MESH:D004298 Chemical @@ -91845,28 +92541,28 @@ surname:Perlmann;given-names:T 68532 alpha-Synuclein-induced down-regulation of Nurr1 disrupts GDNF signaling in nigral dopamine neurons - + 20617 Gene 293 alpha-Synuclein - + 18227 Gene 4509 Nurr1 - + 14573 Gene 433 GDNF - + MESH:D004298 Chemical @@ -91891,27 +92587,27 @@ surname:Romero-Ramos;given-names:M 68632 GDNF fails to exert neuroprotection in a rat alpha-synuclein model of Parkinson's disease - + 25453 Gene 433 GDNF - + 29219 Gene 293 alpha-synuclein - + 10116 Species rat - + MESH:D010300 Disease @@ -91933,26 +92629,26 @@ 289 68724 Lentiviral nigral delivery of GDNF does not prevent neurodegeneration in a genetic rat model of Parkinson's disease - + 25453 Gene 433 GDNF - + 10116 Species rat - + MESH:D019636 Disease neurodegeneration - + MESH:D010300 Disease @@ -91976,25 +92672,19 @@ surname:Riederer;given-names:P 68842 Transforming growth factor-beta-1 levels are elevated in the striatum and in ventricular cerebrospinal fluid in Parkinson's disease - + 21803 Gene 540 Transforming growth factor-beta-1 - + MESH:D010300 Disease Parkinson's disease - - MESH:D065634 - Disease - - ventricular cerebrospinal fluid - surname:Carvey;given-names:P @@ -92012,27 +92702,21 @@ surname:Freed;given-names:WJ 68976 TGF beta 1 and TGF beta 2 concentrations are elevated in Parkinson's disease in ventricular cerebrospinal fluid - + 21803 Gene 540 TGF beta 1 - + 21808 Gene 2432 TGF beta 2 - - MESH:D065634 - Disease - - ventricular cerebrospinal fluid - - + MESH:D010300 Disease @@ -92055,14 +92739,20 @@ surname:Mallet;given-names:J 69090 Transforming growth factor beta 1 overexpression in the nigrostriatal system increases the dopaminergic deficit of MPTP mice - + 21803 Gene 540 Transforming growth factor beta 1 - + + MESH:D009461 + Disease + + deficit + + MESH:D015632 Chemical @@ -92083,7 +92773,7 @@ 2173 69215 Adenovirus-mediated over-expression of TGF beta 1 in the striatum decreases dopaminergic cell survival in embryonic nigral grafts - + 21803 Gene 540 @@ -92103,7 +92793,7 @@ 34 69345 Dual role for TGF-beta 1 in apoptosis - + 21803 Gene 540 @@ -92128,21 +92818,21 @@ surname:Mena;given-names:MA 69383 Dopamine and alpha-synuclein dysfunction in Smad3 null mice - + 20617 Gene 293 alpha-synuclein - + 17127 Gene 55937 Smad3 - + MESH:D004298 Chemical @@ -92164,7 +92854,7 @@ 821 69443 New experimental and clinical links between the hippocampus and the dopaminergic system in Parkinson's disease - + MESH:D010300 Disease @@ -92193,7 +92883,7 @@ surname:Prots;given-names:I 69556 Role of alpha-synuclein in adult neurogenesis and neuronal maturation in the dentate gyrus - + 20617 Gene 293 @@ -92229,24 +92919,19 @@ surname:Maiolino;given-names:F 69647 Extracellular alpha-synuclein oligomers modulate synaptic transmission and impair LTP via NMDA-receptor activation - + 20617 Gene 293 alpha-synuclein - + 109912 Gene LTP - - Chemical - - NMDA - surname:Sanchez-Capelo;given-names:A @@ -92262,7 +92947,7 @@ ref 69762 Smad3 is required for the survival of proliferative intermediate progenitor cells in the dentate gyrus of adult mice - + 17127 Gene 55937 @@ -92283,7 +92968,13 @@ 398 69879 A Drosophila model of Parkinson's disease - + + 7227 + Species + + Drosophila + + MESH:D010300 Disease @@ -92304,26 +92995,33 @@ ref 69923 alpha-Synuclein Transgenic Drosophila As a Model of Parkinson's Disease and Related Synucleinopathies - + 20617 Gene 293 alpha-Synuclein - + 10090 Species Transgenic - + + 7227 + Species + + Drosophila + + MESH:D010300 Disease Parkinson's Disease - + + MESH:D000080874 Disease Synucleinopathies @@ -92348,31 +93046,26 @@ surname:Kowa;given-names:H 70027 Familial Parkinson mutant alpha-synuclein causes dopamine neuron dysfunction in transgenic Caenorhabditis elegans - + 20617 Gene 293 alpha-synuclein - - 10090 - Species - - transgenic + + MESH:D009410 + Disease + + neuron dysfunction in transgenic Caenorhabditis - + MESH:D004298 Chemical dopamine - - Disease - - Caenorhabditis elegans - - + MESH:D010302 Disease @@ -92398,24 +93091,18 @@ surname:Thomas;given-names:JH 70141 Dopaminergic neuronal loss and motor deficits in Caenorhabditis elegans overexpressing human alpha-synuclein - + 6239 Species Caenorhabditis elegans - + 9606 Species human - - MESH:D009410 - Disease - - neuronal loss - surname:Hofstra;given-names:RM @@ -92433,14 +93120,14 @@ surname:Plasterk;given-names:RH 70250 C. elegans model identifies genetic modifiers of alpha-synuclein inclusion formation during aging - + 20617 Gene 293 alpha-synuclein - + 6239 Species @@ -92470,21 +93157,21 @@ surname:Zhao;given-names:C 70348 Silencing alpha-synuclein gene expression enhances tyrosine hydroxylase activity in MN9D cells - + 20617 Gene 293 alpha-synuclein - + 21823 Gene 307 tyrosine hydroxylase - + CVCL:U508 CellLine @@ -92507,14 +93194,14 @@ surname:Perez;given-names:RG 70443 Alpha-synuclein activation of protein phosphatase 2A reduces tyrosine hydroxylase phosphorylation in dopaminergic cells - + 20617 Gene 293 Alpha-synuclein - + 21823 Gene 307 @@ -92538,7 +93225,7 @@ surname:Perez;given-names:RG 70563 Alpha-synuclein inhibits aromatic amino acid decarboxylase activity in dopaminergic cells - + 20617 Gene 293 @@ -92561,14 +93248,14 @@ 377 70653 Dysregulated dopamine storage increases the vulnerability to alpha-synuclein in nigral neurons - + 20617 Gene 293 alpha-synuclein - + MESH:D004298 Chemical @@ -92602,7 +93289,7 @@ surname:Lam;given-names:HA 70748 Parkin-deficient mice exhibit nigrostriatal deficits but not loss of dopaminergic neurons - + 10090 Species @@ -92647,7 +93334,7 @@ surname:Cohen-Salmon;given-names:C 70838 Parkin gene inactivation alters behaviour and dopamine neurotransmission in the mouse - + MESH:D004298 Chemical @@ -92667,13 +93354,13 @@ 2179 70924 Parkin-deficient mice are not a robust model of parkinsonism - + 10090 Species mice - + MESH:D010302 Disease @@ -92706,13 +93393,13 @@ surname:Kitada;given-names:T 70985 Nigrostriatal dopaminergic deficits and hypokinesia caused by inactivation of the familial Parkinsonism-linked gene DJ-1 - + MESH:C564486 Disease - - Parkinsonism-linked + + familial Parkinsonism-linked - + MESH:D018476 Disease @@ -92740,19 +93427,18 @@ surname:Tscherter;given-names:A 71106 Impaired dopamine release and synaptic plasticity in the striatum of PINK1-deficient mice - - 68943 - Gene - 32672 - - PINK1 - - + MESH:D004298 Chemical dopamine + + MESH:C565276 + Disease + + PINK1-deficient + surname:Choi-Lundberg;given-names:D @@ -92779,20 +93465,20 @@ surname:Ho;given-names:WH 71196 Mice lacking alpha-synuclein display functional deficits in the nigrostriatal dopamine system - + 20617 Gene 293 alpha-synuclein - + 10090 Species Mice - + MESH:D004298 Chemical @@ -92822,7 +93508,7 @@ surname:Gottschalk;given-names:W 71290 Synaptic vesicle depletion correlates with attenuated synaptic responses to prolonged repetitive stimulation in mice lacking alpha-synuclein - + 10090 Species @@ -92873,17 +93559,6 @@ alpha-synuclein - Chemical - - DOPAL - - - MESH:D015306 - Chemical - - monoamine - - MESH:D004298 Chemical @@ -92911,26 +93586,20 @@ surname:Wilson;given-names:K 71548 Interplay between cytosolic dopamine, calcium, and alpha-synuclein causes selective death of substantia nigra neurons - + 20617 Gene 293 alpha-synuclein - + MESH:D004298 Chemical dopamine - - MESH:D003643 - Disease - - death - - + MESH:D002118 Chemical @@ -92957,12 +93626,6 @@ surname:Rockenstein;given-names:E 71666 Inclusion formation and neuronal cell death through neuron-to-neuron transmission of alpha-synuclein - - MESH:D003643 - Disease - - death - surname:Freeman;given-names:TB @@ -92980,7 +93643,7 @@ surname:Olanow;given-names:CW 71767 Lewy body-like pathology in long-term embryonic nigral transplants in Parkinson's disease - + MESH:D010300 Disease @@ -93005,24 +93668,19 @@ surname:O'Brien;given-names:P 71859 Pathological alpha-synuclein transmission initiates Parkinson-like neurodegeneration in nontransgenic mice - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease Parkinson-like neurodegeneration - - Chemical - - nontransgenic - surname:Tanik;given-names:SA @@ -93044,18 +93702,18 @@ surname:Riddle;given-names:DM 71966 Exogenous alpha-synuclein fibrils induce Lewy body pathology leading to synaptic dysfunction and neuron death - + 20617 Gene 293 alpha-synuclein - - MESH:D003643 + + MESH:C536122 Disease - - death + + synaptic dysfunction @@ -93081,14 +93739,14 @@ surname:Lee;given-names:S 72076 Intramuscular injection of alpha-synuclein induces CNS alpha-synuclein pathology and a rapid-onset motor phenotype in transgenic mice - + 20617 Gene 293 alpha-synuclein - + 20617 Gene 293 @@ -93117,7 +93775,7 @@ surname:McGarvey;given-names:NH 72210 Amyloidogenic alpha-synuclein seeds do not invariably induce rapid, widespread pathology in mice - + 20617 Gene 293 @@ -93139,14 +93797,14 @@ 98 72307 Extracellular alpha-synuclein-a novel and crucial factor in Lewy body diseases - + 20617 Gene 293 alpha-synuclein - + MESH:D020961 Disease @@ -93178,21 +93836,21 @@ surname:Sotnikova;given-names:TD 72388 Exogenous alpha-Synuclein Decreases Raft Partitioning of Cav2.2 Channels Inducing Dopamine Release - + 20617 Gene 293 alpha-Synuclein - + 12287 Gene 20184 Cav2.2 - + MESH:D004298 Chemical @@ -93212,7 +93870,7 @@ 39 72487 Biophysical characterization of alpha-synuclein and its controversial structure - + 20617 Gene 293 @@ -93235,7 +93893,7 @@ 377 72567 The remarkable conformational plasticity of alpha-synuclein: blessing or curse? - + 20617 Gene 293 @@ -93257,7 +93915,7 @@ 110 72647 alpha-Synuclein occurs physiologically as a helically folded tetramer that resists aggregation - + 20617 Gene 293 @@ -93296,7 +93954,7 @@ surname:Nguyen;given-names:LT 72742 A soluble alpha-synuclein construct forms a dynamic tetramer - + 20617 Gene 293 @@ -93316,14 +93974,14 @@ 299 72803 Mechanistic aspects of Parkinson's disease: alpha-synuclein and the biomembrane - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -93347,14 +94005,14 @@ surname:Citron;given-names:M 72885 alpha-synuclein fibrillogenesis is nucleation-dependent. Implications for the pathogenesis of Parkinson's disease - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -93378,14 +94036,14 @@ surname:Williamson;given-names:RE 73001 Acceleration of oligomerization, not fibrillization, is a shared property of both alpha-synuclein mutations linked to early-onset Parkinson's disease: implications for pathogenesis and therapy - + 20617 Gene 293 alpha-synuclein - + MESH:D010300 Disease @@ -93408,7 +94066,7 @@ surname:Crowther;given-names:RA 73196 Fiber diffraction of synthetic alpha-synuclein filaments shows amyloid-like cross-beta conformation - + 20617 Gene 293 @@ -93430,7 +94088,7 @@ 10744 73296 Evidence for a partially folded intermediate in alpha-synuclein fibril formation - + 20617 Gene 293 @@ -93485,7 +94143,7 @@ surname:Vassallo;given-names:N 73464 Two different binding modes of alpha-synuclein to lipid vesicles depending on its aggregation state - + 20617 Gene 293 @@ -93507,20 +94165,26 @@ 621 73564 alpha-Synuclein oligomers: an amyloid pore? Insights into mechanisms of alpha-synuclein oligomer-lipid interactions - + 20617 Gene 293 alpha-Synuclein - + 20617 Gene 293 alpha-synuclein + + MESH:D008055 + Chemical + + lipid + surname:Nakamura;given-names:K @@ -93542,7 +94206,7 @@ surname:Onoa;given-names:B 73680 Increased expression of alpha-synuclein reduces neurotransmitter release by inhibiting synaptic vesicle reclustering after endocytosis - + 20617 Gene 293 @@ -93565,21 +94229,21 @@ 1349 73815 Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct - + 20617 Gene 293 alpha-synuclein - + 20617 Gene 293 alpha-synuclein - + MESH:D004298 Chemical @@ -93602,25 +94266,25 @@ surname:Fink;given-names:AL 73909 Dopamine and L-dopa disaggregate amyloid fibrils: implications for Parkinson's and Alzheimer's disease - + MESH:D004298 Chemical Dopamine - + MESH:D000544 Disease Alzheimer's disease - + MESH:D010300 Disease Parkinson's - + MESH:D007980 Chemical @@ -93645,7 +94309,7 @@ surname:Trojanowski;given-names:JQ 74014 Reversible inhibition of alpha-synuclein fibrillization by dopaminochrome-mediated conformational alterations - + 20617 Gene 293 @@ -93667,25 +94331,31 @@ 11613 74124 Effect of familial Parkinson's disease point mutations A30P and A53T on the structural properties, aggregation, and fibrillation of human alpha-synuclein - + 9606 Species human - + c.53A>T;RS#:104893877 DNAMutation A53T - + + MESH:D014693 + Disease + + fibrillation + + MESH:D010300 Disease familial Parkinson's disease - + p.A30P;RS#:104893878 ProteinMutation @@ -93711,20 +94381,20 @@ surname:Abbas-Terki;given-names:T 74280 Phosphorylation does not prompt, nor prevent, the formation of alpha-synuclein toxic species in a rat model of Parkinson's disease - + 29219 Gene 293 alpha-synuclein - + 10116 Species rat - + MESH:D010300 Disease @@ -93750,39 +94420,39 @@ surname:Kondrikova;given-names:G 74413 The phosphorylation state of Ser-129 in human alpha-synuclein determines neurodegeneration in a rat model of Parkinson disease - + 6622 Gene 293 alpha-synuclein - + 9606 Species human - + 10116 Species rat - + MESH:D010300 Disease Parkinson disease - + MESH:D019636 Disease neurodegeneration - - MESH:C530429 + + MESH:D012694 Chemical Ser @@ -93802,13 +94472,13 @@ surname:Cenci;given-names:MA 74540 Role of phost-translational modifications in modulating the structure, function and toxicity of a-synuclein: implications for Parkinson's disease pathogenesis and therapies - + MESH:D010300 Disease Parkinson's disease - + MESH:D064420 Disease @@ -93835,7 +94505,7 @@ surname:Bavikar;given-names:SN 74715 Synthetic polyubiquitinated alpha-Synuclein reveals important insights into the roles of the ubiquitin chain in regulating its pathophysiology - + 20617 Gene 293 @@ -93865,18 +94535,12 @@ surname:Evjenth;given-names:R 74858 Proteomics analyses reveal the evolutionary conservation and divergence of N-terminal acetyltransferases from yeast and humans - + 4932 Species yeast - - MESH:D009584 - Chemical - - N - surname:Volker;given-names:W @@ -93897,7 +94561,7 @@ surname:Lorenzen;given-names:I 74985 Differential aggregation properties of alpha-synuclein isoforms - + 20617 Gene 293 @@ -94130,12 +94794,18 @@ paragraph 12951 We used a survey instrument derived from the Group Climate Questionnaire - Short version (GCQ-S), a validated instrument used previously by other researchers. Three items from the GCQ-S were used. In addition, eight items were used from Steele et al in order to create an instrument more useful for our purpose. Using the survey, participants self-scored and self-rated learning, group participation and dynamics using 16 Likert scaled questions. In addition, they were asked how many students were in their group and were asked to report their perceptions of how many students they felt participated fully. (See Figure 1). Students' "in" surveys were then compared to their "out" surveys to assess differences in perceived group process and dynamics. - + 9606 Species participants + + - + Chemical + + GCQ-S + METHODS @@ -94202,7 +94872,7 @@ Tab1.xml 15455 Mean score p Faculty present "In" (n = 114) Faculty absent "Out" (n = 114) Student perception of group dynamics and group learning process The group and its membez Helped each other to express their ideas and feelings.a 4.4 4.6 0.015 Utilized the knowledge of students in the group.a 4.5 4.7 <0.01 The members challenged each other in their efforts to sort things out.a 4.1 4.4 0.012 Utilized the knowledge of the facilitator.a 4.2 3.5 <.001 The members depended upon the faculty leader for direction.a 3.3 2.3 <.001 The group discussion was chaotic and disorganized.a 1.4 1.6 0.06 Faced and accepted differences of opinion expressed by members of the groupa 4.5 4.5 0.058 Worked together efficientlya 4.6 4.6 0.67 Moved toward its goals on the basis of consensusa 4.6 4.6 0.32 Student participation and preference Rate your participation level.b 4.2 4.3 0.03 I felt comfortable speaking during the student discussion.a 4.6 4.6 0.49 I prefer to have faculty present during the entire discussion of this case.a 3.5 3.1 0.0008 Case content Arrived at a reasonable management plan for this case.a 4.6 4.5 0.03 The faculty member was essential to the success of this case discussion.a 3.3 2.5 <.001 I feel I got the most I could out of the discussion of this case.a 4.3 4.1 0.67 Considered psychosocial issues in the management of this case.a 4.4 4.4 0.89 - + MESH:C535569 Disease @@ -94226,7 +94896,7 @@ paragraph 18194 When we assessed case content (the outcome of the discussion), students were less likely to say that they arrived at a reasonable management plan after a case when the faculty was absent. However, when students rated whether they had gotten all they could out of the experience, there was no statistically significant difference between cases with faculty present or absent. Students were more likely to rate the faculty member's presence as "essential" after a case with the faculty present. Finally, the students' assessment of whether psychosocial issues were considered during the discussion was no different whether the faculty member was present or absent. - + MESH:C535569 Disease @@ -94262,7 +94932,7 @@ paragraph 21597 This study had several limitations. Students were recruited throughout the academic year, meaning some of the students in the study were near the end of their third year and more likely to be confident in their clinical knowledge and also had had more experience participating in small group discussions compared with early third-year students. The experiences that were compared using paired t-tests were group discussions within the same group of students and faculty member. However, the cases were different, which may have been partly responsible for some of the differences in scores (for example, a case featuring domestic violence might engender more or less animated discussion than a case featuring diabetes). In addition, faculty members leading groups had different amounts of experience and skill leading small group learning sessions. Because groups met simultaneously, we were not able to compare multiple groups with the same faculty member leading. However, evaluating groups with each faculty member in and out as well as conducting this study over the course of the year may help us address this limitation. Additionally, the cases used as intervention and control were rotated randomly. Finally, while many of the items in our survey had statistically significant differences showing increased discussion and participation when the faculty was absent, some of the absolute differences in scores were small. - + MESH:D003920 Disease @@ -94328,7 +94998,7 @@ footnote 25321 Hoffman M. Xu J. Wilkinson J. Wiecha J. The Effect of Faculty Presence on Small-group Learning/Group Dynamics in a Family Medicine Clerkship. STFM Annual Spring Conference 2010. Vancouver, Canada. April 25, 2010. - + MESH:D012872 Disease diff --git a/data/example/7-sample-full-text-tags.tsv b/data/example/7-sample-full-text-tags.tsv index e70b6d3..9ceba44 100644 --- a/data/example/7-sample-full-text-tags.tsv +++ b/data/example/7-sample-full-text-tags.tsv @@ -1,9 +1,11 @@ pubmed_id type identifier offset end 4272814 Species 28141 16 37 4272814 Species 28141 211 232 -4272814 Chemical C489032 172 179 +4272814 Chemical D011392 172 179 4272814 Species 28141 745 766 4272814 Species 9606 822 828 +4272814 Chemical D014867 1131 1136 +4272814 Chemical D014867 774 779 4272814 Species 562 1235 1251 4272814 Species 562 1525 1532 4272814 Species 28141 1727 1739 @@ -11,56 +13,45 @@ pubmed_id type identifier offset end 4272814 Species 562 1994 2001 4272814 Species 28141 2168 2180 4272814 Species 28141 2211 2223 -4272814 Chemical C015732 1626 1633 -4272814 Chemical C489032 1443 1450 -4272814 Chemical D005998 1455 1462 -4272814 Chemical C041069 2029 2037 +4272814 Chemical D011392 1443 1450 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10926 10933 4272814 Chemical D012965 10961 10965 4272814 Species 562 10996 11003 4272814 Species 562 11195 11202 @@ -70,11 +61,12 @@ pubmed_id type identifier offset end 4272814 Species 562 11810 11817 4272814 Species 562 11906 11913 4272814 Species 562 12121 12128 -4272814 Strain 1239833 11449 11453 -4272814 Strain 1239833 11684 11688 +4272814 Species 1239833 11449 11453 +4272814 Species 1239833 11684 11688 4272814 Chemical D012965 11332 11336 4272814 Chemical D012965 11690 11694 4272814 Chemical D012965 11847 11851 +4272814 Chemical - 11835 11840 4272814 Species 562 12831 12838 4272814 Species 562 12937 12944 4272814 Species 562 13167 13174 @@ -88,10 +80,10 @@ pubmed_id type identifier offset end 4272814 Species 9606 13980 13986 4272814 Species 9606 14066 14073 4272814 Species 28141 14088 14100 -4272814 Disease D054549 14386 14406 4272814 Disease D006849 14206 14219 4272814 Disease D001922 14187 14202 4272814 Disease D009422 14164 14185 +4272814 Disease D004194 14386 14406 4272814 Species 562 14471 14478 4272814 Species 28141 15097 15109 4272814 Species 28141 15270 15282 @@ -99,8 +91,9 @@ pubmed_id type identifier offset end 4272814 Species 562 15668 15675 4272814 Species 562 16048 16055 4272814 Chemical D012965 16186 16190 -4272814 Chemical C489032 15754 15761 +4272814 Chemical D011392 15754 15761 4272814 Chemical D012965 16262 16266 +4272814 Chemical - 16056 16061 4272814 Species 1718 16454 16480 4272814 Species 1423 16492 16509 4272814 Species 562 16747 16754 @@ -112,6 +105,7 @@ pubmed_id type identifier offset end 4272814 Species 28141 18039 18051 4272814 Species 28141 18286 18298 4272814 Chemical D012965 17944 17948 +4272814 Chemical - 16448 16452 4272814 Species 562 18780 18787 4272814 Species 562 18901 18908 4272814 Chemical D012965 18860 18864 @@ -119,24 +113,26 @@ pubmed_id type identifier offset end 4272814 Disease D009101 19360 19362 4272814 Disease D009101 19260 19262 4272814 Disease D001281 19302 19304 +4272814 Chemical D014867 19447 19452 4272814 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3636 4272815 Disease D015179 3350 3360 4272815 Disease D003123 3733 3747 -4272815 Disease D016889 3362 3373 +4272815 Disease D014591 3362 3373 4272815 Disease D003123 3256 3270 4272815 Disease D009369 3384 3391 4272815 Disease D009386 3277 3320 @@ -184,8 +180,7 @@ pubmed_id type identifier offset end 4272815 DNAMutation r.589_592del 4462 4474 4272815 DNAMutation c.791-1G>C;RS#:267607795 4621 4633 4272815 Gene 208 5240 5244 -4272815 Chemical C059568 5472 5483 -4272815 Disease D005770 5402 5437 +4272815 Disease D004067 5402 5437 4272815 Species 9606 5527 5534 4272815 Species 9913 5646 5652 4272815 Species 9606 5746 5754 @@ -193,30 +188,33 @@ pubmed_id type identifier offset end 4272815 DNAMutation c.790+1G>T;RS#:267607789 5947 5961 4272815 DNAMutation c.791-1G>C;RS#:267607795 5786 5798 4272815 DNAMutation c.589-2A>G;RS#:267607767 5772 5784 -4272815 Chemical D002245 5700 5703 +4272815 Chemical - 5700 5703 4272815 DNAMutation c.588+1G>T;RS#:267607772 5756 5770 4272815 DNAMutation c.884G>A;RS#:63750144 5800 5810 -4272815 Disease D015179 5866 5883 4272815 DNAMutation c.1559-2A>T;RS#:267607836 5815 5828 +4272815 Disease D015179 5853 5883 +4272815 Chemical - 5626 5630 4272815 Gene 3908 6982 6987 4272815 DNAMutation c.1G>T 6432 6438 4272815 DNAMutation c.1G>T 6162 6168 -4272815 Chemical C504520 6575 6578 -4272815 DNAMutation c.884G>A;RS#:63750144 6705 6715 4272815 DNAMutation c.589-2A>G;RS#:267607767 6291 6303 4272815 DNAMutation c.1559-2A>T;RS#:267607836 6838 6851 +4272815 DNAMutation c.791-1G>C;RS#:267607795 6564 6576 +4272815 DNAMutation c.884G>A;RS#:63750144 6705 6715 4272815 Gene 128415 7255 7258 4272815 Gene 128415 8049 8052 4272815 Gene 128415 8131 8134 4272815 Species 9606 7683 7690 4272815 Chemical C042345 7553 7562 +4272815 Chemical - 7419 7422 4272815 Chemical D012436 7304 7324 -4272815 Chemical D002725 7497 7514 4272815 Chemical D004492 7467 7471 4272815 Chemical D012965 7438 7442 -4272815 Chemical C014843 7419 7422 +4272815 Chemical D002725 7504 7514 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Disease D015179 20165 20177 -4272815 Disease D015179 20207 20224 +4272815 Disease D015179 20194 20224 4272815 Disease D009369 20369 20375 4272815 Disease D009369 20308 20314 4272815 Disease D009369 20496 20502 @@ -289,23 +291,19 @@ pubmed_id type identifier offset end 4272815 Gene 80218 20601 20606 4272815 Disease D009369 20981 20987 4272815 Species 9606 21154 21159 -4272815 Disease D049290 21254 21264 -4272815 Chemical D000728 21439 21447 4272815 Disease D011471 21495 21510 4272815 Chemical C042345 21517 21526 4272816 Species 8078 69 87 4272816 Species 8078 89 110 -4272816 Chemical D006838 24 40 4272816 Gene 105937229 2063 2067 4272816 Gene 105920361 2223 2227 4272816 Species 8078 1127 1145 4272816 Species 8078 1147 1168 4272816 Species 8078 1184 1193 4272816 Species 8078 1467 1476 -4272816 Chemical D006838 1062 1078 -4272816 Chemical D006838 933 949 +4272816 Chemical D009841 439 455 4272816 Disease D064420 150 158 -4272816 Chemical D006846 1423 1456 +4272816 Chemical D006841 1435 1456 4272816 Gene 105937229 3894 3898 4272816 Species 8078 2515 2533 4272816 Species 8078 2535 2556 @@ -315,23 +313,22 @@ pubmed_id type identifier offset end 4272816 Species 29154 3146 3154 4272816 Species 29278 3690 3700 4272816 Chemical D010100 2791 2797 -4272816 Chemical D006838 3566 3582 +4272816 Chemical D011084 3978 3982 4272816 Chemical D011078 3369 3394 -4272816 Chemical D006846 3327 3360 4272816 Chemical D011078 3664 3668 -4272816 Chemical D013749 3674 3678 +4272816 Chemical D000072317 3440 3444 +4272816 Chemical D011084 3658 3662 +4272816 Chemical D006838 3249 3260 +4272816 Chemical D011084 3316 3320 4272816 Chemical D011078 3396 3400 -4272816 Chemical D006838 3244 3260 4272816 Disease D064420 3711 3719 -4272816 Chemical D006841 3293 3314 -4272816 Chemical D013749 3440 3444 -4272816 Chemical C087686 3658 3662 -4272816 Chemical C087686 3316 3320 -4272816 Chemical D013749 3403 3438 +4272816 Chemical D000072317 3674 3678 +4272816 Chemical D006841 3339 3360 +4272816 Chemical D011084 3281 3314 4272816 Chemical D004147 3447 3453 4272816 Chemical D004147 3466 3472 +4272816 Chemical D000072317 3403 3438 4272816 Chemical D011078 3987 3991 -4272816 Chemical C087686 3978 3982 4272816 Gene 105930482 4416 4433 4272816 Gene 105930482 4435 4440 4272816 Species 8078 4127 4136 @@ -339,7 +336,7 @@ pubmed_id type identifier offset end 4272816 Species 8078 4514 4523 4272816 Species 29278 4623 4633 4272816 Disease D064420 4445 4467 -4272816 Chemical D006838 4385 4401 +4272816 Chemical D060172 4275 4301 4272816 Species 8022 6027 6040 4272816 Species 8090 6042 6048 4272816 Species 1234273 6062 6076 @@ -349,31 +346,44 @@ pubmed_id type identifier offset end 4272816 Species 8078 6643 6652 4272816 Species 8078 6695 6713 4272816 Species 8078 6893 6902 +4272816 Disease C537182 7916 7919 4272816 Chemical C003636 7441 7449 +4272816 Chemical - 7228 7231 +4272816 Disease C537182 7581 7584 4272816 Chemical D017693 7472 7490 +4272816 Chemical C003636 7451 7456 4272816 Gene 105937229 8242 8260 4272816 Species 8078 8698 8707 4272816 Gene 105937229 9223 9227 4272816 Species 29278 9631 9649 4272816 Species 29278 9200 9210 4272816 Species 29278 9521 9531 -4272816 Chemical C017937 9852 9859 +4272816 Chemical D009841 9580 9595 +4272816 Chemical D009841 9430 9446 4272816 Species 8078 10092 10101 4272816 Species 8078 10909 10918 4272816 Species 29154 11664 11672 +4272816 Chemical D012685 10409 10416 +4272816 Chemical D008899 11204 11215 +4272816 Chemical - 10866 10878 4272816 Chemical C049037 11079 11094 -4272816 Chemical C415406 10866 10878 +4272816 Disease D005597 11384 11391 4272816 Chemical C049037 10697 10712 4272816 Species 10090 12609 12614 4272816 Species 3704 12725 12736 4272816 Species 9925 12765 12769 4272816 Species 10090 12775 12780 4272816 Species 29278 12050 12060 -4272816 Chemical C040388 12281 12295 +4272816 Chemical - 12515 12519 +4272816 Chemical C016679 12281 12295 4272816 Chemical D012965 12535 12539 -4272816 Chemical C053576 12515 12519 +4272816 Chemical - 12951 12955 +4272816 Chemical - 13047 13051 +4272816 Chemical D011136 12555 12563 +4272816 Disease D015352 12407 12429 4272816 Chemical D008623 12178 12198 -4272816 Chemical D011136 12555 12560 +4272816 Chemical - 12503 12507 +4272816 Chemical - 14005 14008 4272816 Chemical D004996 14031 14047 4272816 Gene 105937229 14489 14493 4272816 Gene 105920361 14721 14725 @@ -383,6 +393,7 @@ pubmed_id type identifier offset end 4272816 Species 29278 15477 15487 4272816 Species 29278 15573 15583 4272816 Species 29278 15620 15630 +4272816 Chemical C007563 15715 15719 4272816 Gene 105937229 17893 17897 4272816 Gene 105920361 18981 18985 4272816 Gene 105920361 19021 19025 @@ -393,7 +404,6 @@ pubmed_id type identifier offset end 4272816 Gene 105920361 19768 19772 4272816 Gene 105920361 20448 20452 4272816 Gene 105920361 19307 19311 -4272816 Chemical D009584 19111 19112 4272816 Species 8078 21193 21202 4272816 Species 8078 21337 21346 4272816 Species 8078 21597 21606 @@ -409,7 +419,7 @@ 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D011078 27568 27572 4272816 Species 8078 29970 29988 4272816 Species 8078 30060 30069 4272816 Species 8078 31275 31293 4272816 Species 8078 31539 31548 4272816 Gene 105920361 32525 32529 -4272816 Chemical D006838 32488 32504 4272816 Species 8090 32777 32783 4272816 Species 8090 32846 32852 4272816 Species 7159 33011 33024 4272816 Species 8078 33287 33305 4272816 Species 8078 33307 33328 +4272816 Species 7227 33560 33570 4272816 Species 8364 33637 33655 4272816 Species 8078 33692 33700 4272816 Species 71179 33902 33909 4272816 Species 93621 33910 33933 4272816 Chemical D014668 33856 33865 -4272816 Chemical D013256 33870 33877 +4272816 Chemical D013256 33870 33885 4272816 Gene 105937229 33964 33968 4272816 Species 8078 34051 34069 4272816 Species 8078 34071 34092 -4272816 Chemical C023035 34040 34047 +4272816 Chemical D011078 34040 34043 4272816 Chemical D011084 34003 34035 4272816 Species 1234273 34147 34161 4272816 Species 1234273 34163 34186 -4272816 Chemical D055435 34129 34138 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36023 4272816 Gene 105937232 36162 36166 4272816 Gene 105937229 36171 36175 -4272816 Species 8078 36192 36213 -4272816 Chemical D006838 36134 36150 4272816 Species 7719 36365 36383 4272816 Species 8364 36660 36667 4272816 Gene 105938153 36837 36842 4272816 Gene 105926779 36847 36852 -4272816 Chemical D004967 36880 36888 4272816 Species 8078 37054 37075 4272816 Species 8364 37229 37247 4272816 Species 7091 37449 37457 @@ -506,13 +510,13 @@ pubmed_id type identifier offset end 4272816 Species 8078 38087 38108 4272816 Species 29278 38135 38145 4272816 Species 8078 38255 38276 -4272816 Species 8078 38351 38372 4272816 Species 8078 38374 38392 4272816 Chemical D011078 38397 38422 4272816 Chemical D011078 38424 38428 +4272816 Disease D005393 38346 38372 4272816 Species 8364 38451 38469 4272816 Gene 105930542 38701 38705 -4272816 Species 8078 38670 38691 +4272816 Disease C538052 38664 38691 4272816 Species 7656 38758 38768 4272816 Species 9606 38847 38856 4272816 Species 9606 38955 38960 @@ 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16680 +4272823 Disease D009069 16498 16513 +4272823 Disease D010146 16676 16680 4272823 Disease D011471 16390 16405 +4272823 Disease D009069 16429 16443 +4272823 Disease D051271 16276 16290 4272823 Species 9606 17047 17054 4272823 Species 9606 17480 17488 4272823 Species 9606 17939 17947 @@ -1765,7 +1758,12 @@ pubmed_id type identifier offset end 4272823 Species 9606 18376 18384 4272823 Species 9606 18558 18566 4272823 Species 9606 18667 18675 +4272823 Disease D015212 18487 18500 +4272823 Disease D011349 18400 18409 +4272823 Disease D011349 18336 18354 +4272823 Disease D015212 18606 18619 4272823 Disease D011471 18807 18822 +4272823 Disease D051271 19610 19624 4272823 Species 9606 19750 19757 4272823 Disease D011471 19733 19748 4272823 Species 9606 20580 20588 @@ -1773,50 +1771,67 @@ pubmed_id type identifier offset end 4272823 Species 9606 20956 20964 4272823 Species 9606 21258 21266 4272823 Species 9606 21532 21540 +4272823 Disease D015212 20981 20995 +4272823 Disease D051271 21277 21291 +4272823 Disease D051271 20629 20643 +4272823 Disease D051271 21541 21563 +4272823 Disease D015212 20679 20693 4272823 Species 9606 21964 21972 -4272823 Disease D012002 22128 22143 +4272823 Disease D051271 21868 21882 +4272823 Disease D051271 22379 22393 +4272823 Disease D010146 22119 22123 4272823 Disease D064420 21750 21758 -4272823 Disease D011471 21633 21668 4272823 Disease D011471 21825 21840 -4272823 Disease C563475 22112 22123 +4272823 Disease D011349 21633 21668 +4272823 Disease D011349 21796 21805 +4272823 Disease D051271 22246 22260 +4272823 Disease D006470 22135 22143 4272823 Species 9606 23101 23109 -4272823 Disease D001005 22980 23001 -4272823 Disease D012002 22697 22712 -4272823 Disease D012002 23173 23188 +4272823 Disease D006470 22632 22640 +4272823 Disease D006470 22704 22712 4272823 Disease D006484 23005 23015 4272823 Disease D013684 23083 23097 4272823 Disease D009369 23192 23198 -4272823 Disease D006470 22934 22940 -4272823 Disease D012002 22509 22524 -4272823 Disease D012002 22620 22640 -4272823 Disease D012002 22888 22903 +4272823 Disease D006470 22895 22903 +4272823 Disease D001523 22991 23001 +4272823 Disease D064420 22516 22524 +4272823 Disease D006470 23180 23188 4272823 Species 9606 23658 23666 4272823 Species 9606 24017 24025 4272823 Disease D064420 23504 23514 -4272823 Disease D012002 23450 23465 -4272823 Disease D006470 24238 24246 -4272823 Disease D012002 23677 23692 -4272823 Disease D012002 23903 23923 -4272823 Disease D012002 24036 24051 +4272823 Disease D006470 23457 23465 +4272823 Disease D006470 24226 24246 +4272823 Disease D006470 23915 23923 +4272823 Disease D006470 24043 24051 +4272823 Disease D006470 23684 23692 4272823 Disease D011471 24648 24663 4272823 Species 9606 25269 25277 4272823 Species 9606 25777 25785 +4272823 Disease D015212 25429 25442 +4272823 Disease D015212 24875 24884 +4272823 Disease D015212 25318 25331 +4272823 Disease D015212 25025 25034 4272823 Species 9606 26236 26243 4272823 Species 9606 26429 26436 4272823 Species 9606 26572 26579 4272823 Disease D064420 27103 27111 +4272823 Disease D011349 27288 27297 4272823 Disease D064420 27018 27026 4272823 Species 9606 27569 27577 -4272823 Chemical D000728 27876 27884 +4272823 Disease D011349 27385 27394 +4272823 Chemical - 27872 27875 4272823 Disease D064420 27939 27947 -4272823 Disease C564570 28271 28273 +4272823 Chemical - 27984 27987 +4272823 Chemical - 28088 28091 +4272823 Chemical - 28112 28114 +4272823 Chemical - 28189 28192 4272823 Disease D009369 28306 28312 4272823 Disease D029461 28323 28325 +4272823 Disease C536108 28431 28440 4272823 Species 9606 29089 29096 4272823 Species 9606 29197 29205 -4272823 Chemical C441720 29492 29494 4272823 Disease D000544 29484 29486 -4272823 Disease D011471 29858 29886 +4272823 Disease D011472 29858 29886 4272823 Disease D011471 29975 29990 4272823 Disease D011471 30043 30058 4272823 Species 9606 30069 30076 @@ -1824,41 +1839,40 @@ pubmed_id type identifier offset end 4272823 Species 9606 30240 30247 4272823 Disease D011471 30327 30342 4272823 Disease D011471 30385 30400 -4272823 Disease D011832 30343 30364 +4272823 Disease D004194 30343 30364 4272823 Disease D011471 30467 30482 4272823 Species 9606 30560 30568 4272823 Disease D011471 30574 30589 4272823 Species 9606 30659 30667 4272823 Disease D002277 30717 30726 4272823 Disease D011471 30862 30877 -4272823 Disease D064420 30806 30814 -4272823 Disease D002908 30766 30773 +4272823 Disease D012002 30794 30814 4272823 Species 9606 30905 30913 +4272823 Disease D009369 30994 31003 4272823 Disease D064420 30893 30901 4272823 Disease D015212 30919 30945 -4272823 Disease D010386 30987 31003 4272823 Species 9606 31051 31059 -4272823 Disease D064420 31111 31128 +4272823 Disease D006470 31111 31128 4272823 Disease D011471 31035 31050 -4272823 Disease D011349 31143 31162 +4272823 Disease D011349 31153 31162 4272823 Disease D011471 31240 31255 4272823 CellLine D923 31358 31366 4272823 Disease D011471 31318 31333 4272823 Disease D011471 31465 31490 4272823 Disease D011349 31491 31500 -4272823 Disease D012002 31638 31653 +4272823 Disease D064420 31645 31653 4272823 Disease D011471 31538 31553 4272823 Disease D012002 31765 31778 4272823 Disease D064420 31990 31998 4272823 Species 9606 32199 32206 4272823 Species 9606 32245 32252 -4272823 Disease D064420 32332 32342 4272823 Disease D011471 32431 32456 +4272823 Disease D001745 32324 32342 4272823 Disease D064420 32469 32477 4272823 Disease D011471 32643 32658 4272823 Disease D011471 32685 32700 -4272823 Disease D011471 32802 32820 -4272823 Disease D011471 32821 32858 +4272823 Disease D011472 32802 32820 +4272823 Disease D011471 32843 32858 4272823 Disease D011471 32917 32942 4272823 Disease D011471 33016 33041 4272823 Disease D011471 33163 33188 @@ -1868,8 +1882,8 @@ pubmed_id type identifier offset end 4272823 Disease D011471 33730 33745 4272823 Disease D011471 34088 34113 4272823 Disease D011471 34201 34226 -4272823 Disease D012002 34249 34275 -4272823 Disease D011471 34309 34328 +4272823 Disease D011472 34309 34328 +4272823 Disease D009059 34261 34275 4272823 Species 9606 34477 34480 4272823 Disease D011471 34486 34501 4272823 Disease D011471 34372 34387 @@ -1885,78 +1899,87 @@ pubmed_id type identifier offset end 4272824 Species 9544 66 80 4272824 Chemical D003042 39 46 4272824 Species 9544 401 415 -4272824 Chemical D000661 299 312 4272824 Chemical D003042 767 774 4272824 Chemical D003042 518 525 4272824 Chemical D003042 335 342 4272824 Chemical C100294 279 294 4272824 Chemical D003042 227 234 4272824 Chemical D003042 364 371 +4272824 Chemical D003913 299 312 4272824 Chemical D003042 1095 1102 4272824 Chemical D003042 1021 1028 +4272824 Chemical - 977 980 4272824 Chemical D003042 1849 1856 +4272824 Chemical - 1567 1576 4272824 Chemical D003042 1693 1700 -4272824 Chemical D003042 1521 1528 +4272824 Disease D019970 1521 1538 4272824 Chemical D003042 1617 1624 -4272824 Chemical D015306 1567 1576 4272824 Chemical D003042 2760 2767 +4272824 Chemical - 3483 3486 4272824 Chemical D003042 4214 4221 4272824 Chemical D003042 5326 5333 4272824 Chemical D003042 4326 4333 -4272824 Chemical D003042 5076 5083 +4272824 Chemical - 4532 4535 +4272824 Chemical D008748 3893 3895 4272824 Chemical D003042 3716 3723 -4272824 Chemical D003042 4000 4007 4272824 Chemical D003042 4820 4827 +4272824 Chemical D003042 4000 4007 +4272824 Chemical C000604007 4688 4690 4272824 Chemical D003042 3884 3891 4272824 Chemical D003042 5569 5576 +4272824 Chemical D003913 5747 5760 4272824 Chemical C100294 5762 5777 -4272824 Chemical D003042 5936 5943 -4272824 Chemical D015306 5527 5536 +4272824 Chemical - 5527 5536 4272824 Chemical D003042 5693 5700 4272824 Chemical D010633 5783 5796 -4272824 Chemical D000661 5747 5760 -4272824 Chemical D003042 5892 5899 -4272824 Chemical D003042 6205 6212 +4272824 Chemical - 5838 5841 +4272824 Chemical - 5636 5639 4272824 Species 9544 6440 6454 4272824 Species 9544 6456 6470 4272824 Chemical D003042 7688 7695 4272824 Chemical D003042 7985 7992 4272824 Chemical D003042 8470 8477 -4272824 Chemical D003042 8771 8778 4272824 Chemical D003042 8545 8552 4272824 Chemical D003042 8367 8374 -4272824 Chemical D003042 8916 8923 +4272824 Chemical D012965 9324 9330 4272824 Chemical D003042 9533 9540 4272824 Chemical D003042 9401 9408 4272824 Chemical D010633 9238 9255 -4272824 Chemical D000661 9161 9174 4272824 Chemical C100294 9196 9215 +4272824 Chemical D003913 9161 9174 4272824 Chemical D003042 9836 9843 4272824 Chemical D003042 9785 9792 -4272824 Disease D005517 10276 10293 4272824 Chemical D003042 10065 10072 4272824 Chemical D003042 10189 10196 +4272824 Chemical - 11078 11081 4272824 Chemical D003042 10907 10914 4272824 Chemical D003042 10265 10272 +4272824 Chemical - 11046 11049 4272824 Chemical D003042 11232 11239 -4272824 Chemical C027727 11709 11711 -4272824 Chemical C027727 12973 12975 +4272824 Chemical - 11205 11208 +4272824 Chemical C000604007 12463 12465 +4272824 Chemical C000604007 12277 12279 +4272824 Chemical C000604007 12452 12454 +4272824 Chemical C000604007 12478 12480 4272824 Chemical D003042 13405 13412 +4272824 Chemical - 14404 14407 +4272824 Chemical D003913 14224 14237 +4272824 Chemical D003913 14754 14767 4272824 Chemical D003042 14271 14278 -4272824 Chemical D000661 14488 14501 +4272824 Chemical D003913 14488 14501 +4272824 Chemical D003913 14565 14578 4272824 Chemical D003042 14439 14446 -4272824 Chemical D000661 14224 14237 -4272824 Chemical D000661 14380 14393 -4272824 Chemical D000661 14754 14767 -4272824 Chemical D000661 14565 14578 +4272824 Chemical D003913 14380 14393 4272824 Chemical C100294 15555 15574 -4272824 Chemical C100294 15277 15296 -4272824 Chemical D003042 15043 15050 4272824 Chemical C100294 14990 15009 -4272824 Chemical C100294 15419 15438 -4272824 Chemical D003042 15228 15235 4272824 Chemical C100294 15152 15167 +4272824 Chemical D003042 15043 15050 +4272824 Chemical C100294 15277 15296 +4272824 Chemical - 15178 15181 +4272824 Chemical D003042 15228 15235 +4272824 Chemical C100294 15419 15438 4272824 Chemical D010633 16482 16499 +4272824 Chemical - 16115 16118 4272824 Chemical D010633 16092 16105 4272824 Chemical D003042 15983 15990 4272824 Chemical D010633 16214 16231 @@ -1965,57 +1988,64 @@ pubmed_id type identifier offset end 4272824 Chemical D003042 16643 16650 4272824 Chemical D003042 16578 16585 4272824 Chemical D003042 17060 17067 +4272824 Chemical - 16774 16777 4272824 Chemical D003042 16858 16865 4272824 Chemical D003042 16759 16766 -4272824 Chemical D015306 17319 17328 -4272824 Chemical D003042 17669 17676 +4272824 Disease D012640 17729 17733 +4272824 Chemical - 17406 17409 +4272824 Chemical - 17319 17328 +4272824 Chemical - 17627 17630 +4272824 Disease D012640 17657 17661 4272824 Chemical D003042 17994 18001 4272824 Species 9544 18525 18539 4272824 Species 9544 19116 19130 +4272824 Chemical - 19057 19060 4272824 Chemical D003042 19759 19766 4272824 Chemical D003042 21321 21328 +4272824 Chemical - 20491 20494 4272824 Chemical D003042 21276 21283 4272824 Chemical D003042 21373 21380 +4272824 Chemical D012965 21246 21252 4272824 Chemical D003042 21550 21557 4272824 Chemical D003042 22584 22591 +4272824 Chemical - 22244 22247 4272824 Chemical D003042 21826 21833 4272824 Chemical D003042 22452 22459 4272824 Chemical D003042 22361 22368 4272824 Chemical D003042 22517 22524 -4272824 Disease D014397 23211 23227 4272824 Chemical D003042 23075 23082 -4272824 Chemical D000661 22931 22944 -4272824 Chemical D000661 23470 23483 +4272824 Chemical D003913 23470 23483 4272824 Chemical D003042 23358 23365 4272824 Chemical D003042 23169 23176 +4272824 Chemical - 23695 23698 4272824 Chemical D003042 23413 23420 +4272824 Chemical - 22842 22845 4272824 Chemical D003042 23199 23206 -4272824 Chemical D003042 23904 23911 4272824 Chemical D003042 23729 23736 +4272824 Chemical D003913 22931 22944 4272824 Chemical D003042 23322 23329 -4272824 Chemical D003042 23564 23571 +4272824 Chemical - 23529 23532 +4272824 Chemical - 23133 23136 4272824 Chemical D003042 24372 24379 +4272824 Chemical D012965 24136 24142 4272824 Chemical D003042 24013 24020 4272824 Chemical D003042 24593 24600 -4272824 Chemical D003042 25269 25276 -4272824 Chemical D000661 25697 25710 -4272824 Chemical D003042 25653 25660 -4272824 Chemical D003042 25832 25839 +4272824 Chemical D003913 25697 25710 4272824 Chemical D003042 25462 25469 4272824 Species 9606 26637 26643 4272824 Chemical C100294 27069 27084 -4272824 Chemical D000661 27051 27064 4272824 Chemical D003042 25980 25987 +4272824 Chemical D003913 27051 27064 4272824 Chemical C100294 26809 26824 +4272824 Chemical D003913 26791 26804 4272824 Chemical D003042 26325 26332 -4272824 Chemical D000661 26791 26804 4272824 Chemical D003042 26898 26905 4272824 Chemical D003042 28035 28042 -4272824 Disease D010300 28337 28349 +4272824 Disease D010302 28337 28349 4272824 Chemical D000661 28376 28387 4272824 Chemical D003042 28401 28408 4272824 Chemical D009638 28550 28564 -4272824 Chemical D015306 28482 28491 +4272824 Chemical - 28482 28491 4272824 Chemical D003042 28600 28607 4272824 Chemical D012701 28572 28581 4272824 Chemical D004298 28541 28549 @@ -2023,63 +2053,53 @@ pubmed_id type identifier offset end 4272824 Chemical C100294 28907 28922 4272824 Chemical D003042 28941 28948 4272824 Species 9606 29150 29155 -4272824 Chemical C071741 29515 29527 +4272824 Chemical D000077208 29515 29527 4272824 Chemical D003042 29695 29702 4272824 Species 10116 29826 29830 4272824 Chemical D000431 29815 29822 4272824 Chemical D000431 29847 29854 -4272824 Chemical C071741 30185 30197 +4272824 Chemical D000077208 30185 30197 +4272824 Disease D009293 30414 30426 4272824 Chemical D003042 30402 30409 4272824 Chemical D003042 30801 30808 4272824 Chemical D003042 31019 31026 -4272824 Chemical D000661 31073 31086 +4272824 Chemical D003913 31073 31086 4272824 Chemical D003042 31100 31107 4272824 Species 9544 31291 31305 -4272824 Disease D000699 31324 31353 +4272824 Disease D000699 31324 31333 4272824 Chemical D009538 31389 31397 -4272824 Chemical D004298 31572 31580 4272824 Chemical D003042 31702 31709 4272824 Chemical D003042 31813 31820 4272824 Chemical D003042 32336 32343 -4272824 Chemical D003042 32029 32036 4272824 Chemical D003042 32303 32310 4272824 Chemical D003042 32433 32440 -4272824 Chemical D000661 32386 32399 -4272824 Chemical D003042 32634 32641 -4272824 Chemical D000661 32542 32555 -4272824 Chemical D000661 32801 32814 +4272824 Chemical D003913 32801 32814 +4272824 Chemical D003913 32386 32399 +4272824 Chemical D003913 32542 32555 4272824 Chemical D003042 32671 32678 4272824 Chemical C100294 33492 33511 -4272824 Chemical D003042 33325 33332 4272824 Chemical C100294 33065 33084 4272824 Chemical D003042 33362 33369 4272824 Chemical C100294 33227 33246 4272824 Chemical D003042 33118 33125 -4272824 Chemical D003042 34018 34025 4272824 Chemical D010633 33922 33939 4272824 Chemical D010633 34185 34202 4272824 Chemical D010633 33762 33779 4272824 Chemical D003042 34055 34062 4272824 Chemical D003042 33813 33820 4272824 Chemical D003042 34451 34458 -4272824 Chemical D003042 34679 34686 4272824 Chemical D003042 34482 34489 4272824 Chemical D010633 34845 34862 4272824 Chemical D003042 34715 34722 4272824 Chemical D003042 35388 35395 -4272824 Disease D014397 35448 35464 +4272824 Chemical D003913 35105 35118 4272824 Chemical D003042 35151 35158 4272824 Chemical D003042 35436 35443 -4272824 Chemical D000661 35105 35118 -4272824 Chemical C027727 41450 41452 -4272824 Chemical C027727 36768 36770 -4272824 Chemical D000661 36705 36760 -4272824 Chemical C027727 39887 39889 -4272824 Chemical D010633 39824 39879 -4272824 Chemical C027727 38397 38399 -4272824 Chemical C100294 38332 38389 +4272824 Chemical D010633 39820 39837 +4272824 Chemical C000604007 40104 40106 4272825 Gene 55465 72 76 4272825 Disease D003550 80 95 +4272825 Chemical C545203 12 21 4272825 Gene 55465 187 191 4272825 Gene 55465 264 268 4272825 Gene 55465 648 652 @@ -2094,21 +2114,22 @@ pubmed_id type identifier offset end 4272825 Species 9606 618 626 4272825 Species 9606 680 688 4272825 Species 9606 984 989 -4272825 DNAMutation p.508delF 1736 1745 4272825 ProteinMutation p.G551D;RS#:75527207 1123 1128 4272825 Disease D003550 96 111 -4272825 DNAMutation p.508delF 1373 1382 +4272825 Chemical C545203 323 332 +4272825 Chemical C569105 243 249 +4272825 Chemical C569105 231 241 4272825 ProteinMutation p.G551D;RS#:75527207 530 535 4272825 Gene 55465 2132 2136 4272825 Gene 55465 2156 2160 +4272825 Chemical D001639 2178 2182 4272825 Disease D007239 2462 2471 +4272825 Disease D003550 2567 2582 4272825 Disease D003550 1998 2013 4272825 Disease D000402 2431 2449 -4272825 Disease D003550 2554 2582 4272825 Disease D010190 2276 2284 4272825 Disease D007249 2473 2485 4272825 Disease D030342 1901 1936 -4272825 Chemical C504136 2178 2182 4272825 Gene 55465 2661 2665 4272825 Gene 55465 2913 2917 4272825 Gene 55465 2972 2976 @@ -2139,17 +2160,17 @@ pubmed_id type identifier offset end 4272825 Species 9606 5794 5802 4272825 Species 9606 5807 5812 4272825 Species 9606 6022 6030 -4272825 DNAMutation p.508delF 5840 5849 -4272825 ProteinMutation p.G551D;RS#:75527207 3598 3603 -4272825 DNAMutation p.508delF 5850 5859 +4272825 Chemical C569105 4889 4895 4272825 ProteinMutation p.G551D;RS#:75527207 3391 3396 4272825 Chemical D002712 3733 3741 -4272825 DNAMutation p.508delF 6050 6059 -4272825 Chemical D000255 3523 3526 -4272825 ProteinMutation p.G551D;RS#:75527207 3407 3412 +4272825 Chemical C545203 3269 3275 4272825 DNAMutation c.508DELTA>CFTR 5528 5542 -4272825 DNAMutation p.508delF 5661 5670 -4272825 Chemical C545203 3299 3307 +4272825 Chemical C545203 3277 3286 +4272825 Disease D003550 3818 3829 +4272825 Chemical C569105 5035 5041 +4272825 ProteinMutation p.G551D;RS#:75527207 3598 3603 +4272825 ProteinMutation p.G551D;RS#:75527207 3407 3412 +4272825 Chemical C569105 4897 4907 4272825 ProteinMutation p.G551D;RS#:75527207 3951 3956 4272825 Gene 55465 6189 6193 4272825 Gene 55465 6495 6499 @@ -2162,6 +2183,7 @@ pubmed_id type identifier offset end 4272825 Disease D029424 6992 6996 4272825 ProteinMutation p.G551D;RS#:75527207 6803 6808 4272825 Disease D029424 6953 6990 +4272825 Chemical C545203 6870 6876 4272825 Gene 55465 7220 7224 4272825 ProteinMutation p.G551D;RS#:75527207 7214 7219 4272825 Gene 55465 7311 7315 @@ -2173,14 +2195,13 @@ pubmed_id type identifier offset end 4272825 CellLine 0287 7706 7712 4272825 Chemical D005576 8041 8050 4272825 Chemical D005576 8178 8187 -4272825 DNAMutation p.508delF 7510 7519 4272825 ProteinMutation p.G551D;RS#:75527207 7723 7728 4272825 Chemical D005576 8541 8550 +4272825 Chemical C482900 8488 8499 4272825 Chemical D000584 7969 7978 4272825 Chemical D005576 8260 8269 4272825 ProteinMutation p.G551D;RS#:75527207 7654 7659 4272825 Disease D003550 7627 7646 -4272825 DNAMutation p.508delF 7729 7738 4272825 ProteinMutation p.G551D;RS#:75527207 7473 7478 4272825 Gene 55465 8788 8792 4272825 Gene 75079 8965 8991 @@ -2198,22 +2219,26 @@ pubmed_id type identifier offset end 4272825 ProteinMutation DeltaF508/DeltaF508 9772 9791 4272825 DNAMutation c.508DELTA>CFTR 9912 9926 4272825 CellLine 0287 9755 9761 +4272825 Chemical C569105 9558 9564 4272825 Gene 55465 10062 10066 4272825 Gene 55465 10567 10571 4272825 Gene 55465 10650 10654 4272825 Gene 55465 10217 10221 4272825 Gene 55465 10548 10552 4272825 CellLine 0287 10130 10136 -4272825 Chemical D000242 10042 10046 4272825 Chemical D005576 10345 10354 +4272825 Chemical C569105 10316 10322 +4272825 Chemical - 10042 10046 +4272825 Chemical C569105 10176 10182 4272825 Gene 55465 10827 10831 4272825 Gene 55465 11139 11143 4272825 Chemical D014544 11346 11349 +4272825 Chemical C482900 11163 11174 4272825 Chemical D005576 11082 11091 4272825 Gene 55465 11694 11698 4272825 CellLine 0287 11632 11638 4272825 Chemical D005576 11675 11684 -4272825 DNAMutation p.508delF 11538 11547 +4272825 Chemical C569105 11590 11596 4272825 CellLine 0287 11511 11517 4272825 Gene 55465 11965 11969 4272825 Gene 55465 12668 12672 @@ -2230,6 +2255,7 @@ pubmed_id type identifier offset end 4272825 Chemical D003513 13384 13397 4272825 CellLine 0287 12326 12332 4272825 CellLine 0287 12467 12473 +4272825 Chemical C569105 12393 12399 4272825 Gene 55465 13909 13913 4272825 Gene 55465 14097 14101 4272825 Gene 55465 14144 14148 @@ -2246,19 +2272,21 @@ pubmed_id type identifier offset end 4272825 Gene 55465 15632 15636 4272825 Gene 55465 15759 15763 4272825 Gene 55465 15869 15873 +4272825 Chemical C482900 15784 15795 4272825 CellLine 0287 16129 16135 4272825 Chemical D000584 15977 15986 4272825 Chemical D005576 15687 15696 4272825 Chemical D014544 16058 16061 +4272825 Chemical C545203 15926 15932 4272825 Gene 55465 16802 16806 4272825 Gene 75079 16812 16816 4272825 Gene 6267 17531 17534 4272825 Gene 788 17220 17223 4272825 Gene 788 16690 16693 4272825 Chemical D004121 16542 16546 +4272825 Chemical D011090 16976 16983 4272825 Chemical D000584 16745 16754 4272825 Chemical D009761 16964 16972 -4272825 Chemical D011090 16976 16983 4272825 Gene 55465 17626 17630 4272825 ProteinMutation p.G551D;RS#:75527207 17583 17588 4272825 Gene 55465 17679 17683 @@ -2282,7 +2310,9 @@ pubmed_id type identifier offset end 4272825 Gene 55465 19271 19275 4272825 Gene 55465 19388 19392 4272825 Gene 55465 19446 19450 +4272825 Chemical - 18799 18814 4272825 Chemical D000255 19118 19121 +4272825 Chemical - 18578 18593 4272825 ProteinMutation p.G551D;RS#:75527207 19218 19223 4272825 ProteinMutation p.G551D;RS#:75527207 19049 19054 4272825 ProteinMutation p.G551D;RS#:75527207 19382 19387 @@ -2294,7 +2324,6 @@ pubmed_id type identifier offset end 4272825 Gene 55465 20213 20217 4272825 Species 9606 19660 19668 4272825 Species 9606 19761 19769 -4272825 DNAMutation p.508delF 20047 20056 4272825 ProteinMutation p.G551D;RS#:75527207 19699 19704 4272825 Gene 55465 20567 20571 4272825 Gene 55465 20664 20668 @@ -2304,24 +2333,26 @@ pubmed_id type identifier offset end 4272825 Species 9606 20710 20718 4272825 Species 9606 21152 21160 4272825 Chemical D005576 20917 20926 +4272825 Chemical - 20955 20959 4272825 ProteinMutation p.G551D;RS#:75527207 20732 20737 4272825 ProteinMutation p.G551D;RS#:75527207 20300 20305 -4272825 Chemical D000242 20955 20959 +4272825 Chemical - 21056 21060 4272825 ProteinMutation p.G551D;RS#:75527207 21203 21208 4272825 ProteinMutation c.551G>D,D;RS#:75527207 20551 20558 4272825 ProteinMutation p.G551D;RS#:75527207 20392 20397 4272825 ProteinMutation p.G551D;RS#:75527207 21143 21148 -4272825 Chemical D000242 21056 21060 4272825 ProteinMutation c.551G>D,D;RS#:75527207 20784 20791 4272825 ProteinMutation p.G551D;RS#:75527207 20884 20889 4272825 Gene 55465 21432 21436 4272825 Gene 55465 22159 22163 4272825 Gene 55465 22448 22452 4272825 CellLine 0287 21399 21405 +4272825 Chemical C569105 21527 21533 4272825 CellLine 0287 22238 22244 4272825 CellLine 0287 21899 21905 -4272825 DNAMutation p.508delF 22615 22624 4272825 CellLine 0287 22625 22631 +4272825 Chemical C569105 21313 21319 +4272825 Disease D006331 22453 22464 4272825 Gene 55465 22717 22721 4272825 Gene 55465 22951 22955 4272825 Gene 55465 23018 23022 @@ -2342,9 +2373,9 @@ pubmed_id type identifier offset end 4272825 Gene 55465 24588 24592 4272825 Gene 55465 24673 24677 4272825 ProteinMutation p.G551D;RS#:75527207 24292 24297 -4272825 Chemical D000242 24197 24201 +4272825 Chemical - 24351 24355 4272825 ProteinMutation p.G551D;RS#:75527207 24096 24101 -4272825 Chemical D000242 24351 24355 +4272825 Chemical - 24197 24201 4272825 Gene 55465 24838 24842 4272825 Gene 55465 24996 25000 4272825 Gene 55465 25187 25191 @@ -2356,8 +2387,8 @@ pubmed_id type identifier offset end 4272825 Disease D029424 24968 24972 4272825 Disease D029424 24797 24801 4272825 CellLine 0287 25406 25412 +4272825 Chemical C545203 25156 25162 4272825 Disease D029424 25722 25726 -4272825 Chemical D005576 25378 25387 4272825 Disease D029424 24758 24795 4272825 Gene 55465 25763 25767 4272825 Gene 55465 25800 25804 @@ -2365,8 +2396,6 @@ pubmed_id type identifier offset end 4272825 Gene 55465 26807 26811 4272825 Gene 55465 26924 26928 4272825 Species 10116 25876 25879 -4272825 DNAMutation p.508delF 26914 26923 -4272825 DNAMutation p.508delF 26480 26489 4272825 ProteinMutation c.551G>D,D;RS#:75527207 26428 26435 4272825 Gene 55465 27196 27200 4272825 Gene 75079 27247 27251 @@ -2374,8 +2403,8 @@ pubmed_id type identifier offset end 4272825 Gene 75079 27387 27391 4272825 Chemical D014544 27092 27095 4272825 Disease D003550 27141 27160 -4272825 Chemical D014544 27097 27100 4272825 Chemical D014544 27232 27235 +4272825 Chemical D014544 27097 27100 4272825 Gene 55465 27619 27623 4272825 Gene 55465 27963 27967 4272825 Species 9606 27915 27923 @@ -2383,12 +2412,13 @@ pubmed_id type identifier offset end 4272825 Chemical D000584 27495 27504 4272825 Gene 55465 28408 28412 4272825 Species 9606 28842 28850 +4272825 Chemical C545203 29723 29729 +4272825 Chemical C569105 29680 29686 4272825 Gene 55465 30037 30041 4272825 Gene 55465 30657 30661 4272825 Gene 55465 30750 30754 4272825 Gene 55465 30108 30112 4272825 Species 9606 30464 30472 -4272825 DNAMutation p.508delF 30520 30529 4272825 Gene 55465 31372 31376 4272825 Gene 55465 31649 31653 4272825 Gene 55465 31681 31685 @@ -2400,14 +2430,13 @@ pubmed_id type identifier offset end 4272825 Species 10034 31815 31822 4272825 CellLine 0287 31542 31548 4272825 CellLine 1914 31831 31837 -4272825 DNAMutation p.508delF 31857 31866 4272825 Chemical D003513 31886 31899 4272825 Species 9606 32016 32021 4272825 CellLine 0287 32154 32160 -4272825 Chemical D002245 32306 32309 +4272825 Chemical - 32306 32309 4272825 Gene 101831516 32530 32534 +4272825 Chemical - 32477 32480 4272825 CellLine 1914 32384 32390 -4272825 Chemical D002245 32477 32480 4272825 CellLine 1914 32482 32488 4272825 Gene 55465 32687 32691 4272825 Gene 3071 32871 32874 @@ -2415,33 +2444,44 @@ pubmed_id type identifier offset end 4272825 Species 10090 32939 32944 4272825 Species 9925 33002 33006 4272825 Species 10090 33012 33017 -4272825 Chemical C032259 32852 32855 +4272825 Chemical D012967 32852 32855 +4272825 Chemical D012685 32775 32782 4272825 Chemical D003513 33209 33222 4272825 Gene 55465 33314 33318 4272825 CellLine 1914 33273 33279 4272825 Chemical D003513 33428 33441 +4272825 Chemical C569105 33351 33357 +4272825 Disease D011535 33685 33688 +4272825 Chemical D004801 33784 33789 +4272825 Chemical D010232 33759 33767 4272825 Chemical D005557 33723 33731 -4272825 Chemical D004801 33772 33789 +4272825 Chemical D006416 33772 33783 4272825 Gene 55465 34727 34731 4272825 Gene 55465 34733 34737 +4272825 Chemical - 34257 34260 4272825 Chemical D000584 34571 34580 -4272825 Chemical C093415 34186 34191 +4272825 Chemical - 34164 34168 4272825 CellLine 0287 34406 34412 -4272825 Chemical D013481 34251 34253 +4272825 Chemical - 34079 34084 4272825 Chemical D011188 34446 34455 -4272825 Chemical D002245 34257 34260 -4272825 Chemical D002118 34154 34158 +4272825 Chemical D001639 34197 34201 4272825 Chemical D014544 34802 34805 4272825 Chemical D019833 34692 34701 +4272825 Chemical - 34174 34180 4272825 Chemical D005947 34210 34217 -4272825 Chemical D008274 34164 34167 4272825 Chemical D005576 34631 34640 +4272825 Chemical C545203 34654 34660 4272825 Chemical D001639 34085 34096 +4272825 Chemical - 34186 34191 +4272825 Chemical - 34471 34475 +4272825 Chemical D000069285 34154 34158 +4272825 Chemical - 34251 34253 4272825 CellLine 0287 34334 34340 -4272825 Chemical C504136 34197 34201 +4272825 Chemical C482900 34762 34773 +4272825 Chemical C039004 34505 34508 4272825 Chemical D009761 34854 34862 4272825 Chemical D002712 34461 34469 -4272825 Chemical C043243 35723 35727 +4272825 Chemical D014867 35872 35877 4272825 Chemical D000432 35860 35868 4272825 Gene 55465 36354 36358 4272825 Gene 18868 36405 36409 @@ -2469,43 +2509,42 @@ pubmed_id type identifier offset end 4272825 Gene 6267 38312 38315 4272825 Gene 55465 38424 38428 4272825 Gene 6268 38391 38394 -4272825 Chemical C004944 38640 38643 +4272825 Chemical C023025 38835 38838 4272825 CellLine 0287 38751 38757 4272825 ProteinMutation p.G551D;RS#:75527207 38936 38941 -4272825 Chemical C004944 38835 38838 +4272825 Chemical C023025 38640 38643 4272825 Gene 55465 39278 39282 4272825 Disease D003550 39487 39515 4272825 Disease D003550 39543 39558 4272825 Disease D008171 39522 39539 -4272825 Disease D003550 39559 39574 +4272825 Disease D005355 39566 39574 4272825 Disease D003550 39608 39623 4272825 Disease D003550 39669 39684 4272825 Disease D003550 39763 39778 4272825 Gene 55465 39875 39879 4272825 Gene 55465 39926 39930 +4272825 Chemical - 39918 39925 4272825 Gene 55465 40021 40025 4272825 Species 9606 40052 40057 4272825 Disease D003550 40058 40073 4272825 Gene 55465 40166 40170 -4272825 DNAMutation p.508delF 40156 40165 4272825 Gene 55465 40262 40313 +4272825 Chemical - 40255 40261 4272825 Chemical D005897 40362 40371 4272825 Gene 55465 40451 40455 -4272825 Chemical C101426 40393 40403 +4272825 Chemical - 40443 40450 +4272825 Chemical D000068677 40393 40403 4272825 Gene 55465 40503 40554 4272825 Gene 55465 40556 40560 -4272825 Chemical D002712 40562 40570 4272825 Gene 55465 40729 40780 -4272825 Chemical D011720 40629 40656 -4272825 DNAMutation p.508delF 40719 40728 -4272825 Chemical D000242 40801 40805 +4272825 Chemical - 40801 40805 +4272825 Chemical - 40629 40656 4272825 Gene 55465 40904 40955 -4272825 Chemical C025253 40892 40903 -4272825 Chemical D002712 40956 40964 +4272825 Chemical - 40814 40829 4272825 Disease D003550 40985 41000 -4272825 Chemical D001190 40814 40829 4272825 Gene 55465 41070 41074 4272825 DNAMutation p.508delF 41059 41069 +4272825 Chemical C545203 41099 41108 4272825 Gene 55465 41172 41176 4272825 Species 9606 41129 41136 4272825 Disease D003550 41142 41157 @@ -2515,12 +2554,15 @@ pubmed_id type identifier offset end 4272825 Disease D003550 41222 41237 4272825 ProteinMutation p.G551D;RS#:75527207 41246 41251 4272825 Disease D003550 41335 41350 +4272825 Chemical C545203 41305 41314 4272825 Gene 55465 41401 41452 4272825 Gene 55465 41499 41503 4272825 Disease D003550 41458 41473 4272825 Gene 55465 41654 41658 4272825 Disease D003550 41603 41618 -4272825 Disease D005355 41717 41725 +4272825 Chemical - 41646 41653 +4272825 Chemical C545203 41576 41585 +4272825 Disease D003550 41710 41725 4272825 Gene 55465 41766 41770 4272825 Disease D003550 41813 41828 4272825 Gene 55465 41856 41907 @@ -2528,25 +2570,25 @@ pubmed_id type identifier offset end 4272825 Gene 55465 41985 41989 4272825 Gene 55465 42125 42129 4272825 Species 9606 42218 42223 -4272825 Disease D003550 42224 42257 +4272825 Disease D003550 42224 42239 4272825 Gene 55465 42317 42321 4272825 Gene 55465 42402 42406 4272825 Disease D003550 42359 42374 -4272825 Disease D003550 42416 42431 +4272825 Disease D005355 42423 42431 4272825 Disease D003550 42494 42509 4272825 Gene 55465 42570 42574 -4272825 Disease D003550 42595 42631 +4272825 Disease D003550 42595 42610 4272825 Gene 55465 42696 42700 4272825 Disease D003550 42632 42647 4272825 Gene 55465 42792 42843 4272825 Disease D008171 42875 42887 +4272825 Disease D006331 42844 42855 4272825 Gene 55465 42892 42943 4272825 Gene 55465 43006 43010 -4272825 Chemical D000242 43016 43020 -4272825 Chemical D012964 43044 43050 +4272825 Chemical - 43016 43020 4272825 Chemical D009761 43138 43146 4272825 Gene 55465 43229 43233 -4272825 Chemical C119108 43157 43170 +4272825 Chemical D010649 43157 43170 4272825 Gene 55465 43341 43392 4272825 Gene 55465 43394 43398 4272825 Gene 55465 43461 43465 @@ -2561,49 +2603,48 @@ pubmed_id type identifier offset end 4272825 Gene 55465 44249 44300 4272825 DNAMutation p.508delF 44238 44248 4272825 Disease D003550 44400 44415 -4272825 DNAMutation p.508delF 44390 44399 4272825 Gene 55465 44558 44562 4272825 Gene 55465 44505 44556 -4272825 DNAMutation p.508delF 44495 44504 4272825 Gene 55465 44759 44763 4272825 Disease D003550 44805 44820 4272825 Disease D003550 44866 44881 -4272825 DNAMutation p.508delF 44898 44907 4272825 Gene 55465 45152 45156 -4272825 Chemical D015056 45186 45190 4272825 DNAMutation p.508delF 45142 45151 -4272825 Chemical D002712 45199 45207 4272825 DNAMutation p.508delF 45249 45259 4272825 Disease D003550 45260 45275 4272825 ProteinMutation p.G551D;RS#:75527207 45300 45305 +4272825 Chemical C545203 45285 45294 4272825 Disease D003550 45322 45337 4272825 Species 9606 45377 45385 4272825 Disease D003550 45361 45376 +4272825 Chemical C545203 45338 45347 4272825 ProteinMutation p.G551D;RS#:75527207 45395 45400 4272825 Gene 55465 45487 45491 4272825 Gene 55465 45569 45573 4272825 ProteinMutation p.G551D;RS#:75527207 45563 45568 +4272825 Chemical C545203 45529 45538 4272825 Gene 55465 45669 45673 4272825 Gene 55465 45616 45667 4272825 Gene 55465 45869 45920 4272825 Gene 55465 45997 46001 +4272825 Chemical C545203 45984 45993 4272825 Species 9606 46163 46168 -4272825 Chemical D002118 46104 46108 4272825 Disease D003550 46180 46212 +4272825 Chemical D000069285 46104 46108 4272825 Gene 55465 46266 46270 4272825 Species 10090 46227 46233 -4272825 Chemical D002118 46275 46279 +4272825 Chemical D000069285 46275 46279 4272825 Species 10090 46376 46381 4272825 Gene 55465 46447 46451 +4272825 Chemical C545203 46464 46473 4272825 Gene 55465 46538 46542 -4272825 DNAMutation p.508delF 46528 46537 4272825 Species 9606 46585 46590 4272825 Species 9606 46623 46628 4272825 Gene 55465 46763 46767 4272825 Disease D003550 46771 46786 4272825 Species 9606 46793 46798 4272825 Disease D003550 46835 46850 -4272825 Chemical D009584 46868 46869 +4272825 Chemical - 46868 46893 4272825 Disease D003550 46932 46947 4272825 Gene 55465 47039 47043 4272825 Species 9606 47055 47060 @@ -2615,8 +2656,7 @@ pubmed_id type identifier offset end 4272825 ProteinMutation c.551G>D,D;RS#:75527207 47842 47849 4272825 Chemical D005576 47608 47617 4272825 Chemical D005576 47780 47789 -4272825 DNAMutation p.508delF 47356 47365 -4272825 DNAMutation p.508delF 48042 48051 +4272825 Chemical C482900 47804 47815 4272825 Species 9606 48979 48987 4272825 Species 9606 49023 49030 4272825 Chemical D005576 48277 48286 @@ -2645,14 +2685,15 @@ pubmed_id type identifier offset end 4272825 CellLine 0287 50756 50762 4272825 CellLine 0287 50567 50573 4272825 Chemical D005576 50180 50189 +4272825 Chemical C482900 50213 50224 4272825 ProteinMutation DeltaF508/DeltaF508 51170 51189 4272825 CellLine 0287 51754 51760 4272825 CellLine 0287 49807 49813 4272825 CellLine 0287 50960 50966 4272825 Chemical D005576 50883 50892 4272825 ProteinMutation DeltaF508/DeltaF508 50773 50792 -4272825 CellLine 0287 49871 49877 4272825 CellLine 0287 51640 51646 +4272825 CellLine 0287 49871 49877 4272825 CellLine 0287 51225 51231 4272825 CellLine 0287 51153 51159 4272825 ProteinMutation DeltaF508/DeltaF508 51768 51787 @@ -2660,7 +2701,9 @@ pubmed_id type identifier offset end 4272825 Chemical D004121 52058 52062 4272825 Chemical D005576 52221 52230 4272825 Chemical D005576 52194 52203 -4272825 Chemical D005576 52256 52273 +4272825 Chemical C545203 52140 52146 +4272825 Chemical C482900 52283 52294 +4272825 Chemical D005576 52256 52265 4272825 Chemical D006416 52754 52765 4272825 Chemical D004801 52770 52775 4272825 Chemical D009761 53155 53163 @@ -2680,14 +2723,13 @@ pubmed_id type identifier offset end 4272825 Species 9606 54090 54098 4272825 ProteinMutation p.G551D;RS#:75527207 53536 53541 4272825 ProteinMutation c.551G>D,D;RS#:75527207 54100 54107 -4272825 DNAMutation p.508delF 54763 54772 4272825 ProteinMutation p.G551D;RS#:75527207 53477 53482 +4272825 Chemical C545203 53647 53653 4272825 CellLine 0287 53407 53413 4272825 ProteinMutation c.551G>D,D;RS#:75527207 53945 53952 4272825 ProteinMutation c.551G>D,D;RS#:75527207 53566 53573 4272825 ProteinMutation p.G551D;RS#:75527207 54382 54387 4272825 ProteinMutation p.G551D;RS#:75527207 54567 54572 -4272825 DNAMutation p.508delF 54841 54850 4272825 ProteinMutation p.G551D;RS#:75527207 54161 54166 4272826 Gene 31223 0 5 4272826 Gene 121902 7 11 @@ -2720,7 +2762,6 @@ pubmed_id type identifier offset end 4272826 Disease D009369 386 392 4272826 Disease D009369 271 280 4272826 Disease D009369 962 968 -4272826 Disease D013967 551 578 4272826 Disease D008223 775 784 4272826 Gene 7640 1367 1371 4272826 Gene 7640 1447 1451 @@ -2783,9 +2824,9 @@ pubmed_id type identifier offset end 4272826 Species 10090 3904 3919 4272826 Species 10090 3714 3724 4272826 Species 10090 3869 3879 -4272826 Chemical D013754 3593 3605 -4272826 Disease D003643 4172 4177 -4272826 Chemical D013754 3531 3543 +4272826 Chemical D013752 3531 3543 +4272826 Chemical C010349 3482 3485 +4272826 Chemical D013752 3593 3605 4272826 Gene 37561 4292 4296 4272826 Gene 107053 4582 4622 4272826 Gene 31223 4720 4725 @@ -2798,7 +2839,6 @@ pubmed_id type identifier offset end 4272826 Species 10090 4471 4477 4272826 Species 10090 4713 4719 4272826 Disease D008223 4478 4496 -4272826 Chemical C068492 4942 4965 4272826 Gene 7640 5229 5233 4272826 Gene 31223 5418 5423 4272826 Gene 121902 5465 5469 @@ -2819,9 +2859,10 @@ pubmed_id type identifier offset end 4272826 Species 9606 5876 5881 4272826 Species 9986 5949 5955 4272826 Species 10090 6020 6026 -4272826 Chemical C047158 5472 5481 +4272826 Disease D008223 5058 5066 +4272826 Chemical C000469 6184 6187 4272826 Chemical D006416 6236 6247 -4272826 Disease D006327 5058 5073 +4272826 Disease D008223 6664 6672 4272826 Gene 31223 7815 7820 4272826 Gene 11214 7860 7864 4272826 Species 10090 6778 6782 @@ -2833,8 +2874,8 @@ pubmed_id type identifier offset end 4272826 Species 10090 6746 6756 4272826 Species 10090 6890 6900 4272826 Disease D008223 7567 7576 -4272826 Disease D005935 7493 7506 4272826 Disease D008223 7315 7323 +4272826 Disease D016399 7522 7561 4272826 Disease D007938 7071 7080 4272826 Disease D008223 7256 7265 4272826 Disease D008223 7393 7402 @@ -2844,7 +2885,7 @@ pubmed_id type identifier offset end 4272826 Disease D008223 6825 6834 4272826 Disease D008223 7206 7215 4272826 Disease D001523 7416 7433 -4272826 Disease D054218 7522 7561 +4272826 Disease D016399 7493 7506 4272826 Disease D008223 7454 7463 4272826 Gene 31223 7946 7951 4272826 Gene 121902 7953 7957 @@ -2880,20 +2921,20 @@ pubmed_id type identifier offset end 4272826 Species 10090 9510 9515 4272826 Species 10090 9704 9719 4272826 Species 10090 9376 9386 -4272826 Disease D005935 9286 9299 -4272826 Disease D016399 9678 9694 -4272826 Disease D054218 9311 9350 -4272826 Disease D005935 9737 9750 +4272826 Disease D008223 9685 9694 +4272826 Disease D016399 9286 9299 +4272826 Disease D016399 9737 9750 +4272826 Disease D016399 9311 9350 4272826 Gene 31311 10004 10007 4272826 Gene 31311 10615 10618 4272826 Species 1214577 10096 10100 4272826 Species 10090 10176 10186 4272826 Species 10090 10274 10284 -4272826 Disease D054218 10515 10554 -4272826 Disease D054218 10216 10254 +4272826 Disease D016399 10515 10554 4272826 Disease D016399 10354 10369 4272826 Disease D008223 10461 10470 4272826 Disease D009369 10336 10342 +4272826 Disease D016399 10216 10254 4272826 Disease D008223 10674 10682 4272826 Disease D008223 10735 10744 4272826 Disease D008223 10149 10158 @@ -2941,7 +2982,6 @@ pubmed_id type identifier offset end 4272826 Species 10090 12437 12441 4272826 Species 10090 12611 12617 4272826 Species 9606 12633 12639 -4272826 Species 10090 11747 11757 4272826 Species 10090 11840 11850 4272826 Disease D016399 12043 12059 4272826 Disease D009369 12024 12030 @@ -2949,17 +2989,18 @@ pubmed_id type identifier offset end 4272826 Disease D008223 12222 12231 4272826 Disease D008223 12376 12385 4272826 Disease D009369 12267 12273 -4272826 Disease D009369 11758 11764 4272826 Disease D008223 12618 12627 4272826 Disease D008223 11491 11500 4272826 Disease D009369 11583 11589 4272826 Disease D009369 12680 12689 +4272826 Disease D009369 11747 11764 4272826 Disease D008223 12771 12780 4272826 Disease D008223 11949 11958 4272826 Disease D008223 11376 11385 4272826 Disease D009369 11826 11832 4272826 Gene 121902 12925 12929 4272826 Gene 121902 13037 13041 +4272826 Gene 7640 13081 13085 4272826 Gene 121902 13241 13245 4272826 Gene 7640 13299 13303 4272826 Gene 7640 13329 13333 @@ -2970,6 +3011,7 @@ pubmed_id type identifier offset end 4272826 Gene 31223 13589 13594 4272826 Gene 121902 13609 13613 4272826 Gene 121902 13740 13744 +4272826 Gene 7640 13769 13773 4272826 Gene 121902 13832 13836 4272826 Gene 121902 13942 13946 4272826 Gene 7640 14088 14092 @@ -2982,22 +3024,19 @@ pubmed_id type identifier offset end 4272826 Species 9606 13905 13910 4272826 Species 10090 13137 13147 4272826 Gene 7640 13971 13975 -4272826 Disease D054198 12957 12979 4272826 Disease D008223 12878 12887 4272826 Disease D009369 13560 13569 4272826 Disease D008223 13511 13520 4272826 Disease D008223 13667 13676 -4272826 Disease D007938 12980 12989 -4272826 Chemical C060809 14215 14217 -4272826 Disease C563822 13081 13111 -4272826 Disease D016399 13870 13899 4272826 Disease D009369 14183 14188 -4272826 Disease D005935 13999 14012 +4272826 Disease D016399 12950 12989 4272826 Disease D008223 13266 13275 +4272826 Disease D009369 13102 13111 4272826 Disease D008223 13623 13632 +4272826 Disease D016399 13999 14012 +4272826 Disease D016399 13783 13796 4272826 Disease D009369 13718 13724 -4272826 Chemical C513342 13379 13383 -4272826 Disease C563822 13769 13796 +4272826 Disease D016399 13883 13899 4272826 Disease D008223 13911 13920 4272826 Gene 31311 14259 14262 4272826 Gene 7640 14404 14408 @@ -3032,13 +3071,13 @@ pubmed_id type identifier offset end 4272826 Disease D009369 15792 15798 4272826 Disease D008223 15064 15073 4272826 CellLine 4V27 15086 15095 -4272826 Disease D016399 15417 15433 4272826 Disease D008223 15287 15296 4272826 Disease D009369 14418 14427 4272826 Disease D009369 14340 14345 +4272826 Disease D008223 15424 15433 +4272826 Disease D016399 14359 14373 4272826 Disease D009369 15659 15665 4272826 Disease D009369 15746 15755 -4272826 Disease D005935 14361 14373 4272826 Disease D008223 14669 14677 4272826 Disease D008223 14483 14492 4272826 Disease D008223 15029 15038 @@ -3096,7 +3135,6 @@ pubmed_id type identifier offset end 4272826 Species 9606 18073 18078 4272826 Species 9606 18360 18366 4272826 Gene 3854 18231 18235 -4272826 Chemical D009584 18208 18209 4272826 Disease C536227 18323 18336 4272826 Gene 121902 18801 18805 4272826 Gene 121902 19732 19736 @@ -3114,11 +3152,11 @@ pubmed_id type identifier offset end 4272826 Species 10090 19341 19356 4272826 Species 10090 20135 20139 4272826 Disease D009369 20156 20162 -4272826 Disease D008223 19322 19331 4272826 Disease D016399 19066 19080 -4272826 Disease D016399 18884 18900 -4272826 Disease D008223 20067 20076 +4272826 Disease D008223 18891 18900 4272826 Disease D008223 19108 19117 +4272826 Disease D008223 19322 19331 +4272826 Disease D008223 20067 20076 4272826 Gene 31311 20346 20349 4272826 Gene 31311 20660 20663 4272826 Gene 31311 20910 20913 @@ -3132,7 +3170,6 @@ pubmed_id type identifier offset end 4272826 Species 9606 20654 20659 4272826 Species 9606 21233 21239 4272826 Gene 31311 21541 21544 -4272826 Chemical C038361 20461 20468 4272826 Disease D015179 21085 21090 4272826 Disease C536227 21136 21149 4272826 Disease D054218 21817 21850 @@ -3163,12 +3200,11 @@ pubmed_id type identifier offset end 4272826 Disease D054198 22354 22377 4272826 Disease D007951 22519 22539 4272826 Disease D008223 22719 22728 -4272826 Disease D006689 22268 22285 -4272826 Disease D019337 22111 22135 4272826 Disease D015470 22942 22964 4272826 Disease D054198 22884 22906 -4272826 Disease D008232 23074 23108 -4272826 Disease C536227 22094 22107 +4272826 Disease D008232 23072 23108 +4272826 Disease D019337 22094 22135 +4272826 Disease D008228 22264 22285 4272826 Gene 7640 23240 23244 4272826 Gene 31311 24019 24022 4272826 Gene 31223 24185 24190 @@ -3176,19 +3212,18 @@ pubmed_id type identifier offset end 4272826 Gene 31311 24202 24205 4272826 Gene 7640 24244 24248 4272826 Disease D009369 23914 23919 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3856 326 330 2289506 Gene 3856 386 390 @@ -7796,15 +7904,14 @@ pubmed_id type identifier offset end 2289506 Species 10090 767 772 2289506 Species 10090 961 966 2289506 Species 10090 1829 1834 +2289506 Species 9606 1912 1917 2289506 Species 8355 1198 1205 2289506 Disease D007674 1030 1046 -2289506 Disease D015658 1893 1922 -2289506 Disease D008171 945 949 2289506 CellLine 0525 1926 1934 +2289506 CellLine 0030 1918 1922 2289507 Gene 55829 34 53 2289507 Species 9606 28 33 2289507 CellLine 0229 145 149 -2289507 Chemical C102887 79 83 2289507 Gene 55829 562 581 2289507 Gene 55829 878 897 2289507 Species 9606 527 532 @@ -7818,46 +7925,55 @@ pubmed_id type identifier offset end 2289507 CellLine 0229 340 344 2289507 Chemical D015739 1340 1350 2289507 CellLine 0229 1093 1097 -2289507 Chemical C102887 1481 1485 2289507 Chemical D017239 1331 1336 -2289507 Chemical C102887 708 712 -2289507 Chemical D000641 1454 1457 2289508 Disease D009369 63 68 2289508 Gene 55890 435 440 2289508 Gene 55890 645 650 2289508 Gene 8271 1231 1236 2289508 Gene 8271 1253 1294 2289508 Gene 55890 493 498 -2289509 Disease D015823 88 110 +2289508 Chemical D006820 753 763 +2289509 Chemical D010743 36 48 +2289509 Disease D000562 88 110 2289509 Gene 47922 141 159 2289509 Gene 32046 888 897 2289509 Gene 23480 1057 1063 2289509 Gene 23480 980 986 2289509 Gene 23480 669 675 2289509 Gene 23480 403 409 -2289509 Chemical D014280 585 598 -2289509 Chemical C102887 768 772 +2289509 Chemical D008055 543 548 2289509 Chemical D000255 713 716 2289509 Chemical D013261 603 610 -2289509 Disease D015823 824 846 -2289509 Chemical D000641 299 302 -2289509 Disease D015823 223 245 +2289509 Chemical D008055 363 368 +2289509 Disease D000562 824 846 +2289509 Chemical D010743 192 204 +2289509 Chemical D008055 618 623 +2289509 Chemical D010743 517 529 +2289509 Chemical D010743 449 462 +2289509 Disease D000562 223 245 +2289509 Chemical D010743 915 927 +2289509 Chemical - 577 598 +2289510 Chemical D015638 49 63 2289510 Chemical D003078 126 136 2289510 Chemical D003078 1913 1923 -2289510 Chemical D003078 1651 1661 +2289510 Chemical D015638 1928 1930 +2289510 Disease D001259 465 503 +2289510 Chemical D015638 2676 2678 2289510 Chemical D003078 2282 2292 -2289510 Disease D001259 465 478 +2289510 Chemical D015638 330 344 +2289510 Chemical D015638 346 348 +2289510 Chemical D015638 946 948 2289510 Chemical D003078 2390 2400 2289510 Chemical D003078 315 325 +2289510 Chemical D015638 1137 1139 +2289510 Disease C535850 1498 1518 +2289510 Chemical D015638 522 524 +2289510 Chemical D003078 1651 1661 +2289510 Disease C535850 777 797 +2289510 Chemical D015638 2402 2404 2289512 CellLine 0422 66 70 2289512 Chemical D020126 80 91 2289512 Gene 20917 730 740 -2289512 Chemical C524118 402 405 -2289512 Chemical C524118 253 256 -2289512 Chemical C524118 672 675 -2289512 Chemical C524118 192 195 -2289512 Chemical C524118 759 762 -2289512 Chemical C524118 854 857 2289512 CellLine 0422 180 184 2289513 Species 11276 47 73 2289513 Species 11276 255 281 @@ -7865,40 +7981,46 @@ pubmed_id type identifier offset end 2289513 Species 11276 1091 1100 2289514 Species 10090 20 35 2289514 Disease D010009 0 16 -2289514 Chemical D000596 48 61 2289514 Species 10090 152 167 2289514 Species 10090 197 202 2289514 Species 10090 1612 1616 2289514 Species 9606 1680 1685 2289514 Species 10090 803 813 -2289514 Disease D009139 672 692 -2289514 Disease D012128 733 753 +2289514 Disease D000741 607 618 +2289514 Disease D019465 627 660 2289514 Disease D010009 1686 1703 2289514 Disease D006984 1031 1043 +2289514 Disease C535964 1282 1308 +2289514 Disease D009139 672 692 +2289514 Disease D015831 854 867 +2289514 Disease D002357 404 413 2289515 Species 8355 44 58 2289515 Gene 379502 374 385 2289515 Gene 379133 414 417 2289515 Species 8355 138 152 2289515 Species 4565 1121 1126 2289515 Disease D008607 1460 1471 +2289515 Chemical D000117 1215 1234 2289515 Chemical D000255 928 931 -2289515 Chemical C014088 1215 1234 +2289515 Chemical C000615311 191 194 +2289515 Chemical - 967 987 2289517 Gene 40754 39 68 2289517 Gene 40754 70 76 2289517 Gene 40754 118 147 2289517 Gene 40754 149 154 2289517 Gene 1533 285 296 2289517 Gene 40754 2156 2161 -2289517 Chemical D000242 2045 2049 -2289517 Chemical D007295 1744 1763 -2289517 Chemical D002118 1949 1953 -2289517 Chemical D011107 1427 1440 +2289517 Chemical D000069285 1949 1953 2289517 Chemical D002241 2305 2317 -2289517 Chemical C068152 2054 2057 +2289517 Chemical C066524 2054 2057 +2289517 Chemical D007295 1744 1763 +2289517 Chemical - 2045 2049 2289518 Species 6257 82 92 2289518 Species 6257 1161 1171 +2289518 Disease D060725 299 319 2289519 Gene 51880 741 744 2289519 Species 6978 259 280 +2289519 Chemical - 967 972 2289519 Disease D005207 1086 1099 2289520 Gene 136830 140 144 2289520 Gene 137176 149 153 @@ -7924,8 +8046,7 @@ pubmed_id type identifier offset end 2289522 Species 562 512 528 2289522 Species 562 910 917 2289522 Species 562 1213 1220 -2289522 Chemical C478787 547 561 -2289522 Chemical C047902 142 148 +2289522 Chemical - 1276 1281 2289522 ProteinMutation p.S195A 1260 1274 2289524 Gene 1876 0 19 2289524 Gene 1876 643 662 @@ -7935,11 +8056,8 @@ pubmed_id type identifier offset end 2289524 Species 9031 161 168 2289524 Chemical D000242 774 784 2289524 Chemical C020243 223 240 -2289524 Chemical D002701 407 422 2289525 Gene 23480 1683 1689 2289525 Gene 23480 1931 1937 -2289525 Disease D007239 1186 1195 -2289525 Disease D007239 1533 1542 4272812 Gene 293 10 25 4272812 Disease D010300 41 60 4272812 Gene 293 303 318 @@ -7958,22 +8076,23 @@ pubmed_id type identifier offset end 4272812 Species 9606 1232 1237 4272812 Disease D001523 167 178 4272812 Disease D010300 557 559 +4272812 Disease D001342 183 204 4272812 Disease D010300 495 497 +4272812 Chemical D004298 1344 1346 4272812 Disease D010300 61 80 4272812 Disease D010300 1495 1497 4272812 Chemical D004298 1334 1342 4272812 Chemical D012402 944 952 +4272812 Disease D019636 656 673 4272812 Disease D019636 1512 1529 -4272812 Disease D019636 643 673 4272812 Disease D010300 82 84 -4272812 Disease D012735 183 204 4272812 Species 9606 2202 2210 4272812 Disease D010300 2530 2532 4272812 Disease D010300 2199 2201 4272812 Disease D010302 1769 1778 4272812 Disease D010300 2071 2073 +4272812 Disease D009103 1862 1880 4272812 Disease D010300 1625 1627 -4272812 Disease D012598 1871 1880 4272812 Disease D010300 1854 1856 4272812 Disease D010300 1604 1623 4272812 Disease D014202 1932 1938 @@ -7983,31 +8102,33 @@ pubmed_id type identifier offset end 4272812 Disease D019636 2712 2738 4272812 Disease D004409 2822 2830 4272812 Disease D009127 2832 2840 -4272812 Disease D009410 2943 2979 4272812 Gene 1166 3900 3910 4272812 Species 9606 3482 3490 4272812 Disease D007029 3924 3936 +4272812 Disease D004067 4260 4282 4272812 Disease D009410 3637 3650 4272812 Disease D001523 4422 4456 4272812 Disease D007029 4111 4123 4272812 Disease D010300 4473 4475 4272812 Disease D020774 5259 5273 -4272812 Disease D000544 5011 5030 -4272812 Disease D015835 5170 5197 +4272812 Disease D010300 5036 5047 4272812 Disease D019636 4519 4545 -4272812 Disease D019578 4986 5009 -4272812 Disease D012791 5133 5162 -4272812 Disease D010300 4915 4917 4272812 Disease D006211 5104 5130 -4272812 Disease C537240 5199 5220 +4272812 Disease D020961 4811 4824 +4272812 Disease D006211 5072 5102 +4272812 Disease D001284 5002 5009 4272812 Disease D003704 4967 4975 -4272812 Chemical D007501 5072 5082 4272812 Disease D019636 5049 5066 4272812 Disease D005776 5238 5255 +4272812 Disease D000544 5011 5030 +4272812 Disease D010300 4915 4917 +4272812 Disease D020187 5164 5197 +4272812 Disease C537240 5199 5220 +4272812 Disease D012791 5145 5162 4272812 Disease D020388 5987 6001 4272812 Chemical D004801 5506 5511 +4272812 Chemical D006416 5493 5505 4272812 Chemical D004801 5916 5921 -4272812 Chemical C018328 5493 5505 4272812 Gene 293 6252 6261 4272812 Disease D003704 6082 6090 4272812 Gene 293 6790 6799 @@ -8030,7 +8151,7 @@ pubmed_id type identifier offset end 4272812 Gene 37256 9428 9431 4272812 Gene 20414 9749 9752 4272812 Gene 31298 9754 9757 -4272812 Disease D010300 10163 10182 +4272812 Disease D010302 10163 10175 4272812 Disease D010300 9978 9980 4272812 Gene 293 10499 10508 4272812 Gene 55937 10599 10604 @@ -8041,21 +8162,14 @@ pubmed_id type identifier offset end 4272812 Species 10090 10610 10614 4272812 Gene 16473 10569 10572 4272812 Chemical D012402 10580 10588 -4272812 Chemical C045084 10897 10904 -4272812 Chemical D012538 10720 10728 -4272812 Chemical D013910 10939 10943 -4272812 Chemical C045084 10884 10891 -4272812 Disease D009765 10655 10657 -4272812 Chemical C045084 11033 11039 -4272812 Chemical C045084 10845 10852 -4272812 Chemical C045084 10858 10864 +4272812 Chemical C028505 10991 10993 4272812 Gene 293 11470 11479 4272812 Species 9606 11209 11217 4272812 Species 9606 11415 11423 4272812 Disease D010300 11638 11640 4272812 Disease D010300 11206 11208 -4272812 Disease D010300 11524 11532 -4272812 Disease D010300 11219 11227 +4272812 Disease D010300 11225 11227 +4272812 Disease D010300 11530 11532 4272812 Disease D010300 11445 11447 4272812 Gene 293 11661 11670 4272812 Gene 293 12094 12103 @@ -8064,13 +8178,13 @@ pubmed_id type identifier offset end 4272812 Disease D010300 12493 12495 4272812 Species 9606 12536 12544 4272812 Species 9606 12670 12678 -4272812 Disease C537708 12977 13008 4272812 Disease D010300 12941 12943 4272812 Disease D010300 12667 12669 -4272812 Disease D058495 13370 13389 4272812 Disease D010300 13140 13142 4272812 Disease D010300 12550 12552 +4272812 Disease D066263 13370 13389 4272812 Disease D010300 13232 13243 +4272812 Disease D016464 13347 13368 4272812 Gene 293 13535 13544 4272812 Gene 18982 13556 13561 4272812 Gene 293 13704 13708 @@ -8085,14 +8199,15 @@ pubmed_id type identifier offset end 4272812 Species 9606 14506 14514 4272812 Gene 293 14797 14801 4272812 Disease D010300 14475 14477 +4272812 Chemical D015226 14665 14677 4272812 ProteinMutation p.A30P;RS#:104893878 14093 14097 4272812 Disease D010300 14268 14270 4272812 Disease D010300 14089 14091 4272812 Disease D010300 14879 14881 4272812 Disease D010300 14981 14983 4272812 ProteinMutation p.E46K;RS#:104893875 14102 14106 +4272812 Disease D010300 13985 13987 4272812 DNAMutation c.53A>T;RS#:104893877 13898 13902 -4272812 Disease D010300 13976 13987 4272812 Gene 293 14994 14998 4272812 Gene 293 15071 15080 4272812 Gene 293 15382 15386 @@ -8103,11 +8218,12 @@ pubmed_id type identifier offset end 4272812 Gene 293 15984 15988 4272812 DNAMutation c.53A>T;RS#:104893877 15365 15369 4272812 Disease D010300 15683 15685 -4272812 Disease D008569 15528 15539 -4272812 Disease D003704 16092 16100 4272812 Disease D010300 16084 16086 4272812 Disease D019636 15713 15730 +4272812 Chemical D008055 15213 15219 4272812 Disease D058225 15424 15444 +4272812 Disease D003704 16092 16100 +4272812 Disease D009422 15528 15539 4272812 Gene 293 16106 16110 4272812 Gene 74962 16138 16172 4272812 Gene 74962 16174 16178 @@ -8130,9 +8246,9 @@ pubmed_id type identifier offset end 4272812 ProteinMutation p.Y1699C;RS#:35801418 17070 17076 4272812 ProteinMutation p.G2385R;RS#:34778348 17205 17211 4272812 Disease D010300 16660 16662 -4272812 ProteinMutation p.R1441G,C,H;RS#:33939927;34995376 17058 17068 4272812 ProteinMutation p.G2019S;RS#:34637584 17078 17084 4272812 Disease D010300 16731 16761 +4272812 ProteinMutation p.R1441G,C,H;RS#:33939927 17058 17068 4272812 ProteinMutation p.G2019S;RS#:34637584 17235 17241 4272812 Gene 18982 17841 17846 4272812 Gene 18982 17995 18000 @@ -8199,16 +8315,17 @@ pubmed_id type identifier offset end 4272812 Species 9606 23845 23851 4272812 Chemical D015632 23255 23259 4272812 Chemical D012402 23261 23269 -4272812 Disease D010300 23696 23708 -4272812 Chemical C078846 23329 23339 4272812 Chemical D016627 24235 24241 4272812 Chemical D015632 23733 23737 +4272812 Disease D001259 23651 23665 4272812 Disease D010300 23528 23530 4272812 Chemical D015632 24226 24230 +4272812 Chemical D008070 23370 23388 4272812 Chemical D016627 23217 23234 4272812 Disease D010300 23155 23157 4272812 Disease D020258 24242 24252 4272812 Chemical D010269 23274 23282 +4272812 Disease D010302 23696 23708 4272812 Gene 293 24883 24887 4272812 Gene 18982 24891 24896 4272812 Gene 32672 25038 25044 @@ -8225,28 +8342,25 @@ pubmed_id type identifier offset end 4272812 Species 10090 25719 25723 4272812 Species 10090 24834 24844 4272812 Disease D019636 24979 24996 -4272812 Disease D009133 25281 25293 4272812 DNAMutation c.53A>T;RS#:104893877 25682 25686 -4272812 Disease D010300 25202 25204 4272812 Chemical D012402 25614 25622 4272812 Disease D010300 25436 25438 +4272812 Disease D010300 25202 25204 4272812 Disease D010300 24663 24665 4272812 Species 9606 25946 25951 4272812 Gene 293 26558 26567 4272812 Species 9606 26041 26046 4272812 Species 9606 26152 26158 4272812 Disease D010300 26266 26268 -4272812 Disease D003072 26391 26409 4272812 Chemical D015632 26415 26419 +4272812 Disease D020271 26494 26537 4272812 Disease D064420 26455 26463 4272812 Disease D010300 26093 26095 4272812 Disease D003248 26343 26355 +4272812 Disease D003072 26369 26409 4272812 Chemical D015632 25967 26011 -4272812 Disease D019636 26494 26511 4272812 Chemical D015632 26629 26633 4272812 Disease D010302 26136 26148 -4272812 Disease D009422 26516 26537 -4272812 Disease D012893 26369 26386 4272812 Chemical D015632 25961 25965 4272812 Chemical D015632 26699 26703 4272812 Gene 293 27052 27061 @@ -8257,11 +8371,14 @@ pubmed_id type identifier offset end 4272812 Species 10090 26715 26719 4272812 Species 10090 26835 26839 4272812 Species 9606 27591 27596 +4272812 Chemical D011339 27270 27280 4272812 Chemical D015632 26725 26729 4272812 Chemical D015632 27686 27690 4272812 Chemical D015632 28115 28119 +4272812 Chemical D008055 27449 27454 4272812 Chemical D008062 27624 27635 4272812 Chemical D015632 27328 27332 +4272812 Chemical D011339 28135 28145 4272812 Chemical D015632 27238 27242 4272812 Chemical D012402 28265 28273 4272812 Gene 293 28825 28834 @@ -8276,15 +8393,15 @@ pubmed_id type identifier offset end 4272812 Disease D019636 29537 29554 4272812 Chemical D012402 29382 29390 4272812 Disease D010300 28924 28926 +4272812 Chemical D008070 28985 29003 4272812 Chemical D012402 29155 29163 4272812 Chemical D012402 28791 28799 4272812 Disease D010300 29788 29790 4272812 Disease D064420 29391 29399 4272812 Chemical D012402 28282 28290 4272812 Chemical D012402 28564 28572 -4272812 Disease D010300 28521 28523 4272812 Disease D019636 28700 28717 -4272812 Chemical D010269 29860 29868 +4272812 Disease D010300 28521 28523 4272812 Disease D019636 29941 29958 4272812 Gene 37894 30045 30083 4272812 Gene 37894 30085 30091 @@ -8295,8 +8412,7 @@ pubmed_id type identifier offset end 4272812 Species 10116 30702 30706 4272812 Species 10090 30930 30934 4272812 Disease D064420 30182 30190 -4272812 Disease D010300 30096 30107 -4272812 Chemical C078846 30381 30391 +4272812 Disease D010300 30105 30107 4272812 Disease D010300 30845 30847 4272812 Disease D028361 31577 31602 4272812 Gene 293 31977 31986 @@ -8313,15 +8429,14 @@ pubmed_id type identifier offset end 4272812 Species 185869 32113 32120 4272812 Gene 16473 32229 32232 4272812 Gene 16473 32121 32124 -4272812 Disease D009410 32048 32067 4272812 Disease D019636 31704 31721 4272812 Disease D064420 32233 32241 +4272812 Disease C565640 32048 32067 4272812 Disease D010300 31780 31782 4272812 Disease D019636 32544 32561 4272812 Chemical D015632 31882 31886 4272812 Disease D010300 31661 31663 4272812 Chemical D012402 31891 31899 -4272812 Disease D028361 32147 32174 4272812 Gene 293 32731 32740 4272812 Gene 293 32855 32864 4272812 Gene 293 32912 32921 @@ -8344,18 +8459,15 @@ pubmed_id type identifier offset end 4272812 Gene 16473 33876 33879 4272812 ProteinMutation p.A30P;RS#:104893878 32836 32840 4272812 Disease D019636 34571 34588 -4272812 Disease C579880 34393 34404 4272812 Disease D010300 34614 34616 4272812 DNAMutation c.53A>T;RS#:104893877 32830 32834 4272812 Disease D019636 34509 34526 -4272812 Disease D017093 34009 34022 -4272812 Chemical C032259 34285 34288 +4272812 Disease D001259 34009 34022 4272812 Disease D010300 33632 33634 -4272812 Disease D003643 34067 34072 -4272812 Disease D013568 33098 33124 +4272812 Chemical D012967 34285 34288 4272812 Disease D010243 34053 34062 -4272812 Disease D019636 33269 33286 4272812 Disease D010300 33006 33008 +4272812 Disease D019636 33269 33286 4272812 Gene 18982 34647 34652 4272812 Gene 18982 34666 34671 4272812 Gene 293 34842 34851 @@ -8371,6 +8483,7 @@ pubmed_id type identifier offset end 4272812 Species 10090 35366 35370 4272812 Species 185869 35354 35361 4272812 Gene 16473 35362 35365 +4272812 Disease D009461 34709 34716 4272812 ProteinMutation p.R1441C;RS#:33939927 34893 34899 4272812 ProteinMutation p.G2019S;RS#:34637584 35151 35157 4272812 Disease D007674 34773 34786 @@ -8432,14 +8545,14 @@ pubmed_id type identifier offset end 4272812 Species 9606 38500 38508 4272812 Species 10090 38605 38609 4272812 Species 9606 39779 39784 +4272812 Chemical - 39674 39680 4272812 Chemical D015632 38592 38596 -4272812 Disease D014693 38465 38490 -4272812 Chemical C530429 39674 39677 +4272812 Chemical D004298 39131 39133 4272812 Disease D010300 38514 38516 4272812 Disease D010300 38351 38353 4272812 Disease D010300 39006 39008 -4272812 Disease D007027 39991 40010 4272812 Disease D019636 39045 39062 +4272812 Disease D007027 39991 40003 4272812 Gene 293 40717 40726 4272812 Gene 293 40820 40829 4272812 Gene 293 40950 40954 @@ -8468,12 +8581,12 @@ pubmed_id type identifier offset end 4272812 Disease D003704 41071 41079 4272812 Disease D003072 40518 40539 4272812 Disease D010300 41990 41992 -4272812 Disease D002653 40410 40430 4272812 Disease D003072 42079 42099 +4272812 Disease D010300 41185 41187 +4272812 Disease D001523 40410 40430 4272812 Disease D009422 41865 41887 4272812 Disease D010300 40373 40375 4272812 Disease D010300 40553 40555 -4272812 Disease D010300 41176 41187 4272812 Gene 293 42685 42694 4272812 Gene 18982 42931 42936 4272812 Species 9606 42212 42217 @@ -8481,24 +8594,28 @@ pubmed_id type identifier offset end 4272812 Species 9606 42659 42664 4272812 Species 6239 42698 42708 4272812 Species 6239 42868 42878 +4272812 Species 7227 42222 42232 +4272812 Species 7227 42853 42863 4272812 Gene 16473 42979 42982 4272812 Gene 16473 42900 42903 4272812 Gene 16473 42775 42778 4272812 Gene 16473 42348 42351 4272812 Gene 16473 42249 42252 +4272812 Disease D010302 42602 42614 4272812 Disease D010300 42218 42220 -4272812 Disease D019636 42730 42747 -4272812 Disease D010300 42602 42620 -4272812 Disease D009069 42303 42324 4272812 Disease D010300 43099 43101 +4272812 Disease D009069 42303 42324 +4272812 Disease D019636 42730 42747 4272812 Chemical D004298 43188 43196 4272812 Gene 293 43248 43257 4272812 Gene 293 43328 43337 4272812 Gene 307 43347 43367 4272812 Gene 293 43572 43581 4272812 Disease D064420 43560 43568 +4272812 Chemical D004298 43488 43490 +4272812 Chemical D004298 43267 43269 +4272812 Chemical D004298 43516 43518 4272812 Chemical D007980 43478 43484 -4272812 Chemical D065105 43418 43437 4272812 Gene 55937 43587 43592 4272812 Gene 293 43649 43658 4272812 Gene 55937 43686 43691 @@ -8523,12 +8640,17 @@ pubmed_id type identifier offset end 4272812 Species 10090 44916 44920 4272812 Gene 38295 44327 44331 4272812 Disease D009410 44051 44064 +4272812 Chemical D004298 45081 45083 4272812 Disease D010300 44809 44811 -4272812 Disease D006402 45019 45036 +4272812 Chemical D004298 43984 43986 +4272812 Chemical D004298 44484 44486 +4272812 Chemical D004298 44381 44383 +4272812 Chemical D004298 44435 44437 4272812 Disease D010300 45181 45183 -4272812 Chemical D005978 44119 44122 -4272812 Disease D019636 45157 45177 +4272812 Chemical C007430 44963 44968 +4272812 Chemical D004298 45004 45006 4272812 Disease D009410 44866 44879 +4272812 Chemical - 44119 44122 4272812 Chemical C007430 44930 44961 4272812 Gene 293 45413 45422 4272812 Gene 293 45493 45502 @@ -8548,6 +8670,8 @@ pubmed_id type identifier offset end 4272812 Species 10090 46041 46045 4272812 DNAMutation c.53A>T;RS#:104893877 45879 45883 4272812 DNAMutation c.53A>T;RS#:104893877 46029 46033 +4272812 Chemical D004298 46334 46336 +4272812 Chemical D004298 46408 46410 4272812 Disease D010300 45259 45261 4272812 ProteinMutation p.E46K;RS#:104893875 45892 45896 4272812 Gene 293 46651 46660 @@ -8556,15 +8680,15 @@ pubmed_id type identifier offset end 4272812 Gene 293 47212 47221 4272812 Gene 293 47525 47534 4272812 Gene 21195 46983 46986 -4272812 Disease D006402 47483 47500 +4272812 Disease D030342 47227 47237 +4272812 Disease D030342 46963 46973 4272812 Disease C536965 47376 47406 4272812 Gene 293 47862 47871 4272812 Gene 293 47920 47929 4272812 Gene 293 48027 48036 -4272812 Chemical D009584 48093 48094 -4272812 Chemical C489032 48351 48358 -4272812 Chemical C086501 48277 48280 -4272812 Chemical C086501 48176 48179 +4272812 Chemical - 48176 48179 +4272812 Chemical D011392 48351 48358 +4272812 Chemical - 48277 48280 4272812 Gene 293 48442 48451 4272812 Gene 16473 48491 48494 4272812 Gene 16473 48595 48598 @@ -8582,11 +8706,14 @@ pubmed_id type identifier offset end 4272812 Species 185869 49039 49046 4272812 Gene 16473 49047 49050 4272812 Gene 16473 48934 48937 +4272812 Disease D030342 48724 48734 4272812 Chemical D012402 49113 49121 -4272812 Chemical D012402 48888 48898 +4272812 Chemical D012402 48890 48898 +4272812 Disease D030342 48569 48579 4272812 Gene 293 49447 49456 4272812 Gene 293 49785 49794 -4272812 Chemical C086501 50027 50030 +4272812 Disease D030342 49507 49517 +4272812 Chemical - 50027 50030 4272812 Gene 293 50773 50782 4272812 Gene 293 50888 50892 4272812 Gene 293 50981 50990 @@ -8612,12 +8739,14 @@ pubmed_id type identifier offset end 4272812 Gene 293 53592 53601 4272812 Gene 293 53779 53788 4272812 Species 9606 53097 53102 +4272812 Disease D014693 53651 53663 4272812 Disease D064420 52703 52711 4272812 Disease D010300 53421 53432 -4272812 Chemical C047902 52517 52523 -4272812 Chemical C017937 52778 52782 -4272812 Chemical C047902 52650 52656 -4272812 Disease D058495 53485 53504 +4272812 Chemical D012694 52517 52523 +4272812 Disease D014693 52908 52920 +4272812 Chemical D008055 53522 53527 +4272812 Chemical D012694 52650 52656 +4272812 Disease D066263 53485 53504 4272812 Gene 293 53991 54000 4272812 Gene 293 54125 54134 4272812 Gene 293 54267 54276 @@ -8627,6 +8756,7 @@ pubmed_id type identifier offset end 4272812 Chemical D012402 54257 54265 4272812 Disease D019636 54208 54225 4272812 Chemical D004298 54466 54474 +4272812 Disease D030342 54493 54503 4272812 Disease D010300 54541 54543 4272812 Disease D010300 54544 54563 4272812 Disease D010300 54717 54736 @@ -8638,27 +8768,26 @@ pubmed_id type identifier offset end 4272812 Disease D010300 55052 55071 4272812 Disease D010300 55096 55115 4272812 Disease D010300 55183 55213 -4272812 Disease D000544 55309 55318 4272812 Disease D009410 55216 55229 -4272812 Disease D010300 55323 55341 +4272812 Disease D000544 55309 55341 4272812 Disease D010300 55354 55371 4272812 Disease D003704 55342 55350 4272812 Disease D010300 55496 55515 4272812 Gene 1166 55559 55569 4272812 Disease D010300 55592 55611 -4272812 Disease D010300 55644 55663 +4272812 Disease D010300 55626 55663 4272812 Gene 293 55666 55681 4272812 Disease D003704 55702 55710 -4272812 Disease D002659 55796 55843 +4272812 Disease D000080874 55826 55843 4272812 Gene 293 55941 55956 -4272812 Chemical C419142 55973 55980 +4272812 Chemical C419142 55973 55984 +4272812 Disease D020961 55872 55892 4272812 Gene 293 56010 56025 -4272812 Disease D058495 56114 56133 +4272812 Disease D066263 56114 56133 4272812 Disease D010300 56150 56169 4272812 Gene 293 56277 56281 4272812 Gene 293 56282 56297 4272812 Disease D003704 56381 56389 -4272812 Disease C579880 56314 56325 4272812 Disease D010300 56354 56373 4272812 Gene 293 56603 56618 4272812 Gene 293 56634 56649 @@ -8667,12 +8796,12 @@ pubmed_id type identifier offset end 4272812 Disease D010300 56885 56902 4272812 Gene 31353 56919 56940 4272812 Gene 8534 57066 57072 -4272812 Disease D005935 57137 57158 +4272812 Disease D000080874 57143 57158 4272812 Disease D019636 57357 57384 4272812 Gene 293 57398 57413 4272812 Disease D010300 57469 57488 4272812 Disease D010300 57547 57566 -4272812 Disease D004194 57514 57543 +4272812 Disease C537394 57514 57532 4272812 Gene 293 57584 57599 4272812 Disease D010300 57699 57718 4272812 Disease D010300 57765 57784 @@ -8683,18 +8812,18 @@ pubmed_id type identifier offset end 4272812 Disease D010300 57955 57998 4272812 Disease D010300 58031 58074 4272812 Disease D010300 58100 58117 -4272812 Disease D010300 58173 58192 +4272812 Disease D010300 58164 58192 4272812 Disease D010300 58358 58377 4272812 Gene 293 58380 58395 4272812 Disease D010300 58422 58441 -4272812 Disease D010300 58509 58528 +4272812 Disease D010300 58496 58528 4272812 Gene 18982 58544 58549 -4272812 Disease D010300 58556 58587 +4272812 Disease C537176 58556 58587 4272812 Gene 293 58654 58669 4272812 Disease D010300 58673 58692 4272812 Gene 293 58722 58737 -4272812 Disease D010300 58745 58777 4272812 ProteinMutation p.E46K;RS#:104893875 58713 58717 +4272812 Disease D020961 58745 58777 4272812 Gene 293 58854 58869 4272812 Disease D010300 58778 58797 4272812 Gene 293 58903 58918 @@ -8706,7 +8835,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 59181 59185 4272812 DNAMutation c.53A>T;RS#:104893877 59186 59190 4272812 Disease D010300 59236 59264 -4272812 Disease D010300 59200 59219 +4272812 Disease D010300 59191 59219 4272812 Gene 293 59295 59310 4272812 Disease D010300 59341 59358 4272812 Gene 293 59410 59425 @@ -8722,21 +8851,21 @@ pubmed_id type identifier offset end 4272812 Gene 293 59903 59918 4272812 Disease D010300 59856 59882 4272812 Gene 293 60019 60034 -4272812 Disease D010300 59968 59989 +4272812 ProteinMutation p.A53T;RS#:104893877 60010 60018 +4272812 Disease D010302 59968 59989 4272812 Gene 293 60168 60183 -4272812 Disease D058225 60044 60072 4272812 DNAMutation c.53A>T;RS#:104893877 60147 60151 4272812 Disease D020961 60120 60137 4272812 Species 10090 60253 60257 4272812 Species 9606 60274 60279 4272812 Disease D009069 60232 60249 +4272812 Disease D000080874 60204 60219 4272812 DNAMutation c.53A>T;RS#:104893877 60269 60273 -4272812 Disease D009410 60189 60219 4272812 Gene 293 60327 60342 4272812 Chemical D004298 60394 60402 4272812 Gene 293 60493 60508 -4272812 Disease D009386 60523 60546 4272812 Disease D003704 60551 60559 +4272812 Disease D010302 60523 60546 4272812 Gene 74962 60644 60648 4272812 Gene 293 60631 60635 4272812 Disease D010300 60683 60700 @@ -8758,10 +8887,8 @@ pubmed_id type identifier offset end 4272812 Gene 18982 61477 61482 4272812 Disease D007635 61394 61415 4272812 Gene 293 61506 61521 -4272812 Gene 18982 61570 61575 -4272812 Disease D010300 61576 61595 -4272812 ProteinMutation p.G2019S;RS#:34637584 61563 61569 -4272812 Disease D010300 61669 61688 +4272812 Disease D010300 61563 61595 +4272812 Disease D010300 61659 61688 4272812 Disease D010300 61796 61829 4272812 Gene 293 61895 61910 4272812 Gene 293 61921 61936 @@ -8769,7 +8896,7 @@ pubmed_id type identifier offset end 4272812 Gene 32672 62061 62066 4272812 Disease D010300 62104 62121 4272812 Gene 38295 62122 62126 -4272812 Disease OMIM:105500 62141 62200 +4272812 Disease OMIM:105500 62141 62192 4272812 Gene 68040 62228 62246 4272812 Disease D010302 62261 62273 4272812 Gene 68040 62274 62292 @@ -8780,7 +8907,7 @@ pubmed_id type identifier offset end 4272812 Species 10090 62522 62527 4272812 Disease D019636 62538 62564 4272812 Disease D019954 62620 62645 -4272812 Disease D003866 62697 62707 +4272812 Disease D000275 62697 62707 4272812 Disease D010300 62725 62744 4272812 Chemical D007980 62856 62862 4272812 Disease D010300 62832 62851 @@ -8790,6 +8917,7 @@ pubmed_id type identifier offset end 4272812 Species 10090 63008 63018 4272812 Disease D010302 63019 63028 4272812 Gene 293 63093 63108 +4272812 Disease C536122 63186 63206 4272812 Disease D005776 63137 63166 4272812 Gene 48353 63207 63213 4272812 Gene 68040 63241 63264 @@ -8800,7 +8928,6 @@ pubmed_id type identifier offset end 4272812 Disease D010302 63579 63591 4272812 Chemical D015632 63526 63570 4272812 Disease D010300 63658 63677 -4272812 Chemical D015632 63738 63742 4272812 Disease D010302 63743 63755 4272812 Chemical D015632 63778 63822 4272812 Chemical D008694 63834 63849 @@ -8811,14 +8938,13 @@ pubmed_id type identifier offset end 4272812 Disease D010300 63971 63990 4272812 Species 10090 64021 64025 4272812 Disease D010300 64002 64004 -4272812 Chemical D015632 64051 64055 -4272812 Disease D020734 64065 64088 +4272812 Disease D010302 64065 64088 4272812 Chemical D015632 64111 64115 4272812 Chemical D015632 64274 64278 4272812 Disease D020721 64198 64240 +4272812 Chemical D011339 64283 64293 4272812 Gene 293 64409 64424 4272812 Species 10090 64340 64344 -4272812 Chemical D015632 64335 64339 4272812 Disease D009410 64383 64404 4272812 Species 10090 64484 64489 4272812 Chemical D015632 64479 64483 @@ -8831,8 +8957,8 @@ pubmed_id type identifier offset end 4272812 Gene 293 64847 64862 4272812 Chemical D012402 64775 64783 4272812 Gene 293 64940 64955 -4272812 Chemical D004298 64972 64980 4272812 Chemical D012402 65052 65060 +4272812 Chemical D008070 64896 64914 4272812 Gene 293 65117 65132 4272812 Chemical D012402 65137 65145 4272812 Gene 293 65201 65216 @@ -8855,11 +8981,11 @@ pubmed_id type identifier offset end 4272812 Disease D010300 66268 66285 4272812 Disease D010300 66315 66334 4272812 Disease D020258 66286 66296 -4272812 Disease D020178 66342 66350 -4272812 Disease D010302 66417 66426 +4272812 Disease D010300 66417 66434 4272812 Disease D010300 66495 66514 4272812 Species 10090 66545 66549 -4272812 Disease D010302 66517 66541 +4272812 Disease D028361 66555 66582 +4272812 Disease D010302 66529 66541 4272812 Chemical D004298 66583 66591 4272812 Gene 293 66608 66623 4272812 Disease D010300 66669 66688 @@ -8874,33 +9000,31 @@ pubmed_id type identifier offset end 4272812 Gene 293 67141 67156 4272812 Species 9606 67135 67140 4272812 ProteinMutation p.A30P;RS#:104893878 67157 67161 -4272812 Disease D004194 67066 67083 +4272812 Disease D000068079 67066 67083 4272812 Disease D005911 67088 67095 4272812 Gene 293 67198 67213 4272812 Species 10090 67214 67229 4272812 Disease D010300 67178 67197 4272812 Disease D009410 67238 67273 -4272812 Disease D003643 67283 67288 4272812 Gene 18982 67330 67335 4272812 Species 10090 67346 67350 -4272812 Disease D004342 67310 67326 +4272812 Disease D004342 67310 67329 4272812 Chemical D015632 67360 67404 4272812 Chemical D015632 67354 67358 4272812 Gene 18982 67414 67442 4272812 Gene 293 67510 67525 -4272812 Disease D003643 67546 67551 4272812 Gene 18982 67584 67589 4272812 ProteinMutation p.R1441C;RS#:33939927 67565 67571 4272812 Gene 18982 67642 67647 4272812 Species 10090 67718 67723 -4272812 Disease D005935 67693 67714 +4272812 Disease D000080874 67699 67714 4272812 Gene 18982 67730 67758 4272812 Disease D010300 67814 67833 4272812 Species 10090 67954 67969 4272812 ProteinMutation p.G2019S;RS#:34637584 67981 67987 4272812 Disease D009410 67880 67893 -4272812 Disease C536522 67926 67950 4272812 Disease D058225 67895 67921 +4272812 Disease C564093 68001 68025 4272812 Gene 293 68147 68162 4272812 Disease D010300 68110 68129 4272812 Disease D010300 68210 68229 @@ -8923,12 +9047,11 @@ pubmed_id type identifier offset end 4272812 Disease D010300 68820 68839 4272812 Gene 540 68842 68875 4272812 Disease D010300 68954 68973 -4272812 Disease D065634 68919 68950 4272812 Gene 540 68976 68986 4272812 Gene 2432 68991 69001 -4272812 Disease D065634 69056 69087 4272812 Disease D010300 69033 69052 4272812 Gene 540 69090 69123 +4272812 Disease D009461 69194 69201 4272812 Chemical D015632 69205 69209 4272812 Gene 540 69254 69264 4272812 Gene 540 69359 69369 @@ -8940,17 +9063,19 @@ pubmed_id type identifier offset end 4272812 Gene 293 69661 69676 4272812 Gene 109912 69729 69732 4272812 Gene 55937 69762 69767 +4272812 Species 7227 69881 69891 4272812 Disease D010300 69901 69920 4272812 Gene 293 69923 69938 4272812 Species 10090 69939 69949 +4272812 Species 7227 69950 69960 4272812 Disease D010300 69975 69994 +4272812 Disease D000080874 70007 70024 4272812 Gene 293 70053 70068 -4272812 Species 10090 70107 70117 +4272812 Disease D009410 70085 70132 4272812 Chemical D004298 70076 70084 4272812 Disease D010302 70027 70045 4272812 Species 6239 70190 70212 4272812 Species 9606 70228 70233 -4272812 Disease D009410 70154 70167 4272812 Gene 293 70299 70314 4272812 Species 6239 70250 70260 4272812 Gene 293 70358 70373 @@ -8965,27 +9090,24 @@ pubmed_id type identifier offset end 4272812 Chemical D004298 70884 70892 4272812 Species 10090 70941 70945 4272812 Disease D010302 70972 70984 -4272812 Disease C564486 71076 71095 +4272812 Disease C564486 71067 71095 4272812 Disease D018476 70985 71036 -4272812 Gene 32672 71175 71180 4272812 Chemical D004298 71115 71123 +4272812 Disease C565276 71175 71190 4272812 Gene 293 71209 71224 4272812 Species 10090 71196 71200 4272812 Chemical D004298 71274 71282 4272812 Species 10090 71402 71406 4272812 Gene 293 71477 71492 -4272812 Chemical D015306 71507 71516 4272812 Chemical D004298 71539 71547 4272812 Gene 293 71599 71614 4272812 Chemical D004298 71576 71584 -4272812 Disease D003643 71632 71637 4272812 Chemical D002118 71586 71593 -4272812 Disease D003643 71704 71709 4272812 Disease D010300 71837 71856 4272812 Gene 293 71872 71887 4272812 Disease D010300 71911 71943 4272812 Gene 293 71976 71991 -4272812 Disease D003643 72070 72075 +4272812 Disease C536122 72038 72058 4272812 Gene 293 72103 72118 4272812 Gene 293 72131 72146 4272812 Gene 293 72224 72239 @@ -9009,6 +9131,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 73495 73510 4272812 Gene 293 73564 73579 4272812 Gene 293 73636 73651 +4272812 Chemical D008055 73661 73666 4272812 Gene 293 73704 73719 4272812 Gene 293 73844 73859 4272812 Gene 293 73886 73901 @@ -9020,6 +9143,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 74039 74054 4272812 Species 9606 74256 74261 4272812 DNAMutation c.53A>T;RS#:104893877 74188 74192 +4272812 Disease D014693 74240 74252 4272812 Disease D010300 74134 74162 4272812 ProteinMutation p.A30P;RS#:104893878 74179 74183 4272812 Gene 293 74343 74358 @@ -9030,12 +9154,11 @@ pubmed_id type identifier offset end 4272812 Species 10116 74509 74512 4272812 Disease D010300 74522 74539 4272812 Disease D019636 74486 74503 -4272812 Chemical C530429 74442 74445 +4272812 Chemical D012694 74442 74445 4272812 Disease D010300 74666 74685 4272812 Disease D064420 74624 74632 4272812 Gene 293 74743 74758 4272812 Species 4932 74968 74973 -4272812 Chemical D009584 74933 74934 4272812 Gene 293 75024 75039 4272813 Species 9606 7731 7743 4272813 Species 9606 8493 8500 @@ -9044,6 +9167,7 @@ pubmed_id type identifier offset end 4272813 Disease D003920 8585 8593 4272813 Species 9606 12630 12637 4272813 Species 9606 13281 13293 +4272813 Chemical - 13131 13136 4272813 Disease C535569 16780 16792 4272813 Disease C535569 18732 18744 4272813 Disease D003920 22306 22314 diff --git a/data/example/8-hetnet-full-text-tags.tsv b/data/example/8-hetnet-full-text-tags.tsv index 970bc92..b48e5db 100644 --- a/data/example/8-hetnet-full-text-tags.tsv +++ b/data/example/8-hetnet-full-text-tags.tsv @@ -2,17 +2,32 @@ pubmed_id type identifier offset end 2289422 Compound DB00126 514 527 2289422 Compound DB09541 514 527 2289422 Compound DB00755 1544 1557 -2289424 Compound DB00145 590 606 +2289423 Compound DB00134 524 534 +2289423 Compound DB00134 606 616 +2289423 Compound DB00065 1042 1046 +2289423 Compound DB00065 1224 1228 +2289423 Compound DB00065 1331 1337 +2289423 Compound DB00065 1523 1529 +2289423 Compound DB00065 1569 1575 +2289423 Compound DB00065 2016 2020 +2289424 Compound DB00145 590 597 +2289424 Compound DB00135 598 606 +2289424 Compound DB00135 1143 1151 2289424 Compound DB00145 1155 1162 -2289424 Compound DB00145 1679 1695 +2289424 Compound DB00145 1679 1686 +2289424 Compound DB00135 1687 1695 2289425 Gene 7531 297 301 -2289426 Compound DB09152 262 263 2289427 Gene 1877 543 546 2289428 Gene 838 634 667 2289428 Gene 496 681 691 2289428 Gene 3054 1095 1101 2289428 Gene 3054 1417 1423 +2289429 Compound DB00065 26 30 2289429 Disease DOID:2531 450 457 +2289429 Compound DB00065 570 574 +2289429 Compound DB00065 1541 1545 +2289429 Compound DB00065 1630 1634 +2289430 Compound DB03193 994 1006 2289434 Gene 3054 63 69 2289434 Gene 3054 471 477 2289434 Gene 3054 517 523 @@ -69,17 +84,18 @@ pubmed_id type identifier offset end 2289443 Gene 55643 906 909 2289443 Gene 55643 1175 1178 2289445 Gene 8829 95 121 +2289445 Compound DB00065 210 214 2289445 Gene 8829 293 345 2289445 Gene 8829 406 409 2289445 Gene 8829 527 530 2289445 Gene 8829 1041 1044 2289445 Gene 539 1180 1185 2289445 Gene 8829 1438 1441 -2289446 Compound DB03106 774 803 2289446 Gene 4580 999 1004 2289447 Compound DB00171 603 606 2289447 Compound DB00640 603 606 2289447 Gene 2591 820 836 +2289449 Disease DOID:219 259 279 2289449 Compound DB00974 852 856 2289449 Compound DB09153 870 874 2289449 Gene 22999 957 972 @@ -88,7 +104,6 @@ pubmed_id type identifier offset end 2289450 Gene 4502 58 99 2289450 Gene 4502 101 104 2289450 Gene 4502 201 204 -2289450 Gene 4502 329 332 2289450 Gene 4502 433 436 2289450 Gene 4502 527 530 2289450 Compound DB00171 1073 1076 @@ -107,6 +122,7 @@ pubmed_id type identifier offset end 2289453 Compound DB00928 1553 1564 2289453 Gene 1850 1586 1590 2289453 Gene 22999 1685 1700 +2289454 Compound DB00316 108 109 2289454 Gene 539 432 442 2289454 Gene 539 444 473 2289454 Gene 539 475 482 @@ -119,10 +135,6 @@ pubmed_id type identifier offset end 2289454 Gene 539 1061 1072 2289454 Gene 539 1286 1296 2289457 Gene 173 49 56 -2289457 Compound DB09341 69 76 -2289457 Compound DB11280 69 76 -2289457 Compound DB09341 164 171 -2289457 Compound DB11280 164 171 2289457 Gene 173 216 223 2289457 Compound DB09341 235 242 2289457 Compound DB11280 235 242 @@ -134,10 +146,8 @@ pubmed_id type identifier offset end 2289457 Compound DB11280 649 656 2289462 Compound DB00541 1918 1929 2289465 Gene 3908 0 3 -2289465 Compound DB03128 60 73 2289465 Gene 3908 168 198 2289465 Gene 3908 200 203 -2289465 Compound DB03128 252 265 2289465 Gene 3908 416 419 2289465 Gene 3908 571 574 2289465 Gene 3908 871 874 @@ -145,6 +155,29 @@ pubmed_id type identifier offset end 2289465 Gene 3908 1046 1049 2289465 Gene 3908 1254 1257 2289465 Gene 3908 1331 1334 +2289470 Compound DB00065 86 90 +2289470 Compound DB00065 130 134 +2289470 Compound DB00065 207 211 +2289470 Compound DB00065 477 481 +2289470 Compound DB00065 755 759 +2289470 Compound DB00065 893 897 +2289470 Compound DB00065 941 945 +2289470 Compound DB00065 1006 1010 +2289470 Compound DB00065 1208 1212 +2289470 Compound DB00065 1238 1242 +2289470 Compound DB00065 1368 1372 +2289470 Compound DB00065 1404 1408 +2289470 Compound DB00065 1448 1452 +2289470 Compound DB00065 1501 1505 +2289470 Compound DB00065 1574 1580 +2289470 Compound DB00065 1630 1636 +2289470 Compound DB00065 1673 1677 +2289472 Compound DB00065 136 140 +2289472 Compound DB00065 218 222 +2289472 Compound DB00065 399 403 +2289472 Compound DB00065 665 669 +2289472 Compound DB00065 831 835 +2289472 Compound DB00065 1162 1166 2289473 Gene 1876 184 203 2289473 Gene 1876 205 208 2289473 Gene 1876 559 562 @@ -175,6 +208,8 @@ pubmed_id type identifier offset end 2289491 Gene 23480 468 474 2289491 Gene 23480 588 594 2289491 Gene 23480 651 657 +2289491 Gene 23480 772 778 +2289491 Gene 23480 893 899 2289491 Gene 23480 985 991 2289491 Gene 23480 1109 1115 2289491 Compound DB00171 1152 1155 @@ -188,18 +223,14 @@ pubmed_id type identifier offset end 2289491 Gene 23480 1455 1461 2289491 Gene 23480 1646 1652 2289491 Gene 23480 1698 1704 -2289492 Compound DB09341 21 28 -2289492 Compound DB11280 21 28 2289492 Compound DB00171 116 119 2289492 Compound DB00640 116 119 2289492 Gene 173 120 127 2289492 Gene 173 192 199 -2289492 Compound DB09341 231 238 -2289492 Compound DB11280 231 238 2289492 Gene 173 589 596 2289492 Gene 173 832 839 2289492 Gene 173 911 918 -2289492 Gene 173 1017 1024 +2289492 Disease DOID:1826 1006 1024 2289492 Gene 173 1070 1077 2289492 Gene 173 1161 1168 2289492 Compound DB00171 1217 1220 @@ -215,18 +246,23 @@ pubmed_id type identifier offset end 2289492 Compound DB00171 1691 1694 2289492 Compound DB00640 1691 1694 2289492 Gene 173 1709 1716 +2289492 Compound DB09353 1755 1756 2289492 Gene 173 1868 1890 2289492 Gene 173 2034 2041 2289492 Compound DB00171 2117 2120 2289492 Compound DB00640 2117 2120 2289492 Gene 173 2134 2141 -2289492 Gene 173 2380 2387 +2289492 Disease DOID:1826 2369 2387 2289495 Compound DB00171 838 841 2289495 Compound DB00640 838 841 2289495 Compound DB00171 1582 1585 2289495 Compound DB00640 1582 1585 2289496 Gene 22999 46 61 2289496 Gene 22999 149 164 +2289496 Compound DB00172 1561 1568 +2289496 Compound DB00172 1709 1716 +2289496 Compound DB00133 1859 1865 +2289496 Compound DB00135 1870 1878 2289498 Gene 328 59 63 2289498 Gene 328 414 418 2289498 Gene 328 560 564 @@ -236,14 +272,12 @@ pubmed_id type identifier offset end 2289498 Gene 328 1501 1505 2289498 Gene 328 1652 1656 2289498 Gene 328 1907 1911 -2289499 Compound DB09353 384 385 2289499 Compound DB00171 464 467 2289499 Compound DB00640 464 467 2289499 Compound DB09407 662 667 2289499 Compound DB09407 937 942 -2289499 Compound DB00171 1296 1299 -2289499 Compound DB00640 1296 1299 -2289499 Compound DB09353 1306 1307 +2289499 Compound DB00171 1067 1070 +2289499 Compound DB00640 1067 1070 2289500 Compound DB00608 966 977 2289500 Compound DB00608 1102 1113 2289501 Disease DOID:12365 0 19 @@ -253,14 +287,11 @@ pubmed_id type identifier offset end 2289501 Compound DB01109 1186 1193 2289501 Compound DB01109 1581 1588 2289503 Disease DOID:1909 88 96 -2289503 Compound DB01109 137 144 2289503 Gene 2253 214 218 2289503 Disease DOID:1909 242 250 -2289503 Compound DB01109 319 326 2289503 Disease DOID:1909 405 413 2289503 Compound DB01109 466 473 2289503 Disease DOID:1909 500 508 -2289503 Compound DB01109 670 677 2289503 Disease DOID:1909 711 719 2289503 Gene 2253 725 729 2289503 Gene 2253 838 842 @@ -271,7 +302,7 @@ pubmed_id type identifier offset end 2289503 Gene 2253 1325 1329 2289503 Gene 2253 1391 1395 2289503 Gene 2253 1587 1591 -2289503 Disease DOID:1909 1635 1653 +2289503 Disease DOID:1909 1635 1648 2289503 Disease DOID:1909 1740 1748 2289503 Gene 2253 1762 1766 2289503 Gene 2253 1844 1848 @@ -284,6 +315,7 @@ pubmed_id type identifier offset end 2289503 Gene 2253 2247 2251 2289503 Disease DOID:1909 2261 2269 2289503 Disease DOID:1909 2393 2401 +2289504 Compound DB09353 1795 1796 2289505 Disease DOID:1192 66 79 2289505 Disease DOID:1192 176 189 2289505 Compound DB01394 624 634 @@ -310,6 +342,7 @@ pubmed_id type identifier offset end 2289508 Gene 55890 435 440 2289508 Gene 55890 493 498 2289508 Gene 55890 645 650 +2289508 Compound DB08818 753 763 2289509 Gene 23480 403 409 2289509 Gene 23480 669 675 2289509 Compound DB00171 713 716 @@ -322,10 +355,11 @@ pubmed_id type identifier offset end 2289510 Compound DB01394 1913 1923 2289510 Compound DB01394 2282 2292 2289510 Compound DB01394 2390 2400 +2289514 Disease DOID:2355 607 618 2289515 Compound DB00171 928 931 2289515 Compound DB00640 928 931 +2289517 Compound DB00065 1949 1953 2289524 Gene 1876 0 19 -2289524 Compound DB00446 407 422 2289524 Gene 1876 643 662 2289524 Gene 1876 664 667 2289524 Gene 1876 719 722 @@ -347,11 +381,13 @@ pubmed_id type identifier offset end 4272812 Gene 55937 1069 1074 4272812 Gene 293 1203 1212 4272812 Compound DB00988 1334 1342 +4272812 Compound DB00988 1344 1346 4272812 Gene 293 1377 1386 4272812 Disease DOID:14330 1495 1497 4272812 Disease DOID:14330 1604 1623 4272812 Disease DOID:14330 1625 1627 4272812 Disease DOID:14330 1854 1856 +4272812 Disease DOID:2377 1862 1880 4272812 Disease DOID:14330 2071 2073 4272812 Disease DOID:14330 2199 2201 4272812 Disease DOID:14330 2530 2532 @@ -361,8 +397,7 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 4473 4475 4272812 Disease DOID:14330 4915 4917 4272812 Disease DOID:10652 5011 5030 -4272812 Compound DB01592 5072 5082 -4272812 Disease DOID:9835 5170 5197 +4272812 Disease DOID:14330 5036 5047 4272812 Gene 293 6252 6261 4272812 Gene 293 6790 6799 4272812 Gene 293 7067 7076 @@ -373,15 +408,13 @@ pubmed_id type identifier offset end 4272812 Gene 8534 7748 7754 4272812 Disease DOID:10652 8359 8378 4272812 Disease DOID:14330 9978 9980 -4272812 Disease DOID:14330 10163 10182 4272812 Gene 293 10499 10508 4272812 Gene 55937 10599 10604 -4272812 Disease DOID:9970 10655 10657 4272812 Disease DOID:14330 11206 11208 -4272812 Disease DOID:14330 11219 11227 +4272812 Disease DOID:14330 11225 11227 4272812 Disease DOID:14330 11445 11447 4272812 Gene 293 11470 11479 -4272812 Disease DOID:14330 11524 11532 +4272812 Disease DOID:14330 11530 11532 4272812 Disease DOID:14330 11638 11640 4272812 Gene 293 11661 11670 4272812 Gene 293 12094 12103 @@ -396,7 +429,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 13704 13708 4272812 Gene 293 13843 13852 4272812 Gene 293 13854 13858 -4272812 Disease DOID:14330 13976 13987 +4272812 Disease DOID:14330 13985 13987 4272812 Gene 293 14040 14044 4272812 Disease DOID:14330 14089 14091 4272812 Gene 293 14192 14196 @@ -442,7 +475,6 @@ pubmed_id type identifier offset end 4272812 Gene 293 23064 23073 4272812 Disease DOID:14330 23155 23157 4272812 Disease DOID:14330 23528 23530 -4272812 Disease DOID:14330 23696 23708 4272812 Disease DOID:14330 24663 24665 4272812 Gene 293 24883 24887 4272812 Disease DOID:14330 25202 25204 @@ -456,9 +488,11 @@ pubmed_id type identifier offset end 4272812 Gene 293 26558 26567 4272812 Gene 293 27052 27061 4272812 Gene 293 27175 27184 +4272812 Compound DB01032 27270 27280 4272812 Gene 293 27419 27428 4272812 Gene 293 27765 27774 4272812 Gene 293 27962 27971 +4272812 Compound DB01032 28135 28145 4272812 Disease DOID:14330 28521 28523 4272812 Gene 293 28825 28834 4272812 Disease DOID:14330 28924 28926 @@ -467,7 +501,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 29296 29305 4272812 Gene 293 29581 29590 4272812 Disease DOID:14330 29788 29790 -4272812 Disease DOID:14330 30096 30107 +4272812 Disease DOID:14330 30105 30107 4272812 Gene 293 30194 30203 4272812 Disease DOID:14330 30845 30847 4272812 Gene 293 31103 31112 @@ -525,6 +559,7 @@ pubmed_id type identifier offset end 4272812 Gene 55937 38870 38875 4272812 Gene 540 38929 38938 4272812 Disease DOID:14330 39006 39008 +4272812 Compound DB00988 39131 39133 4272812 Gene 55937 39268 39273 4272812 Gene 293 39310 39319 4272812 Gene 293 39439 39448 @@ -541,7 +576,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 40950 40954 4272812 Disease DOID:14330 41063 41065 4272812 Gene 293 41115 41124 -4272812 Disease DOID:14330 41176 41187 +4272812 Disease DOID:14330 41185 41187 4272812 Gene 55937 41209 41214 4272812 Gene 293 41235 41244 4272812 Gene 55937 41299 41304 @@ -551,28 +586,35 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 41990 41992 4272812 Gene 293 42054 42063 4272812 Disease DOID:14330 42218 42220 -4272812 Disease DOID:14330 42602 42620 4272812 Gene 293 42685 42694 4272812 Disease DOID:14330 43099 43101 4272812 Compound DB00988 43188 43196 4272812 Gene 293 43248 43257 +4272812 Compound DB00988 43267 43269 4272812 Gene 293 43328 43337 4272812 Gene 307 43347 43367 4272812 Compound DB00988 43478 43484 4272812 Compound DB01235 43478 43484 +4272812 Compound DB00988 43488 43490 +4272812 Compound DB00988 43516 43518 4272812 Gene 293 43572 43581 4272812 Gene 55937 43587 43592 4272812 Gene 293 43649 43658 4272812 Gene 55937 43686 43691 4272812 Gene 293 43710 43719 4272812 Gene 293 43906 43915 +4272812 Compound DB00988 43984 43986 4272812 Gene 55547 44104 44107 -4272812 Compound DB00143 44119 44122 +4272812 Compound DB00988 44381 44383 +4272812 Compound DB00988 44435 44437 +4272812 Compound DB00988 44484 44486 4272812 Gene 293 44506 44515 4272812 Disease DOID:14330 44809 44811 4272812 Gene 293 44838 44847 4272812 Gene 55937 44900 44905 +4272812 Compound DB00988 45004 45006 4272812 Gene 293 45040 45055 +4272812 Compound DB00988 45081 45083 4272812 Gene 293 45099 45108 4272812 Disease DOID:14330 45181 45183 4272812 Disease DOID:14330 45259 45261 @@ -586,7 +628,9 @@ pubmed_id type identifier offset end 4272812 Gene 293 46129 46138 4272812 Gene 293 46226 46235 4272812 Gene 293 46313 46322 +4272812 Compound DB00988 46334 46336 4272812 Gene 293 46394 46403 +4272812 Compound DB00988 46408 46410 4272812 Gene 293 46436 46445 4272812 Gene 293 46651 46660 4272812 Gene 293 46696 46705 @@ -596,7 +640,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 47862 47871 4272812 Gene 293 47920 47929 4272812 Gene 293 48027 48036 -4272812 Compound DB09152 48093 48094 +4272812 Compound DB00172 48351 48358 4272812 Gene 293 48442 48451 4272812 Gene 55937 48963 48968 4272812 Gene 55937 49068 49073 @@ -617,7 +661,9 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 52087 52089 4272812 Gene 293 52149 52158 4272812 Gene 293 52364 52373 +4272812 Compound DB00133 52517 52523 4272812 Gene 293 52540 52549 +4272812 Compound DB00133 52650 52656 4272812 Gene 293 52729 52738 4272812 Gene 293 52821 52830 4272812 Gene 293 52898 52907 @@ -643,12 +689,11 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 55052 55071 4272812 Disease DOID:14330 55096 55115 4272812 Disease DOID:14330 55183 55213 -4272812 Disease DOID:10652 55309 55318 -4272812 Disease DOID:14330 55323 55341 +4272812 Disease DOID:10652 55309 55341 4272812 Disease DOID:14330 55354 55371 4272812 Disease DOID:14330 55496 55515 4272812 Disease DOID:14330 55592 55611 -4272812 Disease DOID:14330 55644 55663 +4272812 Disease DOID:14330 55626 55663 4272812 Gene 293 55666 55681 4272812 Gene 293 55941 55956 4272812 Gene 293 56010 56025 @@ -671,23 +716,21 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 57955 57998 4272812 Disease DOID:14330 58031 58074 4272812 Disease DOID:14330 58100 58117 -4272812 Disease DOID:14330 58173 58192 +4272812 Disease DOID:14330 58164 58192 4272812 Disease DOID:14330 58358 58377 4272812 Gene 293 58380 58395 4272812 Disease DOID:14330 58422 58441 -4272812 Disease DOID:14330 58509 58528 -4272812 Disease DOID:14330 58556 58587 +4272812 Disease DOID:14330 58496 58528 4272812 Gene 293 58654 58669 4272812 Disease DOID:14330 58673 58692 4272812 Gene 293 58722 58737 -4272812 Disease DOID:14330 58745 58777 4272812 Disease DOID:14330 58778 58797 4272812 Gene 293 58854 58869 4272812 Gene 293 58903 58918 4272812 Gene 293 58951 58966 4272812 Disease DOID:14330 59007 59024 4272812 Gene 293 59181 59185 -4272812 Disease DOID:14330 59200 59219 +4272812 Disease DOID:14330 59191 59219 4272812 Disease DOID:14330 59236 59264 4272812 Gene 293 59295 59310 4272812 Disease DOID:14330 59341 59358 @@ -702,7 +745,6 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 59821 59840 4272812 Disease DOID:14330 59856 59882 4272812 Gene 293 59903 59918 -4272812 Disease DOID:14330 59968 59989 4272812 Gene 293 60019 60034 4272812 Gene 293 60168 60183 4272812 Gene 293 60327 60342 @@ -718,8 +760,8 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 61210 61229 4272812 Gene 293 61457 61472 4272812 Gene 293 61506 61521 -4272812 Disease DOID:14330 61576 61595 -4272812 Disease DOID:14330 61669 61688 +4272812 Disease DOID:14330 61563 61595 +4272812 Disease DOID:14330 61659 61688 4272812 Disease DOID:14330 61796 61829 4272812 Gene 293 61895 61910 4272812 Gene 293 61921 61936 @@ -727,7 +769,6 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 62104 62121 4272812 Disease DOID:14330 62341 62360 4272812 Disease DOID:14330 62419 62438 -4272812 Disease DOID:1595 62697 62707 4272812 Disease DOID:14330 62725 62744 4272812 Disease DOID:14330 62832 62851 4272812 Compound DB00988 62856 62862 @@ -742,6 +783,7 @@ pubmed_id type identifier offset end 4272812 Gene 293 63911 63926 4272812 Disease DOID:14330 63971 63990 4272812 Disease DOID:14330 64002 64004 +4272812 Compound DB01032 64283 64293 4272812 Gene 293 64409 64424 4272812 Disease DOID:14330 64499 64518 4272812 Disease DOID:14330 64565 64584 @@ -749,7 +791,6 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 64740 64759 4272812 Gene 293 64847 64862 4272812 Gene 293 64940 64955 -4272812 Compound DB00988 64972 64980 4272812 Gene 293 65117 65132 4272812 Gene 293 65201 65216 4272812 Disease DOID:14330 65256 65275 @@ -760,6 +801,7 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 66149 66168 4272812 Disease DOID:14330 66268 66285 4272812 Disease DOID:14330 66315 66334 +4272812 Disease DOID:14330 66417 66434 4272812 Disease DOID:14330 66495 66514 4272812 Compound DB00988 66583 66591 4272812 Gene 293 66608 66623 @@ -862,19 +904,28 @@ pubmed_id type identifier offset end 4272812 Disease DOID:14330 74134 74162 4272812 Gene 293 74343 74358 4272812 Disease DOID:14330 74391 74410 +4272812 Compound DB00133 74442 74445 4272812 Gene 293 74459 74474 4272812 Disease DOID:14330 74522 74539 4272812 Disease DOID:14330 74666 74685 4272812 Gene 293 74743 74758 -4272812 Compound DB09152 74933 74934 4272812 Gene 293 75024 75039 4272813 Disease DOID:10763 8551 8563 -4272814 Compound DB00145 1455 1462 +4272814 Compound DB00172 172 179 +4272814 Compound DB00172 1443 1450 +4272814 Compound DB06756 1455 1470 +4272814 Compound DB00149 6340 6347 +4272814 Compound DB00167 6348 6358 4272814 Compound DB09341 7806 7813 4272814 Compound DB11280 7806 7813 +4272814 Compound DB00125 7821 7829 +4272814 Compound DB00167 7837 7847 +4272814 Compound DB00161 7855 7861 4272814 Compound DB09153 8118 8122 +4272814 Compound DB00172 10539 10546 4272814 Compound DB09153 10584 10588 4272814 Compound DB09153 10837 10841 +4272814 Compound DB00172 10926 10933 4272814 Compound DB09153 10961 10965 4272814 Compound DB09153 11167 11171 4272814 Compound DB09153 11305 11309 @@ -885,14 +936,16 @@ pubmed_id type identifier offset end 4272814 Compound DB09153 13116 13120 4272814 Compound DB09153 13355 13359 4272814 Compound DB09153 13527 13531 +4272814 Compound DB00172 15754 15761 4272814 Compound DB09153 16186 16190 4272814 Compound DB09153 16262 16266 4272814 Compound DB09153 17944 17948 4272814 Compound DB09153 18860 18864 4272814 Disease DOID:2531 19260 19262 4272814 Disease DOID:2531 19360 19362 +4272814 Compound DB00172 19678 19685 4272814 Compound DB00583 20252 20261 -4272814 Compound DB00145 20603 20610 +4272814 Compound DB06756 20603 20618 4272815 Gene 208 39 43 4272815 Gene 208 419 433 4272815 Gene 208 435 439 @@ -904,11 +957,11 @@ pubmed_id type identifier offset end 4272815 Gene 208 3945 3949 4272815 Gene 208 3995 3999 4272815 Gene 208 5240 5244 -4272815 Compound DB09157 5700 5703 4272815 Gene 3908 6982 6987 4272815 Compound DB00118 7304 7324 4272815 Compound DB09153 7438 7442 4272815 Compound DB00974 7467 7471 +4272815 Compound DB03255 7497 7503 4272815 Compound DB00898 7527 7534 4272815 Gene 208 8557 8561 4272815 Gene 208 8580 8584 @@ -935,10 +988,8 @@ pubmed_id type identifier offset end 4272816 Compound DB09140 2791 2797 4272816 Compound DB01390 7472 7490 4272816 Compound DB09153 12535 12539 -4272816 Compound DB09152 19111 19112 4272818 Gene 4359 10741 10744 4272818 Gene 4359 10759 10762 -4272818 Compound DB09152 11055 11056 4272818 Gene 4257 18215 18223 4272820 Disease DOID:1826 94 112 4272820 Compound DB00829 290 298 @@ -1040,16 +1091,22 @@ pubmed_id type identifier offset end 4272821 Gene 55665 5716 5720 4272821 Gene 55665 5799 5803 4272821 Compound DB00994 7101 7109 -4272821 Compound DB01174 8933 8940 +4272821 Compound DB03904 9026 9030 +4272821 Compound DB03904 9432 9436 4272821 Gene 1477 10665 10668 4272821 Gene 1854 11060 11068 +4272821 Compound DB00529 11390 11393 4272821 Compound DB09499 11739 11757 4272821 Compound DB03166 11919 11930 4272821 Compound DB03166 12271 12282 +4272821 Compound DB00529 12466 12469 4272821 Compound DB02772 12509 12516 4272821 Gene 8315 13771 13774 4272821 Gene 1521 13798 13803 4272821 Gene 8315 13919 13922 +4272821 Compound DB00529 13978 13981 +4272821 Compound DB03619 13992 14012 +4272821 Compound DB05780 13992 14012 4272821 Compound DB03166 15081 15092 4272821 Compound DB03166 15114 15125 4272821 Compound DB00151 16000 16008 @@ -1066,32 +1123,35 @@ pubmed_id type identifier offset end 4272821 Gene 55665 27253 27257 4272821 Gene 55665 27352 27356 4272821 Gene 55665 27543 27547 +4272821 Compound DB11101 28107 28110 +4272821 Compound DB11101 28136 28139 4272821 Gene 55665 28973 28977 4272821 Gene 55665 29021 29025 4272821 Gene 55665 29158 29162 4272821 Gene 55665 29489 29493 -4272821 Compound DB00148 30507 30515 -4272821 Compound DB00148 30533 30541 -4272821 Compound DB00148 30668 30676 4272821 Gene 55665 35569 35573 4272821 Gene 55665 35678 35682 4272821 Gene 55665 36395 36399 4272821 Gene 7857 36931 36935 -4272821 Compound DB00148 40505 40513 4272821 Gene 55665 40692 40696 4272821 Gene 55665 40798 40802 4272821 Gene 55665 40978 40982 4272821 Gene 55665 41320 41324 4272821 Gene 55665 41523 41527 -4272821 Compound DB00148 43395 43403 +4272821 Compound DB00529 43516 43519 4272821 Gene 7857 46777 46781 4272821 Gene 5144 46791 46796 4272821 Gene 55665 47337 47341 4272822 Gene 3329 14345 14349 4272822 Gene 22999 15000 15005 4272822 Gene 9670 16056 16060 +4272822 Compound DB00065 18261 18265 4272822 Compound DB09152 22416 22424 +4272822 Compound DB00131 23275 23278 +4272822 Compound DB00640 23275 23278 +4272822 Compound DB03904 27005 27009 4272822 Compound DB00145 28092 28099 +4272822 Compound DB03904 28136 28140 4272822 Compound DB04077 28169 28178 4272822 Compound DB09462 28169 28178 4272822 Compound DB00898 28377 28384 @@ -1100,21 +1160,19 @@ pubmed_id type identifier offset end 4272822 Compound DB03843 28939 28951 4272822 Compound DB03843 29137 29149 4272822 Compound DB00145 29208 29215 -4272822 Compound DB09152 30541 30542 4272822 Compound DB00134 30967 30977 -4272822 Compound DB09152 34238 34239 4272822 Disease DOID:9970 35033 35040 4272822 Gene 3329 35193 35197 4272822 Gene 22999 35441 35456 4272822 Gene 22999 35563 35578 4272822 Disease DOID:2994 35856 35865 4272822 Gene 136 35978 35988 -4272822 Disease DOID:986 37009 37024 +4272822 Compound DB00065 36954 36958 4272822 Disease DOID:1324 37265 37291 4272823 Disease DOID:10283 60 75 4272823 Disease DOID:10283 125 140 4272823 Disease DOID:10283 474 489 -4272823 Disease DOID:10283 570 585 +4272823 Disease DOID:10283 560 585 4272823 Disease DOID:10283 2526 2541 4272823 Disease DOID:10283 4997 5012 4272823 Disease DOID:10283 5033 5048 @@ -1131,11 +1189,9 @@ pubmed_id type identifier offset end 4272823 Disease DOID:10283 16390 16405 4272823 Disease DOID:10283 18807 18822 4272823 Disease DOID:10283 19733 19748 -4272823 Disease DOID:10283 21633 21668 4272823 Disease DOID:10283 21825 21840 4272823 Disease DOID:10283 24648 24663 4272823 Disease DOID:10652 29484 29486 -4272823 Disease DOID:10283 29858 29886 4272823 Disease DOID:10283 29975 29990 4272823 Disease DOID:10283 30043 30058 4272823 Disease DOID:10283 30179 30204 @@ -1152,8 +1208,7 @@ pubmed_id type identifier offset end 4272823 Disease DOID:10283 32431 32456 4272823 Disease DOID:10283 32643 32658 4272823 Disease DOID:10283 32685 32700 -4272823 Disease DOID:10283 32802 32820 -4272823 Disease DOID:10283 32821 32858 +4272823 Disease DOID:10283 32843 32858 4272823 Disease DOID:10283 32917 32942 4272823 Disease DOID:10283 33016 33041 4272823 Disease DOID:10283 33163 33188 @@ -1163,7 +1218,6 @@ pubmed_id type identifier offset end 4272823 Disease DOID:10283 33730 33745 4272823 Disease DOID:10283 34088 34113 4272823 Disease DOID:10283 34201 34226 -4272823 Disease DOID:10283 34309 34328 4272823 Disease DOID:10283 34372 34387 4272823 Disease DOID:10283 34486 34501 4272823 Disease DOID:10283 34569 34584 @@ -1175,14 +1229,13 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 39 46 4272824 Compound DB00907 227 234 4272824 Compound DB01579 279 294 -4272824 Compound DB00182 299 312 +4272824 Compound DB01576 299 312 4272824 Compound DB00907 335 342 4272824 Compound DB00907 364 371 4272824 Compound DB00907 518 525 4272824 Compound DB00907 767 774 4272824 Compound DB00907 1021 1028 4272824 Compound DB00907 1095 1102 -4272824 Compound DB00907 1521 1528 4272824 Compound DB00907 1617 1624 4272824 Compound DB00907 1693 1700 4272824 Compound DB00907 1849 1856 @@ -1193,26 +1246,21 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 4214 4221 4272824 Compound DB00907 4326 4333 4272824 Compound DB00907 4820 4827 -4272824 Compound DB00907 5076 5083 4272824 Compound DB00907 5326 5333 4272824 Compound DB00907 5569 5576 4272824 Compound DB00907 5693 5700 -4272824 Compound DB00182 5747 5760 +4272824 Compound DB01576 5747 5760 4272824 Compound DB01579 5762 5777 4272824 Compound DB00830 5783 5796 -4272824 Compound DB00907 5892 5899 -4272824 Compound DB00907 5936 5943 -4272824 Compound DB00907 6205 6212 4272824 Compound DB00907 7688 7695 4272824 Compound DB00907 7985 7992 4272824 Compound DB00907 8367 8374 4272824 Compound DB00907 8470 8477 4272824 Compound DB00907 8545 8552 -4272824 Compound DB00907 8771 8778 -4272824 Compound DB00907 8916 8923 -4272824 Compound DB00182 9161 9174 +4272824 Compound DB01576 9161 9174 4272824 Compound DB01579 9196 9215 4272824 Compound DB00830 9238 9255 +4272824 Compound DB09153 9324 9330 4272824 Compound DB00907 9401 9408 4272824 Compound DB00907 9533 9540 4272824 Compound DB00907 9785 9792 @@ -1223,13 +1271,13 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 10907 10914 4272824 Compound DB00907 11232 11239 4272824 Compound DB00907 13405 13412 -4272824 Compound DB00182 14224 14237 +4272824 Compound DB01576 14224 14237 4272824 Compound DB00907 14271 14278 -4272824 Compound DB00182 14380 14393 +4272824 Compound DB01576 14380 14393 4272824 Compound DB00907 14439 14446 -4272824 Compound DB00182 14488 14501 -4272824 Compound DB00182 14565 14578 -4272824 Compound DB00182 14754 14767 +4272824 Compound DB01576 14488 14501 +4272824 Compound DB01576 14565 14578 +4272824 Compound DB01576 14754 14767 4272824 Compound DB01579 14990 15009 4272824 Compound DB00907 15043 15050 4272824 Compound DB01579 15152 15167 @@ -1248,9 +1296,11 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 16759 16766 4272824 Compound DB00907 16858 16865 4272824 Compound DB00907 17060 17067 -4272824 Compound DB00907 17669 17676 +4272824 Disease DOID:1826 17657 17661 +4272824 Disease DOID:1826 17729 17733 4272824 Compound DB00907 17994 18001 4272824 Compound DB00907 19759 19766 +4272824 Compound DB09153 21246 21252 4272824 Compound DB00907 21276 21283 4272824 Compound DB00907 21321 21328 4272824 Compound DB00907 21373 21380 @@ -1260,34 +1310,29 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 22452 22459 4272824 Compound DB00907 22517 22524 4272824 Compound DB00907 22584 22591 -4272824 Compound DB00182 22931 22944 +4272824 Compound DB01576 22931 22944 4272824 Compound DB00907 23075 23082 4272824 Compound DB00907 23169 23176 4272824 Compound DB00907 23199 23206 4272824 Compound DB00907 23322 23329 4272824 Compound DB00907 23358 23365 4272824 Compound DB00907 23413 23420 -4272824 Compound DB00182 23470 23483 -4272824 Compound DB00907 23564 23571 +4272824 Compound DB01576 23470 23483 4272824 Compound DB00907 23729 23736 -4272824 Compound DB00907 23904 23911 4272824 Compound DB00907 24013 24020 +4272824 Compound DB09153 24136 24142 4272824 Compound DB00907 24372 24379 4272824 Compound DB00907 24593 24600 -4272824 Compound DB00907 25269 25276 4272824 Compound DB00907 25462 25469 -4272824 Compound DB00907 25653 25660 -4272824 Compound DB00182 25697 25710 -4272824 Compound DB00907 25832 25839 +4272824 Compound DB01576 25697 25710 4272824 Compound DB00907 25980 25987 4272824 Compound DB00907 26325 26332 -4272824 Compound DB00182 26791 26804 +4272824 Compound DB01576 26791 26804 4272824 Compound DB01579 26809 26824 4272824 Compound DB00907 26898 26905 -4272824 Compound DB00182 27051 27064 +4272824 Compound DB01576 27051 27064 4272824 Compound DB01579 27069 27084 4272824 Compound DB00907 28035 28042 -4272824 Disease DOID:14330 28337 28349 4272824 Compound DB00182 28376 28387 4272824 Compound DB00907 28401 28408 4272824 Compound DB00988 28541 28549 @@ -1296,61 +1341,64 @@ pubmed_id type identifier offset end 4272824 Compound DB00907 28814 28821 4272824 Compound DB01579 28907 28922 4272824 Compound DB00907 28941 28948 -4272824 Compound DB00899 29515 29527 4272824 Compound DB00907 29695 29702 4272824 Compound DB00898 29815 29822 4272824 Compound DB00898 29847 29854 -4272824 Compound DB00899 30185 30197 4272824 Compound DB00907 30402 30409 4272824 Compound DB00907 30801 30808 4272824 Compound DB00907 31019 31026 -4272824 Compound DB00182 31073 31086 +4272824 Compound DB01576 31073 31086 4272824 Compound DB00907 31100 31107 4272824 Compound DB00184 31389 31397 -4272824 Compound DB00988 31572 31580 4272824 Compound DB00907 31702 31709 4272824 Compound DB00907 31813 31820 -4272824 Compound DB00907 32029 32036 4272824 Compound DB00907 32303 32310 4272824 Compound DB00907 32336 32343 -4272824 Compound DB00182 32386 32399 +4272824 Compound DB01576 32386 32399 4272824 Compound DB00907 32433 32440 -4272824 Compound DB00182 32542 32555 -4272824 Compound DB00907 32634 32641 +4272824 Compound DB01576 32542 32555 4272824 Compound DB00907 32671 32678 -4272824 Compound DB00182 32801 32814 +4272824 Compound DB01576 32801 32814 4272824 Compound DB01579 33065 33084 4272824 Compound DB00907 33118 33125 4272824 Compound DB01579 33227 33246 -4272824 Compound DB00907 33325 33332 4272824 Compound DB00907 33362 33369 4272824 Compound DB01579 33492 33511 4272824 Compound DB00830 33762 33779 4272824 Compound DB00907 33813 33820 4272824 Compound DB00830 33922 33939 -4272824 Compound DB00907 34018 34025 4272824 Compound DB00907 34055 34062 4272824 Compound DB00830 34185 34202 4272824 Compound DB00907 34451 34458 4272824 Compound DB00907 34482 34489 -4272824 Compound DB00907 34679 34686 4272824 Compound DB00907 34715 34722 4272824 Compound DB00830 34845 34862 -4272824 Compound DB00182 35105 35118 +4272824 Compound DB01576 35105 35118 4272824 Compound DB00907 35151 35158 4272824 Compound DB00907 35388 35395 4272824 Compound DB00907 35436 35443 -4272824 Compound DB00182 36705 36760 -4272824 Compound DB01579 38332 38389 -4272824 Compound DB00830 39824 39879 +4272824 Compound DB00830 39820 39837 +4272825 Compound DB08820 12 21 +4272825 Compound DB09280 231 241 +4272825 Compound DB09280 243 249 +4272825 Compound DB08820 323 332 4272825 Disease DOID:1793 2276 2284 -4272825 Compound DB08820 3299 3307 -4272825 Compound DB00171 3523 3526 -4272825 Compound DB00640 3523 3526 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Disease DOID:7147 17876 17898 -4272846 Disease DOID:184 18080 18093 4272847 Disease DOID:3083 24 61 4272847 Disease DOID:3083 166 203 4272847 Disease DOID:3083 300 337 diff --git a/data/temp/batch_0.xml b/data/temp/batch_0.xml new file mode 100644 index 0000000..ddc4e03 --- /dev/null +++ b/data/temp/batch_0.xml @@ -0,0 +1,24065 @@ + + PubTator + + BioC.key + + 2289422 + + 1560033 + 427 + 2 + 1992 + 2289422 + 92218491 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 427-35 + 435 + Descalzi Cancedda F, Gentili C, Manduca P, Cancedda R + 0 + Hypertrophic chondrocytes undergo further differentiation in culture + + MESH:D006984 + Disease + + Hypertrophic + + + + ABSTRACT + abstract + 69 + Conditions have been defined for promoting growth and differentiation of hypertrophic chondrocytes obtained in culture starting from chick embryo tibiae. Hypertrophic chondrocytes, grown in suspension culture as described (Castagnola P., G. Moro, F. Descalzi Cancedda, and R. Cancedda. 1986. J. Cell Biol. 102:2310-2317), when they reached the stage of single cells, were transferred to substrate-dependent culture conditions in the presence of ascorbic acid. Cells showed a change in morphology, became more elongated and flattened, expressed alkaline phosphatase, and eventually mineralized. Type II and X collagen synthesis was halted and replaced by type I collagen synthesis. In addition the cells started to produce and to secrete in large amount a protein with an apparent molecular mass of 82 KD in reducing conditions and 63 KD in unreducing conditions. This protein is soluble in acidic solutions, does not contain collagenous domains, and is glycosylated. The Ch21 protein, a marker of hypertrophic chondrocytes and bone cells, was synthesized throughout the culture. We have defined this additional differentiation stage as an osteoblast-like stage. Calcium deposition in the extracellular matrix occurred regardless of the addition of beta glycerophosphate to the culture medium. Comparable results were obtained both when the cells were plated at low density and when they were already at confluence and maintained in culture without passaging up to 50 d. When retinoic acid was added to the hypertrophic chondrocyte culture between day 1 and day 5 the maturation of the cells to the osteoblast-like stage was highly accelerated. The switch in the collagen secretion was already observed after 2 d and the production of the 63-kD protein after 3 d. Mineralization was observed after 15-20 d. + + 9031 + Species + + chick + + + MESH:D006984 + Disease + + hypertrophic + + + MESH:D014212 + Chemical + + retinoic acid + + + MESH:C031463 + Chemical + + beta glycerophosphate + + + MESH:D006984 + Disease + + Hypertrophic + + + MESH:D002118 + Chemical + + Calcium + + + MESH:D001205 + Chemical + + ascorbic acid + + + MESH:D006984 + Disease + + hypertrophic + + + MESH:D006984 + Disease + + hypertrophic + + + + + 2289423 + + 1313813 + 269 + 2 + 1992 + 2289423 + 92218477 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 269-78 + 278 + Turner MD, Rennison ME, Handel SE, Wilde CJ, Burgoyne RD + 0 + Proteins are secreted by both constitutive and regulated secretory pathways in lactating mouse mammary epithelial cells + + 10090 + Species + + mouse + + + + ABSTRACT + abstract + 120 + Lactating mammary epithelial cells secrete high levels of caseins and other milk proteins. The extent to which protein secretion from these cells occurs in a regulated fashion was examined in experiments on secretory acini isolated from the mammary glands of lactating mice at 10 d postpartum. Protein synthesis and secretion were assayed by following the incorporation or release, respectively, of [35S]methionine-labeled TCA-precipitable protein. The isolated cells incorporated [35S]methionine into protein linearly for at least 5 h with no discernible lag period. In contrast, protein secretion was only detectable after a lag of approximately 1 h, consistent with exocytotic secretion of proteins immediately after passage through the secretory pathway and package into secretory vesicles. The extent of protein secretion was unaffected by the phorbol ester PMA, 8-bromo-cAMP, or 8- bromo-cGMP but was doubled by the Ca2+ ionophore ionomycin. In a pulse- label protocol in which proteins were prelabeled for 1 h before a chase period, constitutive secretion was unaffected by depletion of cytosolic Ca2+ but ionomycin was found to give a twofold stimulation of the secretion of presynthesized protein in a Ca(2+)-dependent manner. Ionomycin was still able to stimulate protein secretion after constitutive secretion had terminated. These results suggest that lactating mammary cells possess both a Ca(2+)-independent constitutive pathway and a Ca(2+)-activated regulatory pathway for protein secretion. The same proteins were secreted by both pathways. No ultrastructural evidence for apocrine secretion was seen in response to ionomycin and so it appears that regulated casein release involves exocytosis. Ionomycin was unlikely to be acting by disassembling the cortical actin network since cytochalasin D did not mimic its effects on secretion. The regulated pathway may be controlled by Ca2+ acting at a late step such as exocytotic membrane fusion. + + 10090 + Species + + mice + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D014238 + Chemical + + TCA + + + MESH:D008715 + Chemical + + methionine + + + MESH:C016276 + Chemical + + 8- bromo-cGMP + + + MESH:D010703 + Chemical + + phorbol ester + + + MESH:D015759 + Chemical + + Ionomycin + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D015124 + Chemical + + 8-bromo-cAMP + + + MESH:D000069285 + Chemical + + Ca(2+) + + + MESH:D015759 + Chemical + + Ionomycin + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D015759 + Chemical + + ionomycin + + + MESH:D015759 + Chemical + + ionomycin + + + MESH:D015759 + Chemical + + ionomycin + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D008715 + Chemical + + methionine + + + MESH:D000069285 + Chemical + + Ca(2+) + + + MESH:D000069285 + Chemical + + Ca(2+) + + + + + 2289424 + + 1560028 + 311 + 2 + 1992 + 2289424 + 92218481 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 311-25 + 325 + Harter C, Mellman I + 0 + Transport of the lysosomal membrane glycoprotein lgp120 (lgp-A) to lysosomes does not require appearance on the plasma membrane + + 100689406 + Gene + + lgp-A + + + + ABSTRACT + abstract + 128 + We have used stably transfected CHO cell lines to characterize the pathway of intracellular transport of the lgp120 (lgp-A) to lysosomes. Using several surface labeling and internalization assays, our results suggest that lgp120 can reach its final destination with or without prior appearance on the plasma membrane. The extent to which lgp120 was transported via the cell surface was determined by two factors: expression level and the presence of a conserved glycine-tyrosine motif in the cytoplasmic tail. In cells expressing low levels of wild-type lgp120, the majority of newly synthesized molecules reached lysosomes without becoming accessible to antibody or biotinylation reagents added extracellularly at 4 degrees C. With increased expression levels, however, an increased fraction of transfected lgp120, as well as some endogenous lgp-B, appeared on the plasma membrane. The fraction of newly synthesized lgp120 reaching the cell surface was also increased by mutations affecting the cytoplasmic domain tyrosine or glycine residues. A substantial fraction of both mutants reached the surface even at low expression levels. However, only the lgp120G----A7 mutant was rapidly internalized and delivered from the plasma membrane to lysosomes. Taken together, our results show that the majority of newly synthesized wild-type lgp120 does not appear to pass through the cell surface en route to lysosomes. Instead, it is likely that lysosomal targeting involves a saturable intracellular sorting site whose affinity for lgp's is dependent on a glycine-tyrosine motif in the lgp120 cytoplasmic tail. + + 100689406 + Gene + + lgp-A + + + 100689316 + Gene + + lgp-B + + + 10029 + Species + + CHO cell lines + + + MESH:D014443 + Chemical + + tyrosine + + + MESH:D005998 + Chemical + + glycine + + + MESH:D005998 + Chemical + + glycine + + + MESH:D005998 + Chemical + + glycine + + + MESH:D014443 + Chemical + + tyrosine + + + - + Chemical + + lgp120 + + + MESH:D014443 + Chemical + + tyrosine + + + + + 2289425 + + 1373145 + 437 + 2 + 1992 + 2289425 + 92218492 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 437-47 + 447 + LaFlamme SE, Akiyama SK, Yamada KM + 0 + Regulation of fibronectin receptor distribution [published erratum appears in J Cell Biol 1992 Jul;118(2):491] + + 2335 + Gene + 1533 + + fibronectin + + + + ABSTRACT + abstract + 111 + To determine the role of each intracellular domain of the fibronectin receptor in receptor distribution, chimeric receptors were constructed containing the human interleukin-2 receptor (gp55 subunit) as the extracellular and transmembrane domains, in combination with either the alpha 5 or beta 1 intracellular domain of the fibronectin receptor as the cytoplasmic domain. These chimeric receptors were transiently expressed in normal fibroblasts, and their localization on the cell surface was determined by immunofluorescence using antibodies to the human interleukin-2 receptor. The alpha 5 chimera was expressed diffusely on the plasma membrane. The beta 1 chimera, however, colocalized with the endogenous fibronectin receptor at focal contacts of cells spread on fibronectin. On cells spread in the presence of serum, the beta 1 chimera colocalized both with the fibronectin receptor at sites of extracellular fibronectin fibrils and with the vitronectin receptor at focal contacts. The beta 1 intracellular domain alone, therefore, contains sufficient information to target the chimeric receptor to regions of the cell where ligand-occupied integrin receptors are concentrated. The finding that the beta 1 chimeric protein behaves like a ligand-occupied receptor, even though the beta 1 chimera cannot itself bind extracellular ligand, suggests an intracellular difference between occupied and unoccupied receptors, and predicts that the distribution of integrin receptors can be regulated by ligand occupancy. We tested this prediction by providing a soluble cell-binding fragment of fibronectin to cells spread on laminin. Under conditions preventing further ligand adsorption to the substrate, this treatment nevertheless resulted in the relocation of diffuse fibronectin receptors to focal contacts. Similarly, a redistribution of diffuse vitronectin receptors to focal contacts occurred on cells spread on laminin after the addition of the small soluble peptide GRGDS. We conclude that the propensity for receptor redistribution to focal contacts driven by the beta 1 cytoplasmic domain alone is suppressed in heterodimeric unoccupied fibronectin receptors, and that ligand occupancy can release this constraint. This redistribution of integrin receptors after the binding of a soluble substrate molecule may provide a direct means of assembling adhesion sites. + + 2335 + Gene + 1533 + + fibronectin + + + 27020 + Gene + 7531 + + gp55 + + + 2335 + Gene + 1533 + + fibronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 7448 + Gene + 532 + + vitronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 7448 + Gene + 532 + + vitronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 2335 + Gene + 1533 + + fibronectin + + + 2335 + Gene + 1533 + + fibronectin + + + + + 2289426 + + 1560026 + 245 + 2 + 1992 + 2289426 + 92218475 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 245-58 + 258 + Wong SH, Low SH, Hong W + 0 + The 17-residue transmembrane domain of beta-galactoside alpha 2,6- sialyltransferase is sufficient for Golgi retention + + + ABSTRACT + abstract + 119 + beta-Galactoside alpha 2,6-sialyltransferase (ST) is a type II integral membrane protein of the Golgi apparatus involved in the sialylation of N-linked glycans. A series of experiments has shown that the 17-residue transmembrane domain of ST is sufficient to confer localization to the Golgi apparatus when transferred to the corresponding region of a cell surface type II integral membrane protein. Lectin affinity chromatography of chimeric proteins bearing this 17-residue sequence suggests that these chimeric proteins are localized in the trans-Golgi cisternae and/or trans-Golgi network. Further experiments suggest that this 17-residue sequence functions as a retention signal for the Golgi apparatus. + + - + Chemical + + N-linked glycans + + + + + 2289427 + + 1313814 + 291 + 2 + 1992 + 2289427 + 92218479 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 291-9 + 299 + Eveleth DD, Bradshaw RA + 0 + Nerve growth factor nonresponsive pheochromocytoma cells: altered internalization results in signaling dysfunction + + MESH:D010673 + Disease + + pheochromocytoma + + + + ABSTRACT + abstract + 115 + Variant rat pheochromocytoma (PC12) cells which fail to respond to nerve growth factor (NGF) (PC12nnr5) (Green, S. H., R. E. Rydel, J. L. Connoly, and L. A. Greene. 1986. J. Cell Biol. 102:830-843) bind NGF at both high and low affinity sites. Although still undefined at the molecular level, these have been referred to as type I (high) and type II (low) receptors. They are apparently composed of two membrane-bound proteins, p75 and the protooncogene trk, both of which bind NGF, and apparently contribute singularly or in concert to the two observed affinities, and to the promotion of the NGF effects. In native PC12 cells, only the high affinity receptors are apparently capable of mediating internalization and degradation. PC12nnr5 cells also display type I binding, but the subsequent internalization is not the same fashion as in the parental cell line, nor is it subjected to lysosomal degradation. Rather it is initially sequestered during the first 15 min, and is eventually released intact into the medium. In contrast, EGF is bound, internalized, and degraded by PC12nnr5 cells, albeit less efficiently than in the parent cells. These observations argue that the defect(s) preventing the PC12nnr5 variants from responding to NGF prevents competent internalization, which in the case of NGF, may be required for the full expression of activity. The absence of trk, as one alteration in PC12nnr5 cells (Loeb, D. M., J. Maragos, D. Martin- Zanca, M. V. Chao, L. F. Parada, and L. A. Greene. 1991. Cell. 66:961- 966), is consistent with this conclusion. + + 24596 + Gene + 1877 + + p75 + + + 59109 + Gene + 1898 + + trk + + + 59109 + Gene + 1898 + + trk + + + 10116 + Species + + rat + + + CVCL:0481 + CellLine + + PC12 + + + MESH:D010673 + Disease + + pheochromocytoma + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + + + 2289428 + + 1560035 + 461 + 2 + 1992 + 2289428 + 92218494 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 461-70 + 470 + van de Wiel-van Kemenade E, van Kooyk Y, de Boer AJ, Huijbens RJ, Weder P, van de Kasteele W, Melief CJ, Figdor CG + 0 + Adhesion of T and B lymphocytes to extracellular matrix and endothelial cells can be regulated through the beta subunit of VLA + + + ABSTRACT + abstract + 127 + Investigating the regulation of very late antigen (VLA)-mediated functions, we found that TS2/16, a mAb directed against the beta chain of the VLA group of integrins, can induce binding of resting peripheral blood lymphocytes, cloned T lymphocytes, and Epstein Barr virus- transformed B cells to extracellular matrix components, fibronectin, laminin, and collagen, but not to fibrinogen. The antibody stimulates VLA-4-, VLA-5-, and VLA-6-mediated binding. Furthermore, it induces VLA- 4-mediated binding to vascular cell adhesion molecule-1 expressed by rTNF-alpha-stimulated endothelial cells, but it does not stimulate homotypic aggregation of cells as described for a number of anti-VLA-4 alpha antibodies (Bednarczyk, J.L., and B. W. McIntyre. 1990. J. Immunol. 144: 777-784; Campanero, M. R., R. Pulido, M. A. Ursa, M. Rodriguez-Moya, M. O. de Landazuri, and F. Sanchez-Madrid. 1990. J. Cell Biol. 110:2157-2165). Therefore, the stimulating activity of this anti-beta 1 antibody clearly contrasts with that of the anti-VLA-4 alpha antibodies, which induce homotypic cell aggregation, but not binding of cells to extracellular matrix components or endothelial cells, indicating that TS2/16 may generate different signals. The observation that also F(ab')2 or Fab fragments of this anti-beta 1 antibody stimulate binding to extracellular matrix components and endothelial cells excludes the possibility that binding requires receptor crosslinking, or is Fc receptor mediated. Induction of this adhesion is cation and energy dependent and requires an intact cytoskeleton. Although changes in the conformation of VLA integrins induced by this antibody may regulate their functional activity, the dependence on metabolic energy indicates that intracellular processes may also play a role. + + 3678 + Gene + 20508 + + VLA-5 + + + 3655 + Gene + 20091 + + VLA-6 + + + 7412 + Gene + 838 + + vascular cell adhesion molecule-1 + + + 24835 + Gene + 496 + + rTNF-alpha + + + 3779 + Gene + 3054 + + beta 1 + + + 2187 + Gene + 51880 + + Fab + + + 3779 + Gene + 3054 + + beta 1 + + + 10376 + Species + + Epstein Barr virus + + + - + Chemical + + Campanero + + + + + 2289429 + + 1373142 + 337 + 2 + 1992 + 2289429 + 92218483 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 337-45 + 345 + Frank JS, Mottino G, Reid D, Molday RS, Philipson KD + 0 + Distribution of the Na(+)-Ca2+ exchange protein in mammalian cardiac myocytes: an immunofluorescence and immunocolloidal gold-labeling study + + 9606 + Species + + mammalian + + + MESH:D000069285 + Chemical + + Ca2+ + + + + ABSTRACT + abstract + 141 + The present study reports on the location of the Na(+)-Ca2+ exchanger in cardiac sarcolemma with immunofluorescence and immunoelectron microscopy. Both polyclonal and monoclonal antibodies to the Na(+)-Ca2+ exchanger were used. The mAb was produced from a hybridoma cell line generated by the fusion of mouse myeloma NS-1 cells with spleen cells from a mouse repeatedly immunized with isolated reconstituted canine cardiac Na(+)-Ca2+ exchanger (Philipson, K. D. S. Longoni, and R. Ward. 1988. Biochim. Biophys. Acta. 945:298-306). The polyclonal antibody has been described previously and reacts with three proteins (70, 120, 160 kD) in cardiac sarcolemma associated with the Na(+)-Ca2+ exchanger (Nicoll, D. A., S. Longoni, and K. D. Philipson. 1990. Science (Wash. DC). 250:562-565). Both the monoclonal and the polyclonal antibodies appear to react with extracellular facing epitopes in the cardiac sarcolemma. Immunofluorescence studies showed labeling of the transverse tubular membrane and patchy labeling of the peripheral sarcolemma. The immunofluorescent labeling clearly delineates the highly interconnected T-tubular system of guinea pig myocytes. This localization of the exchanger to the sarcolemma, with an apparent high density in the transverse tubules, was also seen with immunoelectron microscopy. It is of great interest that the Na(+)-Ca2+ exchanger, as the main efflux route for Ca2+ in heart cells, would be abundantly located in sarcolemma closest to the release of Ca2+. + + 6546 + Gene + 69090 + + Na(+)-Ca2+ exchanger + + + 342538 + Gene + 133972 + + Na(+)-Ca2 + + + 6546 + Gene + 69090 + + Na(+)-Ca2+ exchanger + + + 6546 + Gene + 69090 + + Na(+)-Ca2+ exchanger + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 9615 + Species + + canine + + + 10141 + Species + + guinea pig + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D009101 + Disease + + myeloma + + + MESH:D000069285 + Chemical + + Ca2+ + + + + + 2289430 + + 1560029 + 327 + 2 + 1992 + 2289430 + 92218482 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Apr; 117(2) 327-35 + 335 + Tzen JT, Huang AH + 0 + Surface structure and properties of plant seed oil bodies + + + ABSTRACT + abstract + 58 + Storage triacylglycerols (TAG) in plant seeds are present in small discrete intracellular organelles called oil bodies. An oil body has a matrix of TAG, which is surrounded by phospholipids (PL) and alkaline proteins, termed oleosins. Oil bodies isolated from mature maize (Zea mays) embryos maintained their discreteness, but coalesced after treatment with trypsin but not with phospholipase A2 or C. Phospholipase A2 or C exerted its activity on oil bodies only after the exposed portion of oleosins had been removed by trypsin. Attempts were made to reconstitute oil bodies from their constituents. TAG, either extracted from oil bodies or of a 1:2 molar mixture of triolein and trilinolein, in a dilute buffer were sonicated to produce droplets of sizes similar to those of oil bodies; these droplets were unstable and coalesced rapidly. Addition of oil body PL or dioleoyl phosphatidylcholine, with or without charged stearylamine/stearic acid, or oleosins, to the medium before sonication provided limited stabilization effects to the TAG droplets. High stability was achieved only when the TAG were sonicated with both oil body PL (or dioleoyl phosphatidylcholine) and oleosins of proportions similar to or higher than those in the native oil bodies. These stabilized droplets were similar to the isolated oil bodies in chemical properties, and can be considered as reconstituted oil bodies. Reconstituted oil bodies were also produced from TAG of a 1:2 molar mixture of triolein and trilinolein, dioleoyl phosphatidylcholine, and oleosins from rice (Oryza sativa), wheat (Triticum aestivum), rapeseed (Brassica napus), soybean (Glycine max), or jojoba (Simmondsia chinensis). It is concluded that both oleosins and PL are required to stabilize the oil bodies and that oleosins prevent oil bodies from coalescing by providing steric hindrance. A structural model of an oil body is presented. The current findings on seed oil bodies could be extended to the intracellular storage lipid particles present in diverse organisms. + + 100284535 + Gene + + phospholipase A2 + + + 100284535 + Gene + + Phospholipase A2 + + + 4577 + Species + + maize + + + 4577 + Species + + Zea mays + + + 4530 + Species + + rice + + + 4530 + Species + + Oryza sativa + + + 4565 + Species + + wheat + + + 4565 + Species + + Triticum aestivum + + + 3708 + Species + + Brassica napus + + + 3847 + Species + + soybean + + + 3847 + Species + + Glycine max + + + 3999 + Species + + jojoba + + + 3999 + Species + + Simmondsia chinensis + + + - + Chemical + + oleosins + + + MESH:D009821 + Chemical + + oil + + + MESH:D009821 + Chemical + + oil + + + - + Chemical + + oleosins + + + MESH:C017251 + Chemical + + dioleoyl phosphatidylcholine + + + MESH:C017251 + Chemical + + dioleoyl phosphatidylcholine + + + MESH:D010743 + Chemical + + PL + + + - + Chemical + + seed oil + + + MESH:D009821 + Chemical + + Oil + + + MESH:D014280 + Chemical + + triacylglycerols + + + MESH:D010743 + Chemical + + phospholipids + + + - + Chemical + + oleosins + + + MESH:D009821 + Chemical + + oil + + + MESH:C009317 + Chemical + + stearylamine + + + - + Chemical + + oleosins + + + MESH:D010743 + Chemical + + PL + + + MESH:D009821 + Chemical + + oil + + + MESH:D009821 + Chemical + + oil + + + MESH:D010743 + Chemical + + PL + + + MESH:D008055 + Chemical + + lipid + + + MESH:D009821 + Chemical + + oil + + + MESH:D009821 + Chemical + + oil + + + MESH:C017251 + Chemical + + dioleoyl phosphatidylcholine + + + MESH:D009821 + Chemical + + oil + + + - + Chemical + + oleosins + + + MESH:C031183 + Chemical + + stearic acid + + + - + Chemical + + oleosins + + + - + Chemical + + oleosins + + + + + 2289434 + + 1374069 + 659 + 3 + 1992 + 2289434 + 92242310 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 659-70 + 670 + Arroyo AG, Sánchez-Mateos P, Campanero MR, Martín-Padura I, Dejana E, Sánchez-Madrid F + 0 + Regulation of the VLA integrin-ligand interactions through the beta 1 subunit + + 3779 + Gene + 3054 + + beta 1 + + + + ABSTRACT + abstract + 78 + Integrins from the very late activation antigen (VLA) subfamily are involved in cellular attachment to extracellular matrix (ECM) proteins and in intercellular adhesions. It is known that the interaction of integrin proteins with their ligands can be regulated during cellular activation. We have investigated the regulation of different VLA- mediated adhesive interactions through the common beta 1 chain. We have found that certain anti-beta 1 antibodies strongly enhance binding of myelomonocytic U-937 cells to fibronectin. This beta 1-mediated regulatory effect involved both VLA-4 and VLA-5 fibronectin receptors. Moreover, anti-beta 1 mAb also induced VLA-4-mediated binding to a recombinant soluble form of its endothelial cell ligand VCAM-1. Non- activated peripheral blood T lymphocytes, unable to mediate VLA-4 interactions with fibronectin or VCAM-1, acquired the ability to bind these ligands in the presence of anti-beta 1 mAb. The anti-beta 1- mediated changes in the affinities of beta 1 integrin for their ligands were comparable to those triggered by different lymphocyte activation agents such as anti-CD3 mAb or phorbol ester. Adhesion of melanoma cells to other ECM proteins such as laminin or collagen as well as that of alpha 2-transfected K-562 cells to collagen, was also strongly enhanced by anti-beta 1 mAb. These beta 1-mediated regulatory effects on different VLA-ligand interactions do not involve changes in cell surface membrane expression of different VLA heterodimers. The anti- beta 1-mediated functional effects required an active metabolism, cytoskeleton integrity and the existence of physiological levels of intracellular calcium as well as a functional Na+/H+ antiporter. Beta 1 antibodies not only increased cell attachment but also promoted spreading and cytoplasmic extension of endothelial cells on plates coated with either fibronectin, collagen, or laminin as well as induced the rapid appearance of microspikes in U-937 cells on fibronectin. Moreover, both beta 1 integrin and the cytoskeletal protein talin colocalized in the anti-beta 1 induced microspikes. These results emphasize the central role of the common beta 1 chain in regulating different adhesive functions mediated by VLA integrins as well as cellular morphology. + + 3779 + Gene + 3054 + + beta 1 + + + 2335 + Gene + 1533 + + fibronectin + + + 7412 + Gene + 838 + + VCAM-1 + + + 2335 + Gene + 1533 + + fibronectin + + + 7412 + Gene + 838 + + VCAM-1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3688 + Gene + 22999 + + beta 1 integrin + + + 3779 + Gene + 3054 + + beta 1 + + + 2335 + Gene + 1533 + + fibronectin + + + 2335 + Gene + 1533 + + fibronectin + + + 3688 + Gene + 22999 + + beta 1 integrin + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + Beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 3779 + Gene + 3054 + + beta 1 + + + 2335 + Gene + 1533 + + fibronectin + + + MESH:D008545 + Disease + + melanoma + + + CVCL:0007 + CellLine + + U-937 + + + MESH:D010703 + Chemical + + phorbol ester + + + CVCL:0004 + CellLine + + K-562 + + + CVCL:0007 + CellLine + + U-937 + + + MESH:D002118 + Chemical + + calcium + + + + + 2289435 + + 1315314 + 493 + 3 + 1992 + 2289435 + 92242296 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 493-503 + 503 + Rapiejko PJ, Gilmore R + 0 + Protein translocation across the ER requires a functional GTP binding site in the alpha subunit of the signal recognition particle receptor + + MESH:D006160 + Chemical + + GTP + + + + ABSTRACT + abstract + 140 + The signal recognition particle (SRP)-mediated translocation of proteins across the RER is a GTP dependent process. Analysis of the primary amino acid sequence of one protein subunit of SRP (SRP54), as well as the alpha subunit of the SRP receptor (SR alpha), has indicated that these proteins contain predicted GTP binding sites. Several point mutations confined to the GTP binding consensus elements of SR alpha were constructed by site specific mutagenesis to define a role for the GTP binding site in SR alpha during protein translocation. The SR alpha mutants were analyzed using an in vitro system wherein SR alpha- deficient microsomal membranes were repopulated with SR alpha by in vitro translation of wild-type or mutant mRNA transcripts. SRP receptors containing SR alpha point mutants were analyzed for their ability to function in protein translocation and to form guanylyl-5'- imidodiphosphate (Gpp[NH]p) stabilized complexes with the SRP. Mutations in SR alpha produced SRP receptors that were either impaired or inactive in protein translocation. These SRP receptors were likewise unable to form Gpp(NH)p stabilized complexes with the SRP. One SR alpha point mutant, Thr 588 to Asn 588, required 50- to 100-fold higher concentrations of GTP relative to the wild-type SR alpha to function in protein translocation. This mutant has provided information on the reaction step in protein translocation that involves the GTP binding site in the alpha subunit of the SRP receptor. + + 6729 + Gene + 2362 + + SRP54 + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 6729 + Gene + 2362 + + SRP + + + 4481 + Gene + 12822 + + SR alpha + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + 4481 + Gene + 12822 + + SR alpha + + + 4481 + Gene + 12822 + + SR alpha + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + 6729 + Gene + 2362 + + SRP + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006165 + Chemical + + Gpp(NH)p + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006160 + Chemical + + GTP + + + - + Chemical + + guanylyl-5'- imidodiphosphate + + + p.T588N + ProteinMutation + + Thr 588 to Asn + + + MESH:D009422 + Disease + + impaired + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006160 + Chemical + + GTP + + + MESH:D006160 + Chemical + + GTP + + + + + 2289436 + + 1572896 + 679 + 3 + 1992 + 2289436 + 92242312 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 679-85 + 685 + König A, Bruckner-Tuderman L + 0 + Transforming growth factor-beta stimulates collagen VII expression by cutaneous cells in vitro + + 7040 + Gene + 540 + + Transforming growth factor-beta + + + + ABSTRACT + abstract + 95 + Collagen VII, the major component of cutaneous anchoring fibrils is expressed at a low level by normal human keratinocytes and fibroblasts in vitro. In cocultures of these two cell types, signals from fibroblasts enhance expression of collagen VII by keratinocytes and vice versa. In this study, the effects of a possible mediator of such a stimulation, transforming growth factor-beta (TGF-beta), were investigated. Its effect on the expression and deposition of the highly insoluble collagen VII was assessed in a semiquantitative manner by a newly developed enzyme-linked immunoassay which is based on immunoblotting. In keratinocyte monocultures, 0.5-20 ng/ml of TGF-beta 2 induced a dose-dependent stimulation of collagen VII expression as measured per microgram of DNA. The maximal enhancement was about sevenfold compared to controls. The effect of TGF-beta 2 was observed already after 12 h, with a steady increase at least up to 3 d. As previous studies have implicated, untreated cocultures of keratinocytes and fibroblasts exhibited a higher basic level of collagen VII expression, which could be further stimulated about twofold by TGF-beta 2. Fibroblasts alone synthesized very minor quantities of collagen VII and could be only weakly stimulated by TGF-beta 2. This growth factor seems a specific enhancer of collagen VII since the expression of laminin, collagen IV, as well as total protein was increased to a much lesser extent. Our data suggest that TGF-beta may be an important mediator of epithelial-mesenchymal interactions and may regulate the synthesis of the anchoring fibrils at the skin basement membrane zone. + + 7040 + Gene + 540 + + transforming growth factor-beta + + + 7040 + Gene + 540 + + TGF-beta + + + 7042 + Gene + 2432 + + TGF-beta 2 + + + 7042 + Gene + 2432 + + TGF-beta 2 + + + 7042 + Gene + 2432 + + TGF-beta 2 + + + 7042 + Gene + 2432 + + TGF-beta 2 + + + 7040 + Gene + 540 + + TGF-beta + + + 9606 + Species + + human + + + + + 2289437 + + 1315318 + 629 + 3 + 1992 + 2289437 + 92242308 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 629-41 + 641 + Parvinen M, Pelto-Huikko M, Söder O, Schultz R, Kaipia A, Mali P, Toppari J, Hakovirta H, Lönnerberg P, Ritzén EM + 0 + Expression of beta-nerve growth factor and its receptor in rat seminiferous epithelium: specific function at the onset of meiosis + + 310738 + Gene + 1876 + + beta-nerve growth factor + + + 10116 + Species + + rat + + + + ABSTRACT + abstract + 130 + beta-Nerve growth factor (NGF) is expressed in spermatogenic cells and has testosterone-downregulated low-affinity receptors on Sertoli cells suggesting a paracrine role in the regulation of spermatogenesis. An analysis of the stage-specific expression of NGF and its low affinity receptor during the cycle of the seminiferous epithelium in the rat revealed NGF mRNA and protein at all stages of the cycle. Tyrosine kinase receptor (trk) mRNA encoding an essential component of the high- affinity NGF receptor was also present at all stages. In contrast, expression of low affinity NGF receptor mRNA was only found in stages VIIcd and VIII of the cycle, the sites of onset of meiosis. The low- affinity NGF receptor protein was present in the plasma membrane of the apical Sertoli cell processes as well as in the basal plasma membrane of these cells at stages VIIcd to XI. NGF was shown to stimulate in vitro DNA synthesis of seminiferous tubule segments with preleptotene spermatocytes at the onset of meiosis while other segments remained nonresponsive. We conclude that NGF is a meiotic growth factor that acts through Sertoli cells. + + 310738 + Gene + 1876 + + beta-Nerve growth factor + + + 310738 + Gene + 1876 + + NGF + + + 310738 + Gene + 1876 + + NGF + + + 310738 + Gene + 1876 + + NGF + + + 59109 + Gene + 1898 + + Tyrosine kinase receptor + + + 59109 + Gene + 1898 + + trk + + + 24596 + Gene + 1877 + + NGF receptor + + + 24596 + Gene + 1877 + + NGF receptor + + + 24596 + Gene + 1877 + + NGF receptor + + + 310738 + Gene + 1876 + + NGF + + + 310738 + Gene + 1876 + + NGF + + + 10116 + Species + + rat + + + MESH:D013739 + Chemical + + testosterone + + + + + 2289438 + + 1572893 + 515 + 3 + 1992 + 2289438 + 92242298 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 515-29 + 529 + Duronio RJ, Knoll LJ, Gordon JI + 0 + Isolation of a Saccharomyces cerevisiae long chain fatty acyl:CoA synthetase gene (FAA1) and assessment of its role in protein N- myristoylation + + 854495 + Gene + 137160 + + FAA1 + + + 4932 + Species + + Saccharomyces cerevisiae + + + + ABSTRACT + abstract + 145 + Regulation of myristoylCoA pools in Saccharomyces cerevisiae plays an important role in modulating the activity of myristoylCoA:protein N- myristoyltransferase (NMT), an essential enzyme with an ordered Bi Bi reaction that catalyzes the transfer of myristate from myristoylCoA to greater than or equal to 12 cellular proteins. At least two pathways are available for generating myristoylCoA: de novo synthesis by the multifunctional, multisubunit fatty acid synthetase complex (FAS) and activation of exogenous myristate by acylCoA synthetase. The FAA1 (fatty acid activation) gene has been isolated by genetic complementation of a faal mutant. This single copy gene, which maps to the right arm of chromosome XV, specifies a long chain acylCoA synthetase of 700 amino acids. Analyses of strains containing NMT1 and a faal null mutation indicated that FAA1 is not essential for vegetative growth when an active de novo pathway for fatty acid synthesis is present. The role of FAA1 in cellular lipid metabolism and protein N-myristoylation was therefore assessed in strains subjected to biochemical or genetic blockade of FAS. At 36 degrees C, FAA1 is required for the utilization of exogenous myristate by NMT and for the synthesis of several phospholipid species. This requirement is not apparent at 24 or 30 degrees C, suggesting that S. cerevisiae contains another acylCoA synthetase activity whose chain length and/or temperature optima may differ from Faalp. + + 854495 + Gene + 137160 + + FAA1 + + + 850892 + Gene + 101539 + + NMT1 + + + 854495 + Gene + 137160 + + FAA1 + + + 854495 + Gene + 137160 + + FAA1 + + + 854495 + Gene + 137160 + + FAA1 + + + 4932 + Species + + Saccharomyces cerevisiae + + + 4932 + Species + + S. cerevisiae + + + MESH:D008055 + Chemical + + lipid + + + MESH:D005227 + Chemical + + fatty acid + + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D005227 + Chemical + + fatty acid + + + + + 2289439 + + 1572897 + 687 + 3 + 1992 + 2289439 + 92242313 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 687-94 + 694 + Chen Q, Linsenmayer C, Gu H, Schmid TM, Linsenmayer TF + 0 + Domains of type X collagen: alteration of cartilage matrix by fibril association and proteoglycan accumulation + + MESH:D002357 + Disease + + cartilage + + + + ABSTRACT + abstract + 111 + During endochondral bone formation, hypertrophic cartilage is replaced by bone or by a marrow cavity. The matrix of hypertrophic cartilage contains at least one tissue-specific component, type X collagen. Structurally type X collagen contains both a collagenous domain and a COOH-terminal non-collagenous one. However, the function(s) of this molecule have remained largely speculative. To examine the behavior and functions of type X collagen within hypertrophic cartilage, we (Chen, Q., E. Gibney, J. M. Fitch, C. Linsenmayer, T. M. Schmid, and T. F. Linsenmayer. 1990. Proc. Natl. Acad. Sci. USA. 87:8046-8050) recently devised an in vitro system in which exogenous type X collagen rapidly (15 min to several hours) moves into non-hypertrophic cartilage. There the molecule becomes associated with preexisting cartilage collagen fibrils. In the present investigation, we find that the isolated collagenous domain of type X collagen is sufficient for its association with fibrils. Furthermore, when non-hypertrophic cartilage is incubated for a longer time (overnight) with "intact" type X collagen, the molecule is found both in the matrix and inside of the chondrocytes. The properties of the matrix of such type X collagen-infiltrated cartilage become altered. Such changes include: (a) antigenic masking of type X collagen by proteoglycans; (b) loss of the permissiveness for further infiltration by type X collagen; and (c) enhanced accumulation of proteoglycans. Some of these changes are dependent on the presence of the COOH-terminal non-collagenous domain of the molecule. In fact, the isolated collagenous domain of type X collagen appears to exert an opposite effect on proteoglycan accumulation, producing a net decrease in their accumulation, particularly of the light form(s) of proteoglycans. Certain of these matrix alterations are similar to ones that have been observed to occur in vivo. This suggests that within hypertrophic cartilage type X collagen has regulatory as well as structural functions, and that these functions are achieved specifically by its two different domains. + + MESH:D002357 + Disease + + hypertrophic cartilage + + + MESH:D002357 + Disease + + hypertrophic cartilage + + + MESH:D002357 + Disease + + hypertrophic cartilage type X + + + MESH:D002357 + Disease + + cartilage + + + MESH:C535964 + Disease + + cartilage collagen fibrils + + + MESH:D002357 + Disease + + hypertrophic cartilage + + + MESH:D002357 + Disease + + hypertrophic cartilage + + + MESH:D002357 + Disease + + hypertrophic cartilage + + + + + 2289440 + + 1533397 + 595 + 3 + 1992 + 2289440 + 92242305 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 595-606 + 606 + Ferreira A, Niclas J, Vale RD, Banker G, Kosik KS + 0 + Suppression of kinesin expression in cultured hippocampal neurons using antisense oligonucleotides + + MESH:D009841 + Chemical + + oligonucleotides + + + + ABSTRACT + abstract + 99 + Kinesin, a microtubule-based force-generating molecule, is thought to translocate organelles along microtubules. To examine the function of kinesin in neurons, we sought to suppress kinesin heavy chain (KHC) expression in cultured hippocampal neurons using antisense oligonucleotides and study the phenotype of these KHC "null" cells. Two different antisense oligonucleotides complementary to the KHC sequence reduced the protein levels of the heavy chain by greater than 95% within 24 h after application and produced identical phenotypes. After inhibition of KHC expression for 24 or 48 h, neurons extended an array of neurites often with one neurite longer than the others; however, the length of all these neurites was significantly reduced. Inhibition of KHC expression also altered the distribution of GAP-43 and synapsin I, two proteins thought to be transported in association with membranous organelles. These proteins, which are normally localized at the tips of growing neurites, were confined to the cell body in antisense-treated cells. Treatment of the cells with the corresponding sense oligonucleotides affected neither the distribution of GAP-43 and synapsin I, nor the length of neurites. A full recovery of neurite length occurred after removal of the antisense oligonucleotides from the medium. These data indicate that KHC plays a role in the anterograde translocation of vesicles containing GAP-43 and synapsin I. A deficiency in vesicle delivery may also explain the inhibition of neurite outgrowth. Despite the inhibition of KHC and the failure of GAP- 43 and synapsin I to move out of the cell body, hippocampal neurons can extend processes and acquire as asymmetric morphology. + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 3799 + Gene + 55829 + + KHC + + + 3799 + Gene + 55829 + + KHC + + + 3799 + Gene + 55829 + + KHC + + + 3799 + Gene + 55829 + + KHC + + + 3799 + Gene + 55829 + + KHC + + + 2596 + Gene + 1545 + + GAP-43 + + + 6853 + Gene + 48483 + + synapsin I + + + 2596 + Gene + 1545 + + GAP-43 + + + 6853 + Gene + 48483 + + synapsin I + + + 3799 + Gene + 55829 + + KHC + + + 2596 + Gene + 1545 + + GAP-43 + + + 6853 + Gene + 48483 + + synapsin I + + + 3799 + Gene + 55829 + + KHC + + + 2596 + Gene + 1545 + + GAP- 43 + + + 6853 + Gene + 48483 + + synapsin I + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:C567751 + Disease + + A deficiency in vesicle delivery + + + + + 2289441 + + 1315315 + 505 + 3 + 1992 + 2289441 + 92242297 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 505-13 + 513 + Marquardt T, Helenius A + 0 + Misfolding and aggregation of newly synthesized proteins in the endoplasmic reticulum + + + ABSTRACT + abstract + 86 + As a part of our studies on the folding of glycoproteins in the ER, we analyzed the fate of viral glycoproteins that have misfolded either spontaneously or through inhibition of N-linked glycosylation. Newly synthesized Semliki Forest virus spike glycoproteins E1 and p62 and influenza hemagglutinin were studied in infected and transfected tissue culture cells. Misfolded proteins aggregated in less than 1 min after release from polysomes and aberrant interchain disulfide bonds were formed immediately. When more than one protein was misfolded, mixed aggregates were generated. This indicated that the formation of complexes was nonspecific, random, and not restricted to products from single polysomes. The size of the aggregates varied from small oligomers to complexes of several million daltons. BiP was associated noncovalently with the aggregates and with some of the nonaggregated products. We conclude that aggregation reflects the poor solubility of incompletely folded polypeptide chains. + + 3309 + Gene + 3908 + + BiP + + + 11033 + Species + + Semliki Forest virus + + + MESH:D004220 + Chemical + + disulfide + + + MESH:D009584 + Chemical + + N + + + + + 2289442 + + 1374068 + 607 + 3 + 1992 + 2289442 + 92242306 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 607-16 + 616 + Lasek RJ, Paggi P, Katz MJ + 0 + Slow axonal transport mechanisms move neurofilaments relentlessly in mouse optic axons + + 10090 + Species + + mouse + + + + ABSTRACT + abstract + 87 + Pulse-labeling studies of slow axonal transport in many kinds of axons (spinal motor, sensory ganglion, oculomotor, hypoglossal, and olfactory) have led to the inference that axonal transport mechanisms move neurofilaments (NFs) unidirectionally as a single continuous kinetic population with a diversity of individual transport rates. One study in mouse optic axons (Nixon, R. A., and K. B. Logvinenko. 1986. J. Cell Biol. 102:647-659) has given rise to the different suggestion that a significant and distinct population of NFs may be entirely stationary within axons. In mouse optic axons, there are relatively few NFs and the NF proteins are more lightly labeled than other slowly transported slow component b (SCb) proteins (which, however, move faster than the NFs); thus, in mouse optic axons, the radiolabel of some of these faster-moving SCb proteins may confuse NF protein analyses that use one dimensional (1-D) SDS-PAGE, which separates proteins by size only. To test this possibility, we used a 2-mm "window" (at 3-5 mm from the posterior of the eye) to compare NF kinetics obtained by 1-D SDS-PAGE and by the higher resolution two- dimensional (2-D) isoelectric focusing/SDS-PAGE, which separates proteins both by their net charge and by their size. We found that 1-D SDS-PAGE is insufficient for definitive NF kinetics in the mouse optic system. By contrast, 2-D SDS-PAGE provides essentially pure NF kinetics, and these indicate that in the NF-poor mouse optic axons, most NFs advance as they do in other, NF-rich axons. In mice, greater than 97% of the radiolabeled NFs were distributed in a unimodal wave that moved at a continuum of rates, between 3.0 and 0.3 mm/d, and less than 0.1% of the NF population traveled at the very slowest rates of less than 0.005 mm/d. These results are inconsistent with the proposal (Nixon and Logvinenko, 1986) that 32% of the transported NFs remain within optic axons in an entirely stationary state. As has been found in other axons, the axonal transport system of mouse optic axons moves NFs and other cytoskeletal elements relentlessly from the cell body to the axon tip. + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + MESH:D012967 + Chemical + + SDS + + + MESH:D012967 + Chemical + + SDS + + + MESH:D012967 + Chemical + + SDS + + + MESH:D012967 + Chemical + + SDS + + + MESH:D012967 + Chemical + + SDS + + + + + 2289443 + + 1374067 + 583 + 3 + 1992 + 2289443 + 92242304 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 583-93 + 593 + Omary MB, Baxter GT, Chou CF, Riopel CL, Lin WY, Strulovici B + 0 + PKC epsilon-related kinase associates with and phosphorylates cytokeratin 8 and 18 + + 3856 + Gene + 55643 + + cytokeratin 8 + + + 5581 + Gene + 48343 + + PKC epsilon + + + + ABSTRACT + abstract + 83 + A 40-kD protein kinase C (PKC)epsilon related activity was found to associate with human epithelial specific cytokeratin (CK) polypeptides 8 and 18. The kinase activity coimmunoprecipitated with CK8 and 18 and phosphorylated immunoprecipitates of the CK. Immunoblot analysis of CK8/18 immunoprecipitates using an anti-PKC epsilon specific antibody showed that the 40-kD species, and not native PKC epsilon (90 kD) associated with the cytokeratins. Reconstitution experiments demonstrated that purified CK8 or CK18 associated with a 40-kD tryptic fragment of purified PKC epsilon, or with a similar species obtained from cells that express the fragment constitutively but do not express CK8/18. A peptide pseudosubstrate specific for PKC epsilon inhibited phosphorylation of CK8/18 in intact cells or in a kinase assay with CK8/18 immunoprecipitates. Tryptic peptide map analysis of the cytokeratins that were phosphorylated by purified rat brain PKC epsilon or as immunoprecipitates by the associated kinase showed similar phosphopeptides. Furthermore, PKC epsilon immunoreactive species and CK8/18 colocalized using immunofluorescent double staining. We propose that a kinase related to the catalytic fragment of PKC epsilon physically associates with and phosphorylates cytokeratins 8 and 18. + + 5581 + Gene + 48343 + + PKC)epsilon + + + 3856 + Gene + 55643 + + CK8 + + + 3856 + Gene + 55643 + + CK8 + + + 5581 + Gene + 48343 + + PKC epsilon + + + 5581 + Gene + 48343 + + PKC epsilon + + + 3856 + Gene + 55643 + + CK8 + + + 3875 + Gene + 55448 + + CK18 + + + 5581 + Gene + 48343 + + PKC epsilon + + + 3856 + Gene + 55643 + + CK8 + + + 5581 + Gene + 48343 + + PKC epsilon + + + 3856 + Gene + 55643 + + CK8 + + + 3856 + Gene + 55643 + + CK8 + + + 5581 + Gene + 48343 + + PKC epsilon + + + 3856 + Gene + 55643 + + CK8 + + + 5581 + Gene + 48343 + + PKC epsilon + + + 9606 + Species + + human + + + 10116 + Species + + rat + + + MESH:D010455 + Chemical + + peptide + + + MESH:C537180 + Disease + + brain PKC epsilon + + + + + 2289444 + + 1374066 + 483 + 3 + 1992 + 2289444 + 92242295 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 483-91 + 491 + Olins AL, Olins DE, Bazett-Jones DP + 0 + Balbiani ring hnRNP substructure visualized by selective staining and electron spectroscopic imaging + + 9987 + Gene + 75314 + + hnRNP + + + + ABSTRACT + abstract + 101 + The Balbiani Rings (BR) in the polytene chromosomes of Chironomus salivary glands are intense sites of transcription. The nascent RNPs fold during transcription into 40-50-nm granules, containing in the mature transcript approximately 37-kb RNA. Using a new nucleic acid specific stain, osmium ammine B on Lowicryl sections, in combination with electron energy filtered imaging of sections containing BR granules, we demonstrate a RNA-rich particulate substructure (10-nm particle diameter; 10-12 particles per BR granule). Elemental imaging supports that these particles are enriched in phosphorus. The possible relationship of these RNA-rich particles to ribonucleosomes is discussed, as well as models for their arrangement in the mature BR granules. + + MESH:D010758 + Chemical + + phosphorus + + + MESH:C058639 + Chemical + + osmium ammine B + + + - + Chemical + + Lowicryl + + + + + 2289445 + + 1315317 + 565 + 3 + 1992 + 2289445 + 92242302 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 565-72 + 572 + Kuijpers TW, Hakkert BC, Hart MH, Roos D + 0 + Neutrophil migration across monolayers of cytokine-prestimulated endothelial cells: a role for platelet-activating factor and IL-8 + + 9768 + Gene + 8829 + + platelet-activating factor + + + + ABSTRACT + abstract + 131 + In a previous study we observed that neutrophils respond with a rapid rise in [Ca2+]i during adherence to cytokine-activated endothelial cells (EC), caused by EC membrane-associated platelet-activating factor (PAF). In the present study, we investigated whether this form of PAF was important in neutrophil adherence and migration across monolayers of rIL-1 beta- or rTNF alpha-prestimulated EC. PAF receptor antagonists prevented neutrophil migration across cytokine-pretreated EC by approximately 60% (P less than 0.005) without interfering with the process of adherence. The antagonists WEB 2086 and L-652,731 had no effect on neutrophil migration across resting EC induced by formylmethionyl-leucyl-phenylalanine (FMLP). A murine anti-IL-8 antiserum was found to also partially inhibit the neutrophil transmigration across cytokine-activated EC. When the anti-IL-8 antiserum was used in combination with a PAF receptor antagonist, neutrophil migration across cytokine-pretreated monolayers of EC was completely prevented. During transmigration, LAM-1 and CD44 on the neutrophils were down-modulated; both WEB 2086 and anti-IL-8 antiserum partially prevented this down-modulation caused by cytokine- prestimulated EC. Our results indicate that human neutrophils are activated and guided by EC-associated PAF and EC-derived IL-8 during the in vitro diapedesis in between cytokine-stimulated EC. + + 9768 + Gene + 8829 + + membrane-associated platelet-activating factor (PAF) + + + 9768 + Gene + 8829 + + PAF + + + 3576 + Gene + 47937 + + IL-8 + + + 3576 + Gene + 47937 + + IL-8 + + + 6402 + Gene + 539 + + LAM-1 + + + 960 + Gene + 508 + + CD44 + + + 3576 + Gene + 47937 + + IL-8 + + + 3576 + Gene + 47937 + + IL-8 + + + 9768 + Gene + 8829 + + PAF + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 9768 + Gene + 8829 + + PAF + + + 9768 + Gene + 8829 + + PAF + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D009240 + Chemical + + formylmethionyl-leucyl-phenylalanine + + + + + 2289446 + + 1349305 + 617 + 3 + 1992 + 2289446 + 92242307 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 617-27 + 627 + Rosen CL, Lisanti MP, Salzer JL + 0 + Expression of unique sets of GPI-linked proteins by different primary neurons in vitro + + + ABSTRACT + abstract + 87 + We have surveyed the proteins expressed at the surface of different primary neurons as a first step in elucidating how axons regulate their ensheathment by glial cells. We characterized the surface proteins of dorsal root ganglion neurons, superior cervical ganglion neurons, and cerebellar granule cells which are myelinated, ensheathed but unmyelinated, and unensheathed, respectively. We found that the most abundant proteins are common to all three types of neurons. Reproducible differences in the composition of the integral membrane proteins (enriched by partitioning into a Triton X-114 detergent phase) were detected. These differences were most striking when the expression of glycosylphosphatidyl-inositol (GPI)-anchored membrane proteins by these different neurons was compared. Variations in the relative abundance and degree of glycosylation of several well known GPI- anchored proteins, including Thy-1, F3/F11, and the 120-kD form of the neural cell adhesion molecule (N-CAM), and an abundant 60-kD GPI-linked protein were observed. In addition, we have identified several potentially novel GPI-anchored glycoproteins on each class of neurons. These include a protein that is present only on superior cervical ganglion neurons and is 90 kD; an abundant protein of 69 kD that is essentially restricted in its expression to dorsal root ganglion neurons; and proteins of 38 and 31 kD that are expressed only on granule cell neurons. Finally, the relative abundance of the three major isoforms of N-CAM was found to vary significantly between these different primary neurons. These results are the first demonstration that nerve fibers with diverse ensheathment fates differ significantly in the composition of their surface proteins and suggest an important role for GPI-anchored proteins in generating diversity of the neuronal cell surface. + + 7070 + Gene + 4580 + + Thy-1 + + + 4684 + Gene + 40754 + + neural cell adhesion molecule + + + 4684 + Gene + 40754 + + N-CAM + + + 4684 + Gene + 40754 + + N-CAM + + + MESH:D017261 + Chemical + + glycosylphosphatidyl-inositol + + + MESH:C010615 + Chemical + + Triton X-114 + + + MESH:D017261 + Chemical + + GPI + + + + + 2289447 + + 1572894 + 539 + 3 + 1992 + 2289447 + 92242300 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 539-49 + 549 + Grimes M, Kelly RB + 0 + Intermediates in the constitutive and regulated secretory pathways released in vitro from semi-intact cells + + + ABSTRACT + abstract + 108 + Regulated secretory cells have two pathways that transport secreted proteins from the Golgi complex to the cell surface. To identify carrier vesicles involved in regulated and constitutive secretion, PC12 pheochromocytoma cells were labeled with [35S]sulfate to identify markers for the two secretory pathways, then mechanically permeabilized and incubated in vitro. Small constitutive secretory vesicles, containing mostly sulfated proteoglycans, accumulated during an in vitro incubation with ATP. In the presence of GTP gamma S, the constitutive vesicles became significantly more dense, suggesting that a coated intermediate was stabilized. Larger immature regulated secretory granules, enriched in sulfated secretogranin II, also escaped from the permeabilized cells in vitro. During granule maturation, their density increased and the amount of cofractionating proteoglycans diminished. The data suggest that sorting continues during secretory granule maturation. + + 24765 + Gene + 2591 + + secretogranin II + + + MESH:D010673 + Disease + + pheochromocytoma + + + MESH:D016244 + Chemical + + GTP gamma S + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D000255 + Chemical + + ATP + + + MESH:D013431 + Chemical + + sulfate + + + CVCL:0481 + CellLine + + PC12 + + + + + 2289448 + + 1572895 + 551 + 3 + 1992 + 2289448 + 92242301 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 551-64 + 564 + Zurzolo C, Polistina C, Saini M, Gentile R, Aloj L, Migliaccio G, Bonatti S, Nitsch L + 0 + Opposite polarity of virus budding and of viral envelope glycoprotein distribution in epithelial cells derived from different tissues + + 100616444 + Gene + + envelope glycoprotein + + + + ABSTRACT + abstract + 134 + We compared the surface envelope glycoprotein distribution and the budding polarity of four RNA viruses in Fischer rat thyroid (FRT) cells and in CaCo-2 cells derived from a human colon carcinoma. Whereas both FRT and CaCo-2 cells sort similarly influenza hemagglutinin and vesicular stomatitis virus (VSV) G protein, respectively, to apical and basolateral membrane domains, they differ in their handling of two togaviruses, Sindbis and Semliki Forest virus (SFV). By conventional EM Sindbis virus and SFV were shown to bud apically in FRT cells and basolaterally in CaCo-2 cells. Consistent with this finding, the distribution of the p62/E2 envelope glycoprotein of SFV, assayed by immunoelectronmicroscopy and by domain-selective surface biotinylation was predominantly apical on FRT cells and basolateral on CaCo-2 cells. We conclude that a given virus and its envelope glycoprotein can be delivered to opposite membrane domains in epithelial cells derived from different tissues. The tissue specificity in the polarity of virus budding and viral envelope glycoprotein distribution indicate that the sorting machinery varies considerably between different epithelial cell types. + + 100616444 + Gene + + envelope glycoprotein + + + 100616444 + Gene + + envelope glycoprotein + + + 100616444 + Gene + + envelope glycoprotein + + + 10116 + Species + + rat + + + 9606 + Species + + human + + + 11276 + Species + + vesicular stomatitis virus + + + 11033 + Species + + Semliki Forest virus + + + 11034 + Species + + Sindbis virus + + + 11034 + Species + + Sindbis + + + 11276 + Species + + VSV + + + 11033 + Species + + SFV + + + 11033 + Species + + SFV + + + 11033 + Species + + SFV + + + 100616444 + Gene + + envelope glycoprotein + + + CVCL:0025 + CellLine + + CaCo-2 + + + CVCL:0025 + CellLine + + CaCo-2 + + + CVCL:0025 + CellLine + + CaCo-2 + + + MESH:D015179 + Disease + + colon carcinoma + + + CVCL:0025 + CellLine + + CaCo-2 + + + + + 2289449 + + 1533398 + 671 + 3 + 1992 + 2289449 + 92242311 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 671-8 + 678 + Lee EC, Lotz MM, Steele GD, Mercurio AM + 0 + The integrin alpha 6 beta 4 is a laminin receptor + + + ABSTRACT + abstract + 50 + In this study, the putative laminin receptor function of the alpha 6 beta 4 integrin was assessed. For this purpose, we used a human cell line, referred to as clone A, that was derived from a highly invasive, colon adenocarcinoma. This cell line, which expresses the alpha 6 beta 4 integrin, adheres to the E8 and not to the P1 fragment of laminin. The adhesion of clone A cells to laminin is extremely rapid with half- maximal adhesion observed at 5 min after plating. Adhesion to laminin is blocked by GoH3, and alpha 6 specific antibody (60% inhibition), as well as by A9, a beta 4 specific antibody (30% inhibition). Most importantly, we demonstrate that alpha 6 beta 4 binds specifically to laminin-Sepharose columns in the presence of either Mg2+ or Mn2+ and it is eluted from these columns with EDTA but not with NaCl. The alpha 6 beta 4 integrin does not bind to collagen-Sepharose, but the alpha 2 beta 1 integrin does bind. Clone A cells do not express alpha 6 beta 1 as evidenced by the following observations: (a) no beta 1 integrin is detected in beta 1 immunoblots of GoH3 immunoprecipitates; and (b) no alpha 6 beta 1 integrin is seen in GoH3 immunoprecipitates of clone A extracts that had been immunodepleted of all beta 4 containing integrin using the A9 antibody. These data establish that laminin is a ligand for the alpha 6 beta 4 integrin and that this integrin can function as a laminin receptor independently of alpha 6 beta 1. + + 3688 + Gene + 22999 + + beta 1 integrin + + + 9606 + Species + + human + + + 3688 + Gene + 22999 + + beta 1 integrin + + + 3688 + Gene + 22999 + + beta 1 integrin + + + - + Chemical + + Mg2+ + + + - + Chemical + + Mn2+ + + + MESH:D003110 + Disease + + colon adenocarcinoma + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:D004492 + Chemical + + EDTA + + + MESH:D012965 + Chemical + + NaCl + + + + + 2289450 + + 1315316 + 531 + 3 + 1992 + 2289450 + 92242299 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 531-8 + 538 + Wilson DW, Whiteheart SW, Wiedmann M, Brunner M, Rothman JE + 0 + A multisubunit particle implicated in membrane fusion + + + ABSTRACT + abstract + 54 + The N-ethylmaleimide sensitive fusion protein (NSF) is required for fusion of lipid bilayers at many locations within eukaryotic cells. Binding of NSF to Golgi membranes is known to require an integral membrane receptor and one or more members of a family of related soluble NSF attachment proteins (alpha-, beta-, and gamma-SNAPs). Here we demonstrate the direct interaction of NSF, SNAPs and an integral membrane component in a detergent solubilized system. We show that NSF only binds to SNAPs in the presence of the integral receptor, resulting in the formation of a multisubunit protein complex with a sedimentation coefficient of 20S. Particle assembly reveals striking differences between members of the SNAP protein family; gamma-SNAP associates with the complex via a binding site distinct from that used by alpha- and beta-SNAPs, which are themselves equivalent, alternative subunits of the particle. Once formed, the 20S particle is subsequently able to disassemble in a process coupled to the hydrolysis of ATP. We suggest how cycles of complex assembly and disassembly could help confer specificity to the generalized NSF-dependent fusion apparatus. + + 4905 + Gene + 4502 + + N-ethylmaleimide sensitive fusion protein + + + 4905 + Gene + 4502 + + NSF + + + 4905 + Gene + 4502 + + NSF + + + 4905 + Gene + 4502 + + NSF + + + 4905 + Gene + 4502 + + NSF + + + 4905 + Gene + 4502 + + NSF + + + MESH:D000255 + Chemical + + ATP + + + MESH:D019962 + Disease + + NSF attachment proteins + + + MESH:D008055 + Chemical + + lipid + + + + + 2289451 + + 1533396 + 573 + 3 + 1992 + 2289451 + 92242303 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 573-81 + 581 + Smith EF, Sale WS + 0 + Structural and functional reconstitution of inner dynein arms in Chlamydomonas flagellar axonemes + + + ABSTRACT + abstract + 98 + The inner row of dynein arms contains three dynein subforms. Each is distinct in composition and location in flagellar axonemes. To begin investigating the specificity of inner dynein arm assembly, we assessed the capability of isolated inner arm dynein subforms to rebind to their appropriate positions on axonemal doublet microtubules by recombining them with either mutant or extracted axonemes missing some or all dyneins. Densitometry of Coomassie blue-stained polyacrylamide gels revealed that for each inner dynein arm subform, binding to axonemes was saturable and stoichiometric. Using structural markers of position and polarity, electron microscopy confirmed that subforms bound to the correct inner arm position. Inner arms did not bind to outer arm or inappropriate inner arm positions despite the availability of sites. These and previous observations implicate specialized tubulin isoforms or nontubulin proteins in designation of specific inner dynein arm binding sites. Further, microtubule sliding velocities were restored to dynein-depleted axonemes upon rebinding of the missing inner arm subtypes as evaluated by an ATP-induced microtubule sliding disintegration assay. Therefore, not only were the inner arm dynein subforms able to identify and bind to the correct location on doublet microtubules but they bound in a functionally active conformation. + + MESH:D000255 + Chemical + + ATP + + + MESH:C016679 + Chemical + + polyacrylamide + + + MESH:C048139 + Chemical + + Coomassie blue + + + + + 2289452 + + 1572892 + 473 + 3 + 1992 + 2289452 + 92242294 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 473-82 + 482 + Stochaj U, Silver PA + 0 + A conserved phosphoprotein that specifically binds nuclear localization sequences is involved in nuclear import [published erratum appears in J Cell Biol 1992 Jul;118(1):215] + + + ABSTRACT + abstract + 175 + We have purified proteins of 70 kD from Drosophila, HeLa cells, and Z. mays that specifically bind nuclear localization sequences (NLSs). These proteins are recognized by antibodies raised against a previously identified NLS-binding protein (NBP) from the yeast S. cerevisiae. All NBPs are associated with nuclei and also present in the cytosol. NBPs are phosphorylated and phosphatase treatment abolished NLS binding. The requirement for NBPs in nuclear protein uptake is demonstrated in semipermeabilized Drosophila melanogaster tissue culture cells. Proper import of a fluorescent protein containing the large T antigen NLS requires cytosol and ATP. In the absence of cytosol and/or ATP, NLS- containing proteins are bound to cytosolic structures and the nuclear envelope. Addition of cytosol and ATP results in movement of this bound intermediate into the nucleus. Anti-NBP antibodies specifically inhibited the binding part of this import reaction. These results indicate that a phosphoprotein common to several eukaryotes acts as a receptor that recognizes NLSs before their uptake into the nucleus. + + 4577 + Species + + Z. mays + + + 4932 + Species + + yeast + + + 4932 + Species + + S. cerevisiae + + + 7227 + Species + + Drosophila melanogaster + + + 7227 + Species + + Drosophila + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000255 + Chemical + + ATP + + + CVCL:0030 + CellLine + + HeLa + + + MESH:D000255 + Chemical + + ATP + + + + + 2289453 + + 1315319 + 643 + 3 + 1992 + 2289453 + 92242309 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(3) 643-57 + 657 + Song WK, Wang W, Foster RF, Bielser DA, Kaufman SJ + 0 + H36-alpha 7 is a novel integrin alpha chain that is developmentally regulated during skeletal myogenesis [published erratum appears in J Cell Biol 1992 Jul;118(1):213] + + 104204 + Gene + + H36 + + + + ABSTRACT + abstract + 168 + H36 is a 120,000-D membrane glycoprotein that is expressed during the differentiation of skeletal muscle. H36 cDNA clones were isolated from a lambda UniZapXR rat myotube cDNA library and sequenced. The deduced amino acid sequence demonstrates that H36 is a novel integrin alpha chain that shares extensive homology with other alpha integrins that includes: (a) the GFFKR sequence found in all alpha integrins; (b) a single membrane spanning region; (c) conservation of 18 of 22 cysteines; and (d) a protease cleavage site found in the non-I region integrin alpha chains. The cytoplasmic domain of H36 is unique and additional regions of nonhomology further indicate H36 is distinct from all other alpha chains. In keeping with current nomenclature we designate this alpha chain alpha 7. Northern blots demonstrate that expression of H36-alpha 7 mRNA is regulated both early in the development of the myogenic lineage and later, during terminal differentiation. Detection of H36-alpha 7 mRNA coincides with conversion of H36- myogenic precursor cells to H36+ cells. H36-alpha 7 mRNA is present in replicating myoblasts: expression increases upon terminal differentiation and is markedly reduced in developmentally defective myoblasts. In addition, H36-alpha 7 mRNA is not detected in C3H10T1/2 cells. It is in myotubes derived from myoblasts obtained by treatment of 10T1/2 cells with azacytidine or transfection with MRF4. Immunoblots and immunofluorescence demonstrate that the H36-alpha 7 chain is associated with integrin beta 1. Affinity chromatography demonstrates that H36-alpha 7 beta 1 selectively binds to laminin. The expression of H36-alpha 7 on secondary myoblasts during the development of the limb in vivo corresponds with the appearance of laminin in the limb, with the responsiveness of secondary myoblast proliferation to laminin, and with the onset of increased muscle mass, suggesting that H36-alpha 7 modulates this stage in limb development. We conclude that H36-alpha 7 is a novel alpha integrin laminin binding protein whose expression is developmentally regulated during skeletal myogenesis. + + 104204 + Gene + + H36 + + + 17878 + Gene + 1850 + + MRF4 + + + 16412 + Gene + 22999 + + integrin beta 1 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 10116 + Species + + rat + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + 104204 + Gene + + H36 + + + MESH:D003545 + Chemical + + cysteines + + + MESH:D001374 + Chemical + + azacytidine + + + CVCL:0190 + CellLine + + H10T1/2 + + + + + 2289454 + + 1374413 + 895 + 4 + 1992 + 2289454 + 92250723 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 895-902 + 902 + Foxall C, Watson SR, Dowbenko D, Fennie C, Lasky LA, Kiso M, Hasegawa A, Asa D, Brandley BK + 0 + The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide + + MESH:D009844 + Chemical + + oligosaccharide + + + MESH:D000082 + Chemical + + x + + + + ABSTRACT + abstract + 127 + The selectins (lectin-EGF-complement binding-cell adhesion molecules [LEC-CAMs]) are a family of mammalian receptors implicated in the initial interactions between leukocytes and vascular endothelia, leading to lymphocyte homing, platelet binding, and neutrophil extravasation. The three known selectins, L-selectin (leukocyte adhesion molecule-1 [LECAM-1]), E-selectin (endothelial-leukocyte adhesion molecule-1 [ELAM-1]), and P-selectin (GMP-140) share structural features that include a calcium-dependent lectin domain. The sialyl Lewis(x) carbohydrate epitope has been reported as a ligand for both E- and P-selectins. Although L-selectin has been demonstrated to bind to carbohydrates, structural features of potential mammalian carbohydrate ligand(s) have not been well defined. Using an ELISA developed with a sialyl Lewis(x)-containing glycolipid and an E- selectin-IgG chimera, we have demonstrated the direct binding of the L- selectin-IgG chimera to sialyl Lewis(x). This recognition was calcium dependent, and could be blocked by Mel-14 antibody but not by other antibodies. Recognition was confirmed by the ability of cells expressing the native L-selectin to adhere to immobilized sialyl Lewis(x). These data suggest that the sialyl Lewis(x) oligosaccharide may form the basis of a recognition domain common to all three selectins. + + 6402 + Gene + 539 + + L-selectin + + + 6402 + Gene + 539 + + leukocyte adhesion molecule-1 + + + 6402 + Gene + 539 + + LECAM-1 + + + 6401 + Gene + 389 + + E-selectin + + + 6401 + Gene + 389 + + endothelial-leukocyte adhesion molecule-1 + + + 6401 + Gene + 389 + + ELAM-1 + + + 6403 + Gene + 2260 + + P-selectin + + + 6403 + Gene + 2260 + + GMP-140 + + + 6402 + Gene + 539 + + L-selectin + + + 6402 + Gene + 539 + + L- selectin + + + 6402 + Gene + 539 + + L-selectin + + + 9606 + Species + + mammalian + + + 9606 + Species + + mammalian + + + MESH:D006017 + Chemical + + glycolipid + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002241 + Chemical + + carbohydrate + + + MESH:D002241 + Chemical + + carbohydrates + + + MESH:D002118 + Chemical + + calcium + + + - + Chemical + + (x) oligosaccharide + + + + + 2289455 + + 1577862 + 863 + 4 + 1992 + 2289455 + 92250720 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 863-75 + 875 + Paschke KA, Lottspeich F, Stuermer CA + 0 + Neurolin, a cell surface glycoprotein on growing retinal axons in the goldfish visual system, is reexpressed during retinal axonal regeneration + + 7957 + Species + + goldfish + + + + ABSTRACT + abstract + 144 + The mAb E 21 recognizes a cell surface glycoprotein selectively associated with fish retinal ganglion cell axons that are in a state of growth. All retinal axons and ganglion cells in goldfish embryos stained for E 21. In adult fish, however, E 21 immunoreactivity exhibited a patterned distribution in ganglion cells in the marginal growth zone of the continuously enlarging fish retina and the new axons emerging from these cells in the retina, optic nerve, and optic tract. The E 21 antigen was absent from older axons, except the terminal arbor layer in the tectum, the Stratum fibrosum et griseum superficiale where it was uniformly distributed. Upon optic nerve transection, the previously unlabeled axons reacquired E 21 positivity as they regenerated throughout their path to the tectum. Several months after ONS, however, E 21 staining disappeared from the regenerated axons over most of their lengths but reappeared as in normal fish in the terminal arbor layer. The immunoaffinity-purified E 21 antigen, called Neurolin, has an apparent molecular mass of 86 kD and contains the HNK1/L2 carbohydrate moiety, like several members of the class of cell adhesion molecules of the Ig superfamily. The NH2-terminal amino acid sequence has homologies to the cell adhesion molecule DM-Grasp recently described in the chicken. Thus, retinal ganglion cell axons express Neurolin during their development and are able to reexpress this candidate cell adhesion molecule during axonal regeneration, suggesting that Neurolin is functionally important for fish retinal axon growth. + + 396092 + Gene + 1229 + + DM-Grasp + + + 7957 + Species + + goldfish + + + 9031 + Species + + chicken + + + MESH:D002241 + Chemical + + carbohydrate + + + + + 2289456 + + 1577854 + 745 + 4 + 1992 + 2289456 + 92250710 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 745-55 + 755 + Ghitescu L, Bendayan M + 0 + Transendothelial transport of serum albumin: a quantitative immunocytochemical study + + + ABSTRACT + abstract + 85 + The steady-state distribution of endogenous albumin in mouse diaphragm was determined by quantitative postembedding protein A-gold immunocytochemistry using a specific anti-mouse albumin antibody. Labeling density was recorded over vascular lumen, endothelium, junctions, and subendothelial space. At equilibrium, the volume density of interstitial albumin was 18% of that in circulation. Despite this large difference in albumin concentration between capillary lumen and interstitium, plasmalemmal vesicles labeling was uniformly distributed across the endothelial profile. 68% of the junctions displayed labeling for albumin, which was however low and confined to the luminal and abluminal sides. The scarce labeling of the endothelial cell surface did not confirm the fiber matrix theory. The kinetics of albumin transcytosis was evaluated by injecting radioiodinated and DNP-tagged BSA. At 3, 10, 30, and 60 min, and 3, 5, and 24 h circulation time, blood radioactivity was measured and diaphragms were fixed and embedded. Anti-DNP antibodies were used to map the tracer in aforementioned compartments. A linear relationship between blood radioactivity and vascular labeling density was found, with a detection sensitivity approaching 1 gold particle per DNP-BSA molecule. Tracer presence over endothelial vesicles reached rapidly (10 min) a saturation value; initially localized near the luminal front, it evolved towards a uniform distribution across endothelium during the first hour. An hour was also needed to reach the saturation limit within the subendothelial space. Labeling of the junctions increased slowly, out of phase with the inferred transendothelial albumin fluxes. This suggests that they play little, if any, role in albumin transcytosis, which rather seems to proceed through the vesicular way. + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + MESH:D019297 + Chemical + + DNP + + + + + 2289457 + + 1577853 + 729 + 4 + 1992 + 2289457 + 92250709 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 729-43 + 743 + Piper RC, Tai C, Slot JW, Hahn CS, Rice CM, Huang H, James DE + 0 + The efficient intracellular sequestration of the insulin-regulatable glucose transporter (GLUT-4) is conferred by the NH2 terminus + + 6517 + Gene + 74381 + + GLUT-4 + + + 3630 + Gene + 173 + + insulin + + + + ABSTRACT + abstract + 131 + GLUT-4 is the major facilitative glucose transporter isoform in tissues that exhibit insulin-stimulated glucose transport. Insulin regulates glucose transport by the rapid translocation of GLUT-4 from an intracellular compartment to the plasma membrane. A critical feature of this process is the efficient exclusion of GLUT-4 from the plasma membrane in the absence of insulin. To identify the amino acid domains of GLUT-4 which confer intracellular sequestration, we analyzed the subcellular distribution of chimeric glucose transporters comprised of GLUT-4 and a homologous isoform, GLUT-1, which is found predominantly at the cell surface. These chimeric transporters were transiently expressed in CHO cells using a double subgenomic recombinant Sindbis virus vector. We have found that wild-type GLUT-4 is targeted to an intracellular compartment in CHO cells which is morphologically similar to that observed in adipocytes and muscle cells. Sindbis virus-produced GLUT-1 was predominantly expressed at the cell surface. Substitution of the GLUT-4 amino-terminal region with that of GLUT-1 abolished the efficient intracellular sequestration of GLUT-4. Conversely, substitution of the NH2 terminus of GLUT-1 with that of GLUT-4 resulted in marked intracellular sequestration of GLUT-1. These data indicate that the NH2-terminus of GLUT-4 is both necessary and sufficient for intracellular sequestration. + + 6517 + Gene + 74381 + + GLUT-4 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 3630 + Gene + 173 + + insulin + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6513 + Gene + 68520 + + GLUT-1 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6513 + Gene + 68520 + + GLUT-1 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6513 + Gene + 68520 + + GLUT-1 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6513 + Gene + 68520 + + GLUT-1 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 6513 + Gene + 68520 + + GLUT-1 + + + 6517 + Gene + 74381 + + GLUT-4 + + + 3630 + Gene + 173 + + Insulin + + + 11034 + Species + + Sindbis virus + + + 11034 + Species + + Sindbis virus + + + 3630 + Gene + 173 + + insulin + + + CVCL:0213 + CellLine + + CHO + + + MESH:D005947 + Chemical + + glucose + + + CVCL:0213 + CellLine + + CHO + + + MESH:D005947 + Chemical + + glucose + + + MESH:D005947 + Chemical + + glucose + + + + + 2289458 + + 1577855 + 757 + 4 + 1992 + 2289458 + 92250711 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 757-64 + 764 + Parkos CA, Colgan SP, Delp C, Arnaout MA, Madara JL + 0 + Neutrophil migration across a cultured epithelial monolayer elicits a biphasic resistance response representing sequential effects on transcellular and paracellular pathways + + + ABSTRACT + abstract + 174 + Migration of polymorphonuclear leukocytes across epithelia is a hallmark of many inflammatory disease states. Neutrophils traverse epithelia by migrating through the paracellular space and crossing intercellular tight junctions. We have previously shown (Nash, S., J. Stafford, and J.L. Madara. 1987. J. Clin. Invest. 80:1104-1113), that leukocyte migration across T84 monolayers, a model human intestinal epithelium, results in enhanced tight junction permeability--an effect quantitated by the use of a simple, standard electrical assay of transepithelial resistance. Here we show that detailed time course studies of the transmigration-elicited decline in resistance has two components, one of which is unrelated to junctional permeability. The initial decrease in resistance, maximal 5-13 min after initiation of transmigration, occurs despite inhibition of transmigration by an antibody to the common beta subunit of neutrophil beta 2 integrins, and is paralleled by an increase in transepithelial short-circuit current. Chloride ion substitution and inhibitor studies indicate that the early- phase resistance decline is not attributable to an increase in tight junction permeability but is due to decreased resistance across epithelial cells resulting from chloride secretion. Since T84 cells are accepted models for studies of the regulation of Cl- and water secretion, our results suggest that neutrophil transmigration across mucosal surfaces (for example, respiratory and intestinal tracts) may initially activate flushing of the surface by salt and water. Equally important, these studies, by providing a concrete example of sequential transcellular and paracellular effects on transepithelial resistance, highlight the fact that this widely used assay cannot simply be viewed as a direct functional probe of tight junction permeability. + + 322842 + Species + + L. Madara + + + 9606 + Species + + human + + + MESH:D002712 + Chemical + + chloride + + + MESH:D012492 + Chemical + + salt + + + MESH:D002712 + Chemical + + Chloride + + + MESH:D014867 + Chemical + + water + + + MESH:D014867 + Chemical + + water + + + + + 2289459 + + 1577852 + 717 + 4 + 1992 + 2289459 + 92250708 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 717-28 + 728 + Milgram SL, Johnson RC, Mains RE + 0 + Expression of individual forms of peptidylglycine alpha-amidating monooxygenase in AtT-20 cells: endoproteolytic processing and routing to secretory granules + + + ABSTRACT + abstract + 158 + Peptidylglycine alpha-amidating monooxygenase (PAM: EC 1.14.17.3) is a bifunctional protein which catalyzes the COOH-terminal amidation of bioactive peptides; the NH2-terminal monooxygenase and mid-region lyase act in sequence to perform the peptide alpha-amidation reaction. Alternative splicing of the single PAM gene gives rise to mRNAs generating PAM proteins with and without a putative transmembrane domain, with and without a linker region between the two enzymes, and forms containing only the monooxygenase domain. The expression, endoproteolytic processing, storage, and secretion of this secretory granule-associated protein were examined after stable transfection of AtT-20 mouse pituitary cells with naturally occurring and truncated PAM proteins. The transfected proteins were examined using enzyme assays, subcellular fractionation, Western blotting, and immunocytochemistry. Western blots of crude membrane and soluble fractions of transfected cells demonstrated that all PAM proteins were endoproteolytically processed. When the linker region was present between the monooxygenase and lyase domains, monofunctional soluble enzymes were generated from bifunctional PAM proteins; without the linker region, bifunctional enzymes were generated. Soluble forms of PAM expressed in AtT-20 cells and soluble proteins generated through selective endoproteolysis of membrane-associated PAM were secreted in an active form into the medium; secretion of the transfected proteins and endogenous hormone were stimulated in parallel by secretagogues. PAM proteins were localized by immunocytochemistry in the perinuclear region near the Golgi apparatus and in secretory granules, with the greatest intensity of staining in the perinuclear region in cell lines expressing integral membrane forms of PAM. Monofunctional and bifunctional PAM proteins that were soluble or membrane-associated were all packaged into regulated secretory granules in AtT-20 cells. + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 10090 + Species + + mouse + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + 18484 + Gene + 37369 + + PAM + + + + + 2289460 + + 1577861 + 849 + 4 + 1992 + 2289460 + 92250719 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 849-61 + 861 + Matteoli M, Takei K, Perin MS, Südhof TC, De Camilli P + 0 + Exo-endocytotic recycling of synaptic vesicles in developing processes of cultured hippocampal neurons + + + ABSTRACT + abstract + 103 + In mature neurons synaptic vesicles (SVs) undergo cycles of exo- endocytosis at synapses. It is currently unknown whether SV exocytosis and recycling occurs also in developing axons prior to synapse formation. To address this question, we have developed an immunocytochemical assay to reveal SV exo-endocytosis in hippocampal neurons developing in culture. In this assay antibodies directed against the lumenal domain of synaptotagmin I (Syt I), an intrinsic membrane protein of SVs, are used to reveal exposure of SV membranes at the cell surface. Addition of antibodies to the culture medium of living neurons for 1 hr at 37 degrees C resulted in their rapid and specific internalization by all neuronal processes and, particularly, by axons. Double immunofluorescence and electron microscopy immunocytochemistry indicated that the antibodies were retained within SVs in cell processes and underwent cycles of exo-endocytosis in parallel with SV membranes. In contrast, another endocytotic marker, wheat germ agglutinin, was rapidly cleared from the processes and transported to the cell body. Antibody-labeled SVs were still present in axons several days after antibody loading and became clustered at presynaptic sites in parallel with synaptogenesis. These results demonstrate that SVs undergo multiple cycles of exo-endocytosis in developing neuronal processes irrespective of the presence of synaptic contacts. + + 4565 + Species + + wheat + + + Disease + + SV exocytosis + + + + + 2289461 + + 1577858 + 787 + 4 + 1992 + 2289461 + 92250714 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 787-97 + 797 + Lloyd C, Schevzov G, Gunning P + 0 + Transfection of nonmuscle beta- and gamma-actin genes into myoblasts elicits different feedback regulatory responses from endogenous actin genes + + 11465 + Gene + 74402 + + gamma-actin + + + + ABSTRACT + abstract + 145 + We have examined the role of feedback-regulation in the expression of the nonmuscle actin genes. C2 mouse myoblasts were transfected with the human beta- and gamma-actin genes. In gamma-actin transfectants we found that the total actin mRNA and protein pools remained unchanged. Increasing levels of human gamma-actin expression resulted in a progressive down-regulation of mouse beta- and gamma-actin mRNAs. Transfection of the beta-actin gene resulted in an increase in the total actin mRNA and protein pools and induced an increase in the levels of mouse beta-actin mRNA. In contrast, transfection of a beta- actin gene carrying a single-point mutation (beta sm) produced a feedback-regulatory response similar to that of the gamma-actin gene. Expression of a beta-actin gene encoding an unstable actin protein had no impact on the endogenous mouse actin genes. This suggests that the nature of the encoded actin protein determines the feedback-regulatory response of the mouse genes. The role of the actin cytoskeleton in mediating this feedback-regulation was evaluated by disruption of the actin network with Cytochalasin D. We found that treatment with Cytochalasin D abolished the down-regulation of mouse gamma-actin in both the gamma- and beta sm-actin transfectants. In contrast, a similar level of increase was observed for the mouse beta-actin mRNA in both control and transfected cells. These experiments suggest that the down- regulation of mouse gamma-actin mRNA is dependent on the organization of the actin cytoskeleton. In addition, the mechanism responsible for the down-regulation of beta-actin may be distinct from that governing gamma-actin. We conclude that actin feedback-regulation provides a biochemical assay for differences between the two nonmuscle actin genes. + + 728378 + Gene + 133745 + + beta- and gamma-actin + + + 11465 + Gene + 74402 + + gamma-actin + + + 11461;11465 + Gene + + beta- and gamma-actin + + + 11461 + Gene + 110648 + + beta-actin + + + 11461 + Gene + 110648 + + beta-actin + + + 11461 + Gene + 110648 + + beta- actin + + + 11465 + Gene + 74402 + + gamma-actin + + + 11461 + Gene + 110648 + + beta-actin + + + 11465 + Gene + 74402 + + gamma-actin + + + 11461 + Gene + 110648 + + beta-actin + + + 11465 + Gene + 74402 + + gamma-actin + + + 11461 + Gene + 110648 + + beta-actin + + + 11465 + Gene + 74402 + + gamma-actin + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 11465 + Gene + 74402 + + gamma-actin + + + MESH:D015638 + Chemical + + Cytochalasin D + + + + + 2289462 + + 1349609 + 825 + 4 + 1992 + 2289462 + 92250717 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 825-39 + 839 + Hauft SM, Kim SH, Schmidt GH, Pease S, Rees S, Harris S, Roth KA, Hansbrough JR, Cohn SM, Ahnen DJ + 0 + Expression of SV-40 T antigen in the small intestinal epithelium of transgenic mice results in proliferative changes in the crypt and reentry of villus-associated enterocytes into the cell cycle but has no apparent effect on cellular differentiation programs and does not cause neoplastic transformation + + 10090 + Species + + transgenic mice + + + + ABSTRACT + abstract + 304 + The mouse intestinal epithelium represents a unique mammalian system for examining the relationship between cell division, commitment, and differentiation. Proliferation and differentiation are rapid, perpetual, and spatially well-organized processes that occur along the crypt-to-villus axis and involve clearly defined cell lineages derived from a common multipotent stem cell located near the base of each crypt. Nucleotides -1178 to +28 of the rat intestinal fatty acid binding protein gene were used to establish three pedigrees of transgenic mice that expressed SV-40 large T antigen (TAg) in epithelial cells situated in the uppermost portion of small intestinal crypts and in already committed, differentiating enterocytes as they exited these crypts and migrated up the villus. T antigen production was associated with increases in crypt cell proliferation but had no apparent effect on commitment to differentiate along enterocytic, enteroendocrine, or Paneth cell lineages. Single- and multilabel- immunocytochemical studies plus RNA blot hybridization analyses suggested that the differentiation programs of these lineages were similar in transgenic mice and their normal littermates. This included enterocytes which, based on the pattern of [3H]thymidine and 5-bromo-2'- deoxyuridine labeling and proliferating nuclear antigen expression, had reentered the cell cycle during their migration up the villus. The state of cellular differentiation and/or TAg production appeared to affect the nature of the cell cycle; analysis of the ratio of S-phase to M-phase cells (collected by metaphase arrest with vincristine) and of the intensities of labeling of nuclei by [3H]thymidine indicated that the duration of S phase was longer in differentiating, villus- associated enterocytes than in the less well-differentiated crypt epithelial cell population and that there may be a block at the G2/M boundary. Sustained increases in crypt and villus epithelial cell proliferation over a 9-mo period were not associated with the development of gut neoplasms--suggesting that tumorigenesis in the intestine may require that the initiated cell have many of the properties of the gut stem cell including functional anchorage. + + 10090 + Species + + mouse + + + 9606 + Species + + mammalian + + + 10116 + Species + + rat + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic mice + + + MESH:D001973 + Chemical + + 5-bromo-2'- deoxyuridine + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D005227 + Chemical + + fatty acid + + + - + Chemical + + [3H]thymidine + + + MESH:D014750 + Chemical + + vincristine + + + MESH:D014316 + Chemical + + 3H + + + MESH:D013936 + Chemical + + thymidine + + + + + 2289463 + + 1577857 + 775 + 4 + 1992 + 2289463 + 92250713 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 775-85 + 785 + Schevzov G, Lloyd C, Gunning P + 0 + High level expression of transfected beta- and gamma-actin genes differentially impacts on myoblast cytoarchitecture + + 728378 + Gene + 133745 + + beta- and gamma-actin + + + + ABSTRACT + abstract + 117 + The impact of the human beta- and gamma-actin genes on myoblast cytoarchitecture was examined by their stable transfection into mouse C2 myoblasts. Transfectant C2 clones expressing high levels of human beta-actin displayed increases in cell surface area. In contrast, C2 clones with high levels of human gamma-actin expression showed decreases in cell surface area. The changes in cell morphology were accompanied by changes in actin stress-fiber organization. The beta- actin transfectants displayed well-defined filamentous organization of actin; whereas the gamma-actin transfectants displayed a more diffuse organization of the actin cables. The role of the beta-actin protein in generating the enlarged cell phenotype was examined by transfecting a mutant form of the human beta-actin gene. Transfectant cells were shown to incorporate the aberrant actin protein into stress-fiber-like structures. High level expression of the mutant beta-actin produced decreases in cell surface area and disruption of the actin microfilament network similar to that seen with transfection of the gamma-actin gene. In contrast, transfection of another mutant form of the beta-actin gene which encodes an unstable protein had no impact on cell morphology or cytoarchitecture. These results strongly suggest that it is the nature of the encoded protein that determines the morphological response of the cell. We conclude that the relative gene expression of beta- and gamma-actin is of relevance to the control of myoblast cytoarchitecture. In particular, we conclude that the beta- and gamma-actin genes encode functionally distinct cytoarchitectural information. + + 728378 + Gene + 133745 + + beta- and gamma-actin + + + 728378 + Gene + 133745 + + beta-actin + + + 728378 + Gene + 133745 + + beta-actin + + + 728378 + Gene + 133745 + + beta-actin + + + 728378 + Gene + 133745 + + beta-actin + + + 728378 + Gene + 133745 + + beta-actin + + + 728378 + Gene + 133745 + + beta- and gamma-actin + + + 728378 + Gene + 133745 + + beta- and gamma-actin + + + 9606 + Species + + human + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 9606 + Species + + human + + + + + 2289464 + + 1315783 + 889 + 4 + 1992 + 2289464 + 92250722 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 889-94 + 894 + Uehara Y, Kitamura N + 0 + Expression of a human hepatocyte growth factor/scatter factor cDNA in MDCK epithelial cells influences cell morphology, motility, and anchorage-independent growth + + 9606 + Species + + human + + + CVCL:0422 + CellLine + + MDCK + + + + ABSTRACT + abstract + 163 + The addition of exogenous hepatocyte growth factor (HGF)/scatter factor (SF) to MDCK epithelial cells results in fibroblastic morphology and cell motility. We generated HGF/SF producing MDCK cells by transfection with an expression plasmid containing human HGF/SF cDNA. Production of HGF/SF by these cells induced a change from an epithelial to a fibroblastic morphology and increased cell motility. In addition, the HGF/SF producing cells acquired efficient anchorage-independent growth in soft agar but did not form tumors in nude mice. The morphological change and the stimulation of the anchorage-independent growth were prevented by anti-HGF/SF antibody, suggesting that the factor is secreted and then exerts its effects through cell surface receptors. + + 15234 + Gene + 503 + + SF + + + 403441 + Gene + 503 + + HGF/SF + + + 3082 + Gene + 503 + + HGF/SF + + + 403441 + Gene + 503 + + HGF/SF + + + 15234 + Gene + 503 + + HGF/SF + + + 403441 + Gene + 503 + + HGF/SF + + + 9606 + Species + + human + + + 10090 + Species + + nude mice + + + 403441 + Gene + 503 + + HGF + + + MESH:D009369 + Disease + + tumors + + + CVCL:0422 + CellLine + + MDCK + + + CVCL:0422 + CellLine + + MDCK + + + + + 2289465 + + 1577860 + 841 + 4 + 1992 + 2289465 + 92250718 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 841-7 + 847 + Forsayeth JR, Gu Y, Hall ZW + 0 + BiP forms stable complexes with unassembled subunits of the acetylcholine receptor in transfected COS cells and in C2 muscle cells + + 3309 + Gene + 3908 + + BiP + + + CVCL:0529 + CellLine + + C2 + + + CVCL:0222 + CellLine + + COS + + + + ABSTRACT + abstract + 131 + We have investigated the role of the immunoglobulin-binding protein (BiP) in the folding and assembly of subunits of the acetylcholine receptor (AChR) in COS cells and in C2 muscle cells. Immunoprecipitation in COS cells showed that alpha, beta, and delta subunits are associated with BiP. In the case of the alpha subunit, which first folds to acquire toxin-binding activity and is then assembled with the other subunits to form the AChR, BiP was associated only with a form that is unassembled and does not bind alpha- bungarotoxin. Similar results were found in C2 cells. Although the alpha and beta subunits of the AChR are minor membrane proteins in C2 cells, they were prominent among the proteins immunoprecipitated by antibodies to BiP, suggesting that BiP could play a role in their maturation or folding. In pulse-chase experiments in C2 cells, however, labeled alpha subunit formed a stable complex with BiP that was first detected after most of the alpha subunit had acquired toxin-binding activity and whose amount continued to increase for several hours. These kinetics are not compatible with a role for the BiP complex in the folding or assembly pathway of the AChR, and suggest that BiP is associated with a misfolded form of the subunit that is slowly degraded. + + 3309 + Gene + 3908 + + immunoglobulin-binding protein + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + 3309 + Gene + 3908 + + BiP + + + CVCL:0529 + CellLine + + C2 + + + CVCL:0529 + CellLine + + C2 + + + CVCL:0222 + CellLine + + COS + + + CVCL:0529 + CellLine + + C2 + + + CVCL:0529 + CellLine + + C2 + + + CVCL:0222 + CellLine + + COS + + + + + 2289466 + + 1577856 + 765 + 4 + 1992 + 2289466 + 92250712 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 765-74 + 774 + Coates TD, Watts RG, Hartman R, Howard TH + 0 + Relationship of F-actin distribution to development of polar shape in human polymorphonuclear neutrophils + + 9606 + Species + + human + + + + ABSTRACT + abstract + 106 + Polymerization of actin has been associated with development of polar shape in human neutrophils (PMN). To examine the relation of filamentous actin (F-actin) distribution to shape change in PMN, we developed a method using computerized video image analysis and fluorescence microscopy to quantify distribution of F-actin in single cells. PMN were labeled with fluorescent probe NBD-phallicidin to measure filamentous actin and Texas red to assess cell thickness. We show that Texas red fluorescence is a reasonable measure of cell thickness and that correction of the NBD-phallicidin image for cell thickness using the Texas red image permits assessment of focal F-actin content. Parameters were derived that quantify total F-actin content, movement of F-actin away from the center of the cell, asymmetry of F- actin distribution, and change from round to polar shape. The sequence of change in F-actin distribution and its relation to development of polar shape in PMN was determined using these parameters. After stimulation with chemotactic peptide at 25 degrees C, F-actin polymerized first at the rim of the PMN. This was followed by development of asymmetry of F-actin distribution and change to polar shape. The dominant pseudopod developed first in the region of lower F- actin concentration followed later by polymerization of actin in the end of the developed pseudopod. Asymmetric F-actin distribution was detected in round PMN before development of polar shape. Based upon these data, asymmetric distribution of F-actin is coincident with and probably precedes development of polar shape in PMN stimulated in suspension by chemotactic peptide. + + 9606 + Species + + human + + + - + Chemical + + phallicidin + + + MESH:C034657 + Chemical + + Texas red + + + - + Chemical + + phallicidin + + + MESH:C034657 + Chemical + + Texas red + + + MESH:C034657 + Chemical + + Texas red + + + + + 2289467 + + 1577863 + 903 + 4 + 1992 + 2289467 + 92250724 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 903-14 + 914 + Nakai A, Satoh M, Hirayoshi K, Nagata K + 0 + Involvement of the stress protein HSP47 in procollagen processing in the endoplasmic reticulum + + 871 + Gene + 20331 + + HSP47 + + + + ABSTRACT + abstract + 95 + The 47,000-D collagen-binding glycoprotein, heat shock protein 47 (HSP47), is a stress-inducible protein localized in the ER of collagen- secreting cells. The location and collagen-binding activity of this protein led to speculation that HSP47 might participate in collagen processing. Chemical crosslinking studies were used to test this hypothesis both before and after the perturbation of procollagen processing. The association of procollagen with HSP47 was demonstrated using cleavable bifunctional crosslinking reagents. HSP47 and procollagen were shown to be coprecipitated by the treatment of intact cells with anti-HSP47 or with anticollagen antibodies. Furthermore, several proteins residing in the ER were noted to be crosslinked to and coprecipitated with HSP47, suggesting that these ER-resident proteins may form a large complex in the ER. When cells were heat shocked, or when stable triple-helix formation was inhibited by treatment with alpha,alpha'-dipyridyl, coprecipitation of procollagen with HSP47 was increased. This increase was due to the inhibition of procollagen secretion and to the accumulation of procollagen in the ER. Pulse label and chase experiments revealed that coprecipitated procollagen was detectable as long as procollagen was present in the endoplasmic reticulum of alpha,alpha'-dipyridyl-treated cells. Under normal growth conditions, coprecipitated procollagen was observed to decrease after a chase period of 10-15 min, whereas total procollagen decreased only after 20-25 min. In addition, the intracellular association between HSP47 and procollagen was shown to be disrupted by a change in physiological pH, suggesting that the dissociation of procollagen from HSP47 is pH dependent. These findings support a specific role for HSP47 in the intracellular processing of procollagen, and provide evidence of a new category of "molecular chaperones" in terms of its substrate specificity and the dissociation mechanism. + + 871 + Gene + 20331 + + heat shock protein 47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + 871 + Gene + 20331 + + HSP47 + + + + + 2289468 + + 1349608 + 813 + 4 + 1992 + 2289468 + 92250716 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 813-24 + 824 + Wacker IU, Rickard JE, De Mey JR, Kreis TE + 0 + Accumulation of a microtubule-binding protein, pp170, at desmosomal plaques + + + ABSTRACT + abstract + 76 + The establishment of epithelial cell polarity correlates with the formation of specialized cell-cell junctions and striking changes in the organization of microtubules. A significant fraction of the microtubules in MDCK cells become stabilized, noncentrosomally organized, and arranged in longitudinal bundles in the apical-basal axis. This correlation suggests a functional link between cell-cell junction formation and control of microtubule organization. We have followed the distribution of pp170, a recently described microtubule- binding protein, during establishment of epithelial cell polarity. This protein shows the typical patchy distribution along microtubules in subconfluent fibroblasts and epithelial cells, often associated with the peripheral ends of a subpopulation of microtubules. In contrast to its localization in confluent fibroblasts (A72) and HeLa cells, however, pp170 accumulates in patches delineating the regions of cell- cell contacts in confluent polarizing epithelial cells (MDCK and Caco- 2). Double immunolocalization with antibodies specific for cell-cell junction proteins, confocal microscopy, and immunoelectron microscopy on polarized MDCK cells suggest that pp170 accumulates at desmosomal plaques. Furthermore, microtubules and desmosomes are found in close contact. Maintenance of the desmosomal association of pp170 is dependent on intact microtubules in 3-d-old, but not in 1-d-old MDCK cell cultures. This suggests a regulated interaction between microtubules and desmosomes and a role for pp170 in the control of changes in the properties of microtubules induced by epithelial cell- cell junction formation. + + CVCL:0030 + CellLine + + HeLa + + + CVCL:0422 + CellLine + + MDCK + + + CVCL:0422 + CellLine + + MDCK + + + CVCL:0422 + CellLine + + MDCK + + + CVCL:0422 + CellLine + + MDCK + + + + + 2289469 + + 1577851 + 701 + 4 + 1992 + 2289469 + 92250707 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 701-15 + 715 + Kao CY, Draper RK + 0 + Retention of secretory proteins in an intermediate compartment and disappearance of the Golgi complex in an END4 mutant of Chinese hamster ovary cells + + 10029 + Species + + Chinese hamster + + + + ABSTRACT + abstract + 151 + Mutant V.24.1, a member of the End4 complementation group of temperature-sensitive CHO cells, is defective in secretion at the restrictive temperature (Wang, R.-H., P. A. Colbaugh, C.-Y. Kao, E. A. Rutledge, and R. K. Draper. 1990. J. Biol. Chem. 265:20179-20187; Presley, J. F., R. K. Draper, and D. T. Brown. 1991. J. Virol. 65:1332- 1339). We have further investigated the secretory lesion and report three main findings. First, the block in secretion is not due to aberrant folding or oligomerization of secretory proteins in the endoplasmic reticulum because the hemagglutinin of influenza virus folded and oligomerized at the same rate in mutant and parental cells at the restrictive temperature. Second, secretory proteins accumulated in a compartment intermediate between the ER and the Golgi. Several lines of evidence support this conclusion, the most direct being the colocalization by immunofluorescence microscopy of influenza virus hemagglutinin with a 58-kD protein that is known to reside in an intermediate compartment. Third, at the resolution of fluorescence microscopy, the Golgi complex in the mutant cells vanished at the restrictive temperature. + + 11309 + Species + + influenza virus + + + 11309 + Species + + influenza virus + + + CVCL:0213 + CellLine + + CHO + + + + + 2289470 + + 1577859 + 799 + 4 + 1992 + 2289470 + 92250715 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 799-811 + 811 + Tombes RM, Simerly C, Borisy GG, Schatten G + 0 + Meiosis, egg activation, and nuclear envelope breakdown are differentially reliant on Ca2+, whereas germinal vesicle breakdown is Ca2+ independent in the mouse oocyte + + 10090 + Species + + mouse + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + + ABSTRACT + abstract + 167 + During early development, intracellular Ca2+ mobilization is not only essential for fertilization, but has also been implicated during other meiotic and mitotic events, such as germinal vesicle breakdown (GVBD) and nuclear envelope breakdown (NEBD). In this study, the roles of intracellular and extracellular Ca2+ were examined during meiotic maturation and reinitiation at parthenogenetic activation and during first mitosis in a single species using the same methodologies. Cumulus- free metaphase II mouse oocytes immediately resumed anaphase upon the induction of a large, transient Ca2+ elevation. This resumption of meiosis and associated events, such as cortical granule discharge, were not sensitive to extracellular Ca2+ removal, but were blocked by intracellular Ca2+ chelators. In contrast, meiosis I was dependent on external Ca2+; in its absence, the formation and function of the first meiotic spindle was delayed, the first polar body did not form and an interphase-like state was induced. GVBD was not dependent on external Ca2+ and showed no associated Ca2+ changes. NEBD at first mitosis in fertilized eggs, on the other hand, was frequently, but not always associated with a brief Ca2+ transient and was dependent on Ca2+ mobilization. We conclude that GVBD is Ca2+ independent, but that the dependence of NEBD on Ca2+ suggests regulation by more than one pathway. As cells develop from Ca(2+)-independent germinal vesicle oocytes to internal Ca(2+)-dependent pronuclear eggs, internal Ca2+ pools increase by approximately fourfold. + + 10090 + Species + + mouse + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca(2+) + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:C536875 + Disease + + meiosis I + + + MESH:D000069285 + Chemical + + Ca2+ + + + Disease + + mitosis + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca(2+) + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:C536875 + Disease + + meiosis + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + Disease + + mitosis + + + + + 2289471 + + 1315782 + 877 + 4 + 1992 + 2289471 + 92250721 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 May; 117(4) 877-87 + 887 + Durbec P, Gennarini G, Goridis C, Rougon G + 0 + A soluble form of the F3 neuronal cell adhesion molecule promotes neurite outgrowth + + + ABSTRACT + abstract + 84 + The F3 molecule is a member of the immunoglobulin superfamily anchored to membranes by a glycane-phosphatidylinositol, and is predominantly expressed on subsets of axons of the central and peripheral nervous system. In a previous paper (Gennarini, G., P. Durbec, A. Boned, G. Rougon, and C. Goridis. 1991. Neuron. 6:595-606), we have established that F3 fulfills the operational definition of a cell adhesion molecule and that it stimulates neurite outgrowth when presented to sensory neurons as a surface component of transfected CHO cells. In the present study the question as to whether soluble forms of F3 would be functionally active was addressed in vitro on cultures of mouse dorsal root ganglion neurons. We observed that preparations enriched in soluble F3 had no effect on neuron attachment but enhanced neurite initiation and neurite outgrowth in a dose-dependent manner. By contrast, soluble NCAM-120 does not have any measurable effect on these phenomena. Addition of anti-F3 monovalent antibodies reduced the number of process-bearing neurons and the neuritic output per neuron to control values. Addition of cerebrospinal fluid, a natural source of soluble F3, also stimulated neurite extension, and this effect was partially blocked by anti-F3 antibodies. Our results suggest that the soluble forms of adhesive proteins with neurite outgrowth-promoting properties could act at a distance from their site of release in a way reminiscent of growth and trophic factors. + + 10090 + Species + + mouse + + + MESH:D058225 + Disease + + neurite initiation and neurite outgrowth + + + - + Chemical + + glycane-phosphatidylinositol + + + CVCL:0213 + CellLine + + CHO + + + + + 2289472 + + 1374416 + 1109 + 5 + 1992 + 2289472 + 92250735 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1109-17 + 1117 + Grzesiak JJ, Davis GE, Kirchhofer D, Pierschbacher MD + 0 + Regulation of alpha 2 beta 1-mediated fibroblast migration on type I collagen by shifts in the concentrations of extracellular Mg2+ and Ca2+ [published erratum appears in J Cell Biol 1992 Jul;118(1):219] + + - + Chemical + + Mg2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + + ABSTRACT + abstract + 204 + Extracellular Ca2+ can reverse the Mg(2+)-dependent, alpha 2 beta 1- mediated adhesion of WI38 human fibroblasts to type I collagen substrates. Affinity chromatography data also demonstrate that Ca2+ can specifically elute the fibroblast alpha 2 beta 1 integrin bound to type I collagen-Sepharose in Mg2+. In modified Boyden chamber migration assays, Mg2+ alone supports the alpha 2 beta 1-mediated migration of fibroblasts on type I collagen substrates, while Ca2+ does not. However, a twofold enhancement in migration was observed when combinations of the two cations were used, with optimal migration observed when the Mg2+/Ca2+ ratio was higher than one. Inhibitory mAbs directed against various integrin subunits demonstrate that these observed cation effects appear to be mediated primarily by alpha 2 beta 1. These data, together with reports that under certain physiological conditions significant fluctuations in the concentrations of extracellular Ca2+ and Mg2+ can take place in vivo, suggest that the ratio between these two cations is involved in the up- and downregulation of integrin function, and thus, may influence cell migratory behavior. + + 9606 + Species + + human + + + MESH:D000069285 + Chemical + + Ca2+ + + + - + Chemical + + Mg2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + - + Chemical + + Mg2+ + + + - + Chemical + + Mg(2+) + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D000069285 + Chemical + + Ca2+ + + + - + Chemical + + Mg2+ + + + - + Chemical + + Mg2+ + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:D000069285 + Chemical + + Ca2+ + + + + + 2289473 + + 1577867 + 1085 + 5 + 1992 + 2289473 + 92250732 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1085-92 + 1092 + Troy CM, Greene LA, Shelanski ML + 0 + Neurite outgrowth in peripherin-depleted PC12 cells + + 24688 + Gene + 4559 + + peripherin + + + CVCL:0481 + CellLine + + PC12 + + + + ABSTRACT + abstract + 52 + Peripherin is the major neuronal intermediate filament (IF) protein in PC12 cells and both its synthesis and amount increase during nerve growth factor (NGF) promoted neuronal differentiation. To address the question of the biological function of peripherin in neurite initiation we have used an antisense oligonucleotide complementary to the 5' region of peripherin mRNA to specifically inhibit its transcription. The oligonucleotide blocks both the synthesis of peripherin and its increase in response to NGF. Peripherin was found to be a stable protein with a cellular half-life of approximately 7 d. 6 wk of incubation with the oligonucleotide decreases peripherin to 11% of the level in naive control cells and to 3% of that in NGF-treated control cells. Despite the depletion, NGF elicits apparently normal neurite outgrowth from the oligonucleotide-treated cells. As evaluated by EM, there are few IFs in these cells, either in the cell bodies or neurites. There is no compensatory increase in NF-M, NF-L, or vimentin levels as a result of the inhibition of peripherin synthesis. These findings suggest that peripherin is not required for neurite formation, but is necessary for the formation of a cellular IF network which could be involved in process stability. They also demonstrate the utility of antisense oligonucleotides for the study of proteins with long half- lives. + + 24688 + Gene + 4559 + + Peripherin + + + 24387 + Gene + 1554 + + intermediate filament (IF) + + + 310738 + Gene + 1876 + + nerve growth factor + + + 310738 + Gene + 1876 + + NGF + + + 24688 + Gene + 4559 + + peripherin + + + 24688 + Gene + 4559 + + peripherin + + + 24688 + Gene + 4559 + + peripherin + + + 310738 + Gene + 1876 + + NGF + + + 24688 + Gene + 4559 + + Peripherin + + + 24688 + Gene + 4559 + + peripherin + + + 310738 + Gene + 1876 + + NGF + + + 310738 + Gene + 1876 + + NGF + + + 24588 + Gene + 137211 + + NF-M + + + 83613 + Gene + 4487 + + NF-L + + + 81818 + Gene + 2538 + + vimentin + + + 24688 + Gene + 4559 + + peripherin + + + 24688 + Gene + 4559 + + peripherin + + + 24387 + Gene + 1554 + + IF + + + CVCL:0481 + CellLine + + PC12 + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotide + + + + + 2289474 + + 1577869 + 1119 + 5 + 1992 + 2289474 + 92250736 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1119-33 + 1133 + Yurchenco PD, Cheng YS, Colognato H + 0 + Laminin forms an independent network in basement membranes [published erratum appears in J Cell Biol 1992 Jun;118(2):493] + + + ABSTRACT + abstract + 122 + Laminin self-assembles in vitro into a polymer by a reversible, entropy- driven and calcium-facilitated process dependent upon the participation of the short arm globular domains. We now find that this polymer is required for the structural integrity of the collagen-free basement membrane of cultured embryonal carcinoma cells (ECC) and for the supramolecular organization and anchorage of laminin in the collagen- rich basement membrane of the Engelbreth-Holm-Swarm tumor (EHS). First, low temperature and EDTA induced the dissolution of ECC basement membranes and released approximately 80% of total laminin from the EHS basement membrane. Second, laminin elastase fragments (E4 and E1') possessing the short arm globules of the B1, B2, and A chains selectively acted as competitive ligands that dissolved ECC basement membranes and displaced laminin from the EHS basement membrane into solution. The fraction of laminin released increased as a function of ligand concentration, approaching the level of the EDTA-reversible pool. The smaller (approximately 20%) residual pool of EHS laminin, in contrast, could only be effectively displaced by E1' and E4 if the collagenous network was first degraded with bacterial collagenase. The supramolecular architecture of freeze-etched and platinum/carbon replicated reconstituted laminin gel polymer, ECC, and collagenase- treated EHS basement membranes were compared and found to be similar, further supporting the biochemical data. We conclude that laminin forms a network independent of that of type IV collagen in basement membranes. Furthermore, in the EHS basement membrane four-fifths of laminin is anchored strictly through noncovalent bonds between laminin monomers while one-fifth is anchored through a combination of these bonds and laminin-collagen bridges. + + MESH:D002118 + Chemical + + calcium + + + MESH:D002244 + Chemical + + carbon + + + MESH:D010984 + Chemical + + platinum + + + MESH:D004492 + Chemical + + EDTA + + + MESH:D011108 + Chemical + + polymer + + + MESH:D009369 + Disease + + tumor + + + MESH:D004492 + Chemical + + EDTA + + + MESH:D018236 + Disease + + embryonal carcinoma + + + + + 2289475 + + 1315784 + 1067 + 5 + 1992 + 2289475 + 92250730 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1067-76 + 1076 + Amatruda JF, Cooper JA + 0 + Purification, characterization, and immunofluorescence localization of Saccharomyces cerevisiae capping protein + + 4932 + Species + + Saccharomyces cerevisiae + + + + ABSTRACT + abstract + 112 + Capping protein binds the barbed ends of actin filaments and nucleates actin filament assembly in vitro. We purified capping protein from Saccharomyces cervisiae. One of the two subunits is the product of the CAP2 gene, which we previously identified as the gene encoding the beta subunit of capping protein based on its sequence similarity to capping protein beta subunits in chicken and Dictyostelium (Amatruda, J. F., J. F. Cannon, K. Tatchell, C. Hug, and J. A. Cooper. 1990. Nature (Lond.) 344:352-354). Yeast capping protein has activity in critical concentration and low-shear viscometry assays consistent with barbed- end capping activity. Like chicken capping protein, yeast capping protein is inhibited by PIP2. By immunofluorescence microscopy yeast capping protein colocalizes with cortical actin spots at the site of bud emergence and at the tips of growing buds and shmoos. In contrast, capping protein does not colocalize with actin cables or with actin rings at the site of cytokinesis. + + 420847 + Gene + 101397 + + CAP2 + + + 854545 + Gene + 130567 + + PIP2 + + + 9031 + Species + + chicken + + + 4932 + Species + + Yeast + + + 9031 + Species + + chicken + + + 4932 + Species + + yeast + + + 4932 + Species + + yeast + + + 4932 + Species + + Saccharomyces + + + + + 2289476 + + 1533642 + 1041 + 5 + 1992 + 2289476 + 92250728 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1041-53 + 1053 + Hamaguchi JR, Tobey RA, Pines J, Crissman HA, Hunter T, Bradbury EM + 0 + Requirement for p34cdc2 kinase is restricted to mitosis in the mammalian cdc2 mutant FT210 + + 983 + Gene + 68203 + + p34cdc2 + + + 983 + Gene + 68203 + + cdc2 + + + 9606 + Species + + mammalian + + + Disease + + mitosis + + + + ABSTRACT + abstract + 91 + The mouse FT210 cell line is a temperature-sensitive cdc2 mutant. FT210 cells are found to arrest specifically in G2 phase and unlike many alleles of cdc2 and cdc28 mutants of yeasts, loss of p34cdc2 at the nonpermissive temperature has no apparent effect on cell cycle progression through the G1 and S phases of the division cycle. FT210 cells and the parent wild-type FM3A cell line each possess at least three distinct histone H1 kinases. H1 kinase activities in chromatography fractions were identified using a synthetic peptide substrate containing the consensus phosphorylation site of histone H1 and the kinase subunit compositions were determined immunochemically with antisera prepared against the "PSTAIR" peptide, the COOH-terminus of mammalian p34cdc2 and the human cyclins A and B1. The results show that p34cdc2 forms two separate complexes with cyclin A and with cyclin B1, both of which exhibit thermal lability at the non-permissive temperature in vitro and in vivo. A third H1 kinase with stable activity at the nonpermissive temperature is comprised of cyclin A and a cdc2-like 34-kD subunit, which is immunoreactive with anti-"PSTAIR" antiserum but is not recognized with antiserum specific for the COOH- terminus of p34cdc2. The cyclin A-associated kinases are active during S and G2 phases and earlier in the division cycle than the p34cdc2- cyclin B1 kinase. We show that mouse cells possess at least two cdc2- related gene products which form cell cycle regulated histone H1 kinases and we propose that the murine homolog of yeast p34cdc/CDC28 is essential only during the G2-to-M transition in FT210 cells. + + 12534 + Gene + 68203 + + cdc2 + + + 12534 + Gene + 68203 + + cdc2 + + + 852457 + Gene + 74409 + + cdc28 + + + 12534 + Gene + 68203 + + p34cdc2 + + + 3005 + Gene + 136788 + + histone H1 + + + 983 + Gene + 68203 + + p34cdc2 + + + 890;891 + Gene + + cyclins A and B1 + + + 12534 + Gene + 68203 + + p34cdc2 + + + 12428 + Gene + 55562 + + cyclin A + + + 268697 + Gene + 68982 + + cyclin B1 + + + 12428 + Gene + 55562 + + cyclin A + + + 12534 + Gene + 68203 + + cdc2 + + + 12534 + Gene + 68203 + + p34cdc2 + + + 12534 + Gene + 68203 + + p34cdc2 + + + 12534 + Gene + 68203 + + cdc2 + + + 852457 + Gene + 74409 + + CDC28 + + + 10090 + Species + + mouse + + + 4932 + Species + + yeasts + + + 9606 + Species + + mammalian + + + 9606 + Species + + human + + + 10090 + Species + + mouse + + + 10090 + Species + + murine + + + 4932 + Species + + yeast + + + 3005 + Gene + 136788 + + histone H1 + + + 3005 + Gene + 136788 + + histone H1 + + + 268697 + Gene + 68982 + + cyclin B1 + + + 12428 + Gene + 55562 + + cyclin A + + + + + 2289477 + + 1577865 + 1023 + 5 + 1992 + 2289477 + 92250726 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1023-9 + 1029 + Cao LG, Babcock GG, Rubenstein PA, Wang YL + 0 + Effects of profilin and profilactin on actin structure and function in living cells + + + ABSTRACT + abstract + 84 + Previous studies have yielded conflicting results concerning the physiological role of profilin, a 12-15-kD actin- and phosphoinositide- binding protein, as a regulator of actin polymerization. We have addressed this question by directly microinjecting mammalian profilins, prepared either from an E. coli expression system or from bovine brain, into living normal rat kidney (NRK) cells. The microinjection causes a dose-dependent decrease in F-actin content, as indicated by staining with fluorescent phalloidin, and a dramatic reduction of actin and alpha-actinin along stress fibers. In addition, it has a strong inhibitory effect toward the extension of lamellipodia. However, the injection of profilin causes no detectable perturbation to the cell- substrate focal contact and no apparent depolymerization of filaments in either the nonlamellipodial circumferential band or the contractile ring of dividing cells. Furthermore, cytokinesis of injected cells occurs normally as in control cells. In contrast to pure profilin, high- affinity profilin-actin complexes from brain induce an increase in total cellular F-actin content and an enhanced ruffling activity, suggesting that the complex may dissociate readily in the cell and that there may be multiple states of profilin that differ in their ability to bind or release actin molecules. Our results indicate that profilin and profilactin can function as effective regulators for at least a subset of actin filaments in living cells. + + 9606 + Species + + mammalian + + + 562 + Species + + E. coli + + + 9913 + Species + + bovine + + + 10116 + Species + + rat + + + MESH:D010590 + Chemical + + phalloidin + + + CVCL:3758 + CellLine + + NRK + + + + + 2289478 + + 1315787 + 987 + 5 + 1992 + 2289478 + 92250742 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 987-95 + 995 + Metsikkö K, Hentunen T, Väänänen K + 0 + Local expression and exocytosis of viral glycoproteins in multinucleated muscle cells + + + ABSTRACT + abstract + 86 + We have analyzed the distribution of enveloped viral infections in multinucleated L6 muscle cells. A temperature-sensitive vesicular stomatitis virus (mutant VSV ts045) was utilized at the nonpermissive temperature (39 degrees C). As expected, the glycoprotein (G protein) of this mutant was restricted to the ER when the multinucleated cells were maintained at 39 degrees C. We demonstrate that this G protein remained localized when the infection was performed at low dose. By 4 h after infection the G protein patches spanned an average of 220 microns. The localization was independent of nuclear positions, showing that the ER was a peripheric structure. Thus, the infection did not recognize nuclear domains characteristic of nuclearly encoded proteins. After release of the 39 degrees C block, transport through a perinuclear compartment into a restricted surface domain lying above the internal G protein patch occurred. Accordingly, the transport pathway was locally restricted. After a 16-h infection the G protein spanned 420 microns, while the matrix protein occupied 700-800 microns of the myotube length. Double infection of multinucleated L6 muscle cells with Semliki Forest virus and VSV at high multiplicities showed that the glycoprotein of each virus occupied intracellular domains which were devoid of the other respective glycoprotein. Taken together, these findings indicate that the viral glycoproteins did not range far from their site of synthesis within the ER or other intracellular membrane compartments in these large cells. This result also suggests that relocation of viral RNA synthesis occurred slowly. + + 11276 + Species + + vesicular stomatitis virus + + + 11033 + Species + + Semliki Forest virus + + + MESH:D001102 + Disease + + viral infections + + + + + 2289479 + + 1315786 + 935 + 5 + 1992 + 2289479 + 92250738 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 935-48 + 948 + Klein F, Laroche T, Cardenas ME, Hofmann JF, Schweizer D, Gasser SM + 0 + Localization of RAP1 and topoisomerase II in nuclei and meiotic chromosomes of yeast + + + ABSTRACT + abstract + 85 + Topoisomerase II (topoII) and RAP1 (Repressor Activator Protein 1) are two abundant nuclear proteins with proposed structural roles in the higher-order organization of chromosomes. Both proteins co-fractionate as components of nuclear scaffolds from vegetatively growing yeast cells, and both proteins are present as components of pachytene chromosome, co-fractionating with an insoluble subfraction of meiotic nuclei. Immunolocalization using antibodies specific for topoII shows staining of an axial core of the yeast meiotic chromosome, extending the length of the synaptonemal complex. RAP1, on the other hand, is located at the ends of the paired bivalent chromosomes, consistent with its ability to bind telomeric sequences in vitro. In interphase nuclei, again in contrast to anti-topoII, anti-RAP1 gives a distinctly punctate staining that is located primarily at the nuclear periphery. Approximately 16 brightly staining foci can be identified in a diploid nucleus stained with anti-RAP1 antibodies, suggesting that telomeres are grouped together, perhaps through interaction with the nuclear envelope. + + 4932 + Species + + yeast + + + 4932 + Species + + yeast + + + + + 2289480 + + 1533643 + 1055 + 5 + 1992 + 2289480 + 92250729 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1055-66 + 1066 + Masuda H, Sevik M, Cande WZ + 0 + In vitro microtubule-nucleating activity of spindle pole bodies in fission yeast Schizosaccharomyces pombe: cell cycle-dependent activation in xenopus cell-free extracts + + 4896 + Species + + fission yeast + + + 4896 + Species + + Schizosaccharomyces pombe + + + 8355 + Species + + xenopus + + + + ABSTRACT + abstract + 170 + The spindle pole body (SPB) is the equivalent of the centrosome in fission yeast. In vivo it nucleates microtubules (MTs) during mitosis, but, unlike animal centrosomes, does not act as a microtubule organizing center (MTOC) during interphase. We have studied the MT- nucleating activity of SPBs in vitro and have found that SPBs in permeabilized cells retain in vivo characteristics. SPBs in cells permeabilized during mitosis can nucleate MTs, and are recognized by two antibodies: anti-gamma-tubulin and MPM-2 which recognizes phosphoepitopes. SPBs in cells permeabilized during interphase cannot nucleate MTs and are only recognized by anti-gamma-tubulin. Interphase SPBs which cannot nucleate can be converted to a nucleation competent state by incubation in cytostatic factor (CSF)-arrested Xenopus egg extracts. After incubation, they are recognized by MPM-2, and can nucleate MTs. The conversion does not occur in Xenopus interphase extract, but occurs in Xenopus interphase extract driven into mitosis by preincubation with exogenous cyclin B. The conversion is ATP dependent and inhibited by protein kinase inhibitors and alkaline phosphatase. Purified, active, cdc2 kinase/cyclin B complex in itself is not effective for activation of MT nucleation, although some interphase SPBs are now stained with MPM-2. These results suggest that the ability of SPBs in vitro to nucleate MTs after exposure to CSF- arrested extracts is activated through a downstream pathway which is regulated by cdc2 kinase. + + 4896 + Species + + fission yeast + + + 8355 + Species + + Xenopus + + + 8355 + Species + + Xenopus + + + 8355 + Species + + Xenopus + + + Disease + + mitosis + + + Disease + + mitosis + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + Disease + + mitosis + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:C042995 + Chemical + + SPBs + + + MESH:D000255 + Chemical + + ATP + + + + + 2289481 + + 1374414 + 1077 + 5 + 1992 + 2289481 + 92250731 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1077-84 + 1084 + Linstedt AD, Vetter ML, Bishop JM, Kelly RB + 0 + Specific association of the proto-oncogene product pp60c-src with an intracellular organelle, the PC12 synaptic vesicle + + CVCL:0481 + CellLine + + PC12 + + + + ABSTRACT + abstract + 120 + The protein product of the proto-oncogene c-src is a membrane- associated tyrosine kinase of unknown function. Identification of pp60c- src target membranes may elucidate the function of the c-src protein. The available evidence indicates that pp60c-src associates with distinct membranes within single cell types and has different distributions in different cell types. Our experiments demonstrate targeting of pp60c-src to an isolatable and biochemically identified membrane fraction in the neuroendocrine cell line PC12. The c-src protein was found to be specifically associated with synaptic vesicles since: (a) the pp60c-src immunofluorescent pattern overlapped with a synaptic vesicle marker, synaptophysin; (b) a significant proportion (44%) of the pp60c-src from PC12 but not fibroblast postnuclear supernatants was recovered in a small vesicle fraction; (c) an anti- synaptophysin cytoplasmic domain antibody immunodepleted all of the pp60c-src vesicles in this fraction, and (d) pp60c-src copurified during a 100-fold purification of PC12 synaptic vesicles. These results suggest a role for the c-src protein in the regulation of synaptic vesicle function. + + 6714 + Gene + 21120 + + proto-oncogene c-src + + + 6714 + Gene + 21120 + + c-src + + + 6714 + Gene + 21120 + + c-src + + + 6855 + Gene + 2391 + + synaptophysin + + + 6714 + Gene + 21120 + + c-src + + + 6855 + Gene + 2391 + + synaptophysin + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + + + 2289482 + + 1577868 + 1093 + 5 + 1992 + 2289482 + 92250733 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1093-9 + 1099 + Doherty P, Ashton SV, Skaper SD, Leon A, Walsh FS + 0 + Ganglioside modulation of neural cell adhesion molecule and N-cadherin- dependent neurite outgrowth + + 24586 + Gene + 40754 + + neural cell adhesion molecule + + + 83501 + Gene + 20424 + + N-cadherin + + + MESH:D005732 + Chemical + + Ganglioside + + + + ABSTRACT + abstract + 100 + We have used monolayers of control 3T3 cells and 3T3 cells expressing transfected human neural cell adhesion molecule (NCAM) or chick N- cadherin as a culture substrate for PC12 cells. NCAM and N-cadherin in the monolayer directly promote neurite outgrowth from PC12 cells via a G-protein-dependent activation of neuronal calcium channels. In the present study we show that ganglioside GM1 does not directly activate this pathway in PC12 cells. However, the presence of GM1 (12.5-100 micrograms/ml) in the co-culture was associated with a potentiation of NCAM and N-cadherin-dependent neurite outgrowth. Treatment of PC12 cells with GM1 (100 micrograms/ml) for 90 min led to trypsin-stable increases in both beta-cholera toxin binding to PC12 cells and an enhanced neurite outgrowth response to N-cadherin. The ganglioside response could be fully inhibited by treatment with pertussis toxin. These data are consistent with exogenous gangliosides enhancing neuritic growth by promoting cell adhesion molecule-induced calcium influx into neurons. + + 4684 + Gene + 40754 + + neural cell adhesion molecule + + + 414745 + Gene + 20424 + + N- cadherin + + + 24586 + Gene + 40754 + + NCAM + + + 83501 + Gene + 20424 + + N-cadherin + + + 24586 + Gene + 40754 + + NCAM + + + 83501 + Gene + 20424 + + N-cadherin + + + 83501 + Gene + 20424 + + N-cadherin + + + 4684 + Gene + 40754 + + NCAM + + + 9606 + Species + + human + + + 9031 + Species + + chick + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + CVCL:0481 + CellLine + + PC12 + + + MESH:D058225 + Disease + + neuritic + + + CVCL:0594 + CellLine + + 3T3 + + + CVCL:0481 + CellLine + + PC12 + + + MESH:D005677 + Chemical + + GM1 + + + MESH:D005677 + Chemical + + GM1 + + + MESH:D002118 + Chemical + + calcium + + + CVCL:0594 + CellLine + + 3T3 + + + MESH:D005732 + Chemical + + gangliosides + + + CVCL:0481 + CellLine + + PC12 + + + MESH:D005732 + Chemical + + ganglioside + + + MESH:D005677 + Chemical + + GM1 + + + MESH:D005732 + Chemical + + ganglioside + + + + + 2289483 + + 1577866 + 1031 + 5 + 1992 + 2289483 + 92250727 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1031-40 + 1040 + Chrétien D, Metoz F, Verde F, Karsenti E, Wade RH + 0 + Lattice defects in microtubules: protofilament numbers vary within individual microtubules + + + ABSTRACT + abstract + 91 + We have used cryo-electron microscopy of vitrified specimens to study microtubules assembled both from three cycle purified tubulin (3x- tubulin) and in cell free extracts of Xenopus eggs. In vitro assembled 3x-tubulin samples have a majority of microtubules with 14 protofilaments whereas in cell extracts most microtubules have 13 protofilaments. Microtubule polymorphism was observed in both cases. The number of protofilaments can change abruptly along individual microtubules usually by single increments but double increments also occur. For 3x-tubulin, increasing the magnesium concentration decreases the proportion of 14 protofilament microtubules and decreases the average separation between transitions in these microtubules. Protofilament discontinuities may correspond to dislocation-like defects in the microtubule surface lattice. + + 8355 + Species + + Xenopus + + + MESH:D008274 + Chemical + + magnesium + + + + + 2289484 + + 1577864 + 1007 + 5 + 1992 + 2289484 + 92250725 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1007-22 + 1022 + Lu MH, DiLullo C, Schultheiss T, Holtzer S, Murray JM, Choi J, Fischman DA, Holtzer H + 0 + The vinculin/sarcomeric-alpha-actinin/alpha-actin nexus in cultured cardiac myocytes + + 7414 + Gene + 7594 + + vinculin + + + 87 + Gene + 55553 + + alpha-actinin + + + + ABSTRACT + abstract + 85 + Experiments are described supporting the proposition that the assembly of stress fibers in non-muscle cells and the assembly of myofibrils in cardiac cells share conserved mechanisms. Double staining with a battery of labeled antibodies against membrane-associated proteins, myofibrillar proteins, and stress fiber proteins reveals the following: (a) dissociated, cultured cardiac myocytes reconstitute intercalated discs consisting of adherens junctions (AJs) and desmosomes at sites of cell-cell contact and sub-sarcolemmal adhesion plaques (SAPs) at sites of cell-substrate contact; (b) each AJ or SAP associates proximally with a striated myofibril, and conversely every striated myofibril is capped at either end by an AJ or a SAP; (C) the invariant association between a given myofibril and its SAP is especially prominent at the earliest stages of myofibrillogenesis; nascent myofibrils are capped by oppositely oriented SAPs; (d) the insertion of nascent myofibrils into AJs or into SAPs invariably involves vinculin, alpha-actin, and sarcomeric alpha-actinin (s-alpha-actinin); (e) AJs are positive for A- CAM but negative for talin and integrin; SAPs lack A-CAM but are positive for talin and integrin; (f) in cardiac cells all alpha-actinin- containing structures invariably are positive for the sarcomeric isoform, alpha-actin and related sarcomeric proteins; they lack non-s- alpha-actinin, gamma-actin, and caldesmon; (g) in fibroblasts all alpha- actinin-containing structures are positive for the non-sarcomeric isoform, gamma-actin, and related non-sarcomeric proteins, including caldesmon; and (h) myocytes differ from all other types of adherent cultured cells in that they do not assemble authentic stress fibers; instead they assemble stress fiber-like structures of linearly aligned I-Z-I-like complexes consisting exclusively of sarcomeric proteins. + + 7414 + Gene + 7594 + + vinculin + + + 87 + Gene + 55553 + + alpha-actinin + + + 87 + Gene + 55553 + + alpha-actinin + + + 87 + Gene + 55553 + + alpha-actinin + + + 87 + Gene + 55553 + + alpha-actinin + + + MESH:C580316 + Disease + + myofibrillar proteins + + + + + 2289485 + + 1315785 + 921 + 5 + 1992 + 2289485 + 92250737 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 921-34 + 934 + Shamu CE, Murray AW + 0 + Sister chromatid separation in frog egg extracts requires DNA topoisomerase II activity during anaphase + + + ABSTRACT + abstract + 104 + We have produced metaphase spindles and induced them to enter anaphase in vitro. Sperm nuclei were added to frog egg extracts, allowed to replicate their DNA, and driven into metaphase by the addition of cytoplasm containing active maturation promoting factor (MPF) and cytostatic factor (CSF), an activity that stabilizes MPF. Addition of calcium induces the inactivation of MPF, sister chromatid separation and anaphase chromosome movement. DNA topoisomerase II inhibitors prevent chromosome segregation at anaphase, demonstrating that the chromatids are catenated at metaphase and that decatenation occurs at the start of anaphase. Topoisomerase II activity towards exogenous substrates does not increase at the metaphase to anaphase transition, showing that chromosome separation at anaphase is not triggered by a bulk activation of topoisomerase II. + + 1437 + Gene + 600 + + cytostatic factor + + + 1437 + Gene + 600 + + CSF + + + MESH:D002118 + Chemical + + calcium + + + + + 2289486 + + 1374417 + 959 + 5 + 1992 + 2289486 + 92250740 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 959-73 + 973 + Roitelman J, Olender EH, Bar-Nun S, Dunn WA, Simoni RD + 0 + Immunological evidence for eight spans in the membrane domain of 3- hydroxy-3-methylglutaryl coenzyme A reductase: implications for enzyme degradation in the endoplasmic reticulum + + 25675 + Gene + 30994 + + 3- hydroxy-3-methylglutaryl coenzyme A reductase + + + + ABSTRACT + abstract + 180 + We have raised two monospecific antibodies against synthetic peptides derived from the membrane domain of the ER glycoprotein 3-hydroxy-3- methylglutaryl coenzyme A (HMG-CoA) reductase, the rate limiting enzyme in the cholesterol biosynthetic pathway. This domain, which was proposed to span the ER membrane seven times (Liscum, L., J. Finer- Moore, R. M. Stroud, K. L. Luskey, M. S. Brown, and J. L. Goldstein. 1985. J. Biol. Chem. 260:522-538), plays a critical role in the regulated degradation of the enzyme in the ER in response to sterols. The antibodies stain the ER of cells and immunoprecipitate HMG-CoA reductase and HMGal, a chimeric protein composed of the membrane domain of the reductase fused to Escherichia coli beta-galactosidase, the degradation of which is also accelerated by sterols. We show that the sequence Arg224 through Leu242 of HMG-CoA reductase (peptide G) faces the cytoplasm both in cultured cells and in rat liver, whereas the sequence Thr284 through Glu302 (peptide H) faces the lumen of the ER. This indicates that a sequence between peptide G and peptide H spans the membrane of the ER. Moreover, by epitope tagging with peptide H, we show that the loop segment connecting membrane spans 3 and 4 is sequestered in the lumen of the ER. These results demonstrate that the membrane domain of HMG-CoA reductase spans the ER eight times and are inconsistent with the seven membrane spans topological model. The approximate boundaries of the proposed additional transmembrane segment are between Lys248 and Asp276. Replacement of this 7th span in HMGal with the first transmembrane helix of bacteriorhodopsin abolishes the sterol-enhanced degradation of the protein, indicating its role in the regulated turnover of HMG-CoA reductase within the endoplasmic reticulum. + + 562 + Species + + Escherichia coli + + + 10116 + Species + + rat + + + - + Chemical + + Leu242 + + + MESH:D013261 + Chemical + + sterols + + + - + Chemical + + Thr284 + + + MESH:D013261 + Chemical + + sterol + + + MESH:D013261 + Chemical + + sterols + + + - + Chemical + + Arg224 + + + - + Chemical + + Glu302 + + + - + Chemical + + Asp276 + + + - + Chemical + + Lys248 + + + MESH:D002784 + Chemical + + cholesterol + + + + + 2289487 + + 1577871 + 975 + 5 + 1992 + 2289487 + 92250741 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 975-86 + 986 + Pietrini G, Aggujaro D, Carrera P, Malyszko J, Vitale A, Borgese N + 0 + A single mRNA, transcribed from an alternative, erythroid-specific, promoter, codes for two non-myristylated forms of NADH-cytochrome b5 reductase + + + ABSTRACT + abstract + 147 + Two forms of NADH-cytochrome b5 reductase are produced from one gene: a myristylated membrane-bound enzyme, expressed in all tissues, and a soluble, erythrocyte-specific, isoform. The two forms are identical in a large cytoplasmic domain (Mr approximately 30,000) and differ at the NH2-terminus, which, in the membrane form, is responsible for binding to the bilayer, and which contains the myristylation consensus sequence and an additional 14 uncharged amino acids. To investigate how the two differently targeted forms of the reductase are produced, we cloned a reductase transcript from reticulocytes, and studied its relationship to the previously cloned liver cDNA. The reticulocyte transcript differs from the liver transcript in the 5' non-coding portion and at the beginning of the coding portion, where the seven codons specifying the myristoylation consensus are replaced by a reticulocyte-specific sequence which codes for 13 non-charged amino acids. Analysis of genomic reductase clones indicated that the ubiquitous transcript is generated from an upstream "housekeeping" type promoter, while the reticulocyte transcript originates from a downstream, erythroid- specific, promoter. In vitro translation of the reticulocyte-specific mRNA generated two products: a minor one originating from the first AUG, and a major one starting from a downstream AUG, as indicated by mutational analysis. Both the AUGs used as initiation codons were in an unfavorable sequence context. The major, lower relative molecular mass product behaved as a soluble protein, while the NH2-terminally extended minor product interacted with microsomes in vitro. The generation of soluble reductase from a downstream AUG was confirmed in vivo, in Xenopus oocytes. Thus, differently localized products, with respect both to tissues and to subcellular compartments, are generated from the same gene by a combination of transcriptional and translational mechanisms. + + 8355 + Species + + Xenopus + + + + + 2289488 + + 1577870 + 949 + 5 + 1992 + 2289488 + 92250739 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 949-58 + 958 + Ivessa NE, De Lemos-Chiarandini C, Tsao YS, Takatsuki A, Adesnik M, Sabatini DD, Kreibich G + 0 + O-glycosylation of intact and truncated ribophorins in brefeldin A- treated cells: newly synthesized intact ribophorins are only transiently accessible to the relocated glycosyltransferases + + MESH:D020126 + Chemical + + brefeldin A + + + + ABSTRACT + abstract + 190 + Ribophorins I and II are type I transmembrane glycoproteins of the ER that are segregated to the rough domains of this organelle. Both ribophorins appear to be part of the translocation apparatus for nascent polypeptides that is associated with membrane-bound ribosomes and participate in the formation of a proteinaceous network within the ER membrane that also includes other components of the translocation apparatus. The ribophorins are both highly stable proteins that lack O- linked sugars but each contains one high mannose N-linked oligosaccharide that remains endo H sensitive throughout their lifetimes. We have previously shown (Tsao, Y. S., N. E. Ivessa, M. Adesnik, D. D. Sabatini, and G. Kreibich. 1992. J. Cell Biol. 116:57- 67) that a COOH-terminally truncated variant of ribophorin I that contains only the first 332 amino acids of the luminal domain (RI332), when synthesized in permanent transformants of HeLa cells, undergoes a rapid degradation with biphasic kinetics in the ER itself and in a second, as yet unidentified nonlysosomal pre-Golgi compartment. We now show that in cells treated with brefeldin A (BFA) RI332 molecules undergo rapid O-glycosylation in a multistep process that involves the sequential addition of N-acetylgalactosamine, galactose, and terminal sialic acid residues. Addition of O-linked sugars affected all newly synthesized RI332 molecules and was completed soon after synthesis with a half time of about 10 min. In the same cells, intact ribophorins I and II also underwent O-linked glycosylation in the presence of BFA, but these molecules were modified only during a short time period immediately after their synthesis was completed, and the modification affected only a fraction of the newly synthesized polypeptides. More important, these molecules synthesized before the addition of BFA were not modified by O-glycosylation. The same is true for ribophorin I when overexpressed in HeLa cells although it is significantly less stable than the native polypeptide in control cells. We, therefore, conclude that soon after their synthesis, ribophorins lose their susceptibility to the relocated Golgi enzymes that effect the O-glycosylation, most likely as a consequence of a conformational change in the ribophorins that occurs during their maturation, although it cannot be excluded that rapid integration of these molecules into a supramolecular complex in the ER membrane leads to their inaccessibility to these enzymes. + + MESH:D000116 + Chemical + + N-acetylgalactosamine + + + - + Chemical + + RI332 + + + MESH:D020126 + Chemical + + BFA + + + MESH:D020126 + Chemical + + brefeldin A + + + MESH:D019158 + Chemical + + sialic acid + + + CVCL:0030 + CellLine + + HeLa + + + - + Chemical + + O-linked sugars + + + MESH:D020126 + Chemical + + BFA + + + CVCL:0030 + CellLine + + HeLa + + + MESH:D020126 + Chemical + + BFA + + + - + Chemical + + mannose N-linked oligosaccharide + + + - + Chemical + + O- linked sugars + + + - + Chemical + + RI332 + + + MESH:D005690 + Chemical + + galactose + + + + + 2289489 + + 1374415 + 1101 + 5 + 1992 + 2289489 + 92250734 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 1101-7 + 1107 + Leavesley DI, Ferguson GD, Wayner EA, Cheresh DA + 0 + Requirement of the integrin beta 3 subunit for carcinoma cell spreading or migration on vitronectin and fibrinogen + + 7448 + Gene + 532 + + vitronectin + + + 1934 + Gene + + beta 3 + + + MESH:D002277 + Disease + + carcinoma + + + + ABSTRACT + abstract + 115 + FG human pancreatic carcinoma cells use integrin alpha v beta 5 as their primary vitronectin receptor since they fail to express integrin alpha v beta 3. These cells are unable to form focal contacts, spread, or migrate on vitronectin but readily do so on collagen in a beta 1 integrin-dependent manner. Transfection of FG cells with a cDNA encoding the integrin beta 3 subunit results in the surface expression of a functional integrin alpha v beta 3 heterodimer providing these cells with novel adhesive and biological properties. Specifically, FG cells expressing beta 3 acquire the capacity to attach and spread on vitronectin as well as fibrinogen with beta 3 localization to focal contacts. Moreover, these cells gain the capacity to migrate through a porous membrane in response to either vitronectin or fibrinogen. These results demonstrate that the beta 3 and beta 5 integrin subunits when associated with alpha v, promote distinct cellular responses to a vitronectin extracellular environment. + + 3685 + Gene + 20510 + + integrin alpha v beta 3 + + + 7448 + Gene + 532 + + vitronectin + + + 3690 + Gene + 55444 + + integrin beta 3 + + + 3685 + Gene + 20510 + + integrin alpha v beta 3 + + + 7448 + Gene + 532 + + vitronectin + + + 7448 + Gene + 532 + + vitronectin + + + 2244 + Gene + 3772 + + fibrinogen + + + 1934 + Gene + + beta 3 and beta 5 + + + 7448 + Gene + 532 + + vitronectin + + + 7448 + Gene + 532 + + vitronectin + + + 2244 + Gene + 3772 + + fibrinogen + + + 9606 + Species + + human + + + 1934 + Gene + + beta 3 + + + 1934 + Gene + + beta 3 + + + MESH:C562463 + Disease + + pancreatic carcinoma + + + + + 2289490 + + 1577872 + 997 + 5 + 1992 + 2289490 + 92250743 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(5) 997-1005 + 1005 + Porter GA, Dmytrenko GM, Winkelmann JC, Bloch RJ + 0 + Dystrophin colocalizes with beta-spectrin in distinct subsarcolemmal domains in mammalian skeletal muscle + + 1756 + Gene + 20856 + + Dystrophin + + + 9606 + Species + + mammalian + + + + ABSTRACT + abstract + 106 + Duchenne's muscular dystrophy (DMD) is caused by the absence or drastic decrease of the structural protein, dystrophin, and is characterized by sarcolemmal lesions in skeletal muscle due to the stress of contraction. Dystrophin has been localized to the sarcolemma, but its organization there is not known. We report immunofluorescence studies which show that dystrophin is concentrated, along with the major muscle isoform of beta-spectrin, in three distinct domains at the sarcolemma: in elements overlying both I bands and M lines, and in occasional strands running along the longitudinal axis of the myofiber. Vinculin, which has previously been found at the sarcolemma overlying the I bands and in longitudinal strands, was present in the same three structures as spectrin and dystrophin. Controls demonstrated that the labeling was intracellular. Comparison to labeling of the lipid bilayer and of the extracellular matrix showed that the labeling for spectrin and dystrophin is associated with the intact sarcolemma and is not a result of processing artifacts. Dystrophin is not required for this lattice- like organization, as similar domains containing spectrin but not dystrophin are present in muscle from the mdx mouse and from humans with Duchenne's muscular dystrophy. We discuss the possibility that dystrophin and spectrin, along with vinculin, may function to link the contractile apparatus to the sarcolemma of normal skeletal muscle. + + 1756 + Gene + 20856 + + dystrophin + + + 1756 + Gene + 20856 + + dystrophin + + + 7414 + Gene + 7594 + + Vinculin + + + 1756 + Gene + 20856 + + dystrophin + + + 1756 + Gene + 20856 + + dystrophin + + + 13405 + Gene + 20856 + + Dystrophin + + + 1756 + Gene + 20856 + + dystrophin + + + 7414 + Gene + 7594 + + vinculin + + + 1756 + Gene + 20856 + + Dystrophin + + + 10090 + Species + + mouse + + + 9606 + Species + + humans + + + 1756 + Gene + 20856 + + dystrophin + + + MESH:D020388 + Disease + + Duchenne's muscular dystrophy + + + MESH:D020388 + Disease + + DMD + + + MESH:D020388 + Disease + + Duchenne's muscular dystrophy + + + MESH:D008055 + Chemical + + lipid + + + + + 2289491 + + 1607384 + 1223 + 6 + 1992 + 2289491 + 92299679 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1223-30 + 1230 + Applegate D, Pardee JD + 0 + Actin-facilitated assembly of smooth muscle myosin induces formation of actomyosin fibrils + + 79784 + Gene + 23480 + + myosin + + + + ABSTRACT + abstract + 91 + To identify regulatory mechanisms potentially involved in formation of actomyosin structures in smooth muscle cells, the influence of F-actin on smooth muscle myosin assembly was examined. In physiologically relevant buffers, AMPPNP binding to myosin caused transition to the soluble 10S myosin conformation due to trapping of nucleotide at the active sites. The resulting 10S myosin-AMPPNP complex was highly stable and thick filament assembly was suppressed. However, upon addition to F- actin, myosin readily assembled to form thick filaments. Furthermore, myosin assembly caused rearrangement of actin filament networks into actomyosin fibers composed of coaligned F-actin and myosin thick filaments. Severin-induced fragmentation of actin in actomyosin fibers resulted in immediate disassembly of myosin thick filaments, demonstrating that actin filaments were indispensable for mediating myosin assembly in the presence of AMPPNP. Actomyosin fibers also formed after addition of F-actin to nonphosphorylated 10S myosin monomers containing the products of ATP hydrolysis trapped at the active site. The resulting fibers were rapidly disassembled after addition of millimolar MgATP and consequent transition of myosin to the soluble 10S state. However, reassembly of myosin filaments in the presence of MgATP and F-actin could be induced by phosphorylation of myosin P-light chains, causing regeneration of actomyosin fiber bundles. The results indicate that actomyosin fibers can be spontaneously formed by F-actin-mediated assembly of smooth muscle myosin. Moreover, induction of actomyosin fibers by myosin light chain phosphorylation in the presence of actin filament networks provides a plausible hypothesis for contractile fiber assembly in situ. + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + MESH:D000266 + Chemical + + AMPPNP + + + MESH:D000255 + Chemical + + MgATP + + + MESH:D000255 + Chemical + + MgATP + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000266 + Chemical + + AMPPNP + + + + + 2289492 + + 1607382 + 1181 + 6 + 1992 + 2289492 + 92299676 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1181-96 + 1196 + Robinson LJ, Pang S, Harris DS, Heuser J, James DE + 0 + Translocation of the glucose transporter (GLUT4) to the cell surface in permeabilized 3T3-L1 adipocytes: effects of ATP insulin, and GTP gamma S and localization of GLUT4 to clathrin lattices + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 3630 + Gene + 173 + + insulin + + + MESH:D000255 + Chemical + + ATP + + + MESH:D006160 + Chemical + + GTP + + + + ABSTRACT + abstract + 192 + Insulin stimulates the movement of two glucose transporter isoforms (GLUT1 and GLUT4) to the plasma membrane (PM) in adipocytes. To study this process we have prepared highly purified PM fragments by gently sonicating 3T3-L1 adipocytes grown on glass coverslips. Using confocal laser immunofluorescence microscopy we observed increased PM labeling for GLUT1 (2.3-fold) and GLUT4 (eightfold) after insulin treatment in intact cells. EM immunolabeling of PM fragments indicated that in the nonstimulated state GLUT4 was mainly localized to flat clathrin lattices. Whereas GLUT4 labeling of clathrin lattices was only slightly increased after insulin treatment, labeling of uncoated PM regions was markedly increased with insulin. These data suggest that GLUT4 recycles from the cell surface both in the presence and absence of insulin. In streptolysin-O permeabilized adipocytes, insulin, and GTP gamma S increased PM levels of GLUT4 to a similar extent as observed with insulin in intact cells. In the absence of an exogenous ATP source the magnitude of these effects was considerably reduced. Removal of ATP per se caused a significant increase in cell surface levels of GLUT4 suggesting that ATP may be required for intracellular sequestration of these transporters. When insulin and GTP gamma S were added together, in the presence of ATP, PM GLUT4 levels were similar to levels observed when either insulin or GTP gamma S was added individually. Addition of GTP gamma S was able to overcome this ATP dependence of insulin- stimulated GLUT4 movement. GTP gamma S had no effect on constitutive secretion of adipsin in permeabilized cells. In addition, there was no effect of insulin or GTP gamma S on GLUT4 movement to the PM in noninsulin sensitive streptolysin-O-permeabilized 3T3-L1 fibroblasts overexpressing GLUT4. We conclude that the insulin-stimulated movement of GLUT4 to the cell surface in adipocytes may require ATP early in the insulin signaling pathway and a GTP-binding protein(s) at a later step(s). We propose that the association of GLUT4 with clathrin lattices may be important in maintaining the exclusive intracellular location of this transporter in the absence of insulin. + + 6513 + Gene + 68520 + + GLUT1 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6513 + Gene + 68520 + + GLUT1 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 3630 + Gene + 173 + + insulin + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 1675 + Gene + 20453 + + adipsin + + + 3630 + Gene + 173 + + insulin or GTP gamma S + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 6517 + Gene + 74381 + + GLUT4 + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + Insulin + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin or GTP gamma S + + + 3630 + Gene + 173 + + insulin + + + 3630 + Gene + 173 + + insulin + + + MESH:D006160 + Chemical + + GTP + + + MESH:D016244 + Chemical + + GTP gamma S + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000255 + Chemical + + ATP + + + MESH:D004832 + Disease + + absence of insulin + + + MESH:D013455 + Chemical + + S + + + MESH:D000255 + Chemical + + ATP + + + MESH:D006160 + Chemical + + GTP + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000255 + Chemical + + ATP + + + MESH:D004832 + Disease + + absence of insulin + + + MESH:D006160 + Chemical + + GTP + + + + + 2289493 + + 1607387 + 1251 + 6 + 1992 + 2289493 + 92299682 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1251-61 + 1261 + Brancolini C, Bottega S, Schneider C + 0 + Gas2, a growth arrest-specific protein, is a component of the microfilament network system + + 14453 + Gene + 31301 + + Gas2 + + + + ABSTRACT + abstract + 91 + In this report we analyze the protein product of a growth arrest- specific gene, gas2, by means of an affinity-purified antibody raised against the protein produced in bacteria. The regulation of Gas2 biosynthesis reflects the pattern of mRNA expression (Schneider, C., R. King, and L. Philipson. 1988. Cell. 54:787-793): its relative level is tightly associated with growth arrest. Gas2 seems to be regulated also at the posttranslational level via a phosphorylation mechanism. Gas2 is well conserved during the evolution with the same apparent molecular mass (36 kD) between mouse and human. We also demonstrate that Gas2 is a component of the microfilament system. It colocalizes with actin fiber, at the cell border and also along the stress fiber, in growth- arrested NIH 3T3 cells. The pattern of distribution, detected in arrested cells, can also be observed in growing cells when they are microinjected with the purified GST-Gas2 protein. In none of the analyzed oncogene-transformed NIH 3T3 cell lines was Gas2 expression induced under serum starvation. + + 14453 + Gene + 31301 + + gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 14453 + Gene + 31301 + + Gas2 + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + CVCL:0594 + CellLine + + NIH 3T3 + + + CVCL:0594 + CellLine + + NIH 3T3 + + + + + 2289494 + + 1318883 + 1197 + 6 + 1992 + 2289494 + 92299677 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1197-1209 + 1209 + Lencer WI, Delp C, Neutra MR, Madara JL + 0 + Mechanism of cholera toxin action on a polarized human intestinal epithelial cell line: role of vesicular traffic + + 9606 + Species + + human + + + + ABSTRACT + abstract + 114 + The massive secretion of salt and water in cholera-induced diarrhea involves binding of cholera toxin (CT) to ganglioside GM1 in the apical membrane of intestinal epithelial cells, translocation of the enzymatically active A1-peptide across the membrane, and subsequent activation of adenylate cyclase located on the cytoplasmic surface of the basolateral membrane. Studies on nonpolarized cells show that CT is internalized by receptor-mediated endocytosis, and that the A1-subunit may remain membrane associated. To test the hypothesis that toxin action in polarized cells may involve intracellular movement of toxin- containing membranes, monolayers of the polarized intestinal epithelial cell line T84 were mounted in modified Ussing chambers and the response to CT was examined. Apical CT at 37 degrees C elicited a short circuit current (Isc: 48 +/- 2.1 microA/cm2; half-maximal effective dose, ED50 integral of 0.5 nM) after a lag of 33 +/- 2 min which bidirectional 22Na+ and 36Cl- flux studies showed to be due to electrogenic Cl- secretion. The time course of the CT-induced Isc response paralleled the time course of cAMP generation. The dose response to basolateral toxin at 37 degrees C was identical to that of apical CT but lag times (24 +/- 2 min) and initial rates were significantly less. At 20 degrees C, the Isc response to apical CT was more strongly inhibited (30-50%) than the response to basolateral CT, even though translocation occurred in both cases as evidenced by the formation of A1-peptide. A functional rhodamine-labeled CT-analogue applied apically or basolaterally at 20 degrees C was visualized only within endocytic vesicles close to apical or basolateral membranes, whereas movement into deeper apical structures was detected at 37 degrees C. At 15 degrees C, in contrast, reduction to the A1-peptide was completely inhibited and both apical and basolateral CT failed to stimulate Isc although Isc responses to 1 nM vasoactive intestinal peptide, 10 microM forskolin, and 3 mM 8Br- cAMP were intact. Re-warming above 32 degrees C restored CT-induced Isc. Preincubating monolayers for 30 min at 37 degrees C before cooling to 15 degrees C overcame the temperature block of basolateral CT but the response to apical toxin remained completely inhibited. These results identify a temperature-sensitive step essential to apical toxin action on polarized epithelial cells. We suggest that this event involves vesicular transport of toxin-containing membranes beyond the apical endosomal compartment. + + MESH:D012492 + Chemical + + salt + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D005732 + Chemical + + ganglioside + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D014867 + Chemical + + water + + + MESH:D005677 + Chemical + + GM1 + + + MESH:D012235 + Chemical + + rhodamine + + + MESH:D015124 + Chemical + + 8Br- cAMP + + + - + Chemical + + cAMP + + + + + 2289495 + + 1607378 + 1137 + 6 + 1992 + 2289495 + 92299672 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1137-50 + 1150 + Beckmann RP, Lovett M, Welch WJ + 0 + Examining the function and regulation of hsp 70 in cells subjected to metabolic stress + + 3308 + Gene + 1624 + + hsp 70 + + + + ABSTRACT + abstract + 87 + Members of the heat-shock protein (hsp) 70 family, distributed within various cellular compartments, have been implicated in facilitating protein maturation events. In particular, related hsp 70 family members appear to bind nascent polypeptides which are in the course of synthesis and/or translocation into organelles. We previously reported that in normal, unstressed cells, cytosolic hsp 70 (hsp 72/73) interacted transiently with nascent polypeptides. We suspect that such interactions function to prevent or slow down the folding of the nascent polypeptide chain. Once synthesis is complete, and now with all of the information for folding present, the newly synthesized protein appears to commence along its folding pathway, accompanied by the ATP- dependent release of hsp 72/73. Herein, we examined how these events occur in cells subjected to different types of metabolic stress. In cells exposed to either an amino acid analog or sodium arsenite, two potent inducers of the stress response, newly synthesized proteins bind to but are not released from hsp 70. Under these conditions of metabolic stress, we suspect that the newly synthesized proteins are unable to commence proper folding and consequently remain bound to their hsp 70 chaperone. In cells subjected to heat shock, a large number of both newly synthesized as well as mature proteins are rendered insoluble. Within this insoluble material are appreciable amounts of hsp 72/73. Finally, we show that in cells depleted of ATP, the release of hsp 70 from maturing proteins is inhibited. Thus, in cells experiencing metabolic stress, newly synthesized proteins unable to properly fold, as will as mature proteins which begin to unfold become stably bound to hsp 72/73. As a consequence and over time, the free or available levels of pre-existing hsp 72/73 are reduced. We propose that this reduction in the available levels of hsp 72/73 is the trigger by which the stress response is initiated. + + 3308 + Gene + 1624 + + heat-shock protein (hsp) 70 + + + 3308 + Gene + 1624 + + hsp 70 + + + 3308 + Gene + 1624 + + hsp 70 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3308 + Gene + 1624 + + hsp 70 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3303 + Gene + 74294 + + hsp 72 + + + 3308 + Gene + 1624 + + hsp 70 + + + 3308 + Gene + 1624 + + hsp 70 + + + MESH:D000255 + Chemical + + ATP + + + MESH:C017947 + Chemical + + sodium arsenite + + + MESH:D000255 + Chemical + + ATP + + + MESH:D012769 + Disease + + shock + + + + + 2289496 + + 1376731 + 1321 + 6 + 1992 + 2289496 + 92299688 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1321-30 + 1330 + Reszka AA, Hayashi Y, Horwitz AF + 0 + Identification of amino acid sequences in the integrin beta 1 cytoplasmic domain implicated in cytoskeletal association + + 16412 + Gene + 22999 + + integrin beta 1 + + + + ABSTRACT + abstract + 120 + Wild-type and mutant chicken integrin beta 1 subunit (beta 1c) cDNAs were expressed in NIH 3T3 cells and assayed for localization in focal adhesions of cells plated on fibronectin substrates. Focal adhesion localization in stable transfected cells was assayed by indirect immunofluorescent staining with chicken-specific anti-beta 1c antibodies. Mutant beta 1c integrins containing internal deletions of 13 amino acids adjacent to the membrane, delta 759-771, and 20 centrally located amino acids, delta 771-790, localized in focal adhesions demonstrating that sequences required for direction to focal adhesion structures were not limited to one region of the cytoplasmic domain. Point mutations revealed three clusters of amino acids which contribute to localization in focal adhesions. These three clusters or signals are: cyto-1 (764-774), cyto-2 (785-788), and cyto-3 (797-800). The 11-residue cyto-1 signal is only found on integrin beta subunit sequences, except beta 4. Four residues within this region, D764, F768, F771, and E774, could not be altered without reducing focal adhesion staining intensities, and likely form a signal that occupies one side of an alpha helix. Mutations involving two cyto-1 residues, K770 and F771, also appeared to affect heterodimer affinity and specificity. Cyto-2 (785-788,), NPIY, is an NPXY signal that forms a tight turn motif. Cyto-2 provides a structural conformation, which when perturbed by proline removal or addition, inhibits integrin localization in focal adhesions. Cyto-3 (797-800), NPKY, resembles cyto-2, however, the nonconserved proline residue can be replaced without alteration of the localization phenotype. Cyto-3, therefore, constitutes a unique integrin signal, NXXY. Both serine and tyrosine residues at positions 790 and 788, respectively, which have been implicated in integrin phosphorylation/regulation, were conservatively replaced without detectable effect on focal adhesion localization. However, acidic replacements for these amino acids reduced focal adhesion staining intensities, suggesting that phosphorylation at these sites may negatively regulate integrin function. + + 14268 + Gene + 1533 + + fibronectin + + + 9031 + Species + + chicken + + + 9031 + Species + + chicken + + + 16412 + Gene + 22999 + + integrin beta 1 + + + MESH:D011392 + Chemical + + proline + + + MESH:D011392 + Chemical + + proline + + + CVCL:0594 + CellLine + + NIH 3T3 + + + MESH:D014443 + Chemical + + tyrosine + + + MESH:D012694 + Chemical + + serine + + + + + 2289497 + + 1607389 + 1277 + 6 + 1992 + 2289497 + 92299684 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1277-87 + 1287 + Vallen EA, Hiller MA, Scherson TY, Rose MD + 0 + Separate domains of KAR1 mediate distinct functions in mitosis and nuclear fusion + + 855533 + Gene + + KAR1 + + + Disease + + mitosis + + + + ABSTRACT + abstract + 82 + The yeast KAR1 gene is essential for mitotic growth and important for nuclear fusion. Mutations in KAR1 prevent duplication of the spindle pole body (SPB), and affect functions associated with both the nuclear and cytoplasmic microtubules. The localization of hybrid Kar1-lacZ proteins, described elsewhere (Vallen, E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press), suggest that the protein is associated with the SPB. In this paper, we report a deletion analysis demonstrating that the mitotic and karyogamy functions of KAR1 are separate and independent, residing in discrete functional domains. One region, here shown to be essential for mitosis, coincided with a part of the protein that is both necessary and sufficient to target Karl-lacZ hybrid proteins to the SPB (Vallen, E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press). Complementation testing demonstrated that deletions in this interval did not affect nuclear fusion. A second region, required only for karyogamy, was necessary for the localization of a Kar3-lacZ hybrid protein to the SPB. These data suggest a model for the roles of Kar1p and Kar3p, a kinesin-like protein, in nuclear fusion. Finally, a third region of KAR1 was found to be important for both mitosis and karyogamy. This domain included the hydrophobic carboxy terminus and is sufficient to target a lacZ-Kar1 hybrid protein to the nuclear envelope (Vallen E. A., T. Y. Scherson, T. Roberts, K. van Zee, and M. D. Rose. 1992. Cell. In press). Altogether, the essential mitotic regions of KAR1 comprised 20% of the coding sequence. We propose a model for Kar1p in which the protein is composed of several protein-binding domains tethered to the nuclear envelope via its hydrophobic tail. + + 855533 + Gene + + KAR1 + + + 855533 + Gene + + KAR1 + + + 855533 + Gene + + Kar1 + + + 855533 + Gene + + KAR1 + + + 855533 + Gene + + KAR1 + + + 855533 + Gene + + KAR1 + + + 855533 + Gene + + Kar1 + + + 4932 + Species + + yeast + + + 267260 + Species + + D. Rose + + + 267260 + Species + + D. Rose + + + 267260 + Species + + D. Rose + + + Disease + + mitosis + + + Disease + + mitosis + + + + + 2289498 + + 1607380 + 1161 + 6 + 1992 + 2289498 + 92299674 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1161-9 + 1169 + Dixon JL, Chattapadhyay R, Huima T, Redman CM, Banerjee D + 0 + Biosynthesis of lipoprotein: location of nascent apoAI and apoB in the rough endoplasmic reticulum of chicken hepatocytes + + 396535 + Gene + 328 + + apoB + + + 396536 + Gene + 47900 + + apoAI + + + 9031 + Species + + chicken + + + + ABSTRACT + abstract + 122 + Our previous studies showed that in hepatic RER of young chickens, nascent apoAI is not associated with lipoprotein particles and only becomes part of these lipoprotein structures in the Golgi. In this study, we have used three different methodologies to determine the locations of apoAI and apoB in the RER and compared them to that of albumin. Immunoelectron microscopic examination of the RER cell fractions showed that both apoAI and apoB were associated only with the RER membrane whereas albumin was located both within the lumen and on the limiting membrane of the vesicles. To examine the possibility of membrane integration of nascent apoAI and apoB in the RER, we administered L-[3H]leucine to young chickens for 10 min, isolated RER, treated this cell fraction with buffers of varying pH, and measured the release of radioactive albumin, apoAI, and apoB. The majority of nascent apoAI (64%), nascent apoB (100%), and nascent albumin (97%) was released from RER vesicles at pH 11.2, suggesting that, like albumin, apolipoproteins are not integrated within the membrane. To determine if nascent apoproteins are exposed to the cytoplasmic surface, we administered L-[3H]leucine to young chickens and at various times isolated RER and Golgi cell fractions. Radioactive RER and Golgi cell fractions were treated with exogenous protease and the percent of nascent apoAI and apoB accessible to proteolysis was determined and compared to that of albumin. At 5, 10, and 20 min of labeling, 35-56% of nascent apoAI and 60-75% of apoB in RER were degraded, while albumin was refractive to this treatment. At all times both apolipoproteins and albumin present in Golgi cell fractions were protected from proteolysis. These biochemical and morphological findings indicate that apoAI and apoB are associated with the rough microsomal membrane and are partially exposed to the cytoplasmic surface at early stages of secretion. They may later enter the luminal side of the ER and, on entering the Golgi, form lipoprotein particles. + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396536 + Gene + 47900 + + apoAI + + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 396535 + Gene + 328 + + apoB + + + 9031 + Species + + chickens + + + 9031 + Species + + chickens + + + 9031 + Species + + chickens + + + - + Chemical + + L-[3H]leucine + + + + + 2289499 + + 1607385 + 1231 + 6 + 1992 + 2289499 + 92299680 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1231-9 + 1239 + Yumura S, Kitanishi-Yumura T + 0 + Release of myosin II from the membrane-cytoskeleton of Dictyostelium discoideum mediated by heavy-chain phosphorylation at the foci within the cortical actin network + + 44689 + Species + + Dictyostelium discoideum + + + + ABSTRACT + abstract + 166 + Membrane-cytoskeletons were prepared from Dictyostelium amebas, and networks of actin and myosin II filaments were visualized on the exposed cytoplasmic surfaces of the cell membranes by fluorescence staining (Yumura, S., and T. Kitanishi-Yumura. 1990. Cell Struct. Funct. 15:355-364). Addition of ATP caused contraction of the cytoskeleton with aggregation of part of actin into several foci within the network, but most of myosin II was released via the foci. However, in the presence of 10 mM MgCl2, which stabilized myosin II filaments, myosin II remained at the foci. Ultrastructural examination revealed that, after contraction, only traces of monomeric myosin II remained at the foci. By contrast, myosin II filaments remained in the foci in the presence of 10 mM MgCl2. These observations suggest that myosin II was released not in a filamentous form but in a monomeric form. Using [gamma 32P]ATP, we found that the heavy chains of myosin II released from membrane-cytoskeletons were phosphorylated, and this phosphorylation resulted in disassembly of myosin filaments. Using ITP (a substrate for myosin II ATPase) and/or ATP gamma S (a substrate for myosin II heavy-chain kinase [MHCK]), we demonstrated that phosphorylation of myosin heavy chains occurred at the foci within the actin network, a result that suggests that MHCK was localized at the foci. These results together indicate that, during contraction, the heavy chains of myosin II that have moved toward the foci within the actin network are phosphorylated by a specific MHCK, with the resultant disassembly of filaments which are finally released from membrane- cytoskeletons. This series of reactions could represent the mechanism for the relocation of myosin II from the cortical region to the endoplasm. + + 44689 + Species + + Dictyostelium + + + MESH:D015636 + Chemical + + MgCl2 + + + - + Chemical + + ITP + + + MESH:D015636 + Chemical + + MgCl2 + + + MESH:C000615311 + Chemical + + 32P + + + MESH:C022571 + Chemical + + ATP gamma S + + + MESH:D000255 + Chemical + + ATP + + + MESH:D000255 + Chemical + + ATP + + + + + 2289500 + + 1607383 + 1211 + 6 + 1992 + 2289500 + 92299678 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1211-21 + 1221 + Mallya SK, Partin JS, Valdizan MC, Lennarz WJ + 0 + Proteolysis of the major yolk glycoproteins is regulated by acidification of the yolk platelets in sea urchin embryos + + 7656 + Species + + sea urchin + + + + ABSTRACT + abstract + 118 + The precise function of the yolk platelets of sea urchin embryos during early development is unknown. We have shown previously that the chemical composition of the yolk platelets remains unchanged in terms of phospholipid, triglyceride, hexose, sialic acid, RNA, and total protein content after fertilization and early development. However, the platelet is not entirely static because the major 160-kD yolk glycoprotein YP-160 undergoes limited, step-wise proteolytic cleavage during early development. Based on previous studies by us and others, it has been postulated that yolk platelets become acidified during development, leading to the activation of a cathepsin B-like yolk proteinase that is believed to be responsible for the degradation of the major yolk glycoprotein. To investigate this possibility, we studied the effect of addition of chloroquine, which prevents acidification of lysosomes. Consistent with the postulated requirement for acidification, it was found that chloroquine blocked YP-160 breakdown but had no effect on embryonic development. To directly test the possibility that acidification of the yolk platelets over the course of development temporally correlated with YP-160 proteolysis, we added 3-(2,4-dinitroanilo)-3-amino-N-methyldipropylamine (DAMP) to eggs or embryos. This compound localizes to acidic organelles and can be detected in these organelles by EM. The results of these studies revealed that yolk platelets did, in fact, become transiently acidified during development. This acidification occurred at the same time as yolk protein proteolysis, i.e., at 6 h after fertilization (64-cell stage) in Strongylocentrotus purpuratus and at 48 h after fertilization (late gastrula) in L. pictus. Furthermore, the pH value at the point of maximal acidification of the yolk platelets in vivo was equal to the pH optimum of the enzyme measured in vitro, indicating that this acidification is sufficient to activate the enzyme. For both S. purpuratus and Lytechinus pictus, the observed decrease in the pH was approximately 0.8 U, from 7.0 to 6.2. The trypsin inhibitor benzamidine was found to inhibit the yolk proteinase in vivo. By virtue of the fact that this inhibitor was reversible we established that the activity of the yolk proteinase is developmentally regulated even though the enzyme is present throughout the course of development. These findings indicate that acidification of yolk platelets is a developmentally regulated process that is a prerequisite to initiation of the catabolism of the major yolk glycoprotein. + + 7656 + Species + + sea urchin + + + 7668 + Species + + Strongylocentrotus purpuratus + + + 7653 + Species + + L. pictus + + + 7668 + Species + + S. purpuratus + + + 7653 + Species + + Lytechinus pictus + + + - + Chemical + + DAMP + + + - + Chemical + + 3-(2,4-dinitroanilo)-3-amino-N-methyldipropylamine + + + MESH:D014280 + Chemical + + triglyceride + + + MESH:D002738 + Chemical + + chloroquine + + + MESH:C032157 + Chemical + + benzamidine + + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D019158 + Chemical + + sialic acid + + + MESH:D006601 + Chemical + + hexose + + + MESH:D002738 + Chemical + + chloroquine + + + + + 2289501 + + 1607393 + 1351 + 6 + 1992 + 2289501 + 92299691 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1351-7 + 1357 + Pancake SJ, Holt GD, Mellouk S, Hoffman SL + 0 + Malaria sporozoites and circumsporozoite proteins bind specifically to sulfated glycoconjugates + + MESH:D008288 + Disease + + Malaria sporozoites + + + + ABSTRACT + abstract + 96 + Circumsporozoite (CS) proteins, which densely coat malaria (Plasmodia) sporozoites, contain an amino acid sequence that is homologous to segments in other proteins which bind specifically to sulfated glycoconjugates. The presence of this homology suggests that sporozoites and CS proteins may also bind sulfated glycoconjugates. To test this hypothesis, recombinant P. yoelii CS protein was examined for binding to sulfated glycoconjugate-Sepharoses. CS protein bound avidly to heparin-, fucoidan-, and dextran sulfate-Sepharose, but bound comparatively poorly to chondroitin sulfate A- or C-Sepharose. CS protein also bound with significantly lower affinity to a heparan sulfate biosynthesis-deficient mutant cell line compared with the wild- type line, consistent with the possibility that the protein also binds to sulfated glycoconjugates on the surfaces of cells. This possibility is consistent with the observation that CS protein binding to hepatocytes, cells invaded by sporozoites during the primary stage of malaria infection, was inhibited by fucoidan, pentosan polysulfate, and heparin. The effects of sulfated glycoconjugates on sporozoite infectivity were also determined. P. berghei sporozoites bound specifically to sulfatide (galactosyl[3-sulfate]beta 1-1ceramide), but not to comparable levels of cholesterol-3-sulfate, or several examples of neutral glycosphingolipids, gangliosides, or phospholipids. Sporozoite invasion into hepatocytes was inhibited by fucoidan, heparin, and dextran sulfate, paralleling the observed binding of CS protein to the corresponding Sepharose derivatives. These sulfated glycoconjugates blocked invasion by inhibiting an event occurring within 3 h of combining sporozoites and hepatocytes. Sporozoite infectivity in mice was significantly inhibited by dextran sulfate 500,000 and fucoidan. Taken together, these data indicate that CS proteins bind selectively to certain sulfated glycoconjugates, that sporozoite infectivity can be inhibited by such compounds, and that invasion of host hepatocytes by sporozoites may involve interactions with these types of compounds. + + 5861 + Species + + P. yoelii + + + 5821 + Species + + P. berghei + + + 10090 + Species + + mice + + + MESH:C007045 + Chemical + + cholesterol-3-sulfate + + + MESH:D016264 + Chemical + + dextran sulfate + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:C007789 + Chemical + + fucoidan + + + MESH:D008288 + Disease + + malaria infection + + + MESH:C007789 + Chemical + + fucoidan + + + MESH:D006493 + Chemical + + heparin + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:D006028 + Chemical + + glycosphingolipids + + + MESH:D006497 + Chemical + + heparan sulfate + + + MESH:D016264 + Chemical + + dextran sulfate + + + MESH:D016264 + Chemical + + dextran sulfate + + + MESH:D005732 + Chemical + + gangliosides + + + MESH:D006493 + Chemical + + heparin + + + MESH:D010743 + Chemical + + phospholipids + + + - + Chemical + + galactosyl[3-sulfate]beta 1-1ceramide + + + MESH:D006493 + Chemical + + heparin + + + MESH:D058456 + Disease + + coat malaria + + + - + Chemical + + CS + + + - + Chemical + + CS + + + MESH:C007789 + Chemical + + fucoidan + + + MESH:D010426 + Chemical + + pentosan polysulfate + + + - + Chemical + + CS + + + - + Chemical + + CS + + + MESH:D012685 + Chemical + + Sepharoses + + + MESH:D012685 + Chemical + + Sepharose + + + MESH:C007789 + Chemical + + fucoidan + + + MESH:D002809 + Chemical + + chondroitin sulfate A + + + MESH:D013433 + Chemical + + sulfatide + + + + + 2289502 + + 1607379 + 1151 + 6 + 1992 + 2289502 + 92299673 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1151-9 + 1159 + Baler R, Welch WJ, Voellmy R + 0 + Heat shock gene regulation by nascent polypeptides and denatured proteins: hsp70 as a potential autoregulatory factor + + 3308 + Gene + 1624 + + hsp70 + + + + ABSTRACT + abstract + 118 + Heat shock genes encode proteins (hsp's) that play important structural roles under normal circumstances and are essential to the cells' ability to survive environmental insults. Evidence is presented herein that transcriptional regulation of hsp gene expression is linked with the regulation of overall protein synthesis as well as with the accumulation of proteins denatured by stressful events. The factor that connects the three processes appears to be one of the hsp's, presumably a member(s) of the hsp70 family. Biochemical experiments demonstrate that complexes containing hsp70 and heat shock transcription factor, the specific regulator of hsp gene activity, are formed in the cells. + + 7190 + Gene + + hsp + + + 3308 + Gene + 1624 + + hsp70 + + + 3308 + Gene + 1624 + + hsp70 + + + 7190 + Gene + + hsp + + + 7190 + Gene + + hsp + + + 7190 + Gene + + hsp + + + + + 2289503 + + 1607392 + 1331 + 6 + 1992 + 2289503 + 92299689 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1331-41 + 1341 + Drake SL, Klein DJ, Mickelson DJ, Oegema TR, Furcht LT, McCarthy JB + 0 + Cell surface phosphatidylinositol-anchored heparan sulfate proteoglycan initiates mouse melanoma cell adhesion to a fibronectin-derived, heparin-binding synthetic peptide + + 10090 + Species + + mouse + + + 14268 + Gene + 1533 + + fibronectin + + + MESH:D008545 + Disease + + melanoma + + + MESH:D010716 + Chemical + + phosphatidylinositol + + + + ABSTRACT + abstract + 171 + Cell surface heparan sulfate proteoglycan (HSPG) from metastatic mouse melanoma cells initiates cell adhesion to the synthetic peptide FN-C/H II, a heparin-binding peptide from the 33-kD A chain-derived fragment of fibronectin. Mouse melanoma cell adhesion to FN-C/H II was sensitive to soluble heparin and pretreatment of mouse melanoma cells with heparitinase. In contrast, cell adhesion to the fibronectin synthetic peptide CS1 is mediated through an alpha 4 beta 1 integrin and was resistant to heparin or heparitinase treatment. Mouse melanoma cell HSPG was metabolically labeled with [35S]sulfate and extracted with detergent. After HPLC-DEAE purification, 35S-HSPG eluted from a dissociative CL-4B column with a Kav approximately 0.45, while 35S- heparan sulfate (HS) chains eluted with a Kav approximately 0.62. The HSPG contained a major 63-kD core protein after heparitinase digestion. Polyclonal antibodies generated against HSPG purified from mouse melanoma cells grown in vivo also identified a 63-kD core protein. This HSPG is an integral plasma membrane component by virtue of its binding to Octyl Sepharose affinity columns and that anti-HSPG antibody staining exhibited a cell surface localization. The HSPG is anchored to the cell surface through phosphatidylinositol (PI) linkages, as evidenced in part by the ability of PI-specific phospholipase C to eliminate binding of the detergent-extracted HSPG to Octyl Sepharose. Furthermore, the mouse melanoma HSPG core protein could be metabolically labeled with 3H-ethanolamine. The involvement of mouse melanoma cell surface HSPG in cell adhesion to fibronectin was also demonstrated by the ability of anti-HSPG antibodies and anti-HSPG IgG Fab monomers to inhibit mouse melanoma cell adhesion to FN-C/H II. 35S- HSPG and 35S-HS bind to FN-C/H II affinity columns and require 0.25 M NaCl for elution. However, heparitinase-treated 125I-labeled HSPG failed to bind FN-C/H II, suggesting that HS, and not HSPG core protein, binds FN-C/H II. These data support the hypothesis that a phosphatidylinositol-anchored HSPG on mouse melanoma cells (MPIHP-63) initiates recognition to FN-C/H II, and implicate PI-associated signal transduction pathways in mediating melanoma cell adhesion to this defined ligand. + + 15529 + Gene + 2253 + + HSPG + + + 14268 + Gene + 1533 + + fibronectin + + + 14268 + Gene + 1533 + + fibronectin + + + 70599 + Gene + 4912 + + CS1 + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 14268 + Gene + 1533 + + fibronectin + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 15529 + Gene + 2253 + + HSPG + + + 10090 + Species + + mouse + + + 10090 + Species + + Mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + Mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + MESH:D008545 + Disease + + melanoma + + + MESH:D008545 + Disease + + melanoma + + + MESH:D008545 + Disease + + melanoma + + + MESH:C027176 + Chemical + + Octyl Sepharose + + + MESH:C000615320 + Chemical + + 35S + + + MESH:C027176 + Chemical + + Octyl Sepharose + + + MESH:D013431 + Chemical + + sulfate + + + MESH:D008545 + Disease + + melanoma + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D010716 + Chemical + + phosphatidylinositol + + + MESH:C000615320 + Chemical + + 35S + + + - + Chemical + + CL-4B + + + MESH:D006497 + Chemical + + heparan sulfate + + + MESH:D008545 + Disease + + melanoma + + + - + Chemical + + 3H-ethanolamine + + + MESH:D006493 + Chemical + + heparin + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D006497 + Chemical + + HS + + + MESH:C007369 + Chemical + + DEAE + + + MESH:D008545 + Disease + + melanoma + + + MESH:D008545 + Disease + + melanoma HSPG + + + MESH:D010716 + Chemical + + phosphatidylinositol + + + MESH:D008545 + Disease + + melanoma + + + MESH:D008545 + Disease + + melanoma + + + MESH:C000615320 + Chemical + + 35S + + + MESH:D008545 + Disease + + melanoma + + + MESH:D006497 + Chemical + + HS + + + + + 2289504 + + 1607381 + 1171 + 6 + 1992 + 2289504 + 92299675 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1171-9 + 1179 + Wong DH, Brodsky FM + 0 + 100-kD proteins of Golgi- and trans-Golgi network-associated coated vesicles have related but distinct membrane binding properties + + + ABSTRACT + abstract + 131 + The 100-110-kD proteins (alpha-, beta-, beta'-, and gamma-adaptins) of clathrin-coated vesicles and the 110-kD protein (beta-COP) of the nonclathrin-coated vesicles that mediate constitutive transport through the Golgi have homologous protein sequences. To determine whether homologous processes are involved in assembly of the two types of coated vesicles, the membrane binding properties of their coat proteins were compared. After treatment of MDBK cells with the fungal metabolite Brefeldin A (BFA), beta-COP was redistributed to the cytoplasm within 15 s, gamma-adaptin and clathrin in the trans-Golgi network (TGN) dispersed within 30 s, but the alpha-adaptin and clathrin present on coated pits and vesicles derived from the plasma membrane remained membrane associated even after a 15-min exposure to BFA. In PtK1 cells and MDCK cells, BFA did not affect beta-COP binding or Golgi morphology but still induced redistribution of gamma-adaptin and clathrin from TGN membranes to the cytoplasm. Thus BFA affects the binding of coat proteins to membranes in the Golgi region (Golgi apparatus and TGN) but not plasma membranes. However, the Golgi binding interactions of beta- COP and gamma-adaptin are distinct and differentially sensitive to BFA. BFA treatment did not release gamma-adaptin or clathrin from purified clathrin-coated vesicles, suggesting that their distribution to the cytoplasm after BFA treatment of cells was due to interference with their rebinding to TGN membranes after a normal cycle of disassembly. This was confirmed using an in vitro assay in which gamma-adaptin binding to TGN membranes was blocked by BFA and enhanced by GTP gamma S, similar to the binding of beta-COP to Golgi membranes. These results suggest the involvement of GTP-dependent proteins in the association of the 100-kD coat proteins with membranes in the Golgi region of the cell. + + 281706 + Gene + 3499 + + beta-COP + + + 281706 + Gene + 3499 + + beta-COP + + + 281706 + Gene + 3499 + + beta-COP + + + 281706 + Gene + 3499 + + beta- COP + + + 281706 + Gene + 3499 + + beta-COP + + + CVCL:0422 + CellLine + + MDCK + + + MESH:D020126 + Chemical + + Brefeldin A + + + MESH:D006160 + Chemical + + GTP + + + MESH:D020126 + Chemical + + BFA + + + MESH:D020126 + Chemical + + BFA + + + MESH:D020126 + Chemical + + BFA + + + MESH:D006160 + Chemical + + GTP + + + MESH:D020126 + Chemical + + BFA + + + MESH:D013455 + Chemical + + S + + + MESH:C536528 + Disease + + pits + + + MESH:D020126 + Chemical + + BFA + + + MESH:D020126 + Chemical + + BFA + + + MESH:D020126 + Chemical + + BFA + + + CVCL:0421 + CellLine + + MDBK + + + MESH:D020126 + Chemical + + BFA + + + + + 2289505 + + 1607391 + 1311 + 6 + 1992 + 2289505 + 92299687 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1311-20 + 1320 + Murray BA, Jensen JJ + 0 + Evidence for heterophilic adhesion of embryonic retinal cells and neuroblastoma cells to substratum-adsorbed NCAM + + MESH:D009447 + Disease + + neuroblastoma + + + MESH:D012173 + Disease + + embryonic retinal + + + + ABSTRACT + abstract + 114 + The adhesion of embryonic chicken retinal cells and mouse N2A neuroblastoma cells to purified embryonic chicken retinal NCAM adsorbed on a solid substratum was examined using a quantitative centrifugal adhesion assay. Both cell types adhered to NCAM and the adhesion was specifically inhibited by monovalent anti-NCAM antibody fragments. N2A cell adhesion depended on the amount of NCAM applied to the substratum, was cation independent, and was insensitive to treatment with the cytoskeletal perturbing drugs colchicine and cytochalasin D. These results indicated that the tubulin and actin cytoskeletons were not critically required for adhesion to NCAM and make it unlikely that the cell surface ligand for NCAM is an integrin. Adhesion was however temperature dependent, strengthening greatly after a brief incubation at 37 degrees C. CHO cells transfected with NCAM cDNAs did not adhere specifically to substratum-bound NCAM and pretreatment of N2A cells and retinal cells with anti-NCAM antibodies did not inhibit adhesion to substratum-bound NCAM. These results suggest that a heterophilic interaction between substratum-adsorbed NCAM and a non-NCAM ligand on the surface of the probe cells affects adhesion in this system and support the possibility that heterophilic adhesion may be a function of NCAM in vivo. + + 428253 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 17967 + Gene + 40754 + + NCAM + + + 9031 + Species + + chicken + + + 10090 + Species + + mouse + + + 9031 + Species + + chicken + + + MESH:D009447 + Disease + + neuroblastoma + + + CVCL:G626 + CellLine + + N2A + + + CVCL:0213 + CellLine + + CHO + + + CVCL:G626 + CellLine + + N2A + + + MESH:D003078 + Chemical + + colchicine + + + CVCL:G626 + CellLine + + N2A + + + + + 2289506 + + 1318884 + 1299 + 6 + 1992 + 2289506 + 92299686 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1299-310 + 1310 + Hennemann H, Suchyna T, Lichtenberg-Fraté H, Jungbluth S, Dahl E, Schwarz J, Nicholson BJ, Willecke K + 0 + Molecular cloning and functional expression of mouse connexin40, a second gap junction gene preferentially expressed in lung + + 14613 + Gene + 3856 + + connexin40 + + + 10090 + Species + + mouse + + + + ABSTRACT + abstract + 125 + From a mouse genomic library, a clone has been isolated that codes for a connexin-homologous sequence of 358 amino acids. Because of its theoretical molecular mass of 40.418 kD it is named connexin40 (Cx40). Based on both protein and nucleotide sequence, mouse Cx40 is more closely related to mouse Cx43 (alpha subgroup of connexins) than to mouse Cx32 (beta subgroup). The highest overall homology detected, however, was to chick Cx42 (67% amino acid and 86% nucleotide identity), raising the possibility that Cx40 may be the mouse analogue. The coding region of Cx40 is uninterrupted by introns and is detected as a single copy gene in the mouse genome. High stringency hybridization of Northern blots with the coding sequence of Cx40 identified a single transcript of 3.5 kb that is at least 16-fold more abundant in lung-similar to mouse Cx37-than in other adult tissues (kidney, heart, and skin). In embryonic kidney, skin, and liver the level of the Cx40 transcript is two- to fourfold higher than in the corresponding adult tissues. Microinjection of Cx40 cRNA into Xenopus oocytes induced functional cell-to-cell channels between pairs. These channels show a symmetrical and markedly cooperative closure in response to transjunctional voltage (Boltzmann parameters of Vo = +/- 35 mV; A = 0.32) which is also fast relative to other connexin channels recorded similarly (tau = 580 ms at Vj of +/- 50 mV). Although Cx40- expressing oocytes did not couple efficiently with oocytes expressing endogenous connexins, they did couple well to Cx37-expressing oocytes. The heterotypic channels which formed had voltage-gating properties modified from those of the original homotypic forms. Transfection of mouse Cx40 DNA, under control of the SV-40 early promoter, into coupling-deficient human HeLa or SK-Hep-1 cells resulted in expression of the expected transcript and restoration of fluorescent dye transfer in transfected clones. + + 14613 + Gene + 3856 + + connexin40 + + + 14613 + Gene + 3856 + + Cx40 + + + 14613 + Gene + 3856 + + Cx40 + + + 14611 + Gene + 9670 + + Cx43 + + + 14618 + Gene + 137 + + Cx32 + + + 396502 + Gene + 3856 + + Cx42 + + + 14613 + Gene + 3856 + + Cx40 + + + 14613 + Gene + 3856 + + Cx40 + + + 14613 + Gene + 3856 + + Cx40 + + + 14612 + Gene + 1556 + + Cx37 + + + 14613 + Gene + 3856 + + Cx40 + + + 108707841 + Gene + + Cx40 + + + 14613 + Gene + 3856 + + Cx40 + + + 14612 + Gene + 1556 + + Cx37 + + + 14613 + Gene + 3856 + + Cx40 + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 9031 + Species + + chick + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 8355 + Species + + Xenopus + + + MESH:D007674 + Disease + + embryonic kidney + + + CVCL:0525 + CellLine + + SK-Hep-1 + + + CVCL:0030 + CellLine + + HeLa + + + + + 2289507 + + 1607388 + 1263 + 6 + 1992 + 2289507 + 92299683 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1263-75 + 1275 + Navone F, Niclas J, Hom-Booher N, Sparks L, Bernstein HD, McCaffrey G, Vale RD + 0 + Cloning and expression of a human kinesin heavy chain gene: interaction of the COOH-terminal domain with cytoplasmic microtubules in transfected CV-1 cells + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 9606 + Species + + human + + + CVCL:0229 + CellLine + + CV-1 + + + + ABSTRACT + abstract + 156 + To understand the interactions between the microtubule-based motor protein kinesin and intracellular components, we have expressed the kinesin heavy chain and its different domains in CV-1 monkey kidney epithelial cells and examined their distributions by immunofluorescence microscopy. For this study, we cloned and sequenced cDNAs encoding a kinesin heavy chain from a human placental library. The human kinesin heavy chain exhibits a high level of sequence identity to the previously cloned invertebrate kinesin heavy chains; homologies between the COOH-terminal domain of human and invertebrate kinesins and the nonmotor domain of the Aspergillus kinesin-like protein bimC were also found. The gene encoding the human kinesin heavy chain also contains a small upstream open reading frame in a G-C rich 5' untranslated region, features that are associated with translational regulation in certain mRNAs. After transient expression in CV-1 cells, the kinesin heavy chain showed both a diffuse distribution and a filamentous staining pattern that coaligned with microtubules but not vimentin intermediate filaments. Altering the number and distribution of microtubules with taxol or nocodazole produced corresponding changes in the localization of the expressed kinesin heavy chain. The expressed NH2-terminal motor and the COOH-terminal tail domains, but not the alpha-helical coiled coil rod domain, also colocalized with microtubules. The finding that both the kinesin motor and tail domains can interact with cytoplasmic microtubules raises the possibility that kinesin could crossbridge and induce sliding between microtubules under certain circumstances. + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 3799 + Gene + 55829 + + kinesin heavy chain + + + 3799 + Gene + 55829 + + kinesin heavy chain + + + CVCL:0229 + CellLine + + CV-1 + + + MESH:D015739 + Chemical + + nocodazole + + + CVCL:0229 + CellLine + + CV-1 + + + MESH:D017239 + Chemical + + taxol + + + + + 2289508 + + 1376732 + 1343 + 6 + 1992 + 2289508 + 92299690 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1343-50 + 1350 + Hardwick C, Hoare K, Owens R, Hohn HP, Hook M, Moore D, Cripps V, Austen L, Nance DM, Turley EA + 0 + Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility [published erratum appears in J Cell Biol 1992 Aug;118(3):753] + + MESH:D009369 + Disease + + tumor + + + + ABSTRACT + abstract + 146 + A cDNA encoding a unique hyaluronan receptor has been molecularly cloned from a lambda GT11 3T3 cDNA expression library. Immunoblot analyses of cell lysates, using antibodies to peptides encoded in the cDNA, specifically react with a 58-kD protein. This protein is regulated by the mutant H-ras gene in cells containing a metallothionein promoter H-ras hybrid gene. Further, antibodies to peptide sequences encoded in the cDNA block the increase in locomotion resulting from induction of the mutant H-ras gene in this cell line. In a transblot assay, the bacterially expressed protein binds to biotinylated hyaluronan. Antibodies to peptides encoded in the cDNA react in immunoblot assays with the 58- and 52-kD proteins of a novel hyaluronan receptor complex previously implicated in cell locomotion. Furthermore, antibodies specific to the 58- and 52-kD proteins, which block ras-induced locomotion, also cross-react with the expressed, encoded protein. The gene product described here appears to be a new type of hyaluronan receptor that is involved in cell locomotion. It is named RHAMM, an acronym for receptor for hyaluronan-mediated motility. + + 3265 + Gene + 55890 + + H-ras + + + 3265 + Gene + 55890 + + H-ras + + + 3161 + Gene + 8271 + + RHAMM + + + 3161 + Gene + 8271 + + receptor for hyaluronan-mediated motility + + + 3265 + Gene + 55890 + + H-ras + + + MESH:D006820 + Chemical + + hyaluronan + + + + + 2289509 + + 1607386 + 1241 + 6 + 1992 + 2289509 + 92299681 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1241-9 + 1249 + Doberstein SK, Pollard TD + 0 + Localization and specificity of the phospholipid and actin binding sites on the tail of Acanthamoeba myosin IC + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D000562 + Disease + + Acanthamoeba myosin IC + + + + ABSTRACT + abstract + 111 + We used bacterially expressed beta-galactosidase fusion proteins to localize the phospholipid binding domain of Acanthamoeba myosin IC to the region between amino acids 701 and 888 in the NH2-terminal half of the tail. Using a novel immobilized ligand lipid binding assay, we determined that myosin I can bind to several different acidic phospholipids, and that binding requires a minimum of 5 mol% acidic phospholipid in a neutral lipid background. The presence of di- and triglycerides and sterols in the lipid bilayer do not contribute to the affinity of myosin I for membranes. We confirm that the ATP-insensitive actin binding site is contained in the COOH-terminal 30 kD of the tail as previously shown for Acanthamoeba myosin IA. We conclude that the association of the myosin IC tail with acidic phospholipid head groups supplies much of the energy for binding myosin I to biological membranes, but probably not specificity for targeting myosin I isoforms to different cellular locations. + + 2720 + Gene + 47922 + + beta-galactosidase + + + 4641 + Gene + 32046 + + myosin IC + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + MESH:D008055 + Chemical + + lipid + + + MESH:D000255 + Chemical + + ATP + + + MESH:D013261 + Chemical + + sterols + + + MESH:D008055 + Chemical + + lipid + + + MESH:D000562 + Disease + + Acanthamoeba myosin IA + + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D008055 + Chemical + + lipid + + + MESH:D010743 + Chemical + + phospholipid + + + MESH:D010743 + Chemical + + phospholipids + + + MESH:D000562 + Disease + + Acanthamoeba myosin IC + + + MESH:D010743 + Chemical + + phospholipid + + + - + Chemical + + di- and triglycerides + + + + + 2289510 + + 1607390 + 1289 + 6 + 1992 + 2289510 + 92299685 + front + 117 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jun; 117(6) 1289-98 + 1298 + Dentler WL, Adams C + 0 + Flagellar microtubule dynamics in Chlamydomonas: cytochalasin D induces periods of microtubule shortening and elongation; and colchicine induces disassembly of the distal, but not proximal, half of the flagellum + + MESH:D015638 + Chemical + + cytochalasin D + + + MESH:D003078 + Chemical + + colchicine + + + + ABSTRACT + abstract + 212 + To study the mechanisms responsible for the regulation of flagellar length, we examined the effects of colchicine and Cytochalasin D (CD) on the growth and maintenance of Chlamydomonas flagella on motile wild type cells as well as on pf 18 cells, whose flagella lack the central microtubules and are immobile. CD had no effect on the regeneration of flagella after deflagellation but it induced fully assembled flagella to shorten at an average rate of 0.03 microns-min. Cells remained fully motile in CD and even stubby flagella continued to move, indicating that flagellar shortening did not selectively disrupt machinery necessary for motility. To observe the effects of the drug on individual cells, pf 18 cells were treated with CD and flagella on cells were monitored by direct observation over a 5-hour period. Flagella on control pf 18 cells maintained their initial lengths throughout the experiment but flagella on CD-treated cells exhibited periods of elongation, shortening, and regrowth suggestive of the dynamic behavior of cytoplasmic microtubules observed in vitro and in vitro. Cells behaved individually, with no two cells exhibiting the same flagellar behavior at any given time although both flagella on any single cell behaved identically. The rate of drug-induced flagellar shortening and elongation in pf 18 cells varied from 0.08 to 0.17 microns-min-1, with each event occurring over 10-60-min periods. Addition of colchicine to wild type and pf 18 cells induced flagella to shorten at an average rate of 0.06 microns-min-1 until the flagella reached an average of 73% of their initial length, after which they exhibited no further shortening or elongation. Cells treated with colchicine and CD exhibited nearly complete flagellar resorption, with little variation in flagellar length among cells. The effects of these drugs were reversible and flagella grew to normal stable lengths after drug removal. Taken together, these results show that the distal half to one-third of the Chlamydomonas flagellum is relatively unstable in the presence of colchicine but that the proximal half to two-thirds of the flagellum is stable to this drug. In contrast to colchicine, CD can induce nearly complete flagellar microtubule disassembly as well as flagellar assembly. Flagellar microtubules must, therefore, be inherently unstable, and flagellar length is stabilized by factors that are sensitive, either directly or indirectly, to the effects of CD. + + Disease + + flagella + + + MESH:D003078 + Chemical + + colchicine + + + Disease + + stubby flagella + + + MESH:D015638 + Chemical + + CD + + + MESH:D001259 + Disease + + flagella lack the central microtubules + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + + MESH:D003078 + Chemical + + colchicine + + + Disease + + flagella + + + MESH:D015638 + Chemical + + Cytochalasin D + + + MESH:D015638 + Chemical + + CD + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + + MESH:D003078 + Chemical + + colchicine + + + MESH:D003078 + Chemical + + colchicine + + + MESH:D015638 + Chemical + + CD + + + MESH:C535850 + Disease + + flagellar shortening + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + Disease + + flagella + + + MESH:D003078 + Chemical + + colchicine + + + MESH:C535850 + Disease + + flagellar shortening + + + MESH:D015638 + Chemical + + CD + + + Disease + + flagella + + + + + 2289512 + + 1352300 + 51 + 1 + 1992 + 2289512 + 92317162 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 51-62 + 62 + Low SH, Tang BL, Wong SH, Hong W + 0 + Selective inhibition of protein targeting to the apical domain of MDCK cells by brefeldin A + + CVCL:0422 + CellLine + + MDCK + + + MESH:D020126 + Chemical + + brefeldin A + + + + ABSTRACT + abstract + 92 + Dipeptidyl peptidase IV (DPPIV) is mainly vectorially targeted to the apical surface in MDCK cells. BFA was found to abolish the apical targeting of DPPIV. This BFA effect could be achieved under conditions where the ER to Golgi transport and the total surface expression of DPPIV were essentially unaffected. BFA executed its effect during the transport from the trans-Golgi network (TGN) to the surface. The inhibition of apical targeting resulted in enhanced mis-targeting to the basolateral surface. The mistargeted DPPIV was transcytosed back to the apical domain only after BFA withdrawal. In contrast, the basolateral targeting of uvomorulin was unaffected by BFA. These results established that the apical targeting of DPPIV was selectively abolished by BFA. + + 442858 + Gene + 20917 + + uvomorulin + + + CVCL:0422 + CellLine + + MDCK + + + + + 2289513 + + 1320035 + 43 + 1 + 1992 + 2289513 + 92317161 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 43-50 + 50 + Lotti LV, Torrisi MR, Pascale MC, Bonatti S + 0 + Immunocytochemical analysis of the transfer of vesicular stomatitis virus G glycoprotein from the intermediate compartment to the Golgi complex + + 11276 + Species + + vesicular stomatitis virus + + + + ABSTRACT + abstract + 144 + We performed an immunocytochemical analysis to study the transfer of a marker protein (G glycoprotein coded by vesicular stomatitis virus ts 045 strain) from the intermediate compartment to the Golgi stacks in infected Vero cells. The intermediate compartment seemed to consist of about 30-40 separate units of clustered small vesicles and short tubules. The units contained Rab2 protein and were spread throughout the cytoplasm, with a ratio of about 6:4 in the peripheral versus perinuclear site. Time-course experiments revealed a progressive transfer of G glycoprotein from the intermediate compartment to the Golgi stacks, while the tubulo-vesicular units did not appear to change their intracellular distribution. Moreover, the labeling density of peripheral and perinuclear units decreased in parallel during the transfer. These results support the notion that the intermediate compartment is a station in the secretory pathway, and that a vesicular transport connects this station to the Golgi complex. + + 11276 + Species + + vesicular stomatitis virus + + + 11276 + Species + + vesicular + + + 11276 + Species + + vesicular + + + + + 2289514 + + 1618904 + 203 + 1 + 1992 + 2289514 + 92317158 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 203-12 + 212 + Metsäranta M, Garofalo S, Decker G, Rintala M, de Crombrugghe B, Vuorio E + 0 + Chondrodysplasia in transgenic mice harboring a 15-amino acid deletion in the triple helical domain of pro alpha 1(II) collagen chain + + 10090 + Species + + transgenic mice + + + MESH:D010009 + Disease + + Chondrodysplasia + + + + ABSTRACT + abstract + 134 + We have generated transgenic mice by microinjection of a 39-kb mouse pro alpha 1(II) collagen gene construct containing a deletion of exon 7 and intron 7. This mutation was expected to disturb the assembly and processing of the homotrimeric type II collagen molecule in cartilage. Expression of transgene mRNA at levels equivalent or higher than the endogenous mRNA in the offspring of two founder animals resulted in a severe chondrodysplastic phenotype with short limbs, hypoplastic thorax, abnormal craniofacial development, and other skeletal deformities. The affected pups died at birth due to respiratory distress. Light microscopy of epiphyseal growth plates of transgenic pups demonstrated a marked reduction in cartilaginous extracellular matrix and disruption of the normal organization of the growth plate. The zone of proliferating chondrocytes was greatly reduced whereas the zone of hypertrophic chondrocytes was markedly increased extending deep into the diaphysis suggestive of a defect in endochondral ossification. Electron microscopic examination revealed chondrocytes with extended RER, a very severe reduction in the amount of cartilage collagen fibrils, and abnormalities in their structure. We postulate that the deletion in the alpha 1(II) collagen acts as a dominant negative mutation disrupting the assembly and secretion of type II collagen molecules. The consequences of the mutation include interference with normal endochondral ossification. These mice constitute a valuable model to study the mechanisms underlying human chondrodysplasias and normal bone formation. + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + MESH:D000741 + Disease + + hypoplastic + + + MESH:D019465 + Disease + + abnormal craniofacial development + + + MESH:D010009 + Disease + + chondrodysplasias + + + MESH:D006984 + Disease + + hypertrophic + + + MESH:C535964 + Disease + + cartilage collagen fibrils + + + MESH:D009139 + Disease + + skeletal deformities + + + MESH:D015831 + Disease + + cartilaginous + + + MESH:D002357 + Disease + + cartilage + + + Disease + + chondrodysplastic + + + + + 2289515 + + 1618896 + 1 + 1 + 1992 + 2289515 + 92317148 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 1-9 + 9 + Dargemont C, Kühn LC + 0 + Export of mRNA from microinjected nuclei of Xenopus laevis oocytes + + 8355 + Species + + Xenopus laevis + + + + ABSTRACT + abstract + 67 + Export of mRNA from the nucleus to the cytoplasm was studied in mature Xenopus laevis oocytes. In vitro transcribed, capped 32P-labeled mRNA was microinjected into nuclei, and its appearance in the cytoplasm measured by counting radioactivity or by RNA extraction and gel electrophoresis. Both for a 5.0-kb transferrin receptor mRNA and a 2.0- kb 4F2 antigen heavy chain mRNA we found saturable transport with an apparent Km of 3.6 x 10(8) molecules per oocyte nucleus. Under non- saturating conditions the half-time for mRNA export from the nucleus was approximately 2 min at 20 degrees C. At higher concentrations of injected mRNA this half-time was prolonged, and the maximal transport rate was reached at approximately 1.6 x 10(8) molecules/min. mRNA transport showed properties of an energy-dependent mechanism, since it was inhibited at 4 degrees C or by ATP depletion. Co-injection of the cap dinucleotide m7GpppG blocked the export effectively, suggesting a role for the cap in this process. The export was also inhibited by the pre-injection of wheat germ agglutinin. The effect of the lectin was specific and abolished by co-injection of N-acetylglucosamine. Finally, we found significant competitive inhibition in mRNA export by the presence of tRNA. Our results suggest that mRNA transport is a facilitated process which may share common steps with tRNA transport. Preliminary gel retardation experiments show that injected mRNA associates with endogenous nuclear proteins and suggest an exchange of some of the bound components during the transport to the cytoplasm. + + 379502 + Gene + + transferrin + + + 379133 + Gene + + 4F2 + + + 8355 + Species + + Xenopus laevis + + + 4565 + Species + + wheat + + + MESH:D008607 + Disease + + retardation + + + MESH:D000117 + Chemical + + N-acetylglucosamine + + + MESH:D000255 + Chemical + + ATP + + + MESH:C000615311 + Chemical + + 32P + + + - + Chemical + + dinucleotide m7GpppG + + + + + 2289517 + + 1618903 + 177 + 1 + 1992 + 2289517 + 92317156 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 177-94 + 194 + Frei T, von Bohlen und Halbach F, Wille W, Schachner M + 0 + Different extracellular domains of the neural cell adhesion molecule (N- CAM) are involved in different functions + + 4684 + Gene + 40754 + + neural cell adhesion molecule + + + 4684 + Gene + 40754 + + N- CAM + + + + ABSTRACT + abstract + 114 + The neural cell adhesion molecule (N-CAM) engages in diverse functional roles in neural cell interactions. Its extracellular part consists of five Ig-like domains and two fibronectin type III homologous (type III) repeats. To investigate the functional properties of the different structural domains of the molecule in cell interactions and signal transduction to the cell interior, we have synthesized, in a bacterial expression system, the individual domains and tandem sets of individual domains as protein fragments. These protein fragments were tested for their capacity to influence adhesion and spreading of neuronal cell bodies, promote neurite outgrowth, and influence cellular migration patterns from cerebellar microexplants in vitro. Ig-like domains I and II and the combined type III repeats I-II were most efficient for adhesion of neuronal cell bodies, when coated as substrates. Neurite outgrowth was best on the substrate-coated combined type III repeats I- II, followed by the combined Ig-like domains I-V and Ig-like domain I. Spreading of neuronal cell bodies was best on substrate-coated combined type III repeats I-II, followed by Ig-like domain I and the combined Ig- like domains I-V. The cellular migration pattern from cerebellar microexplant cultures plated on a mixture of laminin and poly-L-lysine was modified by Ig-like domains I, III, and IV, while Ig-like domains II and V and the combined type III repeats I-II did not show significant modifications, when added as soluble fragments. Outgrowth of astrocytic processes from the explant core was influenced only by Ig- like domain I. Metabolism of inositol phosphates was strongly increased by Ig-like domain I and less by the Ig-like domains II, III, IV, and V, and not influenced by the combined type III repeats I-II. Intracellular concentrations of Ca2+ and pH values were increased only by the Ig-like domains I and II. Intracellular levels of cAMP and GMP were not influenced by any protein fragment. These experiments indicate that different domains of N-CAM subserve different functional roles in cell recognition and signal transduction, and are functionally competent without nervous system-derived carbohydrate structures. + + 4684 + Gene + 40754 + + neural cell adhesion molecule + + + 4684 + Gene + 40754 + + N-CAM + + + 2335 + Gene + 1533 + + fibronectin + + + 4684 + Gene + 40754 + + N-CAM + + + MESH:D000069285 + Chemical + + Ca2+ + + + MESH:D002241 + Chemical + + carbohydrate + + + MESH:C066524 + Chemical + + GMP + + + MESH:D007295 + Chemical + + inositol phosphates + + + - + Chemical + + cAMP + + + + + 2289518 + + 1618905 + 23 + 1 + 1992 + 2289518 + 92317159 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 23-32 + 32 + Goday C, González-García JM, Esteban MR, Giovinazzo G, Pimpinelli S + 0 + Kinetochores and chromatin diminution in early embryos of Parascaris univalens + + + ABSTRACT + abstract + 79 + In Parascaris the mitotic chromosomes of gonial germline cells are holocentric and possess a continuous kinetochore along their entire length. By contrast, in meiotic cells, the centromeric activity is restricted to the heterochromatic tips where direct insertion of spindle microtubules into chromatin without any kinetochore plate is seen. In the presomatic cells of early embryos, which undergo heterochromatin elimination, only euchromatin shows kinetic activity. After developing a technique to separate the very resistant egg shell from the embryos, we studied the cell divisions during early embryogenesis by immunochemical and EM approaches. The results reported here show that in presomatic cells microtubules bind only the euchromatin where a continuous kinetochore plate is present. We also report observations suggesting that the binding of the long kinetochores to the mitotic spindle initiates to a limited number of sites and extends along the entire length, during chromosome condensation. The existence of different centromere stages in different cell types, rends Parascaris chromosomes a very good model to study centromere organization. + + 6257 + Species + + Parascaris + + + 6257 + Species + + Parascaris + + + MESH:D060725 + Disease + + heterochromatic tips + + + + + 2289519 + + 1618902 + 163 + 1 + 1992 + 2289519 + 92317155 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 163-76 + 176 + Wang L, Feng Y, Denburg JL + 0 + A multifunctional cell surface developmental stage-specific antigen in the cockroach embryo: involvement in pathfinding by CNS pioneer axons + + + ABSTRACT + abstract + 141 + mAb DSS-8 binds to a 164-kD developmental stage-specific cell surface antigen in the nervous system of the cockroach, Periplaneta americana. The antigen is localized to different subsets of cells at various stages of development. The spatial and temporal distributions of DSS-8 binding were determined and are consistent with this antigen playing multiple roles in the development of the nervous system. Direct identification of some of these functions was made by perturbation experiments in which pioneer axon growth occurs in embryos that are cultured in vitro in the presence of mAb DSS-8 or its Fab fragment. Under these conditions the pioneer axons of the median fiber tract grow but follow altered pathways. In a smaller percentage of the ganglia, the immunoreagents additionally produce defasciculation of a subset of DSS-8 labeled axons. Therefore, direct roles for the DSS-8 antigen in both the guidance of pioneer axons and selective fasciculation have been demonstrated. + + 2187 + Gene + 51880 + + Fab + + + 6978 + Species + + Periplaneta americana + + + - + Chemical + + DSS-8 + + + MESH:D005207 + Disease + + fasciculation + + + Disease + + defasciculation + + + + + 2289520 + + 1618910 + 95 + 1 + 1992 + 2289520 + 92317166 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 95-108 + 108 + Roof DM, Meluh PB, Rose MD + 0 + Kinesin-related proteins required for assembly of the mitotic spindle + + + ABSTRACT + abstract + 70 + We identified two new Saccharomyces cerevisiae kinesin-related genes, KIP1 and KIP2, using polymerase chain reaction primers corresponding to highly conserved regions of the kinesin motor domain. Both KIP proteins are expressed in vivo, but deletion mutations conferred no phenotype. Moreover, kip1 kip2 double mutants and a triple mutant with kinesin- related kar3 had no synthetic phenotype. Using a genetic screen for mutations that make KIP1 essential, we identified another gene, KSL2, which proved to be another kinesin-related gene, CIN8. KIP1 and CIN8 are functionally redundant: double mutants arrested in mitosis whereas the single mutants did not. The microtubule organizing centers of arrested cells were duplicated but unseparated, indicating that KIP1 or CIN8 is required for mitotic spindle assembly. Consistent with this role, KIP1 protein was found to colocalize with the mitotic spindle. + + 852216 + Gene + 136830 + + KIP1 + + + 855948 + Gene + 137176 + + KIP2 + + + 852216 + Gene + 136830 + + kip1 + + + 855948 + Gene + 137176 + + kip2 + + + 852216 + Gene + 136830 + + KIP1 + + + 856648 + Gene + 136907 + + KSL2 + + + 856648 + Gene + 136907 + + CIN8 + + + 852216 + Gene + 136830 + + KIP1 + + + 856648 + Gene + 136907 + + CIN8 + + + 852216 + Gene + 136830 + + KIP1 + + + 856648 + Gene + 136907 + + CIN8 + + + 852216 + Gene + 136830 + + KIP1 + + + 4932 + Species + + Saccharomyces cerevisiae + + + Disease + + mitosis + + + + + 2289521 + + 1618898 + 11 + 1 + 1992 + 2289521 + 92317150 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 11-21 + 21 + Kambach C, Mattaj IW + 0 + Intracellular distribution of the U1A protein depends on active transport and nuclear binding to U1 snRNA + + 6626 + Gene + 3380 + + U1A + + + + ABSTRACT + abstract + 106 + Nuclear transport of the U1 snRNP-specific protein U1A has been examined. U1A moves to the nucleus by an active process which is independent of interaction with U1 snRNA. Nuclear localization requires an unusually large sequence element situated between amino acids 94 and 204 of the protein. U1A transport is not unidirectional. The protein shuttles between nucleus and cytoplasm. At equilibrium, the concentration of the protein in the nucleus and cytoplasm is not, however, determined solely by transport rates, but can be perturbed by introducing RNA sequences that can specifically bind U1A in either the nuclear or cytoplasmic compartment. Thus, U1A represents a novel class of protein which shuttles between cytoplasm and nucleus and whose intracellular distribution can be altered by the number of free binding sites for the protein present in the cytoplasm or the nucleus. + + 26871 + Gene + + U1 snRNA + + + 6626 + Gene + 3380 + + U1A + + + 6626 + Gene + 3380 + + U1A + + + 6626 + Gene + 3380 + + U1A + + + 6626 + Gene + 3380 + + U1A + + + 6626 + Gene + 3380 + + U1A + + + + + 2289522 + + 1618906 + 33 + 1 + 1992 + 2289522 + 92317160 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 33-42 + 42 + Fujishige A, Smith KR, Silen JL, Agard DA + 0 + Correct folding of alpha-lytic protease is required for its extracellular secretion from Escherichia coli + + + ABSTRACT + abstract + 106 + alpha-Lytic protease is a bacterial serine protease of the trypsin family that is synthesized as a 39-kD preproenzyme (Silen, J. L., C. N. McGrath, K. R. Smith, and D. A. Agard. 1988. Gene (Amst.). 69: 237- 244). The 198-amino acid mature protease is secreted into the culture medium by the native host, Lysobacter enzymogenes (Whitaker, D. R. 1970. Methods Enzymol. 19:599-613). Expression experiments in Escherichia coli revealed that the 166-amino acid pro region is transiently required either in cis (Silen, J. L., D. Frank, A. Fujishige, R. Bone, and D. A. Agard. 1989. J. Bacteriol. 171:1320-1325) or in trans (Silen, J. L., and D. A. Agard. 1989. Nature (Lond.). 341:462-464) for the proper folding and extracellular accumulation of the enzyme. The maturation process is temperature sensitive in E. coli; unprocessed precursor accumulates in the cells at temperatures above 30 degrees C (Silen, J. L., D. Frank, A. Fujishige, R. Bone, and D. A. Agard. 1989. J. Bacteriol. 171:1320-1325). Here we show that full- length precursor produced at nonpermissive temperatures is tightly associated with the E. coli outer membrane. The active site mutant Ser 195----Ala (SA195), which is incapable of self-processing, also accumulates as a precursor in the outer membrane, even when expressed at permissive temperatures. When the protease domain is expressed in the absence of the pro region, the misfolded, inactive protease also cofractionates with the outer membrane. However, when the folding requirement for either wild-type or mutant protease domains is provided by expressing the pro region in trans, both are efficiently secreted into the extracellular medium. Attempts to separate folding and secretion functions by extensive deletion mutagenesis within the pro region were unsuccessful. Taken together, these results suggest that only properly folded and processed forms of alpha-lytic protease are efficiently transported to the medium. + + 69 + Species + + Lysobacter enzymogenes + + + 562 + Species + + Escherichia coli + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + - + Chemical + + SA195 + + + p.S195A + ProteinMutation + + Ser 195----Ala + + + + + 2289524 + + 1618900 + 131 + 1 + 1992 + 2289524 + 92317152 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 131-8 + 138 + Gabellini N, Minozzi MC, Leon A, Dal Toso R + 0 + Nerve growth factor transcriptional control of c-fos promoter transfected in cultured spinal sensory neurons + + 396466 + Gene + 1876 + + Nerve growth factor + + + + ABSTRACT + abstract + 109 + High efficiency gene transfer (greater than 90%) in chicken dorsal root ganglion neurons has been obtained by DNA calcium phosphate co- precipitation, hence providing an important tool to study control of gene expression in primary neurons. Transfection with c-fos promoter sequences linked to the chloramphenicol acetyltransferase reporter gene showed that the serum responsive element functions as a strong transcriptional enhancer. Transcription from this element is developmentally regulated, and mediates the genetic response to nerve growth factor (NGF) in developing avian sensory neurons. Furthermore, NGF exerts a negative effect on transcription from the cyclic AMP responsive element, thereby supporting the involvement of tyrosine kinase activation by NGF in primary sensory neurons. + + 396466 + Gene + 1876 + + nerve growth factor + + + 396466 + Gene + 1876 + + NGF + + + 396466 + Gene + 1876 + + NGF + + + 396466 + Gene + 1876 + + NGF + + + 9031 + Species + + chicken + + + MESH:D000242 + Chemical + + cyclic AMP + + + MESH:C020243 + Chemical + + calcium phosphate + + + + + 2289525 + + 1618908 + 71 + 1 + 1992 + 2289525 + 92317164 + front + 118 + This article is distributed under the terms of an Attribution-Noncommercial-Share Alike-No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution-Noncommercial-Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). + TITLE + J. Cell Biol.; 1992 Jul; 118(1) 71-81 + 81 + Tilney LG, DeRosier DJ, Tilney MS + 0 + How Listeria exploits host cell actin to form its own cytoskeleton. I. Formation of a tail and how that tail might be involved in movement + + + ABSTRACT + abstract + 139 + After Listeria is phagocytosed by a macrophage, it dissolves the phagosomal membrane and enters the cytoplasm. The Listeria then nucleates actin filaments from its surface. These actin filaments rearrange to form a tail with which the Listeria moves to the macrophage surface as a prelude to spreading. Since individual actin filaments appear to remain in their same positions in the tail in vitro after extraction with detergent, the component filaments must be cross- bridged together. From careful examination of the distribution of actin filaments attached to the surface of Listeria and in the tail, and the fact that during and immediately after division filaments are not nucleated from the new wall formed during septation, we show how a cloud of actin filaments becomes rearranged into a tail simply by the mechanics of growth. From lineage studies we can relate the length of the tail to the age of the surface of Listeria and make predictions as to the ratio of Listeria with varying tail lengths at a particular time after the initial infection. Since we know that division occurs about every 50 min, after 4 h we would predict that if we started with one Listeria in a macrophage, 16 bacteria would be found, two with long tails, two with medium tails, four with tiny tails, and eight with no tails or a ratio of 1:1:2:4. We measured the lengths of the tails on Listeria 4 h after infection in serial sections and confirmed this prediction. By decorating the actin filaments that make up the tail of Listeria with subfragment 1 of myosin we find (a) that the filaments are indeed short (maximally 0.3 microns in length); (b) that the filament length is approximately the same at the tip and the base of the tail; and (c) that the polarity of these filaments is inappropriate for myosin to be responsible or to facilitate movement through the cytoplasm, but the polarity insures that the bacterium will be located at the tip of a pseudopod, a location that is essential for spreading to an adjacent cell. Putting all this information together we can begin to unravel the problem of how the Listeria forms the cytoskeleton and what is the biological purpose of this tail. Two functions are apparent: movement and pseudopod formation. + + 79784 + Gene + 23480 + + myosin + + + 79784 + Gene + 23480 + + myosin + + + + + 4272812 + + surname:Sanchez-Capelo;given-names:Amelia + surname:Munoz;given-names:Maria Dolores + surname:Antolin-Vallespin;given-names:Monica + 25497491 + surname:Giraldez-Perez;given-names:Rosa Maria + 2014 + 4272812 + 176 + 10.1186/s40478-014-0176-9 + Keywords + front + 176 + 2 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. + TITLE + Acta Neuropathol Commun; 2014 Dec 13; 2176. doi:10.1186/s40478-014-0176-9 + alpha-synuclein Parkinson's disease Lewy body Animal models Smad3 Rotenone + Giráldez-Pérez R, Antolín-Vallespín M, Muñoz M, Sánchez-Capelo A + 0 + Models of alpha-synuclein aggregation in Parkinson's disease + + 20617 + Gene + 293 + + alpha-synuclein + + + MESH:D010300 + Disease + + Parkinson's disease + + + + ABSTRACT + abstract + 61 + Parkinson's disease (PD) is not only characterized by motor disturbances but also, by cognitive, sensory, psychiatric and autonomic dysfunction. It has been proposed that some of these symptoms might be related to the widespread pathology of alpha-synuclein (alpha-syn) aggregation in different nuclei of the central and peripheral nervous system. However, the pathogenic formation of alpha-syn aggregates in different brain areas of PD patients is poorly understood. Most experimental models of PD are valuable to assess specific aspects of its pathogenesis, such as toxin-induced dopaminergic neurodegeneration. However, new models are required that reflect the widespread and progressive formation of alpha-syn aggregates in different brain areas. Such alpha-syn aggregation is induced in only a few animal models, for example perikaryon inclusions are found in rats administered rotenone, aggregates with a neuritic morphology develop in mice overexpressing either mutated or wild-type alpha-syn, and in Smad3 deficient mice, aggregates form extensively in the perikaryon and neurites of specific brain nuclei. In this review we focus on alpha-syn aggregation in the human disorder, its genetics and the availability of experimental models. Indeed, evidences show that dopamine (DA) metabolism may be related to alpha-syn and its conformational plasticity, suggesting an interesting link between the two pathological hallmarks of PD: dopaminergic neurodegeneration and Lewy body (LB) formation. + + 20617 + Gene + 293 + + alpha-synuclein + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 29219 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 6622 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + patients + + + 10116 + Species + + rats + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + MESH:D001523 + Disease + + psychiatric + + + MESH:D010300 + Disease + + PD + + + MESH:D001342 + Disease + + autonomic dysfunction + + + MESH:D010300 + Disease + + PD + + + MESH:D004298 + Chemical + + DA + + + MESH:D010300 + Disease + + Parkinson's disease + + + MESH:D010300 + Disease + + PD + + + MESH:D004298 + Chemical + + dopamine + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_1 + 1560 + Introduction + + + INTRO + paragraph + 1573 + While the first description of Parkinson's disease (PD) may date back to ancient Indian and Chinese texts from 1000 BC, the first clear medical description of this disorder was presented by James Parkinson in 1817. Some years later, in the mid-1800s, Jean-Martin Charcot separated PD from multiple sclerosis and other disorders that are also characterized by tremor, and in 1895 Brissaud formulated the hypothesis that the substantia nigra (SN) is the main brain nucleus pathologically affected in PD. Subsequently, it was Friedrich Lewy who first described the protein aggregates that form in different areas of the brain of PD patients, including the dorsal vagal nucleus, locus coeruleus and globus pallidus. Not long after, Tretiakoff validated Brissaud's hypothesis in 1919 and, by examining post-mortem tissue, he described the protein aggregates in the SN and called them Lewy bodies. Despite this long history, even today the etiology of idiopathic PD remains unknown and given the diversity of the molecular mechanisms proposed, it has been suggested that multiple factors may cause the disease. + + 9606 + Species + + patients + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010302 + Disease + + Parkinson + + + MESH:D010300 + Disease + + PD + + + MESH:D009103 + Disease + + multiple sclerosis + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + Parkinson's disease + + + MESH:D014202 + Disease + + tremor + + + + INTRO + paragraph + 2683 + PD is the second most common neurodegenerative disorder that affects the human brain. It is primarily characterized by motor symptoms like akinesia, rigidity, resting tremor and postural instability, manifestations that are mainly derived from the progressive degeneration of dopaminergic neurons in the SN pars compacta. Non-motor symptoms also develop that are associated with cognitive deficits (ranging from memory impairment to dementia), emotional changes (depression, apathy and anxiety), sleep perturbations, autonomic dysfunction (bladder disturbances, orthostatic hypotension, sweating), sensory symptoms (pain, visual impairment, olfactory deficit, paresthesia, ageusia) and gastrointestinal symptoms (constipation, dribbling of saliva:. + + 9606 + Species + + human + + + MESH:D014202 + Disease + + tremor + + + MESH:D010300 + Disease + + PD + + + MESH:D019636 + Disease + + neurodegenerative disorder + + + MESH:D004409 + Disease + + akinesia + + + MESH:D009127 + Disease + + rigidity + + + + INTRO + paragraph + 3432 + Although the primary motor symptoms are shared by patients, both the full presentation of the disorder and the response to treatment are quite heterogeneous. It must be borne in mind that non-dopaminergic neuronal loss is also detected in some areas of the brain, for example, that of monoaminergic cells in the locus coeruleus and raphe nuclei, cholinergic cells in the nucleus basalis of Meynert and in the pedunculopontine tegmental nucleus, as well as the loss of hypocretin cells in the hypothalamus. Indeed, the other pathological changes observed are widespread, with the appearance of LB inclusions in different areas of the brain (mesostriatal system, cortex, thalamus, hypothalamus, olfactory bulb or brainstem), or alterations in the autonomic system (the spinal cord, sympathetic ganglia and myenteric plexus in the gastrointestinal tract). The widespread nature of this pathology is indicative that the disorder is not just a motor alteration but rather, a sensory, cognitive, psychiatric and autonomic disorder. + + 15171 + Gene + 1166 + + hypocretin + + + 9606 + Species + + patients + + + MESH:D007029 + Disease + + hypothalamus + + + MESH:D004067 + Disease + + gastrointestinal tract + + + MESH:D009410 + Disease + + neuronal loss + + + MESH:D001523 + Disease + + psychiatric and autonomic disorder + + + MESH:D007029 + Disease + + hypothalamus + + + + INTRO + title_1 + 4458 + Lewy bodies in PD + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 4476 + A common neuropathological feature of some neurodegenerative diseases is the presence of proteinaceous inclusion bodies caused by misfolded and intracellular aggregation of proteins in many brain regions. These abnormal protein deposits may provoke LB pathologies that involve the deposition of LBs in cell bodies, or the formation of Lewy neurites (LNs) and Papp-Lantos inclusions. While the presence of LBs is a histological hallmark of PD, they are also associated with disorders such as dementia with LBs, multiple system atrophy, Alzheimer's disease, Down's syndrome, neurodegeneration with brain iron accumulation type I (Hallervorden-Spatz disease), progressive autonomic failure, rapid eye movement sleep disorder, parkinsonism-dementia complex of Guam, Gaucher's disease or Pick's disease. + + MESH:D020774 + Disease + + Pick's disease + + + MESH:D010300 + Disease + + 's syndrome + + + MESH:D019636 + Disease + + neurodegenerative diseases + + + MESH:D006211 + Disease + + Hallervorden-Spatz disease + + + MESH:D020961 + Disease + + Lewy neurites + + + MESH:D006211 + Disease + + brain iron accumulation type I + + + MESH:D001284 + Disease + + atrophy + + + MESH:D003704 + Disease + + dementia + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D005776 + Disease + + Gaucher's disease + + + MESH:D000544 + Disease + + Alzheimer's disease + + + MESH:D010300 + Disease + + PD + + + MESH:D020187 + Disease + + rapid eye movement sleep disorder + + + MESH:C537240 + Disease + + parkinsonism-dementia + + + MESH:D012791 + Disease + + autonomic failure + + + + INTRO + title_2 + 5283 + LB morphology + + + INTRO + paragraph + 5297 + LBs are morphologically heterogeneous, with classic LB arising in the brainstem as cytoplasmic inclusions of 8-30 mum in diameter, with a dense eosinophilic core and a narrow pale stained rim. On haematoxylin/eosin staining, classic LBs are observed as a spherical body with a dense core surrounded by a halo, whereas the cortical LBs present in layers V-VI of the temporal, insular and cingulate cortex have no obvious halo. A third type of LBs are known as pale bodies, that are rounded, pale, eosinophilic granules which lack the eosinophilic core of classic LBs, and that are only weakly and diffusely stained with eosin. These pale bodies are thought to be precursor LBs. In addition, dystrophic LNs are present in axonal processes with both thread-like and spheroid forms. + + MESH:D020388 + Disease + + dystrophic LNs + + + MESH:D004801 + Chemical + + eosin + + + MESH:D006416 + Chemical + + haematoxylin + + + MESH:D004801 + Chemical + + eosin + + + + INTRO + paragraph + 6079 + In dementia with LB disease, inclusions seem to be morphologically homogeneous in the neocortical and paralimbic regions of the brain. In the SN, the LBs immunolabeled with alpha-syn and ubiquitin fall within the spectrum from diffuse to pale bodies, and to classic LBs. Indeed, this morphological diversity in the SN may represent different stages in the formation of LBs. + + 20617 + Gene + 293 + + alpha-syn + + + MESH:D003704 + Disease + + dementia + + + + INTRO + paragraph + 6453 + Electron microscopy has shown that the pale rim and dense core of classic LBs correspond to zones of radially-oriented straight filaments, and zones of circular profiles, respectively. Cortical LBs and LNs also contain filaments. Three-dimensional reconstruction from serial confocal images reflects that the LB core, immunolabeled with alpha-syn and ubiquitin, has a concentric layered structure, with neurofilament encircling these inner layers. Indeed, a frequent continuity between LBs and LNs has been detected, indicating that LNs may evolve into LBs. + + 20617 + Gene + 293 + + alpha-syn + + + + INTRO + title_2 + 7011 + LB composition + + + INTRO + paragraph + 7026 + LBs are thought to be mainly composed of alpha-syn and the morphological characterization of LBs is mostly based on immunochemistry for alpha-syn, ubiquitin and neurofilament. Furthermore, the alpha-syn in LBs undergoes post-translational modifications, such as phosphorylation, ubiquitination and oxidative nitration. However, LBs also contain many different proteins, such as the Leucine-rich repeat kinase 2 (LRRK2:, histone deacetylase 6 (HDAC6: and charged multivesicular body protein 2B (CHMP2B:, all of which are found in the core. Indeed, LRRK2 is associated with the endoplasmic reticulum of dopaminergic neurons, HDAC6 is considered a sensor of proteasomal inhibition that plays a central role in autophagy, and CHMP2B is a component of the endosomal sorting complex involved in protein degradation. The identification of proteins present in LBs may offer molecular clues to the processes that may participate in LBs formation. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + Leucine-rich repeat kinase 2 + + + 66725 + Gene + 18982 + + LRRK2 + + + 15185 + Gene + 31353 + + histone deacetylase 6 + + + 15185 + Gene + 31353 + + HDAC6 + + + 68942 + Gene + 8534 + + charged multivesicular body protein 2B + + + 68942 + Gene + 8534 + + CHMP2B + + + 66725 + Gene + 18982 + + LRRK2 + + + 15185 + Gene + 31353 + + HDAC6 + + + 68942 + Gene + 8534 + + CHMP2B + + + + INTRO + paragraph + 7964 + Nevertheless, the comprehensive molecular composition of LBs is still unclear, although new proteomics approaches based on mass spectrometry provide an interesting approach to discover proteins not yet described in aggregates. However, the success of such approaches will depend on the techniques used to isolate LBs. One study used cortical LB-enriched fractions derived from the LB variant of Alzheimer's disease to identify 40 LB proteins involved in phosphorylation, ubiquitination, oxidative stress or protein trafficking. Laser capture microdissection of cortical LBs from neurons located in the temporal cortex of dementia with LB disease patients, allowed 296 proteins to be detected, including the chaperone HSC71, confirming the presence of chaperone molecules in LBs. + + 3312 + Gene + 68524 + + HSC71 + + + 9606 + Species + + patients + + + MESH:D000544 + Disease + + Alzheimer's disease + + + MESH:D003704 + Disease + + dementia + + + + INTRO + title_2 + 8745 + Histological localization of LBs + + + INTRO + paragraph + 8778 + Stage 1: At the earliest stage, only the dorsal motor nucleus of the vagus nerve (in the lower medulla oblongata) and the anterior olfactory nucleus are affected, suggesting that the brain pathology originates in these structures. + + Disease + + oblongata + + + + INTRO + paragraph + 9009 + Stage 2: LB inclusions appear in the raphe nuclei, magnocellular reticular nuclei and locus coeruleus. + + + INTRO + paragraph + 9112 + Stage 3: The pathology occurs in the basal portions of the midbrain and forebrain, with the SNpc, amygdala, pedunculopontine tegmental nucleus, magnocellular basal forebrain nuclei, tuberomammillary nucleus and spinal cord being affected. + + + INTRO + paragraph + 9351 + Stage 4: Cortical LBs appear in the temporal mesocortex, in the allocortical CA2 of the hippocampus and in thalamic structures. + + 12349 + Gene + 37256 + + CA2 + + + + INTRO + paragraph + 9479 + Stage 5: The anterior cingulate, insular and subgenual mesocortex is affected, and the neocortex appears to be affected for the first time, with LBs in high order sensory association structures and in the prefrontal neocortex. The hippocampus is clearly affected in the CA1, CA3 and entorhinal cortex. + + 12346 + Gene + 20414 + + CA1 + + + 12350 + Gene + 31298 + + CA3 + + + + INTRO + paragraph + 9781 + Stage 6: LBs are present in the primary sensory areas (the auditory field), primary motor field and premotor neocortex. + + + INTRO + paragraph + 9901 + Studies of the topographic distribution of LBs during the course of sporadic PD has enabled a classification of the stages of disease progression to be drawn up. Neurons susceptible to contain LBs have long, thin, unmyelinated or poorly myelinated axons and the parkinsonian brains analyzed can nearly all be categorized into one of six different stages, based on the location of the inclusion bodies: + + MESH:D010302 + Disease + + parkinsonian + + + MESH:D010300 + Disease + + PD + + + + TABLE + Tab1.xml + Tab1 + table_caption + 10303 + Overview of LB-like formation in rodent models. Brain areas with the presence of DA receptors and LB-like and/or LN-like aggregates may suggest an interaction between the dopaminergic system and alpha-syn (see text) + + 20617 + Gene + 293 + + alpha-syn + + + + TABLE + Tab1 + table + Tab1.xml + 10521 + Brain Nucleus Braak's stage PrP-A53T-alpha-syn mice Rotenone in rats Smad3 null mice DA system X-XII 1 +++ LB- LN Yes OB 1 ++ LB-LN Yes sp5 2 ++ LN ++ LN Yes ll 2 ++ LN Yes Sc 2 ++++ LB-LN ++ LB Yes RF/LC 2 +++ LB- LN ++ LN Yes RN 2, 3 +++ LB-LN Yes VTA 3 + LB- LN ++ LN Yes CPu 3 ++ LB- LN +++++ LB- LN ++++ LN Yes SN 3 + LB- LN +++++ LB- LN +++++ LN Yes Hp 3,4 ++++ LB-LN Yes Th 4 +++ LB-LN Yes M1-M2 5,6 ++++ LB-LN Yes Cg 5,6 +++ LN Yes cc-ic-cp -- +++ LN Pn -- ++ LB Yes CB -- ++++ LB-LN ++ LN Yes Pir -- +++ LN Yes IC -- +++ LB-LN SC -- +++ LB-LN Yes + + 25631 + Gene + 55937 + + Smad3 + + + 110027 + Gene + + CPu 3 + + + 185869 + Species + + T-alpha + + + 10090 + Species + + mice + + + 10116 + Species + + rats + + + 10090 + Species + + mice + + + 16473 + Gene + + syn + + + MESH:D012402 + Chemical + + Rotenone + + + MESH:C028505 + Chemical + + Cg + + + + INTRO + paragraph + 11154 + This model reflects the pathological progression in PD patients (Braak PD stage 4-6), with a caudo-rostral gradient in LB deposition from the lower brainstem to the neocortex, and with both dopaminergic and non-dopaminergic areas being affected (Table 1). Some patients do not have clinical PD symptoms, but display alpha-syn deposition at autopsy in areas according to Braak PD stage 1-3. They are referred to as showing incidental LB disease (iLBD), considered a prodromal state of PD. + + 6622 + Gene + 293 + + alpha-syn + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 11644 + Besides neuronal alpha-syn deposition in LBs and LNs, the presence of inclusions in astrocytes has been detected by silver staining. Glial inclusions are widespread in PD, both in areas with neuronal loss and gliosis (substantia nigra, locus coeruleus and dorsal vagal nucleus), and in areas with no clear neurodegeneration or gliosis (cerebral cortex, cerebral white matter, striatum, globus pallidus, thalamus, cerebellum and spinal cord:. Indeed, alpha-syn-immunoreactive (-ir) astrocytes adopt a topographical distribution that closely parallels that of the cortical LBs. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + MESH:D012834 + Chemical + + silver + + + + INTRO + paragraph + 12220 + However, we should consider that this disorder has a very heterogeneous clinical manifestation and as yet, there is no clear significance attributed to the presence of LBs and LNs, as direct determinants of the clinical manifestations and symptoms evident in patients with PD. + + 9606 + Species + + patients + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_2 + 12497 + LB genetics + + + INTRO + paragraph + 12509 + It is considered that most patients with PD develop the idiopathic form of the disease as opposed to that which is genetically inherited. Although, 10-30% of PD patients report a first-degree relative with the disorder, this does not necessarily reflect genetic inheritance as it may correspond with exposure to a common environmental factor. However, it is increasingly clear that genetic factors contribute to the pathogenesis of PD and it is currently considered a multifactorial complex disorder, caused by interactions between genetic and environmental risk factors. Alternatively, only 10% of cases develop the rarer form of PD that follows Mendelian inheritance, representing 2% of late-onset and 50% of early-onset familial PD. Studies into these familial forms have identified several causative genes related to mitochondrial or lysosomal dysfunction, protein aggregation, the proteasome system and kinase signaling. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D066263 + Disease + + protein aggregation + + + MESH:D010300 + Disease + + familial PD + + + MESH:D016464 + Disease + + lysosomal dysfunction + + + + INTRO + paragraph + 13435 + LB deposition is associated with specific mutations in some of these already known genes, including alpha-syn (SNCA: and LRRK2. Moreover, the distinction between the familial and idiopathic form of PD is currently not clear in all cases, as some evidence suggests that SNCA or LRKK2 mutations also participate in the sporadic disease. + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 20617 + Gene + 293 + + SNCA + + + 20617 + Gene + 293 + + SNCA + + + + INTRO + title_3 + 13770 + SNCA + + + INTRO + paragraph + 13775 + Several dominant mutations have been described in the gene encoding alpha-syn, SNCA, with varying penetrance. In 1997, the A53T missense mutation was reported in a large Italian family associated with familial PD. Subsequently, two additional missense mutations in SNCA were also seen to be associated to familial PD, A30P and E46K, although these point mutations are extremely rare. Duplicate and triplicate loci of SNCA of different sizes (from 0.4 to 4.5 Mb) have been seen to give rise to PD, which has been related to the overexpression of the wild-type protein, influencing the age of onset and severity of the disorder. Indeed, SNCA duplications have also been reported in apparently sporadic PD patients and interestingly, patients carrying these mutations present a broad clinical phenotype, even within a given family, suggesting an influence of genetic modifiers. In addition, a dinucleotide repeat polymorphism (Rep1) has been described in the promoter region, 10 Kb upstream of the start codon, which induces SNCA overexpression and that may account for increased risk in 3% of non-familiar PD. Moreover, specific haplotypes, primarily in the 3' UTR region, may also be associated to sporadic PD. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + SNCA + + + 20617 + Gene + 293 + + SNCA + + + 20617 + Gene + 293 + + SNCA + + + 6622 + Gene + 293 + + SNCA + + + 104354 + Gene + + Rep1 + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 20617 + Gene + 293 + + SNCA + + + MESH:D010300 + Disease + + PD + + + MESH:D015226 + Chemical + + dinucleotide + + + p.A30P;RS#:104893878 + ProteinMutation + + A30P + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + p.E46K;RS#:104893875 + ProteinMutation + + E46K + + + MESH:D010300 + Disease + + PD + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + + INTRO + paragraph + 14985 + Although SNCA mutations are very rare, their identification led to the association of alpha-syn with LBs. This protein is predominantly localized in presynaptic nerve terminals and missense mutations may reduce its affinity for lipids, enhancing its propensity to adopt a beta-sheet conformation and promoting self-assemble into oligomers and fibril formation. In this sense, the A53T mutation of SNCA is associated with the predominantly neuritic aggregation of alpha-syn in the human brain and in a mouse model, as well as with severe motor impairments. Alternatively, SNCA overexpression may decrease the density of dopaminergic vesicles and of synaptic contacts. Neuropathological diagnosis of PD requires both dopaminergic neurodegeneration in the SNpc and the presence of LBs, and as described previously, the severity of the clinical manifestation has been correlated with the broad distribution of LBs, which may be related to the number of alleles. Both triplication and point mutations in SNCA are associated with the formation of cortical and subcortical LBs, and a clinical diagnosis of PD with dementia. + + 20617 + Gene + 293 + + SNCA + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + SNCA + + + 6622 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + SNCA + + + 9606 + Species + + human + + + 10090 + Species + + mouse + + + 20617 + Gene + 293 + + SNCA + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D008055 + Chemical + + lipids + + + MESH:D058225 + Disease + + neuritic aggregation + + + MESH:D003704 + Disease + + dementia + + + MESH:D009422 + Disease + + impairments + + + + INTRO + paragraph + 16102 + The SNCA gene and that encoding the microtubule-associated protein tau (MAPT) have consistently been associated in different populations of sporadic PD, mainly those of European origin. However, such an associated was not identified in Japanese population, suggesting that population-specific differences may exist in the genetics of PD. + + 20617 + Gene + 293 + + SNCA + + + 17762 + Gene + 74962 + + microtubule-associated protein tau + + + 17762 + Gene + 74962 + + MAPT + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_3 + 16440 + Leucine-rich repeat kinase 2 (LRRK2) + + 66725 + Gene + 18982 + + Leucine-rich repeat kinase 2 + + + 66725 + Gene + 18982 + + LRRK2 + + + + INTRO + paragraph + 16477 + Point mutations in the LRRK2 gene are frequently associated to PD, and they are found in both late onset familial and sporadic PD. LRRK2 mutations have the highest prevalence rate in PD patients discovered to date, having been found in 10% of cases with autosomal dominant familial PD, in 3.6% of sporadic PD cases and even in 1.8% of healthy controls. The presence of mutations in healthy controls may suggest a reduced and incomplete age-dependent penetrance. However, while over 80 missense variants have been identified, only seven mutations are considered pathogenic (N1437H, R1441G/C/H, Y1699C, G2019S and I2020T), most of them lying in the C-terminal half of the protein, while two others are considered as risk factors (G2385R and R1628P:. Of these, G2019S is the most frequent mutation in the Caucasian population, which explains 1% of PD cases. The prevalence of this mutation is strongly influenced by ethnicity and as such, it is more frequent in patients of Southern European or North African origin, and in Ashkenazi Jews (18-40% PD cases), yet it is very rare in Asians or Northern Europeans. The risk of PD for a person who inherits this mutation is 28% at age 59 years, 51% at 69 years and 74% at 79 years. The G2019S mutation gives rise to a uniform clinical phenotype that resembles sporadic PD, both in homozygous and heterozygous carriers. + + 66725 + Gene + 18982 + + LRRK2 + + + 120892 + Gene + 18982 + + LRRK2 + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + person + + + p.N1437H;RS#:74163686 + ProteinMutation + + N1437H + + + p.R1628P;RS#:33949390 + ProteinMutation + + R1628P + + + p.I2020T;RS#:35870237 + ProteinMutation + + I2020T + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:D010300 + Disease + + PD + + + p.Y1699C;RS#:35801418 + ProteinMutation + + Y1699C + + + p.G2385R;RS#:34778348 + ProteinMutation + + G2385R + + + MESH:D010300 + Disease + + PD + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:D010300 + Disease + + autosomal dominant familial PD + + + p.R1441G,C,H;RS#:33939927 + ProteinMutation + + R1441G/C/H + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + + INTRO + paragraph + 17841 + LRRK2 is a gene with 51 exons and it encodes a large protein with two different enzymatic activities in the same molecule: that of a kinase and a GTPase. LRRK2 also has multiple protein interaction domains suggesting that it may serve as a scaffold for the assembly of other proteins. The G2019S mutation affects the kinase domain of the protein, with other common mutations lying in the GTPase domain or in the protein interaction domains. The function of LRRK2 remains largely elusive but G2019S mutations enhance its kinase activity, which may mediate neural toxicity. LRRK2 is widely expressed in the healthy adult brain, mainly in the endoplasmic reticulum. Interestingly, in sporadic PD patients LRRK2 is found in the core of LBs in the SN and locus coeruleus, suggesting it may contribute to LB formation. Indeed, some PD patients with LRRK2 mutations have LB pathology but this is not always the case, even within a family. How LRRK2 might participate in LB formation remains unknown, although dysfunctional autophagy has been proposed, as well as altered solubility and aggregation of alpha-syn. + + 66725 + Gene + 18982 + + LRRK2 + + + 66725 + Gene + 18982 + + LRRK2 + + + 66725 + Gene + 18982 + + LRRK2 + + + 66725 + Gene + 18982 + + LRRK2 + + + 120892 + Gene + 18982 + + LRRK2 + + + 120892 + Gene + 18982 + + LRRK2 + + + 66725 + Gene + 18982 + + LRRK2 + + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:C564093 + Disease + + dysfunctional autophagy + + + MESH:D010300 + Disease + + PD + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:D010300 + Disease + + PD + + + MESH:D064420 + Disease + + toxicity + + + + INTRO + title_3 + 18946 + Recessive mutations + + + INTRO + paragraph + 18966 + Homozygous or compound heterozygous mutations in the recessive genes Parkin, PINK and DJ-1 are associated to relatively rare forms of familial PD, and these result in early onset PD and nigral dopaminergic neuronal loss. Parkin mutations account for around 50% of familial juvenile and early onset PD, reaching 80% in patients with onset before the age of 20 and decreasing with increasing age at onset, becoming very rare when onset occurs after the age of 50. At least 170 mutations have been described in Parkin, including point mutations, exon rearrangements, "indels" and duplications. Homozygous mutations with loss-of-function predominantly induce neuronal loss in the SN and locus coeruleus, in the absence of LBs. However, compound heterozygous mutations with LB pathology have been described in a minority of patients. This variability may indicate some mutation-specific effect, as it has been described loss-of-function, inactivating and activating mutations. Parkin is a cytosolic E3 ubiquitin ligase that transfers ubiquitin to specific protein substrates for proteasomal and autophagic degradation. Mutations may impair Parkin activity, which might reduce the clearance and aggregation of proteins. As such, these mutations could participate in decreased alpha-synuclein clearance or alternatively, Parkin mutations might decrease its solubility and lead to aggregate formation. + + 57320 + Gene + 38295 + + DJ-1 + + + 20617 + Gene + 293 + + alpha-synuclein + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009410 + Disease + + neuronal loss + + + MESH:D010300 + Disease + + PD + + + MESH:D009410 + Disease + + neuronal loss + + + MESH:D010300 + Disease + + familial PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 20361 + Other recessive mutations are less common, such as PINK-1 and DJ-1, producing clinical manifestations broadly similar to Parkin mutations and in the absence of LBs. + + 68943 + Gene + 32672 + + PINK-1 + + + 57320 + Gene + 38295 + + DJ-1 + + + + INTRO + title_3 + 20526 + GBA mutations + + 14466 + Gene + 68040 + + GBA + + + + INTRO + paragraph + 20540 + Glucocerebrosidase (GBA) mutations were first described in Gaucher's disease, a recessive lysosomal storage disorder that may develop into parkinsonism, and PD patients may have an increased frequency of GBA mutations. Indeed, carriers of GBA mutations exhibit clinical features related to early-onset, levodopa-responsive PD, experiencing hallucinations and symptoms of cognitive decline or dementia, with the abundant presence of LBs in the neocortex. GBA mutations in PD patients confer increased susceptibility to an earlier disease onset, to have affected relatives and to develop atypical clinical manifestations. + + 2629 + Gene + 68040 + + Glucocerebrosidase + + + 2629 + Gene + 68040 + + GBA + + + 2629 + Gene + 68040 + + GBA + + + 14466 + Gene + 68040 + + GBA + + + 2629 + Gene + 68040 + + GBA + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D003072 + Disease + + cognitive decline + + + MESH:D016464 + Disease + + recessive lysosomal storage disorder + + + MESH:D006212 + Disease + + hallucinations + + + MESH:D007980 + Chemical + + levodopa + + + MESH:D010300 + Disease + + PD + + + MESH:D005776 + Disease + + Gaucher's disease + + + MESH:D003704 + Disease + + dementia + + + MESH:D010300 + Disease + + PD + + + MESH:D010302 + Disease + + parkinsonism + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_1 + 21162 + Animal models of alpha-syn aggregation + + 20617 + Gene + 293 + + alpha-syn + + + + INTRO + paragraph + 21201 + Animal models are used in order to study the neurobiological basis of human disorders and to develop new treatments. When examining the validity of an animal model we can consider face validity (similarity between neurological and behavioral phenotypes seen in an animal model and the human patient), construct validity (consistent with a theoretical rational, such that the molecular and cellular changes that result from genetic manipulations should be the same as those that occur in humans), and predictive validity (i.e. it should respond to pharmacological treatment as in humans). Face validity is a major criterion for model evaluation. One can argue that modeling of LB deposition and dopaminergic neurodegeneration may correlate to the human pathology. However, the behavioral phenotypes differ considerably between animals such as mice and humans. Thus, face validity may prove to be an unrealistic criterion for some symptoms of the disease such as motor and cognitive deficits or emotional changes. Although some researchers advocate the primacy of one of these approaches, in practice, the validity of a model should consider all three sources of evidence. + + 9606 + Species + + human + + + 9606 + Species + + human + + + 9606 + Species + + patient + + + 9606 + Species + + humans + + + 9606 + Species + + humans + + + 9606 + Species + + human + + + 10090 + Species + + mice + + + 9606 + Species + + humans + + + MESH:D003072 + Disease + + cognitive deficits + + + MESH:D019636 + Disease + + neurodegeneration + + + + INTRO + paragraph + 22372 + Considering the histopathological hallmarks of PD, models of the disorder should address the progressive loss of dopaminergic neurons in the SNpc, reduced striatal dopamine content and altered catabolism, and it should be age dependent and with progressive alteration of the animal's movement. Furthermore, the Braak staging scheme has strongly influenced what we think would be a good animal model for PD. Indeed, the presence of LBs and LNs, the post-translational modifications observed in the human disorder, and the composition and histological localization of LBs are all central features that an animal model of PD must address. In this review we will focus on those models related to alpha-syn aggregation and LB-like formation. + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + human + + + MESH:D004298 + Chemical + + dopamine + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 23109 + Classical neurotoxin-induced rodent models of PD have been studied in depth and they involve treatment with 6-hydroxydopamine (oxidative stress), MPTP, rotenone and paraquat (mitochondrial complex I inhibition), PSI and epoxomicin (proteasomal inhibition), and lipopolysaccharide (glial activation). Some of these models address the toxic insult hypothesis, whereby pesticide and herbicide use can increase the risk of PD. Most of these chemical models induce degeneration of the dopaminergic neurons in the rodent and primate SN, inducing a motor syndrome that can be modulated by anti-parkinsonian medication. Indeed, the MPTP-treated primate is still the animal model used to test drugs during their selection for clinical trials in humans. These models have been useful to propose pathogenic events that occur in the disease and each mechanism (oxidative stress, mitochondrial complex I inhibition, etc.) is thought to contribute to the pathogenesis of the disorder. While all these models usually display robust nigro-striatal degeneration, the formation of LB inclusions is not a common feature. In this sense, MPTP and 6-OHDA neurotoxic models have face validity related to motor features, although construct (also known as aetiological) validity are limited. Indeed, the poor predictive validity of these models seems to be related to the high failure rate of new treatments in clinical trials. + + 9606 + Species + + humans + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D016627 + Chemical + + 6-OHDA + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D001259 + Disease + + motor syndrome + + + MESH:D010300 + Disease + + PD + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D008070 + Chemical + + lipopolysaccharide + + + MESH:D016627 + Chemical + + 6-hydroxydopamine + + + MESH:D010300 + Disease + + PD + + + MESH:D020258 + Disease + + neurotoxic + + + Disease + + PSI + + + MESH:D010269 + Chemical + + paraquat + + + MESH:D010302 + Disease + + parkinsonian + + + + INTRO + paragraph + 24512 + The discovery of mutations in patients has led to the generation of different genetic models, which may prove to be a more realistic approach to study PD. Many different species and cell models are useful for genetic manipulation, including mice, Drosophila melanogaster and Caenorhabditis elegans. In this sense, several transgenic models have been described that carry SNCA or LRRK2 mutations, and while many of these develop inclusions they fail to display robust neurodegeneration. Recessive models using knockout mice of PINK-1, Parkin or DJ-1 similarly fail to exhibit a nigro-striatal pathology. Animal models carrying mutations in GBA have recently been described in the context of PD, while other animal models focus on altering the intracellular signaling of neurotrophic factors like Smad3 or on the mechanisms of aggregation of alpha-syn. It should be noted that most models do not capture the main hallmarks of PD in the same animal and hence, most models are only suitable to address one particular issue. However, interesting approximations for LB-like formation have been obtained with rotenone treatment, alpha-syn transgenesis with either wild-type or A53T mutation and in Smad3 deficient mice. Thus, the current models do appear to be useful to asses not only motor symptoms but also aetiological and predictive validity of neuroprotective molecules that could halt the pathological progression of the disease. + + 20617 + Gene + 293 + + SNCA + + + 66725 + Gene + 18982 + + LRRK2 + + + 68943 + Gene + 32672 + + PINK-1 + + + 57320 + Gene + 38295 + + DJ-1 + + + 14466 + Gene + 68040 + + GBA + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 9606 + Species + + patients + + + 10090 + Species + + mice + + + 6239 + Species + + Caenorhabditis elegans + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + transgenic + + + MESH:D019636 + Disease + + neurodegeneration + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_2 + 25942 + Non-human primates + + 9606 + Species + + human + + + + INTRO + paragraph + 25961 + MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) is a neurotoxin used in non-human primates as the most relevant animal model of PD, given its ability to induce persistent parkinsonism in humans and on the induction of selective destruction of midbrain dopaminergic neurons. Cardinal motor symptoms of PD are reproduced in this model, as well as other non-motor symptoms such as constipation, salivation, sleep disturbance and cognitive deficits. The MPTP model has limitations as the rapid toxicity results in the acute onset of neurodegeneration and neurological symptoms. Although increased alpha-syn immunoreactivity (-ir) within surviving neurons is detected, MPTP-affected dopaminergic neurons do not develop LBs or LNs. + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + human + + + 9606 + Species + + humans + + + MESH:D010300 + Disease + + PD + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D020271 + Disease + + neurodegeneration and neurological symptoms + + + MESH:D064420 + Disease + + toxicity + + + MESH:D010300 + Disease + + PD + + + MESH:D003248 + Disease + + constipation + + + MESH:D003072 + Disease + + sleep disturbance and cognitive deficits + + + MESH:D015632 + Chemical + + 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D010302 + Disease + + parkinsonism + + + MESH:D015632 + Chemical + + MPTP + + + + INTRO + title_2 + 26691 + Rodents + + + INTRO + title_3 + 26699 + MPTP in mice + + MESH:D015632 + Chemical + + MPTP + + + + INTRO + paragraph + 26712 + In mice, the MPTP model must be induced in specific strains for it to be a consistent model, such as C57 and Swiss Webster mice. The loss of dopaminergic neurons depends on the administration regime (from acute to chronic), ranging from 60% to 90%. Acute or sub-chronic administration does not lead to the formation of inclusions and while alpha-syn inclusions have been detected in the study of some chronic models, this is not always replicated. In this sense, alpha-syn-ir was detected in dopaminergic neurons when chronic MPTP treatment was coupled with probenecid administration to block the rapid clearance of MPTP and its derivates. On examination by electron microscopy, the neurons that accumulate alpha-syn appeared to contain lipid droplets or secondary lysosomes covered by proteins. Although this is quite a distinct morphology to the straight filaments detected in human LBs, it was suggested that lipofuscins may be important for the development of LBs. When MPTP was administered continuously using an osmotic minipump, ubiquitin-ir and alpha-syn-ir nigral inclusions formed, although their co-localization in the same cell or aggregate was not studied. Ultrastructurally, these inclusions appeared as concentric membranes containing alpha-syn that was transformed into fibrillar morphology. However, other studies failed to find such inclusions following acute, semi-chronic or chronic MPTP treatment with probenecid or using osmotic minipumps. Thus, further work is required before this model can be used to study aggregate formation. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + MESH:D011339 + Chemical + + probenecid + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D008055 + Chemical + + lipid + + + MESH:D008062 + Chemical + + lipofuscins + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D011339 + Chemical + + probenecid + + + MESH:D015632 + Chemical + + MPTP + + + + INTRO + title_3 + 28265 + Rotenone in rats + + MESH:D012402 + Chemical + + Rotenone + + + + INTRO + paragraph + 28282 + Rotenone is an insecticide that accumulates in the mitochondria, inhibiting complex I and promoting oxidative stress, as well as inhibiting the ubiquitin-proteasome system in vitro. While this model is one of the most promising models for PD studies, it has several drawbacks since rotenone is not a selective dopaminergic neurotoxin, it produces high rates of mortality (~30%) and there is also strong variability in neurodegeneration (only observed in 50% of rats). Interestingly, peripheral treatment with rotenone induces the formation of alpha-syn inclusions, with a dense core and fibrillar surrounding that resembles those observed in PD. The aggregation of alpha-syn may be enhanced by neonatal lipopolysaccharide (LPS) treatment, suggesting some cooperation between perinatal inflammatory processes and exposure to this pesticide. In vitro studies have shown that rotenone induces a conformational change in alpha-syn and that it accelerates the rate of fibril formation. Indeed, like the human pathology alpha-syn deposits are observed in the myenteric plexus. In order to avoid peripheral rotenone toxicity and hence, the high mortality rate in rats, intracerebral administration of this toxin has been used, showing significant nigro-striatal neurodegeneration but not the appearance of alpha-syn inclusions. Indeed, variability in the results obtained following peripheral administration (subcutaneous versus intravenous infusion) has limited the use of this model as a reliable tool to study PD. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + alpha-syn + + + 29219 + Gene + 293 + + alpha-syn + + + 21898 + Gene + 41317 + + LPS + + + 10116 + Species + + rats + + + 9606 + Species + + human + + + 10116 + Species + + rats + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D010300 + Disease + + PD + + + MESH:D008070 + Chemical + + lipopolysaccharide + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D010300 + Disease + + PD + + + MESH:D064420 + Disease + + toxicity + + + MESH:D012402 + Chemical + + Rotenone + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 29792 + Combined administration of other herbicides and fungicides, such as paraquat and Maneb also produces a high mortality rate and modest nigro-striatal neurodegeneration without LB deposition. + + MESH:D019636 + Disease + + neurodegeneration + + + + INTRO + title_3 + 29982 + Proteasome inhibitors + + + INTRO + paragraph + 30004 + The discovery of mutations in parkin and ubiquitin carboxy terminal hydrolase-1 (UCH-L1) in familial PD focused attention on the ubiquitin-proteasome system, suggesting that the toxicity of alpha-syn was associated with misfolding of the protein, and impaired proteasomal and lysosomal degradation. Indeed, the combined systemic administration of the proteasome inhibitors PSI/epoxomicin produces a loss of nigro-striatal neurons and progressive motor disabilities. Moreover, inclusion bodies were observed not only in the SN but also, in the locus coeruleus, raphe and substantia innominata. However, this model has proved very difficult to reproduce and the data has not been replicated in mice, rats or primates, in which only a partial response to proteasomal inhibitors is obtained. This variability frustrates the use of this model in PD research. More recently, conditional genetic deletion of a proteasomal subunit in mice that disrupts 26S proteasome degradation was shown to induce the loss of dopaminergic neurons and the appearance of inclusions in SN neurons that contain ubiquitin and alpha-syn. Although no biochemical studies of aggregation or double-immunolabeling have been performed to clarify the nature of these inclusions, the inclusions appear to contain mitochondria and double-membraned autophagolysomes at the ultrastructural level. However, there was no resemblance of the radially oriented, straight filaments observed in LBs. Indeed, in this model the accumulation of aberrant mitochondria into inclusions seems to be independent of alpha-syn. + + 22223 + Gene + 37894 + + ubiquitin carboxy terminal hydrolase-1 + + + 22223 + Gene + 37894 + + UCH-L1 + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 10090 + Species + + mice + + + 10116 + Species + + rats + + + 10090 + Species + + mice + + + Disease + + PSI + + + MESH:D064420 + Disease + + toxicity + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_3 + 31577 + Mitochondrial dysfunction + + MESH:D028361 + Disease + + Mitochondrial dysfunction + + + + INTRO + paragraph + 31603 + A loss of mitochondrial complex I is observed in sporadic PD that may lead to selective dopaminergic neurodegeneration due to oxidative stress. Midbrain dopaminergic neurons of PD patients and elderly humans carry high levels of somatic mtDNA mutations. Indeed, neurotoxins like MPTP and rotenone inhibit this mitochondrial complex. Transgenic mice overexpressing wild-type alpha-syn induces mitochondrial fragmentation, which may predispose to neural degeneration, and transgenic mice overexpressing human A53T alpha-syn mutation also display mitochondrial abnormalities that may explain some aspects of the aggregated alpha-syn toxicity. Disruption of the respiratory chain has been modeled in mice by deleting the gene encoding the mitochondrial transcription factor A (TFAM) in dopaminergic neurons, thereby inhibiting the transcription of mtDNA genes. Although these mice have an interesting phenotype, with impaired motor function and neurodegeneration, alpha-syn is not needed to express this phenotype. Indeed, the inclusions found in dopaminergic neurons have no alpha-syn but rather, abnormal mitochondrial membranes. + + 20617 + Gene + 293 + + alpha-syn + + + 21780 + Gene + 31139 + + TFAM + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + patients + + + 9606 + Species + + humans + + + 10090 + Species + + Transgenic mice + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 185869 + Species + + T alpha + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D064420 + Disease + + toxicity + + + MESH:C565640 + Disease + + neural degeneration + + + MESH:D010300 + Disease + + PD + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D010300 + Disease + + PD + + + MESH:D012402 + Chemical + + rotenone + + + + INTRO + title_3 + 32731 + alpha-syn transgenesis + + 20617 + Gene + 293 + + alpha-syn + + + + INTRO + paragraph + 32754 + Different transgenic models have been developed that overexpress wild type, A53T, A30P and truncated alpha-syn. However, despite initial results with mutated alpha-syn in rodents, they have failed to translate into truly effective transgenic models of PD. Attempts to overexpress or to knock-out alpha-syn in rodents have produced a variety of pathological abnormalities, including aggregate formation. However, no clear loss of dopaminergic neurons is observed. On the other hand, viral transduction induces rapid neurodegeneration but it is limited to the region targeted. The use of cell-type promoters, such as tyrosine hydroxylase, is a limitation to transgenic overexpression that prevents the induction of the broad brain pathology observed in humans. However, this localized overexpression of alpha-syn (or truncated forms) may be useful to address specific features of PD. A variety of transgenic models using different promoters that drive broad expression have induced the graded appearance of alpha-syn aggregations with no fibrillar morphology. One particularly interesting model is the transgenic A53T alpha-syn under the control of the mouse prion promoter, which overexpresses the mutant protein in neurons. These mice develop progressive motor failure at 8 months of age leading to paralysis and death. Inclusions have been detected in different brain areas like the SN, raphe, pons, pontine reticular nuclei, locus coeruleus and deep cerebellar nuclei. The biochemical characterization of these inclusions showed SDS-insoluble alpha-syn aggregation into dimers, trimers and multimers, and these LB-like inclusions have a filamentous structure like human LBs. However, as indicated, a major drawback of this model is that no dopaminergic neurodegeneration is detected in the SN. Conversely, there is neurodegeneration in areas not affected in PD, such as among motor neurons. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 21823 + Gene + 307 + + tyrosine hydroxylase + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + alpha-syn + + + 9606 + Species + + humans + + + 10090 + Species + + mouse + + + 36469 + Species + + prion + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 185869 + Species + + T alpha + + + 16473 + Gene + + syn + + + p.A30P;RS#:104893878 + ProteinMutation + + A30P + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D010300 + Disease + + PD + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D001259 + Disease + + motor failure + + + MESH:D010300 + Disease + + PD + + + MESH:D012967 + Chemical + + SDS + + + MESH:D010243 + Disease + + paralysis + + + MESH:D010300 + Disease + + PD + + + MESH:D019636 + Disease + + neurodegeneration + + + + INTRO + title_3 + 34647 + LRRK2 transgenesis + + 66725 + Gene + 18982 + + LRRK2 + + + + INTRO + paragraph + 34666 + LRRK2 null mice have no overt dopaminergic deficit or clear pathology in the brain, although age-dependent renal atrophy is observed that is associated with the aggregation of alpha-syn in renal tubules of the aged kidney. The R1441C mutation in LRRK2 generated by a knock-in strategy in mice does not induce dopaminergic neurodegeneration but rather, DA neurotransmission and D2 receptor dysfunction. However, no obvious accumulation of alpha-syn is detected. Mice overexpressing the G2019S mutation in LRRK2 do not reproduce any obvious gross neuropathological phenotypes nor is alpha-syn aggregation observed, although this LRRK2 mutation may accelerate the pathogenic phenotype of A53T alpha-syn mice. + + 66725 + Gene + 18982 + + LRRK2 + + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 13489 + Gene + 22561 + + D2 receptor + + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + Mice + + + 10090 + Species + + mice + + + 185869 + Species + + T alpha + + + 16473 + Gene + + syn + + + MESH:D009461 + Disease + + deficit + + + p.R1441C;RS#:33939927 + ProteinMutation + + R1441C + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:D007674 + Disease + + renal atrophy + + + MESH:D019636 + Disease + + neurodegeneration + + + + INTRO + title_3 + 35372 + Autophagy-lysosome system + + + INTRO + paragraph + 35398 + Several studies have suggested a role for autophagy in PD. The autophagy-lysosome system is a catabolic pathway involved in protein and organelle degradation. Several types of autophagy have been described, including: microautophagy; macroautophagy, engulfing large structures; and chaperone-mediated autophagy that degrades only soluble proteins in a selective manner. Cellular homeostasis of autophagy is crucial to maintain the balance between healthy and unhealthy cells. In PD and related LB diseases an accumulation of autophagosomes has been described, coupled with a reduction of lysosomal markers in nigral dopaminergic neurons, suggesting a defect in lysosome-mediated clearance of alpha-syn aggregates. Indeed, alpha-syn is physiologically degraded by both the ubiquitin-proteasome and the autophagy-lysosome system (even by the chaperone-mediated mechanism), although the mutant forms of alpha-syn appear to inhibit their own degradation. Hence, these pathways would appear to participate in LB formation. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 36416 + In mice, deletion of the gene essential for macroautophagy (Atg7) in dopaminergic neurons induces progressive moderate dopaminergic loss in the SN and striatal DA depletion at an age of 9 months, although most dopaminergic neurons are resistant to the long-term stress induced by impairing autophagy. These mice have ubiquitinated-SQSTM1 inclusions in dopaminergic neurons but no abnormal aggregation of alpha-syn. In PD patients, the translocation to the nucleus of the TFEB transcription factor (a regulator of autophagy), is dampened in midbrain dopaminergic neurons and it co-localizes with alpha-syn in LBs. Using adeno-associated viral vectors to model alpha-syn transgenesis in rats, under the control of the synapsin-1 promoter and the WPRE enhancer, stimulation of TFEB and Beclin (an activator of autophagy) was shown to rescue nigral DA neurons from alpha-syn toxicity. These results identify interesting new mechanisms, however no clear alpha-syn aggregates have yet been formed by manipulating the autophagy-lysosome system. + + 74244 + Gene + 4662 + + Atg7 + + + 18412 + Gene + 31202 + + SQSTM1 + + + 20617 + Gene + 293 + + alpha-syn + + + 7942 + Gene + 5182 + + TFEB + + + 6622 + Gene + 293 + + alpha-syn + + + 29219 + Gene + 293 + + alpha-syn + + + 24949 + Gene + 48483 + + synapsin-1 + + + 316214 + Gene + 5182 + + TFEB + + + 29219 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 9606 + Species + + patients + + + 10116 + Species + + rats + + + MESH:D010300 + Disease + + PD + + + MESH:D064420 + Disease + + toxicity + + + + INTRO + title_3 + 37454 + Neurotrophic factors: Smad3 deficiency + + 17127 + Gene + 55937 + + Smad3 + + + + INTRO + paragraph + 37493 + GDNF and its close relative Neurturin provide functional rescue of nigro-striatal dopaminergic neurons after 6-OHDA or MPTP treatment. These results led to clinical trials into the use of these molecules in patients with PD, although the use of exogenous GDNF in clinical trials produced inconclusive results. The neuroprotective effect of this neurotrophic factor may not be translated to the clinic as GDNF fails to protect against alpha-syn-induced toxicity, probably due to the blockade of GDNF signaling by alpha-syn. Indeed, transgenesis of alpha-syn induced by viral overexpression in the nigro-striatum, induces dystrophic terminals in the striatum containing alpha-syn aggregates, which are not modified by exogenous administration of GDNF. + + 14573 + Gene + 433 + + GDNF + + + 2668 + Gene + 433 + + GDNF + + + 14573 + Gene + 433 + + neurotrophic factor + + + 14573 + Gene + 433 + + GDNF + + + 20617 + Gene + 293 + + alpha-syn + + + 14573 + Gene + 433 + + GDNF + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 14573 + Gene + 433 + + GDNF + + + 9606 + Species + + patients + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D064420 + Disease + + toxicity + + + MESH:D020388 + Disease + + dystrophic + + + MESH:D016627 + Chemical + + 6-OHDA + + + MESH:D010300 + Disease + + PD + + + + INTRO + paragraph + 38243 + Recently, intracellular TGF-beta1 signaling has been implicated in different pathological events related to PD in a mouse model, including LB-like formation. In humans, TGF-beta1 is up-regulated in the striatum and in the ventricular cerebrospinal fluid of patients with PD. Moreover, active TGF-beta1 overexpression in the nigro-striatal system of MPTP-treated mice using adenoviral vectors produces poorer survival of dopaminergic neurons and higher levels of striatal DA depletion. However, since the effects of TGF-beta1 are dose- and context-dependent, its overexpression may introduce a bias in studies on animal models. Smad3 deficiency, a molecule involved in the intracellular TGF-beta1 signaling cascade, provides us with a new and interesting model of PD, as it promotes selective postnatal neurodegeneration of dopaminergic midbrain neurons, strong MAO-mediated catabolism of DA in the striatum and oxidative stress, as well as dampening the trophic and astrocytic support to dopaminergic neurons. Interestingly, Smad3 deficiency induces the formation of alpha-syn inclusions in selected brain areas, which accumulate with age in a progressive and gene dosage dependent manner. These alpha-syn inclusions appear in both the perikaryon (SN and paralemniscal nucleus) and in neurites (in the motor and cingulate cortices, striatum, corpus callosum and spinal cord). Indeed, these alpha-syn deposits are phosphorylated at Ser129 and ubiquitinated, and they form a core/halo distribution that resembles the deposits observed in human LBs. In other brain areas alpha-syn expression is associated with an irregular morphology, increased alpha-syn-ir staining and neurite thickness (pontine nuclei, cerebellar white matter, cerebral peduncle, diencephalic nuclei and internal capsule). Moreover, alpha-syn inclusions are also detected in glial cells in the cerebellum and spinal cord. Biochemical analyses show the presence of detergent-insoluble dimers, trimers and oligomers of alpha-syn in the ventral midbrain, motor cortex and spinal cord. + + 21803 + Gene + 540 + + TGF-beta1 + + + 7040 + Gene + 540 + + TGF-beta1 + + + 21803 + Gene + 540 + + TGF-beta1 + + + 21803 + Gene + 540 + + TGF-beta1 + + + 17127 + Gene + 55937 + + Smad3 + + + 21803 + Gene + 540 + + TGF-beta1 + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 10090 + Species + + mouse + + + 9606 + Species + + humans + + + 9606 + Species + + patients + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + - + Chemical + + Ser129 + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D004298 + Chemical + + DA + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D019636 + Disease + + neurodegeneration + + + MESH:D007027 + Disease + + diencephalic + + + + INTRO + paragraph + 40293 + There is currently increasing attention being paid to the non-motor symptoms of PD, such as cognitive impairment and behavioral disorders. The hippocampus is implicated in physiological learning and memory, as well as in the cognitive dysfunction seen in some PD patients where there is an interaction with the dopaminergic system. The hippocampus of PD patients is also affected by the presence of LBs and in mouse models, alpha-syn modulates adult neurogenesis in the dentate gyrus. Indeed, the accumulation of extracellular alpha-syn oligomers impairs hippocampal LTP and it enhances basal synaptic transmission. Both triplication and point mutations of SNCA are associated with cortical and subcortical LB accumulation, and such patients are clinically diagnosed as PD with dementia. These data suggest a role for the alpha-syn aggregates in the cognitive impairment observed in demented PD patients. Similarly, Smad3 deficient mice have alpha-syn inclusions in the neuronal layers of the hippocampus. Smad3 is strongly expressed in hippocampal neurons and Smad3 deficiency induces a strong decrease in adult neurogenesis in the dentate gyrus, inducing the apoptosis of early stage and highly proliferative intermediate precursor cells (IPCs). Indeed, Smad3 deficiency abolishes the induction of LTP in the dentate gyrus but not in the CA1, highlighting the specificity of this effect. Both neurogenesis and LTP induction in the adult hippocampus are two aspects of hippocampal brain plasticity related to learning and memory that decline with age, and as a result of neurological disorders. Smad3 deficiency sheds light on a new interesting pathological mechanism and provides a new model of PD to be explored in which dopaminergic dysfunction, widespread alpha-syn inclusions and cognitive impairment co-exist. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 6622 + Gene + 293 + + SNCA + + + 6622 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 17127 + Gene + 55937 + + Smad3 + + + 17127 + Gene + 55937 + + Smad3 + + + 12346 + Gene + 20414 + + CA1 + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 109912 + Gene + + LTP + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 10090 + Species + + mouse + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 10090 + Species + + mice + + + 109912 + Gene + + LTP + + + 109912 + Gene + + LTP + + + MESH:D003072 + Disease + + cognitive impairment + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D003072 + Disease + + cognitive impairment + + + MESH:D003704 + Disease + + dementia + + + MESH:D003072 + Disease + + cognitive dysfunction + + + MESH:D010300 + Disease + + PD + + + MESH:D003072 + Disease + + cognitive impairment + + + MESH:D010300 + Disease + + PD + + + MESH:D001523 + Disease + + behavioral disorders + + + MESH:D009422 + Disease + + neurological disorders + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + + INTRO + title_2 + 42110 + Invertebrate models + + + INTRO + paragraph + 42130 + Several invertebrate models have been developed that recapitulate key features of human PD. Drosophila models of alpha-syn overexpression show loss of dopaminergic neurons, locomotor dysfunction and formation of alpha-syn inclusions. Indeed, these inclusions are observed as a core with peripherally radiating filaments. Despite the simplicity of the nematode model Caenorhabditis elegans, the presence of eight dopaminergic neurons has allowed their evaluation following parkinsonian toxic and genetic insult. Overexpression of human wild-type or mutant alpha-syn on C. elegans induces dopaminergic neurodegeneration and the formation of alpha-syn aggregates with fibrillar morphology. It is interesting to note that both Drosophila and C. elegans do not express alpha-syn and thus, other genes like LRRK2 cannot be studied in the context of alpha-syn inclusion formation. These models also lack the complexity of vertebrates and hence, they are not perfect models of PD but rather, they may be useful for comprehensive genetic analysis and drug screening. + + 6622 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 9606 + Species + + human + + + 6239 + Species + + Caenorhabditis elegans + + + 9606 + Species + + human + + + 6239 + Species + + C. elegans + + + 6239 + Species + + C. elegans + + + 7227 + Species + + Drosophila + + + 7227 + Species + + Drosophila + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + MESH:D010302 + Disease + + parkinsonian + + + MESH:D010300 + Disease + + PD + + + MESH:D010300 + Disease + + PD + + + MESH:D009069 + Disease + + locomotor dysfunction + + + MESH:D019636 + Disease + + neurodegeneration + + + + INTRO + title_1 + 43188 + Dopamine metabolism and alpha-syn + + MESH:D004298 + Chemical + + Dopamine + + + + INTRO + paragraph + 43222 + Several studies associate alpha-syn with the DA metabolism in the presynaptic terminals. It is known that alpha-syn inhibits tyrosine hydroxylase, the rate limiting enzyme of DA biosynthesis, and aromatic amino acid decarboxylase, the enzyme that converts L-Dopa to DA. Indeed, deregulation of DA biosynthesis in the SN induces increased toxicity to alpha-syn. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 21823 + Gene + 307 + + tyrosine hydroxylase + + + 20617 + Gene + 293 + + alpha-syn + + + MESH:D064420 + Disease + + toxicity + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D007980 + Chemical + + L-Dopa + + + + INTRO + paragraph + 43583 + The Smad3 deficient mouse model described earlier also associates alpha-syn and DA metabolism, whereby Smad3 deficiency drives alpha-syn overexpression and aggregation, as well as the deregulation of DA turnover by inducing MAO-dependent DA catabolism, provoking a loss of dopaminergic neurons. Other mouse models, such as alpha-syn, parkin, DJ-1 or PINK1 null mice, also display deregulated striatal DA metabolism, release or re-uptake, although without dopaminergic neuronal loss. Increased extracellular DA, decreased DAT, increased GSH levels and differences in the catabolism of DA have all been described in Parkin deficient mice. However, neither the number of dopaminergic neurons nor the striatal DA levels and turnover are altered in DJ-1 or PINK knock-out mice. Nevertheless, the evoked DA overflow in these mice diminishes due to increased DA uptake or to the decreased quantal release of DA, respectively. The alpha-syn knock-out mice have less striatal DA, enhanced activity-dependent DA release and smaller reserve pools of synaptic vesicles, which may be due to a uninhibited DA vesicular release. All these mutant mice illustrate that impaired presynaptic DA metabolism, release or re-uptake may be common to PD, although the presence of alpha-syn aggregates and DA neuronal loss is only detected in Smad3 deficient mice. Indeed, 3,4-dihydroxyphenylacetaldehyde (DOPAL), a MAO intermediate metabolite of DA, can induce toxic aggregation of alpha-synuclein. These data suggest that DA metabolism and alpha-syn interactions may underlie the susceptibility of SN neurodegeneration in PD. + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 57320 + Gene + 38295 + + DJ-1 + + + 68943 + Gene + 32672 + + PINK1 + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-synuclein + + + 20617 + Gene + 293 + + alpha-syn + + + 13162 + Gene + 55547 + + DAT + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 57320 + Gene + 38295 + + DJ-1 + + + MESH:D009410 + Disease + + neuronal loss + + + MESH:D004298 + Chemical + + DA + + + Disease + + SN neurodegeneration + + + MESH:D010300 + Disease + + PD + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D010300 + Disease + + PD + + + MESH:C007430 + Chemical + + DOPAL + + + MESH:D004298 + Chemical + + DA + + + MESH:D009410 + Disease + + neuronal loss + + + - + Chemical + + GSH + + + MESH:C007430 + Chemical + + 3,4-dihydroxyphenylacetaldehyde + + + + INTRO + paragraph + 45185 + It was recently suggested that the spread of the pathological elements in PD, as described in the Braak stages, can occur by neuron-to-neuron transmission of aggregates to healthy cells. In this hypothesis, preformed fibrils of alpha-syn enter neurons, probably by endocytosis, to recruit soluble endogenous alpha-syn into insoluble LB- and LN-like aggregates. Interestingly, peripheral inoculation of alpha-syn fibrils by intramuscular injections can propagate the pathogenic protein to brain nuclei. However, another study showed that alpha-syn fibrils injected into transgenic mice overexpressing alpha-syn mutations promotes the widespread formation of alpha-syn inclusions in the brain of A53T but not E46K mutant mice, nor in non-transgenic mice. The authors suggest inespecificity with neurofilament of the antibodies used and that only A53T mutant mice have the capacity to induce alpha-syn aggregation upon exogenous administration of alpha-syn. This aggregate transmission may be related to the existence of extracellular forms of alpha-syn that are released from neurons and glia by exocytosis. Indeed, extracellular alpha-syn may induce DA release in the striatum, establishing a new link between alpha-syn and DA metabolism. Furthermore, alpha-syn function may be related to the reorganization of plasma membrane microdomains. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + MESH:D004298 + Chemical + + DA + + + MESH:D004298 + Chemical + + DA + + + MESH:D010300 + Disease + + PD + + + p.E46K;RS#:104893875 + ProteinMutation + + E46K + + + + INTRO + title_1 + 46525 + Conformational plasticity of alpha-syn + + + INTRO + paragraph + 46564 + Advances in cell-free systems and model cell systems have shed light on the process of alpha-syn aggregation. The role proposed for alpha-syn is in the modulation of neurotransmitter release in the presynaptic nerve terminal, as well as influencing DA neurotransmitter biosynthesis, vesicle trafficking and exocytosis. In the cytoplasm and/or vesicle lumen, alpha-syn is present as an intrinsically disordered protein (IDP: i.e. a monomer that lacks a well-organized secondary structure), yet when bound to membranes it adopts several conformations, such as an extended alpha-helix or a broken-helix. Indeed, despite the overwhelming evidence that alpha-syn is a disordered monomer in solution, two recent reports suggest that the native protein exists as a helical tetramer under physiological conditions, with reduced aggregation tendencies, and that the dissociation of the tetramer into monomeric subunits promotes toxic aggregation. Indeed, when monomeric alpha-syn binds to membranes, it changes its conformation to a partially helical form. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 15926 + Gene + 21195 + + IDP + + + MESH:D030342 + Disease + + disordered + + + MESH:D030342 + Disease + + disordered + + + MESH:C536965 + Disease + + reduced aggregation tendencies + + + + INTRO + paragraph + 47612 + Many IDPs are known to interact with a large number of proteins, serving as a nodes or hubs, in a way that IDPs undergo a disorder-to-order transition upon interaction with specific partners. More than 50 proteins have been reported to interact with alpha-syn, although it is unknown the proportion unfolded alpha-syn or that which has adopted a secondary structure as a consequence of binding within the cell. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + + INTRO + paragraph + 48023 + The alpha-syn peptide has 140 amino acids, with 3 distinct regions: a N-terminal (1-60 residues) that contain four imperfect repeats of KTKEGV motifs; a NAC region (61-95 residues), with 3 additional KTKEGV repeats, and the hydrophobic and amyloidogenic NAC region; a C-terminus (96-140 residues) that is enriched in acidic and proline residues, and that facilitates interactions with different proteins. + + 20617 + Gene + 293 + + alpha-syn + + + - + Chemical + + NAC + + + MESH:D011392 + Chemical + + proline + + + - + Chemical + + NAC + + + + FIG + 40478_2014_176_Fig1_HTML.jpg + Fig1 + fig_caption + 48431 + Models of alpha-syn aggregation and LBs formation. A alpha-Syn is present in the vesicle lumen and in the cytoplasma as an intrinsically disordered protein. alpha-Syn bound to membranes has distinct conformation such as an extended alpha-helix or a broken-helix. In the pathological context, disordered monomers may lead to oligomerization and fibril formation, following a nucleation-dependent process, in which monomers are added to existing aggregates. B Rotenone administration in rats, A53T alpha-syn transgenesis in mice and Smad3 deficient mice are interesting models to study LB formation. While A53T alpha-syn transgenesis and Smad3 deficiency can modulate DA metabolism, rotenone and Smad3 deficiency induce oxidative stress, mechanisms that may participate in LBs formation. Indeed, proteasome and autophagy inhibitors may impair degradation of alpha-syn. LRRK2 mutations may participate in LB formation by altering autophagy and alpha-syn solubility. + + 20617 + Gene + 293 + + alpha-syn + + + 16473 + Gene + + Syn + + + 16473 + Gene + + Syn + + + 17127 + Gene + 55937 + + Smad3 + + + 17127 + Gene + 55937 + + Smad3 + + + 17127 + Gene + 55937 + + Smad3 + + + 20617 + Gene + 293 + + alpha-syn + + + 66725 + Gene + 18982 + + LRRK2 + + + 20617 + Gene + 293 + + alpha-syn + + + 14960 + Gene + 123820 + + A alpha + + + 10116 + Species + + rats + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 185869 + Species + + T alpha + + + 185869 + Species + + T alpha + + + 16473 + Gene + + syn + + + 16473 + Gene + + syn + + + MESH:D030342 + Disease + + disordered + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D012402 + Chemical + + Rotenone + + + MESH:D030342 + Disease + + disordered + + + + INTRO + paragraph + 49395 + The major constituent of LBs is a fibrillar form of alpha-syn that adopts a beta-sheet structure and hence, the disordered monomer or helical tetramer is transformed into highly organized fibrils in the course of this pathology. In vitro studies suggest a nucleation-dependent process due to a conformational transition to anti-parallel beta-sheet structures, including a committed step of alpha-syn dimer formation. Nucleation follows a sigmoidal growth profile, with an initial lag phase where the protein changes to a partially folded intermediate to form nuclei with oligomers (Figure 1). This conformational change exposes the NAC domain that can participate in hydrophobic interactions that may initiate the aggregation process. During the growth phase, the nucleus adds monomers to form larger oligomers, polarized protofibrils and finally, fibrils. In the last steady-state phase both the fibrils and monomers appear to be in equilibrium, this model therefore reflecting the addition of monomers to existing aggregates. Recently, a key conformational intermediate was characterized after oligomer formation, with the appearance of stable and compact oligomers that are more damaging to cells (Figure 1). Indeed, it seems that the assembly process can be reversed and that fibrils may disaggregate to form these stable, cytotoxic oligomers. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + MESH:D030342 + Disease + + disordered + + + - + Chemical + + NAC + + + + INTRO + paragraph + 50743 + The pathogenic aggregation of alpha-syn can be modulated by endogenous and exogenous factor, such as metals and pesticides, genetic mutations in SNCA, post-translational modifications and protein-protein interactions. It is not clear how alpha-syn bound to membranes, such as synaptic vesicles or presynaptic plasma membrane, can aggregate. One model proposes that the broken-helix membrane bound state of alpha-syn releases its C-terminal region, converting the protein into a partially helical membrane bound state, which may lead to oligomerization and fibril formation. Alternatively, alpha-helical forms of alpha-syn bound to membranes may inhibit fibril formation, and membrane-bound alpha-syn monomers may be protected from aggregation. Indeed, oligomers have a high propensity to bind to membranes, which may promote permeabilization and disrupt cellular homeostasis. Furthermore, alpha-syn localizes to the nerve terminal, where modest increments in alpha-syn (such as those produced by gene duplication) inhibit neurotransmitter release by reducing synaptic vesicle density and by altering vesicle reclustering after endocytosis. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + SNCA + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + + INTRO + paragraph + 51883 + By contrast, oligomers might be kinetically detained by interactions with small molecules, inducing secondary structures such as annular pores or spheres. The point mutations detected in the SNCA gene of PD patients are also endogenous factors that could accelerate alpha-syn aggregation in vitro, with A53T and G46L forming oligomers and fibrils, and A30P forming oligomers but not fibrils. + + 6622 + Gene + 293 + + SNCA + + + 6622 + Gene + 293 + + alpha-syn + + + 9606 + Species + + patients + + + p.G46L + ProteinMutation + + G46L + + + MESH:D010300 + Disease + + PD + + + p.A30P;RS#:104893878 + ProteinMutation + + A30P + + + c.53A>T;RS#:104893877 + DNAMutation + + A53T + + + + INTRO + title_2 + 52275 + Post-translational modification of alpha-syn + + + INTRO + paragraph + 52320 + Several post-translational modifications of alpha-syn may occur, such as phosphorylation, truncation, ubiquitination, nitration, sumoylation and enzymatic cross-linking. In LBs, phosphorylation at serine 129 is a common alpha-syn modification, although its role is unclear if we consider that overexpression of the phosphorylated serine 129 isoform in animal models does not produce toxicity. The majority of alpha-syn is mono- to tri-ubiquinated, and while poly-ubiquitination serves as a signal for alpha-syn degradation by the proteasome, it does not seem to be required for alpha-syn fibrillation and LB formation. Nevertheless, an interplay between phosphorylation and ubiquitination may render the protein more susceptible to aggregation. It is estimated that 85% of all human proteins undergo Nalpha-acetylation due to the activity of Nalpha-acetyltransferases, probably influencing the subcellular localization of proteins, their rate of synthesis and protein-protein interactions. Tetramers of alpha-syn seem to be Nalpha-acetylated, as is probably are the aggregated alpha-syn isolated from PD deposits. However, Nalpha-acetylation does not seem to alter protein aggregation but more likely, lipid binding. Small amounts of various C-terminal truncated forms of alpha-syn have been detected in LBs, which exhibit greater fibrillation capacity. Furthermore, four alternatively spliced forms add another level of complexity. Compared to the canonical alpha-syn, the three alternative isoforms aggregate significantly less, forming shorter fibrils that are arranged in parallel or with anular structures. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 9606 + Species + + human + + + MESH:D014693 + Disease + + fibrillation + + + MESH:D064420 + Disease + + toxicity + + + MESH:D010300 + Disease + + PD deposits + + + MESH:D012694 + Chemical + + serine + + + MESH:D014693 + Disease + + fibrillation + + + MESH:D008055 + Chemical + + lipid + + + MESH:D012694 + Chemical + + serine + + + MESH:D066263 + Disease + + protein aggregation + + + + INTRO + paragraph + 53932 + Despite the huge amount of research into the properties of alpha-syn aggregation, there is still no coherent picture on the structure, dynamics, and the physiological and pathological roles of alpha-syn. New animal models for PD need to explore LB formation in the context of neurodegeneration and DA metabolism, such as the rotenone, alpha-syn transgenesis and Smad3 deficient mice, in order to clearly understand the pathological mechanism of LB formation as well as to attain effective therapies for this disease. + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 20617 + Gene + 293 + + alpha-syn + + + 17127 + Gene + 55937 + + Smad3 + + + 10090 + Species + + mice + + + MESH:D010300 + Disease + + PD + + + MESH:D012402 + Chemical + + rotenone + + + MESH:D019636 + Disease + + neurodegeneration + + + + ABBR + title + 54449 + Abbreviations + + + ABBR + paragraph + 54463 + DA + + + ABBR + paragraph + 54466 + Dopamine + + MESH:D004298 + Chemical + + Dopamine + + + + ABBR + paragraph + 54475 + IDP + + + ABBR + paragraph + 54479 + Intrinsically disordered protein + + MESH:D030342 + Disease + + disordered + + + + ABBR + paragraph + 54512 + LB + + + ABBR + paragraph + 54515 + Lewy body + + + ABBR + paragraph + 54525 + LN + + + ABBR + paragraph + 54528 + Lewy neurite + + + ABBR + paragraph + 54541 + PD + + MESH:D010300 + Disease + + PD + + + + ABBR + paragraph + 54544 + Parkinson's disease + + MESH:D010300 + Disease + + Parkinson's disease + + + + ABBR + paragraph + 54566 + SN + + + ABBR + paragraph + 54569 + Substantia nigra + + + ABBR + paragraph + 54586 + alpha-syn + + + ABBR + paragraph + 54596 + alpha-synuclein + + + ABBR + footnote + 54612 + Competing interests + + + ABBR + footnote + 54632 + The authors declare that they have no competing interests. + + + REF + title + 54691 + References + + + 1 + REF + surname:Goetz;given-names:CG + a008862 + 22229124 + 2011 + Cold Spring Harb Perspect Med + ref + 54702 + The history of Parkinson's disease: early clinical descriptions and neurological therapies + + MESH:D010300 + Disease + + Parkinson's disease + + + + surname:Lewandowsky;given-names:M + REF + surname:Lewy;given-names:FH + 920 + 1912 + Pathologische anatomie + ref + 933 + 54795 + Paralysis agitans + + MESH:D010300 + Disease + + Paralysis agitans + + + + 37 + surname:Parent;given-names:A + REF + surname:Parent;given-names:M + 313 + 20481265 + 2010 + Can J Neurol Sci + ref + 319 + 54813 + Substantia nigra and Parkinson's disease: a brief history of their long and intimate relationship + + MESH:D010300 + Disease + + Parkinson's disease + + + + surname:Ide;given-names:SE + surname:Leroy;given-names:E + surname:Lavedan;given-names:C + surname:Polymeropoulos;given-names:MH + 9197268 + 1997 + Science + ref + REF + 2047 + 2045 + 276 + surname:Stenroos;given-names:ES + surname:Papapetropoulos;given-names:T + surname:Johnson;given-names:WG + surname:Chandrasekharappa;given-names:S + surname:Athanassiadou;given-names:A + surname:Di Iorio;given-names:G + surname:Golbe;given-names:LI + surname:Lazzarini;given-names:AM + surname:Boyer;given-names:R + surname:Duvoisin;given-names:RC + surname:Rubenstein;given-names:J + surname:Root;given-names:H + surname:Pike;given-names:B + surname:Nussbaum;given-names:RL + surname:Dutra;given-names:A + surname:Dehejia;given-names:A + 54913 + Mutation in the alpha-synuclein gene identified in families with Parkinson's disease + + 20617 + Gene + 293 + + alpha-synuclein + + + MESH:D010300 + Disease + + Parkinson's disease + + + + 30 + REF + surname:Sulzer;given-names:D + 244 + 17418429 + 2007 + Trends Neurosci + ref + 250 + 55000 + Multiple hit hypotheses for dopamine neuron loss in Parkinson's disease + + MESH:D004298 + Chemical + + dopamine + + + MESH:D010300 + Disease + + Parkinson's disease + + + + 8 + surname:Schapira;given-names:AH + REF + surname:Chaudhuri;given-names:KR + 464 + 19375664 + 2009 + Lancet Neurol + ref + 474 + 55074 + Non-motor symptoms of Parkinson's disease: dopaminergic pathophysiology and treatment + + MESH:D010300 + Disease + + Parkinson's disease + + + + 249 + surname:Barker;given-names:RA + surname:Brayne;given-names:C + REF + surname:Foltynie;given-names:T + 138 + 11985378 + 2002 + J Neurol + ref + 145 + 55162 + The heterogeneity of idiopathic Parkinson's disease + + MESH:D010300 + Disease + + idiopathic Parkinson's disease + + + + surname:Chui;given-names:HC + 60 + surname:Mortimer;given-names:JA + surname:Lyness;given-names:SA + REF + surname:Zarow;given-names:C + 337 + 12633144 + 2003 + Arch Neurol + ref + 341 + 55216 + Neuronal loss is greater in the locus coeruleus than nucleus basalis and substantia nigra in Alzheimer and Parkinson diseases + + MESH:D009410 + Disease + + Neuronal loss + + + MESH:D000544 + Disease + + Alzheimer and Parkinson diseases + + + + surname:Weisenbach;given-names:S + surname:Klein;given-names:JC + surname:Thomas;given-names:AV + surname:Hilker;given-names:R + 1716 + 16344512 + 2005 + Neurology + ref + 65 + REF + surname:Heiss;given-names:WD + surname:Herholz;given-names:K + 1722 + surname:Jacobs;given-names:AH + surname:Burghaus;given-names:L + surname:Kalbe;given-names:E + 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Neurotoxic + + + MESH:D015632 + Chemical + + MPTP + + + + surname:Santa Cruz;given-names:K + surname:Petroske;given-names:E + surname:Totterdell;given-names:S + surname:Meredith;given-names:GE + 156 + 12426058 + 2002 + Brain Res + ref + 956 + REF + 165 + surname:Lau;given-names:YS + surname:Callison;given-names:RC;suffix:Jr + 63877 + Lysosomal malfunction accompanies alpha-synuclein aggregation in a progressive mouse model of Parkinson's disease + + 20617 + Gene + 293 + + alpha-synuclein + + + 10090 + Species + + mouse + + + MESH:D010300 + Disease + + Parkinson's disease + + + + surname:Surmeier;given-names:DJ + surname:Potashkin;given-names:JA + surname:Totterdell;given-names:S + surname:Meredith;given-names:GE + S112 + Suppl 2 + 18585085 + 2008 + Parkinsonism Relat Disord + ref + 14 + REF + S115 + 63993 + Modeling PD pathogenesis in mice: advantages of a chronic MPTP protocol + + 10090 + Species + + mice + + + MESH:D010300 + Disease + + PD + + + + surname:Gesi;given-names:M + 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surname:Dawson;given-names:VL + surname:Mandir;given-names:AS + surname:Zhang;given-names:L + surname:Shimoji;given-names:M + 103 + 15790534 + 2005 + Brain Res Mol Brain Res + ref + 134 + REF + 108 + surname:Dawson;given-names:TM + 64436 + Absence of inclusion body formation in the MPTP mouse model of Parkinson's disease + + 10090 + Species + + mouse + + + MESH:D015632 + Chemical + + MPTP + + + MESH:D010300 + Disease + + Parkinson's disease + + + + surname:Seo;given-names:BB + surname:Testa;given-names:CM + surname:Betarbet;given-names:R + surname:Sherer;given-names:TB + 10756 + 14645467 + 2003 + J Neurosci + ref + 23 + REF + surname:Greenamyre;given-names:JT + surname:Matsuno-Yagi;given-names:A + surname:Yagi;given-names:T + 10764 + surname:Miller;given-names:GW + surname:Kim;given-names:JH + surname:Richardson;given-names:JR + 64521 + Mechanism of toxicity in rotenone models of Parkinson's disease + + MESH:D012402 + Chemical + + rotenone + + + MESH:D064420 + Disease + + toxicity + + 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MESH:D004342 + Disease + + hypersensitivity in + + + MESH:D015632 + Chemical + + 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine + + + MESH:D015632 + Chemical + + MPTP + + + + surname:Boyle;given-names:S + surname:Giaime;given-names:E + surname:Yamaguchi;given-names:H + surname:Tong;given-names:Y + 9879 + 20457918 + 2010 + Proc Natl Acad Sci U S A + ref + 107 + REF + 9884 + surname:Shen;given-names:J + surname:Kelleher;given-names:RJ;suffix:3rd + surname:Kopan;given-names:R + 67406 + Loss of leucine-rich repeat kinase 2 causes impairment of protein degradation pathways, accumulation of alpha-synuclein, and apoptotic cell death in aged mice + + 66725 + Gene + 18982 + + leucine-rich repeat kinase 2 + + + 20617 + Gene + 293 + + alpha-synuclein + + + + surname:Karouani;given-names:M + surname:Martella;given-names:G + surname:Pisani;given-names:A + surname:Tong;given-names:Y + 14622 + 19667187 + 2009 + Proc Natl Acad Sci U S A + ref + 106 + REF + 14627 + surname:Shen;given-names:J + surname:Pothos;given-names:EN + surname:Yamaguchi;given-names:H + 67565 + R1441C mutation in LRRK2 impairs dopaminergic neurotransmission in mice + + 66725 + Gene + 18982 + + LRRK2 + + + p.R1441C;RS#:33939927 + ProteinMutation + + R1441C + + + + surname:Hafner;given-names:T + surname:Schweizer;given-names:T + surname:Bidinosti;given-names:M + surname:Herzig;given-names:MC + e36581 + 22615783 + 2012 + PLoS One + ref + 7 + surname:Mayer;given-names:F + REF + surname:van der Putten;given-names:PH + surname:Shimshek;given-names:DR + surname:Schmid;given-names:P + surname:Rovelli;given-names:G + surname:Barske;given-names:C + surname:Stauffer;given-names:D + surname:Vidotto;given-names:N + surname:Danner;given-names:S + surname:Weiss;given-names:A + surname:Stemmelen;given-names:C + 67637 + High LRRK2 levels fail to induce or exacerbate neuronal alpha-synucleinopathy in mouse brain + + 66725 + Gene + 18982 + + LRRK2 + + + 10090 + Species + + mouse + + + MESH:D000080874 + Disease + + synucleinopathy + + + + surname:Wang;given-names:L + surname:Gu;given-names:XL + surname:Parisiadou;given-names:L + surname:Lin;given-names:X + 20064389 + 2009 + Neuron + ref + REF + 827 + 807 + 64 + surname:Chen;given-names:ZZ + surname:Rudow;given-names:G + surname:Troncoso;given-names:JC + surname:Gallant;given-names:PE + surname:Tao-Cheng;given-names:JH + surname:Cai;given-names:H + surname:Liu;given-names:Z + surname:Ding;given-names:J + surname:Li;given-names:Z + surname:Yang;given-names:WJ + surname:Long;given-names:CX + surname:Xie;given-names:C + surname:Sun;given-names:L + surname:Shim;given-names:H + 67730 + Leucine-rich repeat kinase 2 regulates the progression of neuropathology induced by Parkinson's-disease-related mutant alpha-synuclein + + 66725 + Gene + 18982 + + Leucine-rich repeat kinase 2 + + + MESH:D010300 + Disease + + Parkinson's-disease + + + + surname:Stafa;given-names:K + surname:Lin;given-names:BM + surname:Daher;given-names:JP + surname:Ramonet;given-names:D + 21494637 + 2011 + PLoS One + ref + surname:Dawson;given-names:TM + surname:Dawson;given-names:VL + REF + surname:Moore;given-names:DJ + e18568 + 6 + surname:Liu;given-names:Y + surname:Troncoso;given-names:JC + surname:McCaffery;given-names:JM + surname:Swing;given-names:DA + surname:Beal;given-names:MF + surname:Galter;given-names:D + surname:Thomas;given-names:B + surname:Jenkins;given-names:NA + surname:Yang;given-names:L + surname:Copeland;given-names:NG + surname:Glauser;given-names:L + surname:Pletnikova;given-names:O + surname:Westerlund;given-names:M + surname:Lee;given-names:MK + surname:Banerjee;given-names:R + surname:Kim;given-names:J + 67867 + Dopaminergic neuronal loss, reduced neurite complexity and autophagic abnormalities in transgenic mice expressing G2019S mutant LRRK2 + + 10090 + Species + + transgenic mice + + + p.G2019S;RS#:34637584 + ProteinMutation + + G2019S + + + MESH:D009410 + Disease + + neuronal loss + + + MESH:D058225 + Disease + + reduced neurite complexity + + + + surname:Klionsky;given-names:DJ + 451 + surname:Cuervo;given-names:AM + surname:Levine;given-names:B + REF + surname:Mizushima;given-names:N + 1069 + 18305538 + 2008 + Nature + ref + 1075 + 68001 + Autophagy fights disease through cellular self-digestion + + MESH:C564093 + Disease + + Autophagy fights disease + + + + surname:Olanow;given-names:CW + surname:Aebischer;given-names:P + surname:Dodiya;given-names:H + surname:Chu;given-names:Y + 385 + 19505575 + 2009 + Neurobiol Dis + ref + 35 + REF + 398 + surname:Kordower;given-names:JH + 68058 + Alterations in lysosomal and proteasomal markers in Parkinson's disease: relationship to alpha-synuclein inclusions + + 20617 + Gene + 293 + + alpha-synuclein + + + MESH:D010300 + Disease + + Parkinson's disease + + + + surname:Perier;given-names:C + surname:Rodriguez-Muela;given-names:N + surname:Bove;given-names:J + surname:Dehay;given-names:B + 12535 + 20844148 + 2010 + J Neurosci + ref + 30 + REF + 12544 + 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surname:Abbas;given-names:N + surname:Mena;given-names:MA + surname:Ibanez;given-names:P + surname:Itier;given-names:JM + 12915482 + 2003 + Hum Mol Genet + ref + surname:Sanchez;given-names:M + surname:Denefle;given-names:P + REF + surname:Rooney;given-names:TA + surname:Brice;given-names:A + surname:Benavides;given-names:J + surname:Tremp;given-names:G + surname:Garcia de Yebenes;given-names:J + 2291 + 2277 + 12 + surname:Ret;given-names:G + surname:Araujo;given-names:F + surname:Negroni;given-names:J + surname:Joubert;given-names:C + surname:Periquet;given-names:M + surname:Solano;given-names:R + surname:Serrano;given-names:A + surname:Casarejos;given-names:MJ + surname:Pradier;given-names:L + surname:Canals;given-names:S + surname:Corti;given-names:O + surname:Pratt;given-names:J + surname:Laville;given-names:M + surname:Gallego;given-names:E + surname:Bohme;given-names:GA + surname:Cohen-Salmon;given-names:C + 70838 + Parkin gene inactivation alters behaviour and dopamine neurotransmission in the mouse + + MESH:D004298 + Chemical + + dopamine + + + + 102 + surname:Palmiter;given-names:RD + REF + surname:Perez;given-names:FA + 2174 + 15684050 + 2005 + Proc Natl Acad Sci USA + ref + 2179 + 70924 + Parkin-deficient mice are not a robust model of parkinsonism + + 10090 + Species + + mice + + + MESH:D010302 + Disease + + parkinsonism + + + + surname:Vortherms;given-names:TA + surname:Haburcak;given-names:M + surname:Pisani;given-names:A + surname:Goldberg;given-names:MS + 489 + 15721235 + 2005 + Neuron + ref + 45 + surname:Bernardi;given-names:G + REF + surname:Calabresi;given-names:P + surname:Shen;given-names:J + surname:Roth;given-names:BL + surname:Pothos;given-names:EN + surname:Martins;given-names:A + surname:Tscherter;given-names:A + surname:Martella;given-names:G + 496 + surname:Tong;given-names:Y + surname:Costa;given-names:C + surname:Kitada;given-names:T + 70985 + Nigrostriatal dopaminergic deficits and hypokinesia caused by inactivation of the familial Parkinsonism-linked gene DJ-1 + + MESH:C564486 + Disease + + familial Parkinsonism-linked + + + MESH:D018476 + Disease + + Nigrostriatal dopaminergic deficits and hypokinesia + + + + surname:Yamaguchi;given-names:H + surname:Porter;given-names:DR + surname:Pisani;given-names:A + surname:Kitada;given-names:T + 11441 + 17563363 + 2007 + Proc Natl Acad Sci USA + ref + 104 + REF + surname:Shen;given-names:J + surname:Pothos;given-names:EN + surname:Zhang;given-names:C + 11446 + surname:Bonsi;given-names:P + surname:Martella;given-names:G + surname:Tscherter;given-names:A + 71106 + Impaired dopamine release and synaptic plasticity in the striatum of PINK1-deficient mice + + MESH:D004298 + Chemical + + dopamine + + + MESH:C565276 + Disease + + PINK1-deficient + + + + surname:Choi-Lundberg;given-names:D + surname:Farinas;given-names:I + surname:Schmitz;given-names:Y + surname:Abeliovich;given-names:A + 239 + 10707987 + 2000 + Neuron + ref + 25 + surname:Hynes;given-names:M + REF + 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correlates with attenuated synaptic responses to prolonged repetitive stimulation in mice lacking alpha-synuclein + + 10090 + Species + + mice + + + + 27 + surname:Shorter;given-names:J + REF + surname:Gitler;given-names:AD + 2433 + 17344380 + 2007 + J Neurosci + ref + 2434 + 71431 + Prime time for alpha-synuclein + + + surname:Panneton;given-names:WM + surname:Pandey;given-names:N + surname:Kumar;given-names:VB + surname:Burke;given-names:WJ + 193 + 17965867 + 2008 + Acta Neuropathol + ref + 115 + surname:Galvin;given-names:JE + REF + surname:Chung;given-names:HD + surname:Pan;given-names:Y + surname:Li;given-names:SW + 203 + surname:O'Dell;given-names:M + surname:Franko;given-names:MW + surname:Gan;given-names:Q + 71462 + Aggregation of alpha-synuclein by DOPAL, the monoamine oxidase metabolite of dopamine + + 20617 + Gene + 293 + + alpha-synuclein + + + MESH:D004298 + Chemical + + dopamine + + + + surname:Phillips;given-names:KA + surname:Kanter;given-names:E + 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alpha-Synuclein reveals important insights into the roles of the ubiquitin chain in regulating its pathophysiology + + 20617 + Gene + 293 + + alpha-Synuclein + + + + surname:Helsens;given-names:K + surname:Polevoda;given-names:B + surname:Van Damme;given-names:P + surname:Arnesen;given-names:T + 8157 + 19420222 + 2009 + Proc Natl Acad Sci U S A + ref + 106 + surname:Gevaert;given-names:K + REF + surname:Sherman;given-names:F + surname:Lillehaug;given-names:JR + surname:Vandekerckhove;given-names:J + 8162 + surname:Varhaug;given-names:JE + surname:Colaert;given-names:N + surname:Evjenth;given-names:R + 74858 + Proteomics analyses reveal the evolutionary conservation and divergence of N-terminal acetyltransferases from yeast and humans + + 4932 + Species + + yeast + + + + surname:Volker;given-names:W + surname:Regulin;given-names:A + surname:Appenzeller;given-names:S + surname:Bungeroth;given-names:M + 1913 + 24629672 + 2014 + Neurobiol Aging + ref + 35 + REF + surname:Kuhlenbaumer;given-names:G + 1919 + surname:Pendziwiat;given-names:M + surname:Grotzinger;given-names:J + surname:Lorenzen;given-names:I + 74985 + Differential aggregation properties of alpha-synuclein isoforms + + 20617 + Gene + 293 + + alpha-synuclein + + + + + 4272813 + + surname:Wiecha;given-names:John + surname:Xu;given-names:Jin + surname:Wilkinson;given-names:Joanne E + 25491129 + surname:Hoffman;given-names:Miriam + 2014 + 4272813 + 258 + 10.1186/s12909-014-0258-1 + Keywords + front + 258 + 14 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + BMC Med Educ; 2014 Dec 10; 14258. doi:10.1186/s12909-014-0258-1 + Small-group learning Group dynamics Faculty presence Medical education + Hoffman M, Wilkinson JE, Xu J, Wiecha J + 0 + The perceived effects of faculty presence vs. absence on small-group learning and group dynamics: a quasi-experimental study + + + ABSTRACT + abstract_title_1 + 125 + Background + + + ABSTRACT + abstract + 136 + Medical education increasingly relies on small-group learning. Small group learning provides more active learning, better retention, higher satisfaction, and facilitates development of problem-solving and team-working abilities. However, less is known about student experience and preference for different small groups teaching models. We evaluated group educational dynamics and group learning process in medical school clerkship small group case-based settings, with a faculty member present versus absent. + + + ABSTRACT + abstract_title_1 + 645 + Methods + + + ABSTRACT + abstract + 653 + Students completed surveys after cases when the faculty was present ("in") or absent ("out") for the bulk of the discussion. 228 paired surveys (114 pairs) were available for paired analysis, assessing group dynamics, group learning process, student preference, and participation through self-report and self-rating of group behaviors tied to learning and discussion quality. + + + ABSTRACT + abstract_title_1 + 1037 + Results + + + ABSTRACT + abstract + 1045 + Ratings of group dynamics and group learning process were significantly higher with the faculty absent vs. present (p range <0.001 to 0.015). Students also reported higher levels of participation when the faculty member was absent (p = 0.03). Students were more likely to express a preference for having the faculty member present after "in" case vs. "out" case discussions. (p < 0.001). There was no difference in reported success of the case discussion after "in" vs. "out" cases (p = 0.67). + + + ABSTRACT + abstract_title_1 + 1567 + Conclusions + + + ABSTRACT + abstract + 1579 + Student groups without faculty present reported better group dynamics, group learning processes, and participation with faculty absent. Students reported that they feel somewhat dependent on faculty, especially when the faculty is present, though there was no significant difference in students reporting that they obtained the most they could from the discussion of the case after both "in" and "out" cases. + + + INTRO + title_1 + 1996 + Background + + + INTRO + paragraph + 2007 + Medical school educators are increasingly using small group, problem-based learning strategies, particularly in the pre-clinical years. Small group learning appears to be associated with better retention of material, more active learning, and higher learner satisfaction. Students who engage in small group learning also appear to have higher satisfaction with their education, better performance on both written exams and objective standardized clinical exams (OSCEs), especially when students arrive for small group work prepared, trained and knowledgeable. Small group learning may also help overcome cultural barriers and promote unified, collaborative learning among diverse student groups. + + + INTRO + paragraph + 2703 + While the potential benefits of small group learning are many, it is unclear exactly why small group learning is successful. The engagement of students in the group process and group dynamics appears important, though few students and faculty have a clear idea of what that means. Researchers have had mixed results defining successful models of group learning by examining the respective roles of students and faculty in both student-led and faculty-led groups. Some researchers have demonstrated that an active role for the faculty leads to improved student satisfaction, increased USMLE scores, and increased perception by students of the faculty's importance, particularly when the faculty member is well-versed in the subject matter and positively influences group functioning. + + + INTRO + paragraph + 3488 + However, another study noted that faculty members were responsible for 63% of the interactions in faculty-led groups of veterinary students, who also followed a case-based curriculum. High numbers of faculty-led interactions can result in a teacher-dominated discussion. Though the faculty member presumably has a much stronger command of the subject matter than students, teacher-dominated discussions may impede student leadership development and limit opportunities for interactive learning. In addition, students' critical thinking may be impeded by the presence of an authority figure who is assumed to have information that they lack. These finding are inconsistent with the goal of engaging students in the group process. However, another study noted that student-led groups were more likely than faculty-led groups to use shortcuts, which may undermine the development of complex problem-solving skills. Additional studies reported either strong or no relationships between the skills of the faculty leader and the groups' interaction and performance, suggesting that the effect of faculty leadership on small group learning may be highly variable. + + + INTRO + paragraph + 4649 + Students' perspectives on small-group learning are similarly varied. A majority of students ranked the role of the faculty member as least important in the group process of small-group learning, and several researchers have found that students prefer peer-led groups. Student behavior and group functioning appears to be influenced by their perception of the learning situation, which suggests that students might do better in small group situations led by peers. Group functioning, in turn, is correlated with academic performance. + + + INTRO + paragraph + 5184 + In these reports, key characteristics of the faculty-led or student-led groups (i.e., the students involved and the nature of the group members' interactions) were not described. Adult Learning Theory describes the benefits of active learner involvement in all steps of learning, and as described in Self Directed Learning Theory, students are more engaged and active when they are responsible for their learning, leading to better learning outcomes. Therefore, identifying teaching strategies consistent with these theories will promote learner engagement and outcomes. Our faculty made anecdotal observations that student discussion appeared to have increased in their absence when they stepped out of the room and then returned. Faculty noted that they often returned to more robust discussion than before they had left. Therefore, we decided to design a study to analyze student report and perceptions of group dynamics and group learning process within the same small group of students, comparing student report and perceptions of group dynamics and group learning process with the faculty member present and absent from the discussion. Examining these results will add to and strengthen what is known about the role of the faculty member in small group discussions, particularly how their presence or absence affects group learning and group dynamics. + + + METHODS + title_1 + 6544 + Methods + + + METHODS + title_2 + 6552 + Student population + + + METHODS + paragraph + 6571 + All students rotating through the Boston University Family Medicine Clerkship between 7/21/06 and 5/15/07 were asked to participate in the study. Students were not required to participate, and clerkship faculty members were blinded as to which students completed surveys. Students who participated were told to label both their surveys with a random code that contained no information that might identify them. Students were read a standard informed consent informing them that they were being asked to participate in a survey related to group dynamics that might be used to improve instructional quality in the future. They were also clearly informed that participation was voluntary, anonymous and would not influence their grade. + + + METHODS + paragraph + 7304 + During the academic year in which this study was conducted, there were 139 students who rotated through the Family Medicine clerkship, all of whom were invited to participate. Students were at various points in their third year, as this study was conducted over the course of a year, with students constantly rotating through this clerkship. The class was 46% male and 53% female; racial/ethnic data is unavailable, and survey participants were not asked to identify their gender, ethnicity or race on their survey. All students were enrolled at Boston University; no one was from out of town or participating in visiting clerkships. + + 9606 + Species + + participants + + + + METHODS + title_2 + 7938 + Setting and curriculum + + + METHODS + paragraph + 7961 + Boston University School of Medicine requires a six week clerkship in Family Medicine, which all students rotate through at some point during their third year. During the Family Medicine clerkship, students return to the medical school for four days for didactics. The didactic curriculum is structured around two simulated families, with students managing and following multiple family members over time through the use of case studies related to various members of the fictional families. These case studies illustrate common out-patient issues routinely seen in family medicine, such as hypertension, family planning and diabetes. Case studies are explored during a series of small group sessions, in which students are randomly divided into three small groups by an administrative assistant, each with a faculty member. All small groups meet simultaneously. In the small groups, students are given paper cases for each family member's doctor visits, which include history, physical exam, and laboratory data. Students work through the cases in a structured format: first they discuss the patient's concerns and then they discuss the physician's concerns. They then generate an assessment and plan for each visit. One student volunteers to scribe the group's thoughts and plans on the blackboard. No other student roles are assigned. Faculty review the students' assessment and plan, as written by the students on the blackboard, and direct the group to correct any mistakes. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D006973 + Disease + + hypertension + + + MESH:D003920 + Disease + + diabetes + + + + METHODS + paragraph + 9450 + During the course of the six week clerkship, each of the three small groups of students meet four times, with 4 to 5 cases discussed per session. Small group sessions last 3-4 hours. Student small groups are comprised of 6-8 students each, and the group composition remains constant. Cases are discussed in the same order in each group, as faculty members leading the groups are instructed to follow the order given in a standardized faculty guide. + + + METHODS + paragraph + 9903 + There are four small group sessions during each clerkship block, with three small groups of students meeting simultaneously during each of those small group sessions. An administrative assistant randomly assigned which small group sessions in each block were to be studied. All small group sessions and all small groups of students were assigned to be studied at some point over the course of the study. In each of those small group sessions, students completed two surveys:one after a case when the faculty was in the room ("in") and one after a case when the faculty had left the room for the bulk of the case discussion ("out"). Before leaving the room, the faculty gave the students scripted instructions, and left the room for a standardized amount of time. We rotated which cases were used as interventions ("out") and controls ("in"). + + + METHODS + paragraph + 10763 + We used the small group discussion structure that was explained to students prior to their first small group session and used in all small group case discussions, during cases that were studied. Students were told at the start of group work that they were being asked to participate in a voluntary, anonymous study that might be used to improve educational experiences in the future. This study design was quasi-experimental, as the same students participated and evaluated both formats of case discussions. Students were further informed that participation would not affect their grade. Students were not made aware that they were participating in a study of faculty "in" vs "out" of the room. Faculty members were given scripted language to use when they stepped out of the room explaining that they needed to step out for about 15 minutes (no reason for the absence was given) and instructing the students to continue with case discussion as before. Faculty left the room at the beginning of the case discussion. Each case is typically discussed for 20-40 minutes, varying based on the nature of the case and the speed of each particular small group of students. + + + METHODS + paragraph + 11940 + Surveys were randomly distributed to each group at different times. An administrative assistant with no other role in the study was instructed to randomly include a survey packet in with the general instructional packets she distributed to each group at the start of each session. Groups would then discuss the cases already slated to be discussed that day, in the pre-determined order used by all groups. Each group was surveyed twice. While cases were not used in this study at the same time, over the course of the 8 block year, each case was discussed with faculty in and with faculty out at least once. (Certain cases include a role play where the faculty member plays the part of the patient. These cases require the presence of faculty and as such were excluded from the study. Students were not aware of the differentiation of these cases, and no groups were surveyed about these "faculty-required" cases). The difficulty level of all cases is felt to be equivalent. + + 9606 + Species + + patient + + + + METHODS + title_2 + 12919 + Instrument + + + FIG + 12909_2014_258_Fig1_HTML.jpg + Fig1 + fig_caption + 12930 + Survey instrument. + + + METHODS + paragraph + 12951 + We used a survey instrument derived from the Group Climate Questionnaire - Short version (GCQ-S), a validated instrument used previously by other researchers. Three items from the GCQ-S were used. In addition, eight items were used from Steele et al in order to create an instrument more useful for our purpose. Using the survey, participants self-scored and self-rated learning, group participation and dynamics using 16 Likert scaled questions. In addition, they were asked how many students were in their group and were asked to report their perceptions of how many students they felt participated fully. (See Figure 1). Students' "in" surveys were then compared to their "out" surveys to assess differences in perceived group process and dynamics. + + 9606 + Species + + participants + + + - + Chemical + + GCQ-S + + + + METHODS + title_2 + 13716 + Analysis + + + METHODS + paragraph + 13725 + We performed a paired t-test analysis for each item to assess the change between the "in" and "out" score. SAS was used for all analyses (version 9.2, SAS Institute, Inc, Cary, NC). As there were 16 Likert-scaled items on the scale, we also performed a Bonferroni adjustment for multiple comparisons, dividing .05 by 16 and yielding a significance level of 0.0031. The Bonferroni adjustment is extremely conservative and led to some results being described as having a 'trend' toward significance (with p values less than .04 but more than .0031). + + + METHODS + paragraph + 14285 + This study was reviewed and approved by the Boston University research ethics Institutional Review Board (IRB). + + + RESULTS + title_1 + 14397 + Results + + + FIG + 12909_2014_258_Fig2_HTML.jpg + Fig2 + fig_caption + 14405 + Recruitment and enrollment. + + + RESULTS + paragraph + 14435 + During the academic year in which this study was conducted, there were 139 students who rotated through the Family Medicine clerkship, all of whom were invited to participate. We collected 283 surveys in total. Of these, 228 had a pair, leading to 114 paired surveys, which were analyzed. In addition, we collected 55 surveys that did not have a pair, and so were excluded from analysis. We think this is the result of non-matching identifiers (i.e. student completed 2 surveys but failed to use the same identifier on each) as well as the possibility that some students chose to only complete one survey. (See Figure 2). + + + TABLE + table + 15058 + aItems are on a 5 point Likert scale as follows: 1 = strongly disagree; 2 = disagree; 3 = neutral; 4 = agree; 5 = strongly agree. + + + TABLE + table + 15209 + bItems are on a 5 point Likert scale as follows: 1 = Minimal; 5 = Most possible. + + + TABLE + Tab1.xml + Tab1 + table_caption + 15299 + Students' perception of group dynamics/learning process, their participation, and case outcomes with faculty in and out of the discussion (n = 114) + + + TABLE + Tab1 + table + Tab1.xml + 15455 + Mean score p Faculty present "In" (n = 114) Faculty absent "Out" (n = 114) Student perception of group dynamics and group learning process The group and its membez Helped each other to express their ideas and feelings.a 4.4 4.6 0.015 Utilized the knowledge of students in the group.a 4.5 4.7 <0.01 The members challenged each other in their efforts to sort things out.a 4.1 4.4 0.012 Utilized the knowledge of the facilitator.a 4.2 3.5 <.001 The members depended upon the faculty leader for direction.a 3.3 2.3 <.001 The group discussion was chaotic and disorganized.a 1.4 1.6 0.06 Faced and accepted differences of opinion expressed by members of the groupa 4.5 4.5 0.058 Worked together efficientlya 4.6 4.6 0.67 Moved toward its goals on the basis of consensusa 4.6 4.6 0.32 Student participation and preference Rate your participation level.b 4.2 4.3 0.03 I felt comfortable speaking during the student discussion.a 4.6 4.6 0.49 I prefer to have faculty present during the entire discussion of this case.a 3.5 3.1 0.0008 Case content Arrived at a reasonable management plan for this case.a 4.6 4.5 0.03 The faculty member was essential to the success of this case discussion.a 3.3 2.5 <.001 I feel I got the most I could out of the discussion of this case.a 4.3 4.1 0.67 Considered psychosocial issues in the management of this case.a 4.4 4.4 0.89 + + MESH:C535569 + Disease + + psychosocial + + + + RESULTS + paragraph + 16876 + Students completed surveys at the completion of each case. They rated the entire case discussion - including time when the faculty was out of the room and time when the faculty was back in the room. Students self-rated the questions assessing and quantifying various elements of group dynamics/group learning process more highly during cases when faculty members were absent, stating that they felt the discussions were more participatory, used the knowledge of students within the group, and involved the members of the group challenging one another (Table 1). There was a trend towards rating the discussion as more "chaotic and disorganized" when faculty members were absent (p = 0.06). They were less likely to respond that they used the facilitator's knowledge or relied upon the facilitator when the faculty member was absent, suggesting that faculty-absent groups might be more likely to motivate students towards the kind of critical thinking and responsibility needed by medical professionals. + + + RESULTS + paragraph + 17892 + Regarding student participation, the students also rated their participation level and their comfort in participating more highly when the faculty member was absent. They were less likely to say that they preferred having the faculty member in the room for the entire case following an "out" case. + + + RESULTS + paragraph + 18194 + When we assessed case content (the outcome of the discussion), students were less likely to say that they arrived at a reasonable management plan after a case when the faculty was absent. However, when students rated whether they had gotten all they could out of the experience, there was no statistically significant difference between cases with faculty present or absent. Students were more likely to rate the faculty member's presence as "essential" after a case with the faculty present. Finally, the students' assessment of whether psychosocial issues were considered during the discussion was no different whether the faculty member was present or absent. + + MESH:C535569 + Disease + + psychosocial + + + + DISCUSS + title_1 + 18865 + Discussion + + + DISCUSS + paragraph + 18876 + This study examined student report of group dynamics and group learning process with the faculty member present or absent from the discussion. We found that students have higher self-rated group dynamics, group learning process, and participation with the faculty member absent. Results on several domains (preference for faculty presence, arriving at a reasonable management plan for the case) suggest that students feel somewhat dependent on the faculty member, more so after a case when the faculty was present. However, after cases when the faculty was absent, students felt that the faculty member was not essential to the success of the case. And after cases when the faculty was present, students were only slightly in agreement that the faculty member was essential (3.3 on the same Likert scale). Additionally, there was no difference in how much they felt they got out of the case based on faculty presence or absence. + + + DISCUSS + paragraph + 19805 + Given the goal of maximizing student learning, skill acquisition, and group experience, it is critical to understand the processes at play in the small group learning setting. The cognitive processes involved in small group learning situations can lead to better activation of prior knowledge, information recall, concept and theory building, and collaborative learning. Likewise group discussion has been shown to motivate students and increase their interest in the subject matter further enhancing student engagement and learning. Some studies suggest that higher social networking and peer interactions, both formal and informal, can increase learning as well. Additionally, it has been shown that educational interventions regarding group dynamics and group function can improve small group effectiveness. + + + DISCUSS + paragraph + 20616 + With the current emphasis on active learning, retention of material, and learner satisfaction associated with small group learning, the structure, process, and composition of the small group must also be better understood to promote these goals. Prior studies of the role of the faculty member in small group learning in medical education have shown mixed results with some showing better test scores, OSCE performance, and student satisfaction with an active faculty member, and others showing that a more active faculty member impedes the group learning process. While many of the previous studies looked at faculty-led or student-led groups, this study compared self-reports from the same students in both faculty-led and student-led groups. Comparing self-reports from the same students in different kinds of groups showed that small groups without a faculty member present leads to better perceived group dynamics, group learning process, and increased student participation. + + + DISCUSS + paragraph + 21597 + This study had several limitations. Students were recruited throughout the academic year, meaning some of the students in the study were near the end of their third year and more likely to be confident in their clinical knowledge and also had had more experience participating in small group discussions compared with early third-year students. The experiences that were compared using paired t-tests were group discussions within the same group of students and faculty member. However, the cases were different, which may have been partly responsible for some of the differences in scores (for example, a case featuring domestic violence might engender more or less animated discussion than a case featuring diabetes). In addition, faculty members leading groups had different amounts of experience and skill leading small group learning sessions. Because groups met simultaneously, we were not able to compare multiple groups with the same faculty member leading. However, evaluating groups with each faculty member in and out as well as conducting this study over the course of the year may help us address this limitation. Additionally, the cases used as intervention and control were rotated randomly. Finally, while many of the items in our survey had statistically significant differences showing increased discussion and participation when the faculty was absent, some of the absolute differences in scores were small. + + MESH:D003920 + Disease + + diabetes + + + + DISCUSS + paragraph + 23024 + This study implies that while we assume that students may have a tendency or desire to be more dependent on faculty, the critical activity of discussion and debate of clinical cases may necessitate structuring our curricula, methods, faculty development and/or educational design of our sessions to further promote and enable student participation and discussion. Next steps from this study include assessing learning outcomes such as test scores or clinical performance. Future study could build on these findings by obtaining student permission to record conversation in faculty-present and faculty-absent groups. This would enable a comparison of student participation and content using qualitative and quantitative methods. + + + CONCL + title_1 + 23752 + Conclusions + + + CONCL + paragraph + 23764 + In this study, student small groups without faculty present reported better group dynamics, group learning processes, and participation with faculty absent. Our respondents reported that they feel somewhat dependent on faculty, especially when the faculty is present, though they report similar case success after both "in" and "out" cases. Further research into best practices and effective teaching methodology for small groups is called for in order to maximize student satisfaction and learning. As a result of these findings, we now encourage faculty to use stepping out of the room as a tool to increase small group discussion. This study has made us devote more faculty development time and discussion to this and other techniques to enhance small group participation. These results can guide changes to teaching modalities and methodologies as well as lead to redistribution of faculty time and resources in response to self-reported student needs. Clerkships may choose to have more small group sessions without faculty presence, or with intermittent faculty presence. + + + CONCL + footnote + 24850 + Competing interests + + + CONCL + footnote + 24870 + The authors declare that they have no competing interests. + + + CONCL + footnote + 24929 + Authors' contributions + + + CONCL + footnote + 24954 + MH co-conceived the study, designed the methods and directed the study and manuscript preparation; JEW performed statistical analyses; JX created initial draft of manuscript; JW co-conceived the study and contributed to design and write-up of results. All authors read and approved the final manuscript. + + + CONCL + footnote + 25258 + Authors' information + + + CONCL + footnote + 25281 + Previous presentation of this material: + + + CONCL + footnote + 25321 + Hoffman M. Xu J. Wilkinson J. Wiecha J. The Effect of Faculty Presence on Small-group Learning/Group Dynamics in a Family Medicine Clerkship. STFM Annual Spring Conference 2010. 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surname:Lucid;given-names:Alan + surname:Johnston;given-names:Christopher D + 25530808 + surname:Feeney;given-names:Audrey + 2014 + 4272814 + 46 + 10.1186/s13099-014-0046-9 + front + 46 + 6 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + Gut Pathog; 2014; 6(1) 46. doi:10.1186/s13099-014-0046-9 + surname:Sleator;given-names:Roy D + surname:Lucey;given-names:Brigid + surname:Coffey;given-names:Aidan + Feeney A, Johnston CD, Lucid A, O Mahony J, Coffey A, Lucey B, Sleator RD + 0 + The role of the Cronobacter sakazakii ProP C-terminal coiled coil domain in osmotolerance + + 28141 + Species + + Cronobacter sakazakii + + + + ABSTRACT + abstract_title_1 + 90 + Background + + + ABSTRACT + abstract + 101 + We investigate the role of the C-terminal coiled coil of the secondary proline porter ProP in contributing to Cronobacter sakazakii osmotolerance. + + 28141 + Species + + Cronobacter sakazakii + + + MESH:D011392 + Chemical + + proline + + + + ABSTRACT + abstract_title_1 + 248 + Findings + + + ABSTRACT + abstract + 257 + The extended C-terminal domain of ProP1 (encoded by ESA_02131) was spliced onto the truncated C-terminal end of ProP2 (encoded by ESA_01706); creating a chimeric protein (ProPc) which exhibits increased osmotolerance relative to the wild type. + + + ABSTRACT + abstract_title_1 + 501 + Conclusions + + + ABSTRACT + abstract + 513 + It appears that the C-terminal coiled coil domain tunes ProP at low osmolality, whereas ProP transporters lacking the coiled coil domain are more active at a higher osmolality range. + + + INTRO + title_1 + 696 + Introduction + + + INTRO + paragraph + 709 + Survival of the foodbourne pathogen Cronobacter sakazakii in low water activity (aw) environments, e.g. powdered infant formula (PIF), is largely attributed to the accumulation of organic compounds termed osmolytes or compatible solutes. Synthesised de novo, or transported from the bathing solution, compatible solutes function to increase cell turgor thereby counterbalancing the external osmotic upshift and preventing water loss from the cell, which if left unchecked can lead to plasmolysis and ultimately cell death. + + 28141 + Species + + Cronobacter sakazakii + + + 9606 + Species + + infant + + + MESH:D014867 + Chemical + + water + + + MESH:D014867 + Chemical + + water + + + + INTRO + paragraph + 1232 + In Escherichia coli, a model organism for the study of bacterial osmoadaptation, the transmembrane protein ProP is perhaps the best characterised compatible solute uptake system; facilitating the uptake of both proline and glycine betaine. A member of the major facilitator superfamily (MFS), E. coli ProP is a 500-amino acid protein comprising of 12 transmembrane domains and a characteristic carboxy-terminal extension. In a previous in silico study we identified seven ProP homologues on the C. sakazakii BAA-894 genome; one of which, ESA_02131, encodes a protein exhibiting 90% identity to E. coli ProP. While the remaining six homologues encode proteins exhibiting features of classic secondary transporters, they are all 60-70 amino acids shorter than the E. coli ProP; lacking the extended carboxyl tail. Notwithstanding the lack of structural consistency, particularly at the C-terminal end, we have shown that six of the seven C. sakazakii proP homologues contribute to C. sakazakii osmotolerance, albeit to varying degrees. + + 562 + Species + + Escherichia coli + + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + 28141 + Species + + C. sakazakii + + + MESH:D011392 + Chemical + + proline + + + MESH:D001622 + Chemical + + glycine betaine + + + + INTRO + paragraph + 2268 + Culham et al. first described the E. coli ProP as harbouring unusual structural features which appeared unique within the transporter superfamily. This study predicted the formation of an alpha helical coiled coil resulting from the presence of the carboxyl terminal extension. Indeed, a synthetic polypeptide corresponding to the C-terminal extension of ProP formed a dimeric alpha helical coiled coil. Interestingly, when amino acid changes were introduced to the coiled coil, ProP required a larger osmotic upshift to become activated, suggesting that the C-terminal domain likely plays a role in osmosensing. Furthermore, a derivative of ProP which lacked the 26 amino acid C-terminal domain was expressed, but inactive. In contrast, despite the structural degeneracy observed between the homologues, C. sakazakii ProP homologues lacking the C-terminal extension do contribute to osmotolerance, albeit to a lesser extent than the extended ProP (which we designate Prop1) encoded by ESA_02131. + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + - + Chemical + + ProP + + + + INTRO + paragraph + 3265 + While several studies have focused on elucidating the role of the carboxyl extension in E. coli, little is known about the role, if any, of the ProP1 carboxyl extension in the far more osmotolerant C. sakzakii. Herein, we investigate the role of the C-terminal coiled coil of ProP1 in contributing to C. sakazakii osmotolerance, by creating a chimeric protein (ProPc) in which the extended C-terminal domain of ProP1 (encoded by ESA_02131) is spliced onto the truncated C-terminal end of ProP2 (encoded by ESA_01706). + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + + METHODS + title_1 + 3783 + Material and methods + + + METHODS + title_2 + 3804 + Bacterial strains and growth conditions + + + TABLE + table + 3844 + Ampr. This strain is resistant to ampicillian. + + - + Chemical + + ampicillian + + + + TABLE + Tab1.xml + Tab1 + table_caption + 3891 + Bacterial strains and plasmids + + + TABLE + Tab1 + table + Tab1.xml + 3924 + Strain or plasmid Relevant genotype or characteristics Source or reference Plasmids pUC18 Ampr, lacZ', pMB9 replicon pUC18: ESA_02131 pUC18 harboring ESA_02131 gene under control of the native promoter pUC18: ESA_01706 pUC18 harboring ESA_01706 gene under control the native promoter pUC18: ESA_01706CTE pUC18 harboring chimeric ESA_01706 with fused C-terminal extension (ESA_02131) under control of the native promoter This work Strains Cronobacter sakazakii BAA-894 C.sakazakii strain isolated from powdered formula associated with neonatal intensive care unit Escherichia coli DH5alpha Intermediate cloning host.supE44 DeltalacU169(80lacZDeltaM15)R17 recA1 endA1 gyrA96 thi-1 relA1 Invitrogen MKH13 MC4100Delta(putPA)101Delta(proP)2Delta(proU) MKH13 pUC18:ESA_02131+ Host strain harbouring pUC18: ESA_02131 plasmid. Ampr MKH13 pUC18:ESA_01706+ Host strain harbouring pUC18: ESA_01706 plasmid. Ampr MKH13 pUC18:ESA_01706CTE Host strain harbouring pUC18: ESA_01706CTE plasmid. Ampr This work + + 28141 + Species + + Cronobacter sakazakii + + + 28141 + Species + + C.sakazakii + + + 668369 + Species + + Escherichia coli DH5alpha + + + + METHODS + paragraph + 4945 + Bacterial strains and plasmids used in this study are listed in Table 1. + + + METHODS + title_2 + 5019 + Creation of the chimeric ProPc protein + + + TABLE + Tab2.xml + Tab2 + table_caption + 5058 + Primers + + + TABLE + Tab2 + table + Tab2.xml + 5068 + Primer name Primer sequence (5' to 3') Length Characteristics ESA_02131 F CATCGGCCGACAGGCCAGTCAATGAATGATGC 32 EagI cut site R CATTCTAGAGAGTACAACGGAATGCGGGG 29 XbaI cut site ESA_01706 F CATTCTAGAGTCGGGCGGCTCTTTATCTGG 30 XbaI cut site R CATGGATCCTTGACCAGATGACGCAGTCTTTC 32 BamHI cut site Chimeric-01706 F AATAAGCTTGTGGCTTTTTATGCCGGGCTGC 31 HindIII cut site R CAGGCCAGTAATCAGCGCCGCGCCCATGAC 30 3' SOEing overhang Chimeric-02131CTE F CGCGGCGCTGATTACTGGCCTGACGATGAAAG 32 5' SOEing overhang R AATGGATCCTTACTCGTTAATACGAGGATGCTGG 34 BamHI cut site pUC18 MCS Check F CATTAGCTCACTCATTAGGCACC 23 pUC18 insert check R CATTGTAAAACGACGGCCAGTG 22 pUC18 insert check + + + METHODS + paragraph + 5734 + PCR primers (Table 2) were designed for each proP homologue based on C. sakazakii strain BAA-894 sequence data available from the NCBI database (NC_009778.1). The formation of the chimeric ProP protein (ProPc), which consists of the extended coiled coil region of ProP1 (amino acid position 422 to 505) fused to the C-terminus of ProP2 (encoded by ESA_01706), was performed using a modified SOEing (Splicing by overlap extension) technique. In silico comparative analysis of the native ProP1 and ProP2 sequences, revealed a point of amino acid homology within the twelfth predicted transmembrane domain, a leucine/isoleucine/threonine triplet (LIT) at position 422-424 and 437-439 respectively, which was selected as the splice site. Briefly, the fusion was performed using three separate PCR reactions: the first PCR (primer set Chimeric-01706) resulted in an ESA_01706 (proP2) amplicon lacking the C-terminal extension but containing a 15-bp 3'overhang corresponding to the LIT triplet of the ProP1 C- terminal extension. The second PCR (primer set Chimeric-02131CTE) formed an amplicon of 210-bp encoding ProP1 C-terminal extension with a 5'-overhang, also corresponding to the LIT triplet. The final PCR (primer set Chimeric-01706-F Chimeric-02131tail-R) was performed with the two previous amplicons as template; resulting in a final product of 1,623-bp, representing the ESA_01706 native promoter and modified coding region (encoding the fused ProP1 C-terminal extension after the LIT triplet). This product was digested with restriction enzymes BamHI and HindIII and ligated to a similarly digested pUC18 vector forming pUC18:ESA_01706CTE (C-Termini Extension). The integrity of the chimeric sequence was confirmed by sequencing (MWG Operon, Germany and GATC, Germany) and transformed into E. coli MKH13. + + 28141 + Species + + C. sakazakii + + + 562 + Species + + E. coli + + + MESH:D007930 + Chemical + + leucine + + + MESH:D007532 + Chemical + + isoleucine + + + MESH:D013912 + Chemical + + threonine + + + + METHODS + title_2 + 7555 + Osmotolerance assay + + + METHODS + paragraph + 7575 + Overnight cultures of E. coli MKH13 clones expressing the wild-type and chimeric ProP proteins (ProP1, ProP2 and ProPC respectively) were grown at 37 C with shaking at 200 rpm in either 10 ml LB or M9 minimal media containing 0.5% glucose, 0.04% arginine, 0.04% isoleucine, 0.04% valine (Sigma-Aldrich Co.). Cells were pelleted by centrifugation at 5,000 g, washed and re-suspended in 200 mul 1/4 strength Ringer's solution. The cell suspension was added to the appropriate filter sterilized media with varying concentrations (0-10%) of added NaCl. Growth was monitored in the relevant media over a 48 hour period, with optical density (OD600) readings being taken every hour. Triplicate readings were taken and graphs were plotted using SigmaPlot version 11.0. E. coli MKH13 harbouring the empty pUC18 plasmid was used as a negative control. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 1239833 + Species + + at 5 + + + MESH:D001120 + Chemical + + arginine + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D014633 + Chemical + + valine + + + MESH:D007532 + Chemical + + isoleucine + + + - + Chemical + + MKH13 + + + MESH:D005947 + Chemical + + glucose + + + + RESULTS + title_1 + 8426 + Results + + + RESULTS + title_2 + 8434 + C. sakazakii ProP structures + + 28141 + Species + + C. sakazakii + + + + FIG + 13099_2014_46_Fig1_HTML.jpg + Fig1 + fig_caption + 8463 + Predicted transmembrane and tertiary structures of A) ProP2 encoded by ESA_01706, B) ProP1 encoded by ESA_02131 and C) ProPc. + + + RESULTS + paragraph + 8591 + Based on sequence similarity to the E. coli ProP protein, we identified ProP1 (the product of ESA_02131) as the most likely ProP homolog in C. sakazakii; exhibiting 90% amino acid sequence identity and structural features characteristic of E. coli ProP. Indeed, further analysis using TMHMM and TexTopo software predicted ProP1 to be a membrane protein with 12-transmembrane domains, an extended central hydrophilic loop and carboxy terminal extension (Figure 1). While the remaining five ProP homologues on the C. sakazakii BAA-894 genome were also predicted to encode proteins with 12 transmembrane domains and an extended central hydrophilic loop, they each lacked the extended carboxy-terminal domain identified in ProP1, a feature which likely affects the final protein structure and function. + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + + RESULTS + paragraph + 9391 + Figure 1B illustrates the tertiary structure for ProP1 (predicted using the I-TASSER server). Most notably the presence of a coiled coil domain is evident as a result of the extended carboxy-terminal identified by sequence analysis. The coiled coil domain likely protrudes into the intracellular cytoplasm of the organism where its function remains unclear. By contrast, the tertiary structure of ProP2 (Figure 1A), representative of the remaining 6 ProP homologues and exhibiting 40% identity to E. coli ProP and 49% identity to ProP1, lacks the coiled coil domain at the carboxy-terminal end. + + 562 + Species + + E. coli + + + + RESULTS + title_2 + 9988 + Chimeric protein (ProPc) expression in E. coli MKH13 + + 562 + Species + + E. coli + + + + RESULTS + paragraph + 10041 + The osmoprotective properties of ProP1, ProP2 and ProPc were measured and compared in E. coli MKH13; an osmosensitive mutant incapable of growth in high osmolality environments (>=4%). The pUC18 plasmid containing each gene of interest was transformed to E. coli MKH13. Transformation efficiencies of 60 CFU/mug DNA were achieved, with successful transformation being confirmed by colony PCR, followed by sequencing. Transformants were screened for osmotolerance on media (both LB and M9 plus 1 mM proline) containing between 4% and 10% added NaCl. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + MESH:D011392 + Chemical + + proline + + + MESH:D012965 + Chemical + + NaCl + + + + RESULTS + title_2 + 10593 + Assessment of osmotolerance + + + RESULTS + paragraph + 10621 + To determine the effect of each ProP homologues, both native (ProP1 and ProP2) and chimeric (ProPc), on the osmotolerance of E. coli MKH13, each of the strains was grown in media containing varying concentrations of NaCl. Growth was monitored over a 48 hour period in minimal media supplemented with 1 mM proline and containing 0-10% added NaCl. + + 562 + Species + + E. coli + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D011392 + Chemical + + proline + + + MESH:D012965 + Chemical + + NaCl + + + + FIG + 13099_2014_46_Fig2_HTML.jpg + Fig2 + fig_caption + 10969 + Physiological analysis of E. coli MKH13 expressing ProP1, ProP2, ProPc and the empty pUC18 plasmid. A) Optical density was measured over a 48 hour period in media supplemented with 0-10% added NaCl. B) The growth rate of E. coli MKH13 expressing each of the proteins of interest was measured in media supplemented with 0-10% added NaCl. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + + RESULTS + paragraph + 11312 + Media containing 5% NaCl yielded the most discriminatory results. While E. coli MKH13 expressing the empty pUC18 vector showed no growth at 5%, each of the other three strains tested conferred some degree of osmotolerance on the host (Figure 2A). The strain expressing ProP1 was the most osmotolerant, with a maximum optical density (OD600) of 0.326 after 37 hours growth at 5% NaCl. The strain possessing ProPc grew to an OD significantly higher than the strains expressing either ProP1 or ProP2. E. coli MKH13 expressing ProP2 in 5% NaCl grew to a maximum OD of 0.111 after 48 hours, whereas E. coli MKH13 containing ProPc grew to a final OD of 0.189 at the same time point. Interestingly, growth of the stains expressing ProP2 and the chimeric protein continued to increase up to 48 hours, while growth of E. coli MKH13 expressing ProP1 reached maximum OD after only 37 hours. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 1239833 + Species + + at 5 + + + 1239833 + Species + + at 5 + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + - + Chemical + + ProP2 + + + + TABLE + Tab3.xml + Tab3 + table_caption + 12197 + Growth rate and optical density @ 600 nm + + + TABLE + Tab3 + table + Tab3.xml + 12241 + Protein expressed Gene locus tags Name 5% 6% 7% 8% 9% 10% Max. OD Growth rate (hr -1 ) Max. OD Growth rate (hr -1 ) Max. OD Growth rate (hr -1 ) Max. OD Growth rate (hr -1 ) Max. OD Growth rate (hr -1 ) Max. OD Growth rate (hr -1 ) Native ESA_02131 ProP1 0.326 0.009 0.144 0.005 0.152 0.004 0.177 0.004 0.181 0.004 0.204 0.004 Native ESA_01706 ProP2 0.111 0.003 0.096 0.002 0.083 0.002 0.074 0.002 0.127 0.002 0.171 0.002 Chimeric ESA_02131 ESA_01706 ProPc 0.189 0.004 0.130 0.003 0.113 0.003 0.117 0.003 0.124 0.003 0.141 0.003 + + + RESULTS + paragraph + 12826 + Each E. coli MKH13 strain expressing a proP gene of interest conferred osmotolerance (Figure 2B). As expected, E. coli MKH13 demonstrated a significant reduction in growth rate as NaCl concentrations increased, with a final growth rate of 0.0004 hr-1 recorded in media supplemented with 4% NaCl and no subsequent growth recorded thereafter. E. coli MKH13 expressing ProP1 demonstrated the highest osmotolerance of all the strains tested with growth rates of 0.009 hr-1 to 0.004 hr-1 recorded in media supplemented with 5% to 10% NaCl respectively. The next most osmotolerant strain was that expressing ProPc. Growth rates of 0.004 hr-1 to 0.003 hr-1 were recorded in media supplemented with 5% to 10% NaCl respectively (Table 3). This was higher than the growth rates observed when E. coli MKH13 expressing the native ESA_01706 gene (ProP2) was grown in a high osmolality environment, suggesting an important role for the C. sakazakii ProP C-terminal extension in osmotolerance. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D012965 + Chemical + + NaCl + + + + DISCUSS + title_1 + 13822 + Discussion + + + DISCUSS + paragraph + 13833 + A unique feature of the neonatal pathogen C. sakazakii is its ability to survive for prolonged periods in environments of low aW, such as powdered infant formula (PIF), making it a significant cause for concern. Indeed, up to 80% of infants infected with C. sakazakii die within days of birth, while survivors often suffer delayed neurological symptoms, brain abscesses or hydrocephalus. However, despite this, little is known about the molecular mechanisms that allow this organism to survive in environments such as PIF where it is subject to extreme hyper-osmotic stress. + + 28141 + Species + + C. sakazakii + + + 9606 + Species + + infant + + + 9606 + Species + + infants + + + 28141 + Species + + C. sakazakii + + + MESH:D006849 + Disease + + hydrocephalus + + + MESH:D001922 + Disease + + brain abscesses + + + MESH:D009422 + Disease + + neurological symptoms + + + MESH:D004194 + Disease + + hyper-osmotic stress + + + + DISCUSS + paragraph + 14408 + In a previous in silico study we identified seven copies of an E. coli proP homolog on the BAA-894 genome. Physiological analysis confirmed that six of the proP homologues identified played a role in osmotolerance. The availability of osmolytes in the media also had an effect on the osmotolerance of the host, with growth rates varying depending on the type or variety of compatible solutes present. While all six ProP proteins exhibited features characteristic of classic secondary transporters, five of the proteins were between 60-70 amino acids shorter than ProP1; lacking the characteristic C-terminal cytoplasmic extension Previous studies in our lab have demonstrated that the six C. sakazakii ProP homologues lacking the C-terminal coiled coil are significantly less osmoprotective than ProP1, suggesting an important role for this domain in modulating C. sakazakii osmotolerance. + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + 28141 + Species + + C. sakazakii + + + + DISCUSS + paragraph + 15300 + In the current study, the C. sakazakii ProP2 (encoded by ESA_01706) was chosen as the prototypical ProP homologue to study the role of the alpha helical coiled coil in osmotolerance. Genetic splicing yielded a chimeric protein structure (ProPc) possessing the native ProP2 domains in addition to the C-terminal alpha helical coiled coil domain from ProP1 (Figure 1C). E. coli MKH13 expressing ProPc grew to a higher OD in minimal media supplemented with proline, when compared to the native protein ProP2 which lacked the extended coiled coil domain (Figure 2A). These data demonstrate that the addition of the coiled coil domain from ProP1 to ProP2 results in a protein with an increased osmoprotective effect on the usually osmotically sensitive E. coli MKH13. However, as the osmolality of the medium increased, this trend appeared to reverse with OD readings becoming similar at 9% NaCl and the chimeric protein growing to a higher OD than the native in 10% NaCl, suggesting that the extent of osmotic pressure also has a role to play in the activity of the proteins. + + 28141 + Species + + C. sakazakii + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D011392 + Chemical + + proline + + + MESH:D012965 + Chemical + + NaCl + + + - + Chemical + + MKH13 + + + + DISCUSS + paragraph + 16374 + The role of the C-terminal domain of other osmolyte transporters, such as BetP (Corynebacterium glutamicum) and OpuA (Bacillus subtilis), was demonstrated to be important for the activation of these proteins during an increase in the osmolality of the surrounding medium. Furthermore, Culham et al. created a synthetic polypeptide corresponding to the C-terminal domain of E. coli ProP which formed a dimeric alpha helical coiled coil structure, similar to the coiled coil structure of ProP1 (illustrated in Figure 1A). In the same study ProP proteins from both E. coli and Agrobacterium tumefaciens, possessing the characteristic alpha helical coiled coil, were activated at a lower osmolality than orthologues lacking the coiled coil structure. C. glutamicum possesses a ProP protein which lacks the C-terminal alpha helical coiled coil domain and, presumably as a result of this, requires a higher osmolality for activation. E. coli ProP variants lacking the coiled coil or with an amino acid substitution disrupting the formation of the alpha helical coiled coil, also require a larger osmotic upshift than the wild-type transporter. This study demonstrates that the activity of these ProP orthologues is dependent on the osmolality of the surrounding medium, and the alpha helical coiled coil is believed to tune the transporter to osmoregulate the cell over a low osmolality range. These data may therefore offer an explanation for the increased growth observed in E. coli MKH13 expressing ProP2, which lacks the coiled coil domain, in media supplemented with 10% NaCl relative to either ProP1 or ProPc (Figure 2). It is likely that the coiled coil domain of C. sakazakii ProP1 has a similar tuning function. Furthermore, the presence of multiple ProP porters lacking the C-terminal coiled coil domain, and therefore only active at a higher osmolality, may well explain the extreme osmotolerance unique to C. sakazakii; allowing the pathogen to survive in environments like PIF. The ProP1 protein, on the other hand possessing the coiled coil, may be the only osmolyte transporter required to respond to low or moderate hyperosmotic challenge. + + 1718 + Species + + Corynebacterium glutamicum + + + 1423 + Species + + Bacillus subtilis + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 358 + Species + + Agrobacterium tumefaciens + + + 1718 + Species + + C. glutamicum + + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + 28141 + Species + + C. sakazakii + + + 28141 + Species + + C. sakazakii + + + MESH:D012965 + Chemical + + NaCl + + + - + Chemical + + BetP + + + + CONCL + title_1 + 18526 + Conclusion + + + CONCL + paragraph + 18537 + The addition of the coiled coil domain from ProP1 to ProP2 resulted in a chimeric protein (ProPc) which demonstrated higher osmotolerance compared to the native ProP2 (under moderate osmotic stress conditions). Furthermore, the growth rate of E. coli MKH13 expressing ProP2 increased in minimal media supplemented with 10% NaCl; suggesting that, as is the case in E. coli, the coiled coil domain tunes ProP at low osmolality, whereas ProP transporters lacking the coiled coil domain are more active at a higher osmolality range. + + 562 + Species + + E. coli + + + 562 + Species + + E. coli + + + MESH:D012965 + Chemical + + NaCl + + + + CONCL + footnote + 19066 + Audrey Feeney and Christopher D Johnston are contributed equally to this work. + + + CONCL + footnote + 19145 + Competing interests + + + CONCL + footnote + 19165 + The authors declare that they have no competing interests. + + + CONCL + footnote + 19224 + Authors' contributions + + + CONCL + footnote + 19249 + AF, CJ and AL carried out the the experimental work. AF drafted the manuscript together with together with CJ, AL and RDS. 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The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + Clin Epigenetics; 2014; 6(1) 32. doi:10.1186/s13148-014-0032-6 + Lynch syndrome Colorectal cancer Nucleosome Splice site Splicing Acceptor Donor Exon + surname:Ward;given-names:Robyn L + Sloane MA, Hesson LB, Nunez AC, Thompson BA, Ward RL + 0 + Nucleosome positioning is unaltered at MLH1 splice site mutations in cells derived from Lynch syndrome patients + + 17350 + Gene + 208 + + MLH1 + + + MESH:D003123 + Disease + + Lynch syndrome + + + + ABSTRACT + abstract_title_1 + 112 + Background + + + ABSTRACT + abstract + 123 + Splicing is more efficient when coupled with transcription and it has been proposed that nucleosomes enriched in exons are important for splice site recognition. Lynch syndrome is a familial cancer syndrome that can be caused by the autosomal dominant inheritance of splice site mutations in the MutL homolog 1 (MLH1) gene. To better understand the role of nucleosomes in splicing, we used MLH1 splice site mutations in Lynch syndrome cases as a model to investigate if abnormal splicing was associated with altered nucleosome positioning at exon-intron boundaries. + + 4292 + Gene + 208 + + MutL homolog 1 + + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + MESH:D003123 + Disease + + Lynch syndrome + + + MESH:D003123 + Disease + + Lynch syndrome + + + MESH:D009386 + Disease + + familial cancer syndrome + + + + ABSTRACT + abstract_title_1 + 689 + Findings + + + ABSTRACT + abstract + 698 + Nucleosome Occupancy and Methylome sequencing (NOMe-seq) was used to determine the allele-specific positioning of nucleosomes around heterozygous splice site mutations in lymphoblastoid cells lines (LCLs) derived from six Lynch syndrome patients. These mutations were previously shown to cause exon skipping in five of the six patients. Allele-specific high-resolution nucleosome mapping across exons and exon-intron boundaries revealed high levels of nucleosomes across all regions examined. Alleles containing donor or acceptor splice site mutations showed no consistent alteration in nucleosome positioning or occupancy. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + donor + + + MESH:D003123 + Disease + + Lynch syndrome + + + + ABSTRACT + abstract_title_1 + 1322 + Conclusion + + + ABSTRACT + abstract + 1333 + Nucleosomes were enriched at MLH1 exons in LCLs derived from Lynch syndrome patients, and in this model system the positioning of nucleosomes was unaltered at exon-intron boundaries containing splice site mutations. Thus, these splice site mutations alone do not significantly change the local organisation of nucleosomes. + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + patients + + + MESH:D003123 + Disease + + Lynch syndrome + + + + RESULTS + title_1 + 1656 + Findings + + + RESULTS + title_2 + 1665 + Background + + + RESULTS + paragraph + 1676 + Splicing removes introns from a longer pre-cursor RNA molecule to produce a final processed mRNA. Splice donor and acceptor sites located at the junctions between exons and introns contain conserved sequence elements that are necessary for splicing. Although in vitro splicing can occur, splicing is more efficient when coupled with transcription, suggesting that factors additional to the nucleic acid sequence are important for splice site recognition. It has been proposed that the positioning of nucleosomes at exons aids in splice-site recognition. + + 9606 + Species + + donor + + + + RESULTS + paragraph + 2230 + Nucleosome positioning describes the precise location of a given nucleosome, whereas nucleosome occupancy refers to the proportion of molecules bearing a nucleosome at a specific location, at any given instant. It is hypothesised that nucleosomes positioned within exons, especially those with weak splice sites, cause RNA Polymerase II (RNAPII) to pause, enabling an interaction with the spliceosome and more efficient splicing. In support of this, nucleosome occupancy is enriched across exons, which have a high GC content that favours nucleosome assembly, and the average size of an exon within the body of a human gene is 151 bp, that is similar in length to nucleosomal DNA (approximately 147 bp). Splicing factors associate with the C-terminal domain of RNA Polymerase II (RNAPII), while the histone modification H3K36me3 is enriched in exons, where it may act as a scaffold to recruit splicing factors. Taken together this indicates there is interplay between nucleosomes, the splicing machinery and the DNA sequence. + + 9606 + Species + + human + + + + RESULTS + paragraph + 3256 + Lynch syndrome is an autosomal dominant familial cancer syndrome characterised by early onset colorectal, endometrial and other cancers. It is most commonly caused by the inheritance of heterozygous loss-of-function mutations, including splice site mutations, in the DNA mismatch repair (MMR) genes MLH1 and MSH2. In this study we used MLH1 splice site mutations in Lynch syndrome as a model to better understand the role of nucleosomes in splicing. We investigated cells from Lynch syndrome patients with splice site mutations to determine whether splicing aberrations were associated with altered nucleosome positioning at exon-intron boundaries. + + 4292 + Gene + 208 + + MLH1 + + + 4436 + Gene + 210 + + MSH2 + + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + patients + + + MESH:D003123 + Disease + + Lynch syndrome + + + MESH:D015179 + Disease + + colorectal + + + MESH:D003123 + Disease + + Lynch syndrome + + + MESH:D014591 + Disease + + endometrial + + + MESH:D003123 + Disease + + Lynch syndrome + + + MESH:D009369 + Disease + + cancers + + + MESH:D009386 + Disease + + autosomal dominant familial cancer syndrome + + + + METHODS + title_1 + 3905 + Materials and methods + + + METHODS + title_2 + 3927 + Identification of MLH1 splice site mutations + + 4292 + Gene + 208 + + MLH1 + + + + TABLE + Tab1.xml + Tab1 + table_caption + 3972 + Molecular features of MLH1 splice site mutations in six individuals with Lynch syndrome + + 4292 + Gene + 208 + + MLH1 + + + MESH:D003123 + Disease + + Lynch syndrome + + + + TABLE + Tab1 + table + Tab1.xml + 4062 + Mutation Mutation location (GRC37, Feb 2009) Affected splice site Nucleosome occlusion of splice site Splicing error Allele Proportion of alleles c.588 + 1G > T Chr. 3: 37,053,354 Donor Wild-type (G) 19/20 Exon 7 skipping (r.546_588del) First bp of intron 7 Mutant (T) 21/22 Acceptor Homozygous 36/42 c.589-2A > G Chr. 3: 37,053,500 Acceptor Wild-type (A) 17/20 4 bp deletion in cDNA (r.589_592del) 2nd last bp of intron 7 Mutant (G) 19/22 c.790 + 1G > T Chr. 3: 37,056,036 Donor Wild-type (G) 33/34 Unknown 1st bp of intron 9 Mutant (T) 21/24 c.791-1G > C Chr. 3: 37,058,996 Acceptor Wild-type (G) 30/30 Exon 10 skipping (r.791_884del) Last bp of intron 9 Mutant (C) 12/18 c.884G > A Chr. 3: 37,059,090 Donor Wild-type (G) 42/42 Exon 10 skipping (r.791_884del) Last bp of exon 10 Mutant (A) 24/30 c.1559-2A > T Chr. 3: 37,081,675 Acceptor Wild-type (A) 17/20 Two aberrant transcripts (Exon 14 skipping or exon 14 and 15 skipping - r.[1559_1667del, 1559_1731del]) Second last bp of intron 13 Mutant (T) 16/16 + + 9606 + Species + + Donor + + + 9606 + Species + + Donor + + + 9606 + Species + + Donor + + + c.1559-2A>T;RS#:267607836 + DNAMutation + + c.1559-2A > T + + + r.[1559_1667del + DNAMutation + + r.[1559_1667del + + + r.791_884del + DNAMutation + + r.791_884del + + + c.1G>T + DNAMutation + + 1G > T + + + r.791_884del + DNAMutation + + r.791_884del + + + c.1559_1731del] + ProteinMutation + + 1559_1731del] + + + c.589-2A>G;RS#:267607767 + DNAMutation + + c.589-2A > G + + + r.546_588del + DNAMutation + + r.546_588del + + + c.884G>A;RS#:63750144 + DNAMutation + + c.884G > A + + + c.1G>T + DNAMutation + + 1G > T + + + r.589_592del + DNAMutation + + r.589_592del + + + c.791-1G>C;RS#:267607795 + DNAMutation + + c.791-1G > C + + + + METHODS + paragraph + 5123 + Bioinformatic analysis and in vitro assays previously showed that genetic mutations at exon-intron boundaries in the MLH1 gene generate aberrantly spliced transcripts (see Table 1). All mutations were classified as pathogenic (Class 5) according to the International Society for Gastrointestinal Hereditary Tumours Variant Interpretation Committee (InSiGHT VIC). + + 4292 + Gene + 208 + + MLH1 + + + MESH:D004067 + Disease + + Gastrointestinal Hereditary Tumours + + + + METHODS + title_2 + 5487 + Cell culture + + + METHODS + paragraph + 5500 + LCLs were established from patient blood by transformation with Epstein-Barr Virus (as described previously), and cultured in RPMI with 10% fetal bovine serum (Gibco, Life Technologies) at 37 C in 5% CO2. Lymphocytes for transformation from five patients (c.588 + 1G > T, c.589-2A > G, c.791-1G > C, c.884G > A and c.1559-2A > T) were obtained from the Australasian Colorectal Cancer Family Registry (ACCFR). Lymphocytes from the patient with the c.790 + 1G > T mutation were obtained from the MCO collection. + + 9606 + Species + + patient + + + 9913 + Species + + bovine + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + c.790+1G>T;RS#:267607789 + DNAMutation + + c.790 + 1G > T + + + c.791-1G>C;RS#:267607795 + DNAMutation + + c.791-1G > C + + + c.589-2A>G;RS#:267607767 + DNAMutation + + c.589-2A > G + + + - + Chemical + + CO2 + + + c.588+1G>T;RS#:267607772 + DNAMutation + + c.588 + 1G > T + + + c.884G>A;RS#:63750144 + DNAMutation + + c.884G > A + + + c.1559-2A>T;RS#:267607836 + DNAMutation + + c.1559-2A > T + + + MESH:D015179 + Disease + + Australasian Colorectal Cancer + + + - + Chemical + + RPMI + + + + METHODS + title_2 + 6043 + NOMe-seq + + + TABLE + Tab2.xml + Tab2 + table_caption + 6052 + NOMe-seq primer sequences and amplification conditions + + + TABLE + Tab2 + table + Tab2.xml + 6109 + Region NOMe-seq primers Annealing ( C) c.588 + 1G > T F:TTGATATTTAGTGTGTGTTTTTGGYAAT F/R = 54 C R:CACATAATATCTTAAAAAATTCCAAAATAATA F/RN = 56 C RN:ATACCRACTAACARCATTTCCAAAAATAA c.589-2A > G F:TTAGGTATTTAGTATATAATGYAGG F/R = 51 C R:CACTATAAATATTTTCAAAACTAAAACCTTA F/RN = 52 C RN:CACAAAATCTAAAAAATTACATACACCTAA c.790 + 1G > T F:TAGGYATAGGAGGATTTTAAATGGATTAAGTT F/R = 52 C R:CAATTTCTTTAATAACAATRCCTATACCTAAA F/RN = 52 C RN:TTACTCRTAAAAACTCTAACACCATCAA c.791-1G > C F:GATGTGATGTGYATATTATTATAGAAATGTT F/R = 55 C R:TATCARCACCTCCTAATAAAATGAARCATA F/RN = 56 C RN:ATCCTTTTRCCAATAATATATAAAATTCACTCTA c.884G > A F:GGATGTGATGTGYATATTATTATAGAAATGTT F/R = 58 C R:CTATTATARCTTCCCAACTAACCCCARCAA F/RN = 58 C RN:CTACAARCTATCARCACCTCCTAATAA c.1559-2A > T F:TATTAGGAGGYTTAATTTAGGYTTTTTTGYTTAT F/R = 58 C R:ACCCTCACCACCTAATTCACAACATTTATAA F/RN = 57 C RN:ACTAARCAACTACCAAAAACTAARCTTCTTA HSPA5 F: GAGAAGAAAAAGTTTAGATTTTATA F/R = 56 C R: AAACACCCCAATAAATCAATC + + 3309 + Gene + 3908 + + HSPA5 + + + c.1G>T + DNAMutation + + 1G > T + + + c.1G>T + DNAMutation + + 1G > T + + + c.589-2A>G;RS#:267607767 + DNAMutation + + c.589-2A > G + + + c.1559-2A>T;RS#:267607836 + DNAMutation + + c.1559-2A > T + + + c.791-1G>C;RS#:267607795 + DNAMutation + + c.791-1G > C + + + c.884G>A;RS#:63750144 + DNAMutation + + c.884G > A + + + + METHODS + paragraph + 7153 + NOMe-seq was performed as described previously. Briefly, intact nuclei were treated with 200 to 300 U GpC methyltransferase M.CviPl and 160 to 320 muM S-adenosylmethionine for 15 min at 37 C followed by termination of the reaction with an equal volume of 20 mM Tris HCl pH 7.9, 600 mM NaCl, 1% (w/v) SDS and 10 mM EDTA. DNA was extracted using phenol chloroform followed by ethanol precipitation and bisulfite modified using the EZ DNA Methylation-Gold Kit (Zymo Research). Regions incorporating the splice site mutation in each patient were amplified from 40 ng of bisulfite treated DNA using a nested PCR with the primers and annealing temperatures described in Table 2. Single molecule sequencing of PCR amplicons was performed as described previously. Wild-type and mutant alleles were distinguished using the splice site sequence alteration. The M.CviPI enzyme methylates accessible DNA at GpC sites, whereas nucleosome bound DNA is inaccessible and remains refractory to GpC methylation. Regions of M.CviPI inaccessibility of >=150 bp (the length of DNA wrapped around a single nucleosome) within a single molecule were considered to represent regions of nucleosome occupancy. In addition, NOMe-seq retains the endogenous methylation status of the DNA allowing nucleosome positions and DNA methylation to be determined on each molecule. + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 9606 + Species + + patient + + + MESH:C042345 + Chemical + + bisulfite + + + - + Chemical + + HCl + + + MESH:D012436 + Chemical + + S-adenosylmethionine + + + MESH:D004492 + Chemical + + EDTA + + + MESH:D012965 + Chemical + + NaCl + + + MESH:D002725 + Chemical + + chloroform + + + MESH:C042345 + Chemical + + bisulfite + + + - + Chemical + + Tris + + + MESH:D012967 + Chemical + + SDS + + + MESH:D019800 + Chemical + + phenol + + + MESH:D000431 + Chemical + + ethanol + + + + RESULTS + title_1 + 8502 + Results + + + FIG + 13148_2014_32_Fig1_HTML.jpg + Fig1 + fig_caption + 8510 + Allele-specific positioning of nucleosomes at MLH1 variants. (A) The MLH1 gene with arrows showing the location of the splice site mutations investigated in the LCLs derived from the six patients. The numbers correspond to the NOMe-seq assays shown in panel B (exon, vertical bar; intron, horizontal bar with arrows). (B) Each panel shows the nucleosome occupancy on the wild-type allele (upper) and the allele harbouring the indicated splice site mutation (lower). Blue box = exon; blue arrows = intron and direction of transcription; yellow diamond = location of splice site mutation; vertical black bars below line = GpC sites; vertical black bars above line = CpG sites; the pink bar represents a single nucleosome (150 bp, drawn to scale); pink shading indicates the location of nucleosomes on individual DNA molecules as determined by GpC methyltransferase inaccessibility; black circles = methylated GpC/accessible to M.CviPI; white circles = unmethylated GpC/inaccessible to M.CviPI. CpG sites are depicted by triangles. Black triangle = methylated CpG; white triangle = unmethylated CpG. + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 9606 + Species + + patients + + + + FIG + 13148_2014_32_Fig2_HTML.jpg + Fig2 + fig_caption + 9644 + Nucleosome occupancy and endogenous CpG methylation at the HSPA5 gene. (A) Schematic showing the location of the NOMe-seq assay (black bar) in relation to the HSPA5 promoter. The arrow represents the transcriptional start site. (B) NOMe-seq was used to map the position of nucleosomes on individual DNA molecules at the HSPA5 gene in LCLs derived from the six individuals with splice site mutations in MLH1. Vertical black bars below line = GpC sites; vertical black bars above line = CpG sites; pink bars = the location of nucleosomes on individual DNA molecules as determined by GpC methyltransferase inaccessibility; black circles = methylated GpC/accessible to M.CviPI; white circles = unmethylated GpC/inaccessible to M.CviPI. All endogenous CpG sites were unmethylated and triangles are not shown to provide better clarity of GpC sites. + + 3309 + Gene + 3908 + + HSPA5 + + + 3309 + Gene + 3908 + + HSPA5 + + + 3309 + Gene + 3908 + + HSPA5 + + + 4292 + Gene + 208 + + MLH1 + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + 2995 + Gene + 128415 + + GpC + + + + RESULTS + paragraph + 10508 + We determined the positioning of nucleosomes across exons 7, 8, 9, 10 or 14 of the MLH1 gene in LCLs derived from six Lynch syndrome patients. Splice site mutations within or near these exons affected splice donor or acceptor sites (Table 1). NOMe-seq and single molecule sequencing allowed us to distinguish mutant from wild-type alleles using the splice site sequence mutation. At all sites analysed the majority of molecules were inaccessible at GpC sites and methylated at CpG sites (Figure 1). NOMe-seq analysis of the control gene HSPA5, a constitutively active gene that maintains a nucleosome-depleted region at the transcription start site, showed that the CpG island was unmethylated (data not shown) and confirmed that GpC inaccessibility was due to nucleosome occupancy rather than incomplete M.CviPI treatment (Figure 2). This shows that the majority of molecules at the sites analysed are occupied by nucleosomes in these cells. + + 4292 + Gene + 208 + + MLH1 + + + 2995 + Gene + 128415 + + GpC + + + 3309 + Gene + 3908 + + HSPA5 + + + 2995 + Gene + 128415 + + GpC + + + 9606 + Species + + patients + + + 9606 + Species + + donor + + + MESH:D003123 + Disease + + Lynch syndrome + + + + RESULTS + paragraph + 11454 + Separation of alleles by presence or absence of the splice site mutation showed no overall difference in either nucleosome positioning or occupancy between wild-type and mutant alleles (Figure 1 and Table 1). In the six splice site mutations analysed, three were located within donor splice sites and three were located within acceptor splice sites (Table 1). Though some difference in precise positioning between mutant and wild-type alleles was observed across some exons (for example, exons 7 and 10, Figure 1B) this was confined to a small subset of molecules, rather than the majority of molecules as would be expected if the mutations affected nucleosome positioning. Our study therefore shows that nucleosome occupancy is unaltered at exon-intron boundaries containing these splice site mutations. + + 9606 + Species + + donor + + + + DISCUSS + title_1 + 12263 + Discussion + + + DISCUSS + paragraph + 12274 + In this study, we show that nucleosome positioning is unaltered at exon-intron boundaries containing splice site mutations. This finding suggests that the positioning of nucleosomes at exon-intron boundaries containing splice site mutations does not play a role in the splicing defect. + + + DISCUSS + paragraph + 12560 + NOMe-seq has been used to measure nucleosome occupancy at gene promoters, CTCF binding sites and regulatory elements but this is the first study that has utilised the technique to measure nucleosome occupancy at splice sites. NOMe-seq previously showed that the active CpG island (CGI) promoter of HSPA5 has a nucleosome-depleted region (NDR) upstream of the transcriptional start site. This region was included as a control in each assay, and in all cases the NDR was present. This indicated that the absence of GpC methylation observed in MLH1 was caused by nucleosome occlusion, not an artefact caused by a defective M.CviPI enzyme. In addition, CpG methylation was absent from the HSPA5 NDR which is a feature of active CGI promoters. Finally, gene body methylation is a feature of human cells and the majority of CpG sites within the MLH1 exons and introns were methylated. Together, these controls indicated that the NOMe-seq assays captured the true state of nucleosome positioning at MLH1 splice sites. + + 10664 + Gene + 4786 + + CTCF + + + 3309 + Gene + 3908 + + HSPA5 + + + 2995 + Gene + 128415 + + GpC + + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + human + + + 3309 + Gene + 3908 + + HSPA5 + + + + DISCUSS + paragraph + 13571 + Alternative splicing of MLH1 occurs in a range of normal tissues, but several pieces of evidence indicate that the transcripts observed in our study are caused by the mutation, rather than being normal splicing events. First, the splicing aberrations were predicted with bioinformatic splicing software, and second, although one of the aberrant transcripts (Delta10) has been reported among naturally occurring MLH1 splice transcripts, the clinical data associated with these variants are also indicative of pathogenicity as demonstrated by the results of previous multifactorial likelihood analyses. + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + c.del10 + DNAMutation + + Delta10 + + + + DISCUSS + paragraph + 14172 + One strength of our study was the ability to determine the allele-specific position of nucleosomes by incorporating a heterozygous single nucleotide variant into each NOMe-seq assay. Small changes in positioning were observed but these molecules constituted approximately the same small proportion of total molecules on both alleles. Subtle changes in nucleosome positioning have been considered important in regulating the expression of cell cycle-dependent genes and enhancer accessibility, but in those studies the subtle changes were observed on a significant number of molecules that resulted in an overall change in nucleosome occupancy. There was no consistent alteration in nucleosome occupancy between the wild-type and mutant alleles at a variety of donor and acceptor sites, which would be expected if nucleosome positioning at these sites played a role in mediating the effects of splice site mutations. Together our data shows that alleles containing a splice site mutation show no differences in nucleosome occupancy to wild-type alleles. + + 9606 + Species + + donor + + + + DISCUSS + paragraph + 15225 + A limitation of the approach was that in most of the assays (assays 2 to 6) it was only possible to measure nucleosome positions at either the donor site or the acceptor site. Bisulfite conversion causes fragmentation of genomic DNA and it is technically challenging to amplify fragments greater than 500 bp in length. The donor and acceptor sites were separated by more than 2 kb of intronic sequence, making it impossible to concurrently determine the allele-specific position of nucleosomes at the donor and acceptor site on the same molecule. Although an independent NOMe-seq assay could be performed at the other donor or acceptor site, it would provide no information on allele-specificity. The data from one assay (c.588 + 1 G > T), however, in which both splice sites were present in the one amplicon, showed no significant change in nucleosome occupancy at the donor or acceptor site in intron 7. + + 9606 + Species + + donor + + + 9606 + Species + + donor + + + 9606 + Species + + donor + + + 9606 + Species + + donor + + + 9606 + Species + + donor + + + c.588+1G>T;RS#:267607772 + DNAMutation + + c.588 + 1 G > T + + + MESH:C042345 + Chemical + + Bisulfite + + + + DISCUSS + paragraph + 16139 + Previous studies have utilised genome-wide datasets of micrococcal nuclease (MNase) digested chromatin to investigate nucleosome occupancy. Here we used NOMe-seq to map the position of nucleosomes relative to exons and splice sites in a single gene at single molecule resolution. This approach provides the most accurate possible measurement of nucleosome positioning and enables allele-specific mapping of nucleosomes. In agreement with previous reports in humans, Caenorhabditis elegans and Drosophila melanogaster, we observed high nucleosome occupancy at exons. Thus, the findings from genome-wide studies were supported by our independent, single-molecule approach at the MLH1 gene. + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + humans + + + Disease + + Caenorhabditis elegans and Drosophila melanogaster + + + + DISCUSS + paragraph + 16827 + A key finding of our study was that nucleosome positioning was not significantly affected by mutations at the MLH1 splice sites investigated. Recent work with an in vitro IKBKAP mini-gene system found that alternative splicing changed chromatin organisation, with splice site strength and factors needed for splicing, such as U1 snSNP, playing a role in the regulation of nucleosome occupancy in exons. This and an earlier study indicate that splice site strength is an important determinant of nucleosome occupancy in exons. Exons with stronger polypyrimidine tracts (PPT; one of the conserved DNA elements located at 3' splice sites) have increased nucleosome occupancy compared to immediately adjacent introns, suggesting that nucleosomes act as a barrier that help to define the physical location of the 3' splice site. Although we investigated an equal number of donor and acceptor splice site mutations, nucleosome positioning was not significantly different between exons and the immediate 5' and 3' intronic sequence on the wild-type or mutant allele. The discrepancy between earlier studies and ours may in part relate to the strength of the MLH1 splice sites investigated. Alternatively, changes to chromatin organisation induced by splice mutations may be different to those caused by normal splicing events. Co-transcriptional splicing involves a complex interplay between RNAPII, the spliceosome and histone modifications and alterations in these factors, in addition to DNA sequence changes may be needed to alter nucleosome occupancy at exons. + + 4292 + Gene + 208 + + MLH1 + + + 8518 + Gene + 2699 + + IKBKAP + + + 6863 + Gene + 2394 + + PPT + + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + donor + + + - + Chemical + + polypyrimidine + + + + DISCUSS + paragraph + 18394 + In summary this study used NOMe-seq to determine the allele-specific position of nucleosomes in relation to MLH1 splice site mutations. We conclude that splice site mutations that cause aberrant splicing of MLH1 do not alone significantly affect local nucleosome positioning in LCLs from Lynch syndrome patients. + + 4292 + Gene + 208 + + MLH1 + + + 4292 + Gene + 208 + + MLH1 + + + 9606 + Species + + patients + + + MESH:D003123 + Disease + + Lynch syndrome + + + + DISCUSS + footnote + 18707 + Competing interests + + + DISCUSS + footnote + 18727 + The authors declare that they have no competing interests. + + + DISCUSS + footnote + 18786 + Authors' contributions + + + DISCUSS + footnote + 18811 + MAS and LBH designed NOM-seq assays. ACN and MAS performed NOMe-seq assays. BAT provided LCL samples. RLW and LBH conceived the study, participated in its coordination and helped to draft the manuscript. 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It may also be used consistent with the principles of fair use under the copyright law. + TITLE + Aquat. Toxicol.; 2015 Jan; 158192-201. doi:10.1016/j.aquatox.2014.11.016 + non-model organisms gene knock-outs mummichog adaptation zinc finger nucleases CRISPR-Cas9 + 201 + surname:Hahn;given-names:Mark E. + NIHMS645550 + surname:Champlin;given-names:Denise + Aluru N, Karchner SI, Franks DG, Nacci D, Champlin D, Hahn ME + 0 + Targeted mutagenesis of aryl hydrocarbon receptor 2a and 2b genes in Atlantic killifish (Fundulus heteroclitus) + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + Fundulus heteroclitus + + + + ABSTRACT + abstract + 112 + Understanding molecular mechanisms of toxicity is facilitated by experimental manipulations, such as disruption of function by gene targeting, that are especially challenging in non-standard model species with limited genomic resources. While loss-of-function approaches have included gene knock-down using morpholino-modified oligonucleotides and random mutagenesis using mutagens or retroviruses, more recent approaches include targeted mutagenesis using zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 technology. These latter methods provide more accessible opportunities to explore gene function in non-traditional model species. To facilitate evaluations of toxic mechanisms for important categories of aryl hydrocarbon pollutants, whose actions are known to be receptor mediated, we used ZFN and CRISPR-Cas9 approaches to generate aryl hydrocarbon receptor 2a (AHR2a) and AHR2b gene mutations in Atlantic killifish (Fundulus heteroclitus) embryos. This killifish is a particularly valuble non-traditional model for this study, with multiple paralogs of AHR whose functions are not well characterized. In addition, some populations of this species have evolved resistance to toxicants such as halogenated aromatic hydrocarbons. AHR-null killifish will be valuable for characterizing the role of the individual AHR paralogs in evolved resistance, as well as in normal development. We first used five-finger ZFNs targeting exons 1 and 3 of AHR2a. Subsequently, CRISPR-Cas9 guide RNAs were designed to target regions in exon 2 and 3 of AHR2a and AHR2b. We successfully induced frameshift mutations in AHR2a exon 3 with ZFN and CRISPR-Cas9 guide RNAs, with mutation frequencies of 10% and 16%, respectively. In AHR2b, mutations were induced using CRISPR-Cas9 guide RNAs targeting sites in both exon 2 (17%) and exon 3 (63%). We screened AHR2b exon 2 CRISPR-Cas9-injected embryos for off-target effects in AHR paralogs. No mutations were observed in closely related AHR genes (AHR1a, AHR1b, AHR2a, AHRR) in the CRISPR-Cas9-injected embryos. Overall, our results demonstrate that targeted genome-editing methods are efficient in inducing mutations at specific loci in embryos of a non-traditional model species, without detectable off-target effects in paralogous genes. + + 105937229 + Gene + + AHR2 + + + 105920361 + Gene + + AHRR + + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + Fundulus heteroclitus + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + MESH:D009841 + Chemical + + oligonucleotides + + + MESH:D064420 + Disease + + toxicity + + + MESH:D006841 + Chemical + + aromatic hydrocarbons + + + + INTRO + title_1 + 2495 + 1. INTRODUCTION + + + INTRO + paragraph + 2511 + The Atlantic killifish (Fundulus heteroclitus) is one of the most ecologically and environmentally important estuarine fish distributed along the East coast of the United States. Their ability to tolerate wide changes in environmental conditions, including temperature, salinity, oxygen and pH, have made them an ideal model species to investigate the biochemical, physiological and evolutionary basis of environmental adaptation. Some populations of killifish are also valuable models for understanding the mechanisms of evolved resistance to toxicants. Populations of killifish inhabiting contaminated coastal waters along the North Atlantic U.S. coast have evolved resistance to some contaminants representing major categories of aryl hydrocarbon pollutants, such as polynuclear aromatic hydrocarbons (PAHs), and halogenated aromatic hydrocarbons such as polychlorinated biphenyls (PCBs), 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD, 'dioxin') and other dioxin-like compounds (DLCs). This evolved resistance involves alterations in signaling through the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor through which some PAHs, PCBs, and TCDD alter gene expression and cause toxicity. Killifish express four AHR paralogs (AHR1a, AHR1b, AHR2a and AHR2b), the products of distinct loci. Although the respective functions of these AHRs are not well understood, AHR2 proteins appear to play a major role in mediating the developmental effects of PAHs and PCBs in fish. + + 105937229 + Gene + + AHR2 + + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + Fundulus heteroclitus + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 8078 + Species + + Killifish + + + 29154 + Species + + Atlantic + + + 29278 + Species + + expression + + + MESH:D010100 + Chemical + + oxygen + + + MESH:D011084 + Chemical + + PAHs + + + MESH:D011078 + Chemical + + polychlorinated biphenyls + + + MESH:D011078 + Chemical + + PCBs + + + MESH:D000072317 + Chemical + + TCDD + + + MESH:D011084 + Chemical + + PAHs + + + MESH:D006838 + Chemical + + hydrocarbon + + + MESH:D011084 + Chemical + + PAHs + + + MESH:D011078 + Chemical + + PCBs + + + MESH:D064420 + Disease + + toxicity + + + MESH:D000072317 + Chemical + + TCDD + + + MESH:D006841 + Chemical + + aromatic hydrocarbons + + + MESH:D011084 + Chemical + + polynuclear aromatic hydrocarbons + + + MESH:D004147 + Chemical + + dioxin + + + MESH:D004147 + Chemical + + dioxin + + + MESH:D000072317 + Chemical + + 2,3,7,8-tetrachlorodibenzo-p-dioxin + + + MESH:D011078 + Chemical + + PCBs + + + + INTRO + paragraph + 4005 + While there is a great deal of understanding about the physiological and biochemical basis of environmental adaptation in killifish, very little is known about its genetic basis, mainly due to the lack of genetic tools to evaluate gene function. Recently, the antisense morpholino oligonucleotide (MO)-based gene knockdown method was adapted to killifish to determine the role of aryl hydrocarbon receptors and cytochrome P4501A (CYP1A) in developmental toxicity, and to study the role of aquaporins in adult killifish. However, MOs are effective only for a limited period of time and do not completely eliminate gene expression. In order to conclusively demonstrate the function of any protein, gene knockouts are essential. + + 105930482 + Gene + + cytochrome P4501A + + + 105930482 + Gene + + CYP1A + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 29278 + Species + + expression + + + MESH:D064420 + Disease + + developmental toxicity + + + MESH:D060172 + Chemical + + morpholino oligonucleotide + + + + INTRO + paragraph + 4731 + In the past decade, several targeted genome-editing methods have been successfully employed to manipulate genes in a variety of model and non-model organisms. They include zinc-finger nucleases (ZFNs), transactivator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system (Fig. 1). ZFN and TALENs utilize engineered nucleases consisting of a sequence-specific DNA-binding domain fused to a nonspecific DNA cleavage module. The DNA-binding domains can be customized to recognize any sequence of interest. Distinct from the site-specific nucleases, CRISPR-Cas9 is an RNA-guided DNA nuclease (RGN), where a small guide RNA (sgRNA) complementary to a 20 base target DNA sequence guides Cas9, a bacterially derived nuclease, to induce a double-strand break (DSB) at the target site. Because the DSB repair process by non-homologous end-joining (NHEJ) is error prone, it leaves insertions or deletions (indels) at the target site. The relative ease of generating target-specific mutations using these techniques has opened up new avenues for conducting functional studies in any organism of interest. Several studies have demonstrated the efficacy of ZFNs, TALENs and RGNs in producing heritable mutations in non-model organisms such as rainbow trout, medaka, tilapia and yellow catfish (Table 1). Mutagenesis rates of up to 95% have been observed with CRISPR-Cas9. So far, there are no studies describing the generation of gene knock-outs in killifish or of targeted AHR mutants in any non-mammalian vertebrate. In this study, we utilized ZFN and CRISPR-Cas9 approaches to induce mutations in paralogous killifish AHR2a and AHR2b genes, and we observed indels in the injected embryos. We discuss strategies for the application of this method to knock out closely related paralogs in a species with a long generation time. + + 8022 + Species + + rainbow trout + + + 8090 + Species + + medaka + + + 1234273 + Species + + yellow catfish + + + 8078 + Species + + killifish + + + 9606 + Species + + mammalian + + + 8078 + Species + + killifish + + + Disease + + tilapia + + + + METHODS + title_1 + 6613 + 2. MATERIALS AND METHODS + + + METHODS + title_2 + 6638 + 2.1. Killifish adults and embryos + + 8078 + Species + + Killifish + + + + METHODS + paragraph + 6672 + Mature male and female Atlantic killifish were collected from Scorton Creek (Sandwich, MA) during full moon and new moon in Spring and Summer of each year (2011-2014) using minnow traps, as described previously. Although killifish populations are known to vary dramatically in their sensitivity to DLCs, this source population is known to be relatively sensitive to DLCs, characteristic of other populations of this species resident to similarly uncontaminated sites. Fish were maintained in tanks with continuous flow-through seawater (SW) at 18-20 C and 14h:10h light/dark photoperiod conditions. The animal husbandry practices were approved by the Animal Care and Use Committee of the Woods Hole Oceanographic Institution. Female fish were lightly anesthetized with Tricaine (MS222; buffered with sodium bicarbonate, Sigma-Aldrich, St. Louis, MO, USA) and oocytes were obtained for in vitro fertilization (IVF) by gently squeezing the abdomen. Oocytes were collected in glass petri dishes with filtered SW (25 parts per thousand; ppt). Milt was obtained by euthanizing mature males in MS222, dissecting out the gonads, and chopping them with a scalpel blade in seawater. A few drops of milt were added to the oocytes for fertilization. Each IVF experiment included a pool of oocytes stripped from at least 2-3 females and milt from 1 or 2 males. Approximately 20 minutes after the addition of milt, embryos were rinsed with filtered SW to remove any excess sperm. Fertilized embryos were maintained at 23 C until further use. + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + killifish + + + MESH:C537182 + Disease + + IVF + + + MESH:C003636 + Chemical + + Tricaine + + + - + Chemical + + 14h + + + MESH:C537182 + Disease + + IVF + + + MESH:D017693 + Chemical + + sodium bicarbonate + + + MESH:C003636 + Chemical + + MS222 + + + + METHODS + title_2 + 8209 + 2.2. Zinc Finger Nuclease design + + + METHODS + paragraph + 8242 + AHR2a exon 2 and 3 (Fig. 2) were each targeted with a five-finger ZFN pair using the CompoZr custom ZFN service (Sigma-Aldrich, St. Louis, MO, USA) (Table 2; Fig. 2). Left- and right-ZFN plasmids were in vitro transcribed and polyadenylated using the mMessage mMachine T7 kit (Life Technologies, Carlsbad, CA, USA). ZFN mRNA from each half-site was diluted to a concentration of 100-400 ng/microl and 2.5 nl of each was co-injected into 1 or 2-cell stage killifish embryos, as described below. + + 105937229 + Gene + + AHR2a exon 2 and 3 + + + 8078 + Species + + killifish + + + + METHODS + title_2 + 8737 + 2.3. CRISPR-Cas guide RNA design and construction + + + METHODS + paragraph + 8787 + Guide RNAs (gRNA) targeting AHR2a and AHR2b were designed using the ZiFiT Targeter website (http://zifit.partners.org/ZiFiT_Cas9). Exons 1, 2 and 3 of AHR2a and AHR2b were searched for target sites in order to obtain the shortest truncated mutant protein. A single target site was found in each exon. The AHR2a exon 1 target site did not begin with GG residues, which prevented it from being cloned into the gRNA expression vector. The AHR2b exon 1 target site was not optimal due to repetitive sequences. Therefore, the target sites for exons 2 and 3 of AHR2a and AHR2b were selected for gRNA synthesis (Table 2). PAGE-purified complementary oligonucleotides (Eurofins Genomics, Huntsville, AL, USA) were annealed and cloned into an expression vector following a published protocol. The gRNA oligonucleotide sequences are provided in Table 2. Expression vectors for guide RNA (pDR274) and Cas9 endonuclease (MLM3613) were obtained from Addgene (https://www.addgene.org/). The gRNA was transcribed using the MAXIscript T7 kit, and the Cas9 mRNA was transcribed and poly(A)-tailed with the mMESSAGE mMACHINE T7 ULTRA kit (Life Technologies, Carlsbad, CA, USA). gRNA and Cas9 mRNA were mixed at a ratio of 1:12 for the microinjections. + + 105937229 + Gene + + AHR2 + + + 29278 + Species + + Expression vectors + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + MESH:D009841 + Chemical + + oligonucleotide + + + MESH:D009841 + Chemical + + oligonucleotides + + + + METHODS + title_2 + 10021 + 2.4. Microinjection of ZFN mRNA and CRISPR-Cas9 gRNA + + + METHODS + paragraph + 10074 + Microinjection of killifish embryos was performed following a previously established protocol with some modifications. Briefly, prior to microinjection, 1-cell stage embryos were rolled on wet paper towels to prevent the filaments on the chorion from adhering to the microinjection needle. They were subsequently placed on custom-made agarose microinjection embryo trays and oriented so that the cell directly faces the microinjection needle. The microinjection setup consisted of a Zeiss Stemi 2000-C stereomicroscope, Narishige IM-300 microinjector and a micromanipulator (Narishige International, East Meadow, NY, USA). Aluminosilicate glass tubes (1.0 mm outer diameter, 0.64 mm inner diameter) were used to prepare microinjection needles. This type of needle was found to be superior to borosilicate needles for microinjection of killifish embryo, which have a thick chorion. A P-30 vertical micropipette puller (Sutter Instruments, Novato, CA, USA) at a heater output setting of 97 was used to pull aluminosilicate glass tubing into microinjection needles. The injection volume was calibrated by injecting the solution into mineral oil and measuring the diameter of the droplet using a stage micrometer. The microinjection volume was approximately 5 nL per embryo. To compensate for variations in needle opening size, injection time and pressure were altered during microinjection. Injected embryos were maintained in filtered SW at 23 C. Some of the injected embryos were transferred to the fish-rearing facility at the National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, Environmental Protection Agency, Narragansett, RI, to be raised to adulthood. The numbers of injected fish that are being raised for identifying founders are provided in Supplementary Table 1. + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 29154 + Species + + Atlantic + + + MESH:D012685 + Chemical + + agarose + + + MESH:D008899 + Chemical + + mineral oil + + + - + Chemical + + borosilicate + + + MESH:C049037 + Chemical + + aluminosilicate + + + MESH:D005597 + Disease + + opening + + + MESH:C049037 + Chemical + + Aluminosilicate + + + + METHODS + title_2 + 11885 + 2.5. ZFN protein expression + + + METHODS + paragraph + 11913 + ZFN-injected and uninjected embryos were sampled at 3, 6, 9 and 24 hours post-injection to determine the temporal profile of ZFN protein expression. At each time point, one pool of 10 embryos was sampled. Embryos were homogenized in 1x sample treatment buffer with beta-mercaptoethanol and boiled for 5 minutes to denature the protein. Samples were separated on an 8% polyacrylamide gel using the discontinuous buffer system. The proteins were transferred onto a nitrocellulose membrane with a semi-dry transfer unit (BioRad, Waltham, MA, USA), and the membrane was blocked with 5% milk in TBST (20 mM Tris pH 7.5, 300 mM NaCl and 0.1% (v/v) Tween 20) overnight. ZFN protein was detected using a mouse monoclonal anti-Flag antibody (Sigma, St. Louis, MO, USA) at 1:5000 dilution. The secondary antibody used was horseradish peroxidase (HRP)-conjugated goat anti-mouse antibody (Santa Cruz Biotechnology, Dallas, TX, USA) at 1:5000 dilution. The membranes were incubated in the primary antibody for 60 min at room temperature, washed with TBST twice (10 min each), incubated with secondary antibody for 60 min, and finally washed with TBST (15 min). Detection was done using the ECL Prime Western blotting reagent (GE Healthcare, Pittsburgh, PA, USA). + + 10090 + Species + + mouse + + + 3704 + Species + + horseradish + + + 9925 + Species + + goat + + + 10090 + Species + + mouse + + + 29278 + Species + + expression + + + - + Chemical + + Tris + + + MESH:C016679 + Chemical + + polyacrylamide + + + MESH:D012965 + Chemical + + NaCl + + + - + Chemical + + TBST + + + - + Chemical + + TBST + + + MESH:D011136 + Chemical + + Tween 20 + + + MESH:D015352 + Disease + + semi-dry transfer unit + + + MESH:D008623 + Chemical + + beta-mercaptoethanol + + + - + Chemical + + TBST + + + + METHODS + title_2 + 13164 + 2.6. Surveyor endonuclease assay + + + METHODS + paragraph + 13197 + Somatic mutations were determined using the Surveyor mutation detection kit (Transgenomic Inc., Omaha, NE, USA) following the manufacturer's instructions. Embryos (pool of 5 embryos per replicate) were sampled at 48 hours post-injection and genomic DNA was isolated using the proteinase K digestion method. The AHR2a and AHR2b target regions were amplified by polymerase chain reaction (PCR) using Advantage polymerase (Clontech, Mountain View, CA, USA). The primer sequences are given in Table 2. PCR cycling conditions were 94 C for 1 min, followed by 35 cycles of [94 C for 5 s, 68 C for 1 min], followed by 72 C for 7 min. The PCR products were melted and re-annealed to form hetero-duplex DNA and digested with Surveyor nuclease for 20 min at 42 C. The resulting products were electrophoresed on a 15% TBE gel and visualized by ethidium bromide staining. + + - + Chemical + + TBE + + + MESH:D004996 + Chemical + + ethidium bromide + + + + METHODS + title_2 + 14066 + 2.7. DNA sequencing + + + METHODS + paragraph + 14086 + PCR products from samples that were positive in the Surveyor endonuclease assay were cloned into the pGEM-T easy vector (Promega, Madison, WI, USA) and sequenced in 96-well plates (Functional Biosciences, Madison, WI, USA). Insertions and deletions were identified by aligning the sequences with the wildtype AHR sequence (Sequencher 5.1, Gene Codes Corp., Ann Arbor, MI, USA). Off-target screening for AHR2b CRISPR-Cas9 exon2 was done by direct-sequencing of PCR products. The amplification reactions were purified using the GeneClean kit (MP Biomedicals, Santa Ana, CA, USA). Direct-sequencing of the amplicons for AHR1a, AHR2a, and AHRR was done with primers Fh2a-ex2rev, 2b-1aF, and RR-ex2fwd, respectively (Table 2). The chromatograms were visually inspected for regions of double peaks indicating insertions or deletions. + + 105937229 + Gene + + AHR2 + + + 105920361 + Gene + + AHRR + + + + RESULTS + title_1 + 14914 + 3. RESULTS + + + RESULTS + title_2 + 14925 + 3.1. ZFN-induced AHR2a mutations + + + RESULTS + paragraph + 14958 + We tested different concentrations of ZFN mRNA on killifish embryos by injecting 500, 1000 and 2000 pg per embryo. No overt toxic effects or phenotypic abnormalities were observed at any of the concentrations as determined by microscopic observation of embryos during development. Hence, we selected 2000 pg to inject in subsequent experiments. ZFN protein expression was confirmed in the embryos injected with 2000 pg ZFN mRNA. Embryos were sampled at 3, 6, 9 and 24 hours post-microinjection. The maximum ZFN protein expression was observed at 6 hours post-microinjection (Fig. 3A). At 3 and 9 hours, ZFN protein expression was much lower, and at 24 hours, no expression was detected. These results suggest that the maximum efficiency of ZFNs in inducing DSBs in killifish is around 6 hours post-injection. + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + MESH:C007563 + Chemical + + DSBs + + + + RESULTS + paragraph + 15767 + Approximately 400 embryos were injected with the ZFN pair targeting exon 2 of AHR2a, and all were screened for mutations using the Surveyor mutation detection assay. We did not detect any insertions or deletions in these embryos; therefore, we next tested the ZFN pair targeting exon 3. Using the Surveyor assay, we analyzed 14 pools of ZFN-injected embryos (5 embryos per pool), and detected the presence of mutations in 4 pools (Fig. 3B). Endonuclease digestion resulted in fragments of approximately 240 bp and 95 bp (Fig. 2B, 3B). The PCR product from one of the embryo pools that was positive in the Surveyor assay was cloned into the pGEM-T Easy vector. Sequencing of multiple clones revealed 6 mutant sequences out of 59 (10%), represented by four different types of deletions at the target site (2, 4, 5 and 28-base pair deletions) (Fig. 3C). We did not observe any cuts with the Surveyor endonuclease in the 2 pools of uninjected embryos. The remaining fish have been raised to adulthood, and their progeny are being screened to identify potential founders (Supplemental Table 1). + + + RESULTS + title_2 + 16857 + 3.2. CRISPR-Cas9-induced AHR2a mutations + + + RESULTS + paragraph + 16898 + Microinjection of the CRISPR-Cas9 guide RNA targeting exon 2 of AHR2a was not successful in generating mutations (20 pools of 5 embryos/pool were analyzed). However, for the guide RNA targeting exon 3, Surveyor mutation detection assay results were positive in 2 out of 6 pools of injected embryos analyzed (30 embryos total; 5 embryos per pool) (Fig. 4A). The mutated target sites resulted in fragments of approximately 160 bp and 95 bp (Fig. 2B, 4A). The PCR products from the two positive samples (10 embryos total) were combined, cloned into the pGEM-T Easy vector, and sequenced. We observed 13 deletions in 81 sequences (16%). The 13 deletions varied in length from 1 to 58 bp, representing 10 distinct deletions (Fig. 4B). + + + RESULTS + title_2 + 17628 + 3.3. CRISPR-Cas9-induced AHR2b mutations + + + RESULTS + paragraph + 17669 + CRISPR-Cas9 guide RNAs targeting exon 2 and 3 of the AHR2b locus were both efficient at generating mutations in the microinjected embryos. Surveyor mutation detection assay showed the presence of mutations in all 4 pools of AHR2b exon 2 CRISPR-Cas-injected embryos analyzed (20 embryos total; 5 embryos per pool) (Fig. 5A). The PCR products from these 4 pools were combined and cloned into the pGEM-T Easy vector. Sequencing results revealed that 16 sequences out of 70 (23%) had indels at the target site, represented by 8 distinct mutations (5 deletions and 3 insertions) (Fig. 5B). + + 105937229 + Gene + + AHR2 + + + + RESULTS + paragraph + 18254 + Similarly, injection of guide RNA targeting exon 3 of AHR2b induced mutations as observed with Surveyor mutation detection assay (Fig. 5C). All 4 pools of injected embryos showed the presence of mutations (5 embryos per pool). PCR products from two of the samples were combined, cloned into the pGEM-T easy vector and sequenced. We observed 55 sequences with indels out of 87 (63%). There were 20 different deletions and 7 different insertion mutants (Fig. 5D). + + + RESULTS + title_2 + 18716 + 3.4. Screening for CRISPR-Cas9 off-target effects + + + RESULTS + paragraph + 18766 + In order to assess potential off-target effects of the CRISPR-Cas9 experiments, we chose one of the guide RNAs that gave positive results (exon 2 of AHR2b) to screen for possible mutations in other AHR paralogs and AHRR. The multiple AHR paralogs and the AHRR are closely related genes and share substantial sequence identity, especially in the N-terminus. Alignment of the AHR2b CRISPR-Cas9 target sequence with the other genes is shown in Fig. 6A. The numbers of mismatches between the 20 bp AHR2b target site and AHR1a, AHR2a, AHR1b, and AHRR are 4, 4, 6, and 3, respectively. These corresponding regions were amplified from the uninjected and AHR2b CRISPR-Cas9-injected embryo genomic DNA samples in which AHR2b mutations were observed. The PCR products were analyzed for mutations using the Surveyor endonuclease assay. The AHR1a, AHR2a, and AHRR samples contained small fragments indicating endonuclease digestion of the PCR products (Fig. 6B). These fragments were also present in the AHR2a and AHRR reactions from the uninjected embryos. These amplification reactions were purified and analyzed by direct-sequencing. The chromatograms of the AHR1a and AHR2a sequencing reactions contained double peaks at previously characterized SNP locations (#141 and #159 for AHR1a, and #156 for AHR2a). Surveyor endonuclease is sensitive to even a single nucleotide mismatch between the two DNA strands, therefore it will cleave DNA at the SNP sites. The sizes of the observed Surveyor endonuclease fragments in these samples (Fig. 6B) can be explained by these SNPs, thereby suggesting that they do not represent off-target mutations. On the other hand, we did not detect a SNP in the AHRR sequences that would explain the fragments observed in the Surveyor assay. However the presence of these fragments in both uninjected and injected samples suggest that they are rare SNPs that may not be detected by direct sequencing. + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + + DISCUSS + title_1 + 20687 + 4. DISCUSSION + + + DISCUSS + paragraph + 20701 + In this study, we demonstrated the feasibility of targeted mutagenesis approaches in inducing mutations in an environmental model species with limited genomic and genetic resources. The ease with which ZFN and CRISPR-Cas9 methods can be utilized to cleave specific genomic DNA sequences has led to the generation of gene knockouts in species not traditionally used for conducting genetic and functional studies (Table 1). Using these methods, we produced mutations in AHR2a and AHR2b loci in killifish embryos. These embryos are currently being raised to adulthood, and their progeny will be screened to identify potential founders. As killifish have a long generation time (2 years minimum) and lack established breeding techniques, the time it takes to generate homozygous mutants is relatively long. Here, we propose some strategies to expedite the process of generating homozygous mutants in killifish. AHR-null killifish will be valuable for characterizing the role of the individual AHR paralogs in evolved resistance, as well as in normal development. + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + + DISCUSS + paragraph + 21760 + We initially adopted the ZFN approach for generating mutations, as their design and activity are well characterized. In addition, ZFNs were the first targeted mutagenesis approach that was successfully used in generating mutations in a number of model and non-model organisms. ZFNs are assembled using zinc finger motifs, each of which recognizes a unique 3-bp DNA sequence. Initial studies with ZFNs used 3-finger zinc-finger proteins (ZFPs), but recent studies have used up to six fingers per ZFP to increase specificity and target rare cleavage sites. We used five-finger zinc finger proteins that enabled ZFN dimers to specifically target a 30-bp region in exon 2 and exon 3 of AHR2a (Table 2). + + + DISCUSS + paragraph + 22459 + In order to determine the translational efficiency of the injected ZFN mRNA, we measured ZFN protein expression in embryos using an anti-FLAG antibody. Our results demonstrated that maximal ZFN protein expression is observed at 6 hours post-injection, suggesting that ZFN protein binding to the target site and creating DSBs occurs around this time. In comparison, in zebrafish embryos injected with a ZFN targeting the aryl hydrocarbon receptor repressor (ahrra) gene, we observed maximal ZFN protein expression at 3 hours post-injection. These species-specific differences in ZFN protein expression could be linked to the developmental rate in the two species. The developmental period for zebrafish from fertilization to hatching is 2 days, whereas killifish hatch at 15 days post-fertilization. Another factor that could contribute to the differences in protein expression is the water temperature at which embryos are incubated. Zebrafish embryos are reared at a higher temperature (28 C) than killifish (23 C), which may affect the rate of transcription and translation. + + 641291 + Gene + 123929 + + ahrra + + + 7955 + Species + + zebrafish + + + 7955 + Species + + zebrafish + + + 8078 + Species + + killifish + + + 7955 + Species + + Zebrafish + + + 8078 + Species + + killifish + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + MESH:D014867 + Chemical + + water + + + + DISCUSS + paragraph + 23538 + We were successful in obtaining mutations in the AHR2a gene with a ZFN targeting exon 3. Sequencing results revealed the presence of 4 different deletions at the target site in 10% of the sequenced clones. These results are comparable to the frequencies observed in other species. The CRISPR-Cas9 method has been reported to result in higher mutation frequencies in comparison to ZFNs or TALENs. Therefore, we used the CRISPR-Cas9 approach to target AHR2a and observed indels in 16% of the sequences from embryos microinjected with a gRNA targeting exon 3. We detected mutations in the CRISPR-Cas9 injected embryos in the very first trial compared to several ZFN trials, even though the proportion of indels showed only a modest increase with the CRISPR-Cas9 method. Similarly, microinjection of gRNAs targeting exon 2 and exon 3 of AHR2b produced indels in 17% and 63% of the sequences. We are currently raising the ZFN-AHR2a, CRISPR-Cas9-AHR2a and -AHR2b fish to reproductive maturity so that we can screen for founders (Supplemental Table 1). To our knowledge, this is the first study comparing the effectiveness of two different targeted genome-editing methods (protein and gRNA based) in a species with limited genomic resources. Additional comparisons with other target loci will be necessary to confirm our observations. + + + DISCUSS + paragraph + 24866 + Genome-editing methods have become a powerful resource for generating mutants in non-model species, but there are some concerns with regard to their off-target effects. ZFNs, as well as CRISPR-Cas9, have been associated with cytotoxicity, presumably due to binding and cleavage at non-target sites, although toxic effects could also result from the target gene disruption. Screening the genome for off-target effects a priori by blast searches with the target sequence is not feasible for species that do not have sequenced genomes. However, paralogs and closely related genes that have been characterized can be screened for mutations, as these represent likely targets. We checked for off-target effects of CRISPR-Cas9 in all AHR paralogs, as well as AHRR, a gene closely related to AHR (Fig. 6). Based on the Surveyor assay and sequencing results, no off-target effects were detected, even though the number of sequence mismatches among these genes was as low as 3 residues. If genomic resources are available, several tools exist for in silico identification of potential off-target sites. Alternatively, potential off-target sites can be searched using the fuzznuc algorithm (http://embossgui.sourceforge.net/demo/manual/fuzznuc.html) in RNAseq data, if available. + + 105920361 + Gene + + AHRR + + + MESH:D064420 + Disease + + cytotoxicity + + + + DISCUSS + paragraph + 26136 + In ZFN and CRISPR-Cas9 injected embryos we did not observe any developmental toxicity, including the highest concentration of ZFN (2000 pg of ZFN mRNA/embryo). These results are in contrast to studies in other fish species, in which overt developmental phenotypes were observed at these concentrations. One reason for the lack of toxicity could be the relative size of the killifish embryos in comparison to zebrafish embryos, in which most of these methods have been optimized. Freshly fertilized killifish embryos are 2 mm in diameter, and are larger than zebrafish (0.7 mm), where ZFNs above 100 pg per embryo have been shown to cause embryo toxicity. Similarly, in medaka with an egg diameter (1.2 mm), 160 pg of ZFN caused embryo toxicity. In addition to the size of the embryo, rapid turnover of the ZFN protein might be responsible for the lack of embryo toxicity in killifish. Our results suggest that the ZFN protein is expressed transiently around 6 hours post-injection and is degraded by 9 hours (Fig. 3A), suggesting faster degradation of the injected mRNA in killifish. We cannot rule out the possibility of additional factors, yet to be determined, contributing to the low toxicity of injected mRNA in killifish. + + 8078 + Species + + killifish + + + 7955 + Species + + zebrafish + + + 8078 + Species + + killifish + + + 7955 + Species + + zebrafish + + + 8090 + Species + + medaka + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + MESH:D064420 + Disease + + toxicity + + + MESH:D064420 + Disease + + toxicity + + + MESH:D064420 + Disease + + toxicity + + + MESH:D064420 + Disease + + toxicity + + + MESH:D064420 + Disease + + toxicity + + + MESH:D064420 + Disease + + toxicity + + + + DISCUSS + paragraph + 27364 + Our results demonstrate the utility of the ZFN and CRISPR-Cas9 approaches in generating mutations in an ecologically relevant fish species that have successfully adapted to environments contaminated with PCBs and PAHs. Generating AHR knockout mutants will provide a powerful tool to functionally characterize the role of these proteins in evolved resistance to PCBs and PAHs. We are currently raising the ZFN- and CRISPR-Cas9-injected fish to adults. The numbers of injected animals that are being raised and the breeding strategy that will be followed are provided in the supplemental information (Supp. Table 1 and Supp. Fig. 1). Killifish mature in 1 year; however, larger fish, producing sufficient offspring per pair for testing, require > 2 years of laboratory rearing. Most organisms in which these approaches are extensively used to generate mutants have short life history cycles (e.g. 2-3 months). However, in the past few years targeted mutagenesis approaches have been successfully used in species with longer life cycles such as yellow catfish, rainbow trout and Atlantic salmon. In order to expedite the process of obtaining homozygous mutants, several strategies can be followed. During certain steps of the homozygous mutant breeding, fin-clips from juvenile fish can be screened by direct-sequencing to identify mutants (Supp. Fig. 1). Direct-sequencing allows screening a single DNA sample per fish, as opposed to cloning the PCR product and screening multiple clones. Another possible approach to accelerate the identification of founders is to screen gametes stripped from juveniles. Once the founder animals are identified, it is possible to artificially induce precocious maturation of gametes by treating the founders with gonadotropins, a common practice in breeding aquaculture species. + + 8078 + Species + + Killifish + + + 1234273 + Species + + yellow catfish + + + 8022 + Species + + rainbow trout + + + 8030 + Species + + Atlantic salmon + + + MESH:D011078 + Chemical + + PCBs + + + MESH:D011084 + Chemical + + PAHs + + + MESH:D011084 + Chemical + + PAHs + + + MESH:D011078 + Chemical + + PCBs + + + + DISCUSS + paragraph + 29178 + Other strategies to consider while generating mutants in species with long life history cycles is to generate at least two mutant lines targeting different sites in each gene. As it is time consuming to raise embryos to reproductive maturity, targeting multiple regions of the same gene confirms the specificity of the observed phenotypes associated with the gene knockout. In addition, if a frameshift mutation is not produced in one of the targets, the other target will serve as an alternative. We followed these approaches and designed CRISPR-Cas9 guide RNAs against AHR2a, in addition to the ZFN method. Similarly, we designed two different gRNAs against AHR2b, and observed mutations at both target sites. Another important consideration in target site selection in species such as the Atlantic killifish is to avoid regions with known single nucleotide polymorphisms (SNPs). Killifish have high genetic diversity, and if the target site has multiple mismatches due to SNPs, the efficacy of the ZFN proteins or guide RNAs will be reduced. In addition, SNPs interfere with the mutation detection assays such as Surveyor endonuclease by creating cuts at polymorphic sites. An additional strategy to consider is optimizing injection conditions to maximize the chances for mutating both alleles at the target locus simultaneously. Such bi-allelic mutations in CRISPR-Cas9-injected embryos have been demonstrated previously. The presence of bi-allelic mutations in the germline will expedite the process of generating homozygous individuals. + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + Killifish + + + + DISCUSS + paragraph + 30721 + Overall, targeted mutagenesis approaches have provided new opportunities for conducting functional studies in non-traditional model organisms. As a proof of concept, we and others (Table 1) have demonstrated that it is feasible to generate mutants in non-traditional model species. The relative ease of these methods, as well as the open-source sharing of reagents, have made these methods accessible to a wide range of laboratories. + + + CONCL + title_1 + 31155 + 5. CONCLUSIONS + + + CONCL + paragraph + 31170 + In this study we demonstrated the utility of targeted gene-editing methods for generating ahr mutants in Atlantic killifish. We observed higher frequency of mutations with CRISPR-Cas9 compared to ZFN mRNA injected embryos. We are currently raising the injected embryos to identify potential germ line mutants. Some of the challenges in generating homozygous mutants in killifish include long generation time and lack of established breeding methods. We have proposed some strategies to overcome these challenges and expedite the process of identifying the founders and generating ahr-null mutants. The strategies proposed here are applicable to any fish species with limited aquaculture resources, providing an opportunity to conduct functional studies. + + 8078 + Species + + Atlantic killifish + + + 8078 + Species + + killifish + + + + SUPPL + title_1 + 31924 + Supplementary Material + + + SUPPL + footnote + 31947 + This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final citable form. 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designing CRISPR sgRNA and evaluating potential off-target cleavage sites + + + surname:Guiguen;given-names:Y + 16 + surname:Jouanno;given-names:E + surname:Nicol;given-names:B + REF + surname:Yano;given-names:A + 243 + 24085607 + 2014 + Mar Biotechnol (NY) + ref + 250 + 41156 + Heritable targeted inactivation of the rainbow trout (Oncorhynchus mykiss) master sex-determining gene using zinc-finger nucleases + + 8022 + Species + + rainbow trout + + + 8022 + Species + + Oncorhynchus mykiss + + + + surname:Sakuma;given-names:T + surname:Hozumi;given-names:A + surname:Treen;given-names:N + surname:Yoshida;given-names:K + 431 + 24619765 + 2014 + Genesis + ref + 52 + REF + 439 + surname:Sasakura;given-names:Y + surname:Yamamoto;given-names:T + 41287 + Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases + + 7719 + Species + + Ciona intestinalis + + + + surname:Ankoudinova;given-names:I + surname:Doyon;given-names:Y + surname:Cherone;given-names:JM + surname:Young;given-names:JJ + 7052 + 21471457 + 2011 + Proc Natl Acad of Sci U S A + ref + 108 + surname:Zhang;given-names:L + REF + surname:Urnov;given-names:FD + surname:Harland;given-names:RM + surname:Rebar;given-names:EJ + surname:Gregory;given-names:PD + surname:Zeitler;given-names:B + surname:Miller;given-names:JC + surname:Paschon;given-names:DE + surname:Guschin;given-names:DY + 7057 + surname:Ngo;given-names:C + surname:Lee;given-names:AH + surname:Faraji;given-names:FM + 41362 + Efficient targeted gene disruption in the soma and germ line of the frog Xenopus tropicalis using engineered zinc-finger nucleases + + 8364 + Species + + Xenopus tropicalis + + + Disease + + soma + + + + surname:Naruse;given-names:K + surname:Chen;given-names:J + surname:Guan;given-names:G + surname:Zhang;given-names:X + 125 + 24149659 + 2014 + Mar Biotechnol (NY) + ref + 16 + REF + 134 + surname:Hong;given-names:Y + 41493 + Parameters and efficiency of direct gene disruption by zinc finger nucleases in medaka embryos + + 8090 + Species + + medaka + + + + FIG + nihms645550f1.jpg + F1 + fig_caption + 41588 + Outline of the targeted mutagenesis techniques currently used in generating mutants. A. Zinc finger nucleases B. TALENs and C. CRISPR-Cas system. + + + FIG + nihms645550f2.jpg + F2 + fig_caption + 41734 + AHR target sites. A. Functional domain structure of the AHR protein. The location of the targeted exons 2 and 3 in relation to the functional domains is shown as red bars. DBD: DNA-binding domain, LBD: ligand-binding domain, TAD: transcriptional activation domain. B. Schematic representations of the ZFN and CRISPR-Cas target regions in AHR2a and AHR2b loci. Arrows represent the primers used in the PCR amplification of genomic DNA for screening. Surveyor nuclease fragments are shown below the target regions. + + + FIG + nihms645550f3.jpg + F3 + fig_caption + 42247 + ZFN-mediated mutagenesis of killifish AHR2a. A. ZFN protein expression in killifish embryos. Uninjected (-) or ZFN-injected (+) embryos were sampled at 3, 6, 9, and 24 hours after micro-injection. Homogenates were resolved on SDS-PAGE and probed with a Flag-tag antibody. Lysate from COS-7 cells transfected with the ZFN expression plasmid was run as a positive control. B. Surveyor nuclease detection of mutations in the ZFN target region of AHR2a. Each lane represents a pool of 5 embryos from which a 335 bp genomic DNA fragment was amplified. U: uninjected control, ZFN: injected embryos. Lanes 1 and 2 are two representative samples that were positive in the mutation detection assay. Approximate sizes of the Surveyor nuclease-digested fragments containing the deletions are shown (240 and 95 bp). The full-length PCR product from sample 2 was cloned and sequenced. C. AHR2a exon 3 sequence surrounding the ZFN target site (in red). Four types of deletion mutants were observed among the sequenced clones (deletions of 2, 4, 5, and 28 nucleotides). + + 8078 + Species + + killifish + + + 8078 + Species + + killifish + + + 29278 + Species + + expression + + + 29278 + Species + + expression + + + CVCL:0224 + CellLine + + COS-7 + + + MESH:D012967 + Chemical + + SDS + + + + FIG + nihms645550f4.jpg + F4 + fig_caption + 43304 + CRISPR-Cas9-mediated mutagenesis of killifish AHR2a targeting exon 3. A. Surveyor nuclease detection of mutations in the CRISPR-Cas9 target region of AHR2a exon 3. Each lane represents a pool of 5 embryos from which a 255 bp genomic DNA fragment was amplified. U: uninjected, lanes 1 and 2: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (160 and 95 bp). The full-length PCR products from samples 1 and 2 were pooled, cloned, and sequenced. B. AHR2a exon 3 sequence surrounding the CRISPR-Cas9 target site (in red). Ten different deletion mutants were observed among the sequenced clones. + + 8078 + Species + + killifish + + + + FIG + nihms645550f5.jpg + F5 + fig_caption + 43970 + CRISPR-Cas9-mediated mutagenesis of killifish AHR2b targeting exon 2 and exon 3. A. Surveyor nuclease detection of mutations in the AHR2b CRISPR-Cas9 target region. Each lane represents a pool of 5 embryos from which a 309 bp genomic DNA fragment was amplified. U: uninjected control, lanes 1-4: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (175 and 135 bp). The full-length PCR products from samples 1-4 were pooled, cloned, and sequenced. B. AHR2b exon 2 sequence surrounding the CRISPR-Cas9 target site (in red). Five types of deletion mutants and 3 types of insertions were observed among the sequenced clones. C. Surveyor nuclease detection of mutations in the CRISPR-Cas9 target region of AHR2b exon 3. Each lane represents a pool of 5 embryos from which a 198 bp genomic DNA fragment was amplified. U: uninjected, lanes 1 and 2: CRISPR-Cas9-injected embryos. Approximate sizes of the digested fragments containing the deletions and insertions are shown (110 and 90 bp). The full-length PCR products from samples 1 and 2 were pooled, cloned, and sequenced. D. AHR2b exon 3 sequence surrounding the CRISPR-Cas9 target site (in red). Twenty different types of deletion mutants and seven types of insertion mutants were observed among the sequenced clones. + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 8078 + Species + + killifish + + + + FIG + nihms645550f6.jpg + F6 + fig_caption + 45313 + AHR2b CRISPR-Cas9 off-target analysis. A. Nucleotide sequence alignment of the AHR2b CRISPR-Cas9 target sequence with the corresponding regions of closely related genes. AHR1a, AHR2a, AHR1b, and AHRR have 4, 4, 6, and 3 mismatches to AHR2b in this region, respectively. B. Surveyor nuclease detection of mutations in AHR1a, AHR2a, AHR1b, and AHRR in the AHR2b CRISPR-Cas9 target region. Digested fragments are indicated by a *. Each lane represents a pool of 5 embryos from which genomic DNA fragments were amplified. U: uninjected control, 1-4: CRISPR-Cas9 injected embryos. AHR1a, AHR2a, and AHRR PCR products from uninjected and injected embryos were sequenced. + + 105937229 + Gene + + AHR2 + + + 105937229 + Gene + + AHR2 + + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + 105937229 + Gene + + AHR2 + + + 105920361 + Gene + + AHRR + + + + TABLE + T1.xml + T1 + table_caption + 45980 + Published studies employing gene targeting in non-traditional model species. + + + TABLE + T1 + table + T1.xml + 46057 + Gene-editingapproach Species Reference ZFN Monarch butterfly (Danaus plexippus)Two-spotted cricket (Gryllus bimaculatus)Sea urchin (Hemicentrotus pulcherrimus)Ascidian (Ciona intestinalis)Western clawed frog (Xenopus tropicalis)Yellow catfish (Pelteobagrus fulvidraco)Medaka (Oryzias melastigma)Rainbow trout (Oncorhynchus mykiss) TALEN Nematode (Pristionchus pacificus)Marine polycheate (Platyneries dumerilii)Culicine mosquito (Aedes aegypti)Silk worm (Bombyx mori)Sea urchin (Hemicentrotus pulcherrimus)Ascidian (Ciona intestinalis)Western clawed frog (Xenopus tropicalis)Iberian ribbed newts (Pleurodeles waltl)Tilapia (Oreochromis niloticus)Medaka (Oryzias melastigma)Yellow catfish (Pelteobagrus fulvidraco) CRISPR-Cas9 Western clawed frog (Xenopus tropicalis)Axolotl (Ambystoma mexicanum)Atlantic Salmon (Salmo salar)Tilapia (Oreochromis mossambicus) (Ji-Feng et al., 2014) + + 13037 + Species + + Monarch butterfly + + + 13037 + Species + + Danaus plexippus + + + 6999 + Species + + Two-spotted cricket + + + 6999 + Species + + Gryllus bimaculatus + + + 7656 + Species + + Sea urchin + + + 7650 + Species + + Hemicentrotus pulcherrimus + + + 7719 + Species + + Ciona intestinalis + + + 8364 + Species + + Western clawed frog + + + 8364 + Species + + Xenopus tropicalis + + + 1234273 + Species + + Yellow catfish + + + 1234273 + Species + + Pelteobagrus fulvidraco + + + 8090 + Species + + Medaka + + + 54126 + Species + + Pristionchus pacificus + + + 7159 + Species + + Aedes aegypti + + + 7091 + Species + + Bombyx mori + + + 7656 + Species + + Sea urchin + + + 7650 + Species + + Hemicentrotus pulcherrimus + + + 7719 + Species + + Ciona intestinalis + + + 8364 + Species + + Western clawed frog + + + 8364 + Species + + Xenopus tropicalis + + + 8319 + Species + + Iberian ribbed newts + + + 8319 + Species + + Pleurodeles waltl + + + 1234273 + Species + + Pelteobagrus fulvidraco + + + 8364 + Species + + Western clawed frog + + + 8364 + Species + + Xenopus tropicalis + + + 8296 + Species + + Axolotl + + + 8127 + Species + + Oreochromis mossambicus + + + Disease + + Oryzias melastigma)Rainbow trout (Oncorhynchus mykiss + + + MESH:D015004 + Disease + + Tilapia (Oreochromis niloticus)Medaka (Oryzias melastigma)Yellow catfish + + + + TABLE + T2.xml + T2 + table_caption + 47232 + AHR2a and AHR2b target regions and oligonucleotides used in ZFN and CRISPR-Cas9-based mutagenesis. The ZFN target site is shown in capital letters. The sequence flanking the target sites are recognized by the ZFN proteins. + + MESH:D009841 + Chemical + + oligonucleotides + + + + TABLE + T2 + table + T2.xml + 47455 + Name Description Sequence AHR2a exon 2 ZFN target site ctgcgcctcagcgtgGGATAcctgagggtcaagag AHR2a exon 3 ZFN target site gactcattcagcttcTCTGAaggagagctgctgct AHR2a exon 2 CRISPR-Cas9 target site GGAGCGAATCTCATCAGGGA AHR2a exon 3 CRISPR-Cas9 target site GGGTGGCTCCTGGAGTCAAC AHR2b exon2 CRISPR-Cas9 target site GGACCGTCTGAACGGCGAGC AHR2b exon3 CRISPR-Cas9 target site GGGCGAAGGTCTGAGCGGTC 2-ex3Fwd AHR2a ZFN pair3 screening TGTTTCACCTCCTGCACAGCTTCC 1XRR1 AHR2a ZFN pair3 screening TACTAGCTAAACGTGACCTGTCG 2a-ex3CrF AHR2a CRISPR-Cas exon3 screening CTGTTCTATCAGGTGGAGAGTCC 2a-ex3CrR AHR2a CRISPR-Cas exon3 screening GTTGTTGGATGAGTATGCAGGTGG AHR2b-CrF AHR2b CRISPR-Cas9 exon2 screening AATCTTTGATCCGGTCTGATCTGG AHR2b-CrR AHR2b CRISPR-Cas9 exon2 screening AGACACAACCAAGCAGCAACACG 2bex3-Fwd AHR2b CRISPR-Cas9 exon3 screening ATCGCCCAGCCTTAGCTTCCAGTCG 2bex3-Rev AHR2b CRISPR-Cas9 exon3 screening GCGTTTTTGCTTTGTCTGCTGTCGG Fh2A-ex3-CR1 AHR2a exon3 gRNA oligo1 TAGGGTGGCTCCTGGAGTCAAC Fh2A-ex3-CR2 AHR2a exon3 gRNA oligo2 AAACGTTGACTCCAGGAGCCAC FhAHR2b-CR1 AHR2b exon2 gRNA oligo1 TAGGACCGTCTGAACGGCGAGC FhAHR2b-CR2 AHR2b exon2 gRNA oligo2 AAACGCTCGCCGTTCAGACGGT Fh2B-ex3-CR1 AHR2b exon3 gRNA oligo1 TAGGGCGAAGGTCTGAGCGGTC Fh2B-ex3-CR2 AHR2b exon3 gRNA oligo2 AAACGACCGCTCAGACCTTCGC 2b-1aF AHR1a off-target screening GCTTAATCCTGCTTTCGTCTCTGC Fh1a-ex2rev AHR1a off-target screening AAGAAGTGTTTGGCTCGTAGGTAGC 2b-2aF AHR2a off-target screening TTTTACCCCTCACGGAAGAAGATGG Fh2a-ex2rev AHR2a off-target screening GTAGCTCTTGACCCTCAGGTATCC 2b-1bF AHR1b off-target screening TTTCCTCCTCTCCAGAGCCAAACC Fh1b-ex2rev AHR1b off-target screening AAGAAGCTCTTGGTGCGCAGGTAGC RR-ex2fwd AHRR off-target screening CTCATCCCTTCTGCCGTCTCGTCC RR-ex2rev AHRR off-target screening AAGAAGCTTTTGACGCGGAGGTAGG + + 105920361 + Gene + + AHRR + + + 105920361 + Gene + + AHRR + + + 2498238 + Species + + CR2 + + + 2498238 + Species + + CR2 + + + 2498238 + Species + + CR2 + + + + REF + title_caption + 51064 + Highlights + + + REF + paragraph + 51075 + AHR2 genes were mutated without detectable off-target effects in paralogs. + + 105937229 + Gene + + AHR2 + + + + REF + paragraph + 51150 + CRISPR-Cas9 method was more efficient in inducing mutations compared to ZFN. + + + REF + paragraph + 51227 + We suggest strategies for generating knockouts in non-traditional fish models. + + + REF + paragraph + 51306 + AHR knockouts will be valuable for understanding evolved resistance to toxicants. + + + + 4272818 + + surname:Haynes;given-names:R Brian + surname:Law;given-names:Mary + surname:Chesworth;given-names:Bert M + 25495467 + surname:Shi;given-names:Qiyun + 2014 + 4272818 + 263 + 10.1186/s12909-014-0263-4 + Keywords + front + 263 + 14 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + BMC Med Educ; 2014 Dec 14; 14263. doi:10.1186/s12909-014-0263-4 + Evidence-based Scale Self-reported Validation Clinician + surname:MacDermid;given-names:Joy C + Shi Q, Chesworth BM, Law M, Haynes RB, MacDermid JC + 0 + A modified evidence-based practice- knowledge, attitudes, behaviour and decisions/outcomes questionnaire is valid across multiple professions involved in pain management + + MESH:D010146 + Disease + + pain + + + + ABSTRACT + abstract_title_1 + 170 + Background + + + ABSTRACT + abstract + 181 + A validated and reliable instrument was developed to knowledge, attitudes and behaviours with respect to evidence-based practice (EBB-KABQ) in medical trainees but requires further adaptation and validation to be applied across different health professionals. + + + ABSTRACT + abstract_title_1 + 441 + Methods + + + ABSTRACT + abstract + 449 + A modified 33-item evidence-based practice scale (EBP-KABQ) was developed to evaluate EBP perceptions and behaviors in clinicians. An international sample of 673 clinicians interested in treatment of pain (mean age = 45 years, 48% occupational therapists/physical therapists, 25% had more than 5 years of clinical training) completed an online English version of the questionnaire and demographics. Scaling properties (internal consistency, floor/ceiling effects) and construct validity (association with EBP activities, comparator constructs) were examined. A confirmatory factor analysis was used to assess the 4-domain structure EBP knowledge, attitudes, behavior, outcomes/decisions). + + MESH:D010146 + Disease + + pain + + + + ABSTRACT + abstract_title_1 + 1144 + Results + + + ABSTRACT + abstract + 1152 + The EBP-KABQ scale demonstrated high internal consistency (Cronbach's alpha = 0.85), no evident floor/ceiling effects, and support for a priori construct validation hypotheses. A 4-factor structure provided the best fit statistics (CFI =0.89, TLI =0.86, and RMSEA = 0.06). + + + ABSTRACT + abstract_title_1 + 1435 + Conclusions + + + ABSTRACT + abstract + 1447 + The EBP-KABQ scale demonstrates promising psychometric properties in this sample. Areas for improvement are described. + + + ABSTRACT + abstract_title_1 + 1566 + Electronic supplementary material + + + ABSTRACT + abstract + 1600 + The online version of this article (doi:10.1186/s12909-014-0263-4) contains supplementary material, which is available to authorized users. + + + INTRO + title_1 + 1740 + Background + + + INTRO + paragraph + 1751 + Evidence-based practice (EBP) is defined as the integration of the best research evidence with patients' interests and clinical circumstances in decision making. As EBP is associated with improved clinical decision-making and patient care, health professional organizations have advocated for increased training in EBP for all health care professionals at all levels of education. Understanding how EBP is understood and implemented across different health professionals can identify educational needs and outcomes, and predict where new research evidence is more likely to be implemented. As such, a validated and reliable instrument is required to evaluate an individual's perceptions of EBP. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + + INTRO + paragraph + 2450 + A systematic review which studied 104 instruments on EBP suggested that evaluation of EBP could be divided into the following definable components: EBP knowledge, attitudes toward EBP, application/use of EBP and practitioners' EBP behaviors in the clinical setting. Knowledge about EBP means that clinicians have knowledge of fundamental EBP concepts and terminology and concepts related to quality or levels of evidence. It also includes the ability to search the literature and critically appraise the evidence for its validity, impact and applicability. Attitude toward EBP includes the intuitive appeal of EBP, the likelihood of adopting EBP given professional requirements to do so, openness to new practices, and the perceived divergence between research-based/academically developed interventions versus current practice. Application and use of EBP refers to whether health professionals are able to apply their EBP knowledge to the specific clinical scenarios. This includes: capability to generate clinical question(s) regarding disease prevention, diagnosis and management as well as implementation of evidence with integrity of clinical circumstances. EBP behaviors refer to practitioners' performance of the instrumental activities associated with EBP such as searching and obtaining higher quality evidence in their own practice. + + MESH:D005597 + Disease + + openness + + + + INTRO + paragraph + 3797 + Although the rise of EBP awareness has led to the development of instruments to assess its integration into clinical practice, there are gaps in the evidence supporting these tools. There is a lack of empirical data that can be applied to a wider range of experience and types of clinicians, in particular nurses and allied health professionals. Moreover, as most scales have targeted samples with minimal experience in clinical practice, the questionnaires may not accurately reflect the perception of EBP by clinicians who have been practicing in different clinical settings. + + + INTRO + paragraph + 4375 + Among available scales, one that has taken a multi-dimensional approach and shown early promise is the The knowledge, attitude and behavior questionnaire (KAB) originally developed by Johnson and colleagues. The KAB scale was designed to evaluate EBP teaching and learning in the undergraduate medical education setting. With permission from the developers, two study authors (JMD and ML) developed a modified KAB scale (EBP-KABQ), to be applicable to health professionals other than physicians using expert review and pilot testing. This process resulted in removal of items that were perceived by users as redundant or unclear. + + MESH:C537366 + Disease + + ML + + + Disease + + JMD + + + + INTRO + paragraph + 5005 + The goal of this study was to validate the modified scale (EBP-KABQ) for use in a multidisciplinary group of clinicians by determining: (1) Scaling properties- internal consistency, floor/ceiling effects, and (2) Construct validity- based on predetermined hypotheses on the relationship of subcomponents of EBP, and (3) Structural validity: the integrity of a 4-domain structure based on confirmatory factor analysis. + + + METHODS + title_1 + 5423 + Methods + + + METHODS + paragraph + 5431 + The EBP-KABQ incorporates 33 items in four domains of EBP: knowledge (8 items, 6 ordinal items), attitudes (14 items, 14 ordinal items), behaviour (8 items, 5 ordinal items) and outcomes/decisions (3 items, 3 ordinal items) (KABQ). The knowledge items retain a 7-point Likert scale with lower scores indicating a lower level of EBP knowledge. The Attitudes towards EBP items retain a 7-point Likert scale. High scores indicate positive attitude after several items were reversely scored. For EBP behaviour, lower scores indicate a lower frequency of using EBP in current practice. A 6-point Likert scale is used for responses to the items in the outcomes/decisions domain. Lower scores indicate unfavorable patient outcomes and poor clinical evidence-based decision making. Detail of the EBP-KABQ scale and a summary of the changes to original scale are presented in Additional files 1 and 2. + + 9606 + Species + + patient + + + + METHODS + title_2 + 6324 + Subject recruitment and data collection + + + METHODS + paragraph + 6364 + All participants were recruited from a clinical trial assessing use of pain research evidence about pain. Eligible practitioners were (1) physicians, nurses, occupational therapists (OTs), physical therapists (PTs), or psychologists who were currently working in clinical practice at least one day/week; (2) fluent in English; (3) able to access a computer at home or at work that provided unrestricted access to the World Wide Web; (4) possessed an active email account;(5) consent to participate in this research studyA total of 870 clinicians met the inclusion criteria and were invited to participate. From August 2011 to February 2013, 673 clinicians (physicians, nurses, OTs/PTs, psychologists etc.) completed an online EBP-KABQ scale prior to receiving new pain information. Demographic and practice characteristics were also obtained. The study received Ethics Approval from the McMaster University Research Ethics Board. + + 9606 + Species + + participants + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + METHODS + title_2 + 7294 + Data analysis + + + METHODS + paragraph + 7308 + Quality checks, descriptive statistics and checks for normality were completed prior to analysis. Item 33 "I don't use evidence-based practice for another reason (specify)" was removed from the analyses because the specified reason varied across respondents, making it a nonstandard item. Therefore, 27 ordinal items across the following four domains of EBP were analyzed in this study: knowledge (n = 6 items), attitudes (n = 13 items), behavior (n = 5 items) and outcomes/decisions (n = 3 items). + + + METHODS + title_2 + 7829 + Scaling properties (internal consistency and floor/ceiling effects) + + + METHODS + paragraph + 7897 + Internal consistency reliability scores were assessed for both the full EBP-KABQ scale and its corresponding 4 subscales using Cronbach's alpha, where >0.7 was considered as minimum and >0.9 was desirable. Scaling properties such as floor/ceiling effects, which was observed in >15% of scores at minimum or maximum scale/subscale were also assessed. + + + METHODS + title_2 + 8249 + Construct validation + + + METHODS + paragraph + 8270 + Four hypotheses were tested to assess the construct validation of EBP-KABQ scale. First, we hypothesized that the mean item score in "knowledge" would be higher than those in "behaviour", "outcomes/decisions" and "attitude" domains because knowledge is considered a necessary precursor, but not a sufficient guarantee, for changes in practice and outcomes. Secondly, we hypothesized that the domain of "outcomes/decisions" would be more strongly correlated to the other 3 domains since it focuses on how EBP influences the decision making process. Thirdly, we hypothesized that EBP-KABQ subscale scores would be correlated with corresponding EBP activities assessed by relevant open ended questions. For example, the frequency that clinicians search for evidence should be correlated with subtotal score of "behaviour" to a greater extent than other domains such as "knowledge" or "EBP outcomes/decisions". Finally, we hypothesized that following demographic variables would be associated with total EBP-KABQ scale score in the multivariate modeling: age, highest level of education, and possession of advanced clinical training since these have been suggested in the literature on EBP. Details of all construct validity testing and a priori hypotheses are provided in the Results section. + + + METHODS + title_2 + 9592 + Structural validity + + + METHODS + paragraph + 9612 + Confirmatory factor analysis (CFA, maximum likelihood estimation) was conducted to examine our proposed 4-domain model. Four conceptual domains of EBP (knowledge, attitudes, behavior and outcomes/decisions) were tested as second-order factors (latent variables) based on the originally defined conceptual framework. We evaluated the model fit with a number of goodness-of-fit statistics including Root Mean Square Error of Approximation (RMSEA) <0.06 (ideal) and <0.08 (acceptable), comparative fit index (CFI) >=0.90-0.95 (acceptable), Tucker Lewis Index (TLI) >=0.90-0.95 (acceptable) and Chi-square test (P > 0.05, acceptable). We considered RMSEA, CFI and TLI as primary statistics because Chi-square is vulnerable to a large sample size (sample size > 300). We also examined modification indices to identify the potential to improve the model fit. We modified our model when it was indicated by theoretical and statistical findings. We considered standardized coefficients (i.e., factor loadings) >=0.30 (p < 0.05) as 'representing' a hypothesized dimension. + + + METHODS + paragraph + 10699 + All analyses except CFA were conducted by SAS (version 9.3, SAS Institute Inc, Cary, NC, USA). We used IBM SPSS v20 Amos statistical software for CFA. + + 6302 + Gene + 4359 + + SAS + + + 6302 + Gene + 4359 + + SAS + + + + RESULTS + title_1 + 10850 + Results + + + RESULTS + title_2 + 10858 + Sample characteristics + + + TABLE + table + 10881 + Years of clinical experience: Mean = 17.96 years (SD = 11.23 years; range = 0-52). + + + TABLE + Tab1.xml + Tab1 + table_caption + 10980 + Characteristics of 673 participants of EBP-KABQ study + + 9606 + Species + + participants + + + + TABLE + Tab1 + table + Tab1.xml + 11036 + Characteristics N (%) Age 20-35 178 (26.4) 36-45 158 (23.4) 46-55 221 (32.8) 56+ 116 (17.2) Clinical designation MD 131 (19.5) OT/PT 326 (48.4) RN 127 (18.8) RPsych or CPsych 52 (7.7) Others 37 (5.5) Highest education level Diploma/BA 234 (34.8) MA/MSC 222 (33.0) MD 122 (18.1) Ph.D. 95 (14.1) Received advanced clinical certifications 364 (54.1) Years of clinical training Less than 2 years 190 (28.2) 2-5 years 295 (43.8) Above 5 years 188 (27.9) Location of practice Urban 463 (68.8) Rural 101 (15.0) Both 109 (16.2) + + Disease + + Diploma + + + + RESULTS + paragraph + 11588 + In total, 673 health professionals completed EBP-KABQ questionnaire. The description of demographic characteristics is presented in Table 1. Half of participants were age 45 or younger. Nearly half of clinicians were OTs or PTs, while 1/4 were nurses and 1/5 were physicians. One quarter of the sample had more than 5 years of clinical training; and they had a mean time in clinical practice of almost 18 years. Most participants practiced in an urban setting, while 15% were in a rural practice area. + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + + RESULTS + title_2 + 12093 + Scaling properties (internal consistency and floor/ceiling effects) + + + RESULTS + paragraph + 12161 + Overall, EBP-KABQ scale achieved acceptable satisfactory internal consistency (Cronbach's alpha alpha = 0.85) although the subscale of "knowledge" still showed marginal acceptable internal consistency with Cronbach's alpha = 0.66 after removal of item 3. However, this was improved compared to the original 6-item "knowledge" subscale (Cronbach's alpha = 0.56). This finding supported the decision to remove item 3 ("Clinical trials and observational methods are equally valid in establishing treatment effectiveness"). + + + TABLE + table + 12708 + Bold indicated floor or ceiling effect. Item 3 was removed from the scale based on factor structure. + + + TABLE + Tab2.xml + Tab2 + table_caption + 12809 + Descriptive statistics of the EBP-KABQ scale, scaling properties and internal consistency (n = 673) + + + TABLE + Tab2 + table + Tab2.xml + 12915 + Scale Item Item mean(SD) Median Floor% Ceiling% Subscale mean(SD) Floor% Ceiling% Cronbach's alpha at subscale/total level Knowledge-5 items EBP-KABO1 5.79 (1.02) 6.00 0.1 23.8 29.57(3.62) 0.1 1.5 0.66 EBP-KABO2 6.01 (0.99) 6.00 0.3 34.0 EBP-KABO4 5.44 (1.41) 6.00 1.6 25.6 EBP-KABO5 6.08 (1.06) 6.00 0.6 41.0 EBP-KABO6 6.25 (1.12) 7.00 0.6 55.0 Behaviour-5 items EBP-KABO9 3.14 (1.16) 3.00 1.2 15.6 11.22(4.28) 0.1 0.3 0.77 EBP-KABO10 2.02 (1.15) 2.00 17.5 3.1 EBP-KABO11 2.42 (1.22) 2.00 7.0 5.8 EBP-KABO12 1.66 (1.11) 2.00 25.6 1.6 EBP-KABO13 1.98 (1.31) 2.00 34.3 5.9 Outcome/Decision-3 items EBP-KABO17 4.56 (0.94) 5.00 1.0 12.0 12.56(2.52) 0.3 0.4 0.83 EBP-KABO18 4.11 (1.04) 4.00 1.9 4.8 EBP-KABO19 3.88 (0.93) 4.00 0.3 3.4 Attitude-13 items EBP-KABO20 4.17 (0.69) 4.00 1.3 32.1 64.58(8.99) 0.1 0.3 0.75 EBP-KABO21 5.20 (1.53) 5.00 0.9 25.7 EBP-KABO22 3.94 (1.59) 4.00 3.9 4.2 EBP-KABO23 4.18 (1.56) 4.00 3.6 8.5 EBP-KABO24 4.88 (1.59) 5.00 1.8 18.6 EBP-KABO25 4.81 (1.39) 5.00 0.9 11.9 EBP-KABO26 6.22 (1.02) 7.00 0.7 50.1 EBP-KABO27 4.72 (1.51) 5.00 2.5 9.4 EBP-KABO28 5.77 (0.99) 6.00 0.3 25.3 EBP-KABO29 5.99 (0.98) 6.00 0.3 36.4 EBP-KABO30 3.80 (1.51) 4.00 10.1 2.1 EBP-KABO31 5.26 (1.66) 6.00 1.9 31.4 EBP-KABO32 5.66 (1.41) 6.00 0.6 38.0 MEBP-26 items Full version ----- 118.00 ----- ----- 117.93(15.10) 0.1 0.1 0.85 + + + RESULTS + paragraph + 14305 + Table 2 presents a summary of the item-level properties of EBP-KABQ. The mean and median total score of EBP-KABQ scale was 117.93 (SD: 15.10) and 118 respectively, with no floor/ceiling effects detected. The mean scores of four subscales ranged from 11.22 to 64.58. Similarly, no obvious floor/ceiling effects were observed in all four subscales although some individual items particularly in "knowledge" presented a ceiling effect. + + + RESULTS + title_2 + 14743 + Construct validity + + + TABLE + table + 14762 + *P < 0.05. + + + TABLE + table + 14777 + Q 1: How often do you now look up evidence immediately before, or during patient treatment visit per week? + + 9606 + Species + + patient + + + + TABLE + table + 14884 + Q 2: How many hours do you spend looking up evidence per week? + + + TABLE + table + 14947 + Q 3: How many hours do you spend reading new research evidence per week? + + + TABLE + Tab3.xml + Tab3 + table_caption + 15020 + Results of construct validity against a series of theoretical constructs + + + TABLE + Tab3 + table + Tab3.xml + 15095 + Theoretical constructs A priori hypotheses Results 1. EBP knowledge is more easily affected than other other aspects of EBP Mean item score in "knowledge" > other domains Knowledge: 5.91 Behaviour: 2.24 Outcome/Decision : 4.18 Attitude: 4.96 2. "Outcome/Decision" is correlated to other 3 domains Correlation coefficients between "outcome" and "knowledge"/"application"/"attitude" > other correlation coefficients. routcome-knowledge=0.54*, routcome-behaviour=0.40*, routcome-attitude=0.57*; rattitude-knowledge=0.41*, rknowledge-application=0.33*, rapplication-attitude=0.26*; 3. MEBP subscale scores are correlated with corresponding EBP activities Correlation coefficients between "application" and 3 external questions evaluating EBP application > other correlation coefficients. rapplication-Q1=0.32*, rknowledge-Q1=0.19*, routcome-Q1=0.28*; rattitude-Q1=0.19*; rapplication-Q2=0.41*, rknowledge-Q2=0.24*, routcome-Q2=0.30*; rattitude-Q2=0.19*; rapplication-Q3=0.35*, rknowledge-Q3=0.24*, routcome-Q3=0.26*; rattitude-Q3=0.16*; 4. Demographic variables would be associated with total MEBP scale score Age, highest education level, possession of advanced clinical training are significant factors are associated with in multivariate modeling Adjusted beta coefficients of following variable: Age: beta = -0.32 Higher education level (ref: diploma/BA): beta =4.63* Years of clinical training (ref: less than 2 years): beta =2.36* Advanced clinical training (ref: No): beta =4.37* Practice setting (ref: urban): beta =1.87* + + - + Chemical + + routcome + + + Disease + + diploma + + + + TABLE + table + 16664 + *P < 0.05, variables were selected if p value <0.2 in univariate model. + + + TABLE + Tab4.xml + Tab4 + table_caption + 16740 + Unadjusted and adjusted linear regression coefficients for EBP-KABQ total score + + + TABLE + Tab4 + table + Tab4.xml + 16822 + Unadjusted Adjusted Characteristics B SD P value B SD P value Age (years) -0.32 0.59 0.60 -- -- -- Education (ref: diploma/BA) 5.08 0.73 <0.01* 4.63 0.73 <0.01* Clinical designation (ref: MD) 0.06 0.06 0.27 -- -- -- Years of clinical training (ref: less than 2 years) 3.57 0.73 <0.01* 2.36 0.81 <0.01* Advanced clinical training (ref: No) 5.68 1.23 <0.01* 4.37 1.21 <0.01* Practice setting (ref: urban) 1.24 0.82 0.13 1.87 0.79 0.02* Constant -- -- -- 106.2 2.43 <0.01* + + Disease + + diploma + + + + RESULTS + paragraph + 17331 + Details of the construct validity testing and a priori hypotheses were provided in Table 3. As we expected, mean item score in "knowledge" was 5.91, significantly higher than the rest of the domains (p < 0.05). Our constructed hypotheses were supported in that the correlation coefficients between "outcomes/decision" and "knowledge", "behaviour" and "attitude" were 0.54, 0.40 and 0.57 respectively, which were higher correlations than observed between other subscales. Construct validity was also supported in that there was a significant relationship between the frequency of searching reported by clinicians and the "behaviour" score, with correlation coefficient ranges from 0.32 to 0.41 (hypothesis 3). Regression analyses supported our a priori hypothesis that health professionals who had higher levels of education (beta = 4.63, P < 0.01), longer years in clinical training (beta = 2.36, P < 0.01) and possession of advanced clinical training (beta = 4.37, P < 0.01) were more likely to use EBP (Table 4). Although younger age was related to EBP practice in the direction anticipated, it did not reach statistical significance (beta = -0.32, P = 0.06). + + 10242 + Gene + 4257 + + beta = 2 + + + 10381 + Gene + 68503 + + beta = 4 + + + 10381 + Gene + 68503 + + beta = 4 + + + + RESULTS + title_2 + 18549 + Structural validity + + + FIG + 12909_2014_263_Fig1_HTML.jpg + Fig1 + fig_caption + 18569 + Standardized parameter estimates for the refined EBP-KABQ factor structure model. Rectangles represent the scale items and ellipses represent the proposed factor constructs. Values on the single-headed arrows leading from the factors are standardized factor loadings. Values on the curved double-headed arrows between rectangles are correlations between error terms. Values on the curved double-headed arrows between ellipses are correlations between latent variables. + + 301928 + Species + + ellipses + + + 301928 + Species + + ellipses + + + MESH:D000069451 + Disease + + error terms + + + + RESULTS + paragraph + 19039 + The Initial second-order model demonstrated poor model fit (x2 = 1838.24, df = 269, P < 0.001, CFI = 0.73, TLI = 0.70, RMSEA = 0.093). Modification indices suggested overall model fit would be improved by adding the correlation of six pairs of error terms (item 4 & 5 within "knowledge", 12 & 13 in "application", 21 & 24, 23 & 31, 27 & 30, and 31 & 32 in "attitude"). After the modification was executed, statistical fit of the model was improved to as follows: x2 = 1205.20, df = 312, P < 0.001, CFI = 0.86, TLI = 0.84, RMSEA = 0.065. Although the overall fit improved, model fit indices especially CFI and TLI were still inadequate. We observed factor loading (beta = 0.05) of the item 3 ("Clinical trials and observational methods are equally valid in establishing treatment effectiveness") was significantly lower than the other five items on the dimension of knowledge. After removing this item from the scale, goodness-of-fit statistics improved to x2 = 1056.65, df = 287, P < 0.001, CFI = 0.89, TLI = 0.86, RMSEA = 0.06 (Figure 1) which was very close to our a priori threshold (CFI/TLI >= 0.90, RMSEA < 0.08). + + MESH:D000069451 + Disease + + error terms + + + + DISCUSS + title_1 + 20260 + Discussion + + + DISCUSS + paragraph + 20271 + This study provided support for the use of a modified EBQ-KABQ questionnaire to understand different aspects of EBP knowledge, attitudes, behavior and outcomes/decisions in a variety of healthcare professionals with respect to EBP. We confirmed that the 26 ordinal items in the modified EBP-KABQ exhibit a four-domain construct consistent with the proposed four aspects of EBP. Our scale was modified based on our need to change wording to make the scale more broadly applicable to different disciplines since the original version targeted medical students. We also made changes based our experiences in pilot testing the measure since an expert committee and pilot users found some items to be redundant or difficult to understand. Our work builds on that of the developers who targeted medical trainees by providing a more broadly applicable and validated version. The newly proposed subscale construct of "outcomes/decisions" contains the items previously termed "future use" in the original scale. Outcomes/decisions more accurately reflect the item content and the targeting of the EBP-KABQ. Whereas, as the original instrument was focused on trainees who might be responding about future use, experienced clinicians will be reporting how they use EBP in current clinical decision-making and whether they attribute better outcomes to their evidence-based decisions. This domain is considered an important aspect of self-reported EBP since its focuses on the impact on practice and outcomes. We found the "outcomes/decisions" domain was moderately correlated with the other three domains, suggesting it played a role in perception of EBP. The shorter measure has improved measurement characteristics, retains conceptual domains and may be save administration time. + + MESH:D008569 + Disease + + wording + + + + DISCUSS + paragraph + 22052 + We found the EBP-KABQ scale demonstrates promising psychometric properties when measuring EBP in practicing health professionals because our analysis supported hypotheses posed for construct validity, and we found appropriate scaling properties. The overall Cronbach's alpha (0.85) was superior to that of the original KAB scale (0.75) which may be attributed to deletion of problematic items. + + + DISCUSS + paragraph + 22448 + The correlation between the knowledge and attitude/application domains was relatively weak. This suggests that these are relatively distinct domains. One explanation for this low correlation may be that increased focus on EBP in entry-level and post-professional education may have had more impact on knowledge than on attitudes and application of EBP. However, measurement error may also have contributed. We observed lower internal consistency of the "knowledge" domain compared to other subscales and compared to the original KAB. Low internal consistency suggested that the six items within the construct of "knowledge" were not adequately correlated. As item 3 (Clinical trials and observational methods are equally valid in establishing treatment effectiveness) demonstrated low factor loading to domain of "knowledge", we questioned the content validity of this item. One explanation for this misfit item could be that clinicians might have confused the words "observational study" with "clinical observation". However, we suspect that controversy over the "level of evidence" or "quality" of observational studies may have contributed to misfit on this item. In fact, more recent trends in evidence rating have acknowledged large observational studies as offering high quality evidence. Respondents may value large observational studies more than small trials and not endorse this item despite strong knowledge of EBP. Since this item does not appear to reflect the domain of "knowledge", and did not fit in CFA, we proposed removal. We suggest caution when using the "knowledge" subscale on its own to evaluate EBP knowledge, as further investigation is warranted to improve this sub-scale. + + + DISCUSS + paragraph + 24184 + We found items in EBP knowledge skewed to the high extreme, whereas the others subscales did not demonstrate this. As evidence-based practice has become accepted around the world, it is now commonly integrated in the clinical training of many professionals. Hence, knowledge about what evidence-based practice is, becomes prevalent over time. Our finding may be explained by the fact that traditional evidence-based training focuses on providing knowledge to help practitioners enhance their techniques and skill level when searching and appraising evidence but less consistently focuses on implementation behaviours for integrating EBP into daily clinical activities nor resolving attitudinal barriers towards EBP. For instance, clinicians may enhance their knowledge of methods to find and appraise evidence, including the importance of systematic reviews in the evidence-based practice paradigm, but not be willing to able to incorporate this into their day-to-day clinical decision-making. Continuing medical education events often focus on providing content knowledge rather than active approaches, although the latter is more effective in promoting behavior change. This may contribute to the findings observed in the study. + + + DISCUSS + paragraph + 25415 + We found several factors were associated with better uptake of EBP. People with a higher level of education, more years of training, completion of advanced clinical training and those practicing in rural areas reported a greater willingness to implement EBP in their daily practice. Our findings were consistent with other studies that also found health professionals with a higher level of education were more willing to adopt evidence-based practice. On the other hand, our finding that age was not a factor influencing EBP is in contrast to the literature that shows recent graduates are more likely to accept EBP than clinicians who are older. Our findings were narrowly insignificant (p < 0.06) suggesting a small effect of age may not have reached significance. However, age may be less important over time as EBP spreads through post-professional training. + + 9606 + Species + + People + + + + DISCUSS + paragraph + 26283 + Out findings suggest clinicians who practices in rural areas are more amenable to EBP which was an unexpected finding. This may be explained by several reasons. First, clinicians in rural areas are more likely to seek evidence because they have fewer colleagues in their work environment to discuss clinical issues when questions emerge in day-to-day practice. As a consequence, they would be more accustomed to going to the Internet looking for online evidence as a medical resource. Secondly, geography is no longer a barrier for clinicians to acquire evidence based education. McColl reported only 16% of physicians in England received official education regarding literature search techniques. Therefore, clinicians in rural areas may have access to gaining skills in EBP during their professional training, or through other avenues and be motivated to use these skills to solve their clinical questions. + + + DISCUSS + paragraph + 27192 + Our study has some limitations. While it was a strength that we had different professions and a geographically diverse sample, we were unable to explore how contextual factors contributed to our findings. Local differences regarding the EBP training, culture and language among these participants were not captured in our data collection and we could not test for the influence of many potential covariates and limited covariate testing to factors suggested as important in the literature. However, h a broader sample improves the generalizability of our findings. Since the survey was only offered in English, our findings may not represent contexts where English was not a common language. A further consideration is that the data were self-reported. We have no external criterion to examine whether the self-reported evidence-based practice behaviors are consistent with actual practice. The impact of EBP decisions on patient outcomes may be overestimated if physicians overestimate their ability to improve outcome. Studies of EBP that measure patient outcomes by patient-report or objective measures are preferable indicators of the impact of EBP, but can be challenging to measure. We had to make decisions about deletion of items based on expert review and statistical performance. Studies of the reasons for poor item performance that included qualitative techniques such as cognitive interviewing may have identified ways to reform problematic items or captured new concepts. However, since our goal was to stay true to the original KABQ, if possible, our approach was reasonable. Finally, since our sample was derived from clinicians interested in pain, it may not reflect all. Since pain is the most common patient complaint and one relevant across different professions it represented an ideal context to test the EBP-KABQ across professions and contexts. + + 9606 + Species + + participants + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + CONCL + title_1 + 29061 + Conclusion + + + CONCL + paragraph + 29072 + This study provides evidence in a large sample of experienced clinicians from a range of professions interested in pain management that the EBP-KABQ can be used to assess four domains of EBP: Knowledge, attitude, behavior, outcomes/decisions. + + MESH:D010146 + Disease + + pain + + + + CONCL + title_1 + 29315 + Additional files + + + ABBR + title + 29332 + Abbreviations + + + ABBR + paragraph + 29346 + CFA + + - + Chemical + + CFA + + + + ABBR + paragraph + 29350 + Confirmatory factor analysis + + + ABBR + paragraph + 29379 + CFI + + + ABBR + paragraph + 29383 + Comparative fit index + + + ABBR + paragraph + 29405 + EBP + + + ABBR + paragraph + 29409 + Evidence-based practice + + + ABBR + paragraph + 29435 + KAB + + + ABBR + paragraph + 29439 + Knowledge, attitudes, behavior + + + ABBR + paragraph + 29470 + RMSEA + + + ABBR + paragraph + 29476 + Root mean square error of approximation + + + ABBR + paragraph + 29516 + TLI + + + ABBR + paragraph + 29520 + Tucker Lewis index + + + ABBR + footnote + 29539 + Competing interests + + + ABBR + footnote + 29559 + The authors declare that they have no competing interests. + + + ABBR + footnote + 29618 + Authors' contributions + + + ABBR + footnote + 29643 + JCM, ML and RBH conceived the study. 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It may also be used consistent with the principles of fair use under the copyright law. + TITLE + Prehosp Emerg Care; 2014 Jul-Sep; 18(3) 368-74. doi:10.3109/10903127.2013.869642 + emergency medical services benzodiazepines temperature + 374 + surname:Silbergleit;given-names:Robert + surname:Zaleski;given-names:Erin + NIHMS644369 + surname:Spaite;given-names:Daniel + McMullan JT, Jones E, Barnhart B, Denninghoff K, Spaite D, Zaleski E, Silbergleit R + 0 + Degradation of Benzodiazepines after 120 Days of EMS Deployment + + MESH:D001569 + Chemical + + Benzodiazepines + + + + ABSTRACT + abstract_title_1 + 64 + Introduction + + + ABSTRACT + abstract + 77 + EMS treatment of status epilepticus improves outcomes, but the benzodiazepine best suited for EMS use is unclear, given potential high environmental temperature exposures. + + MESH:D013226 + Disease + + status epilepticus + + + MESH:D001569 + Chemical + + benzodiazepine + + + + ABSTRACT + abstract_title_1 + 249 + Objective + + + ABSTRACT + abstract + 259 + To describe the degradation of diazepam, lorazepam, and midazolam as a function of temperature exposure and time over 120 days of storage on active EMS units. + + MESH:D008874 + Chemical + + midazolam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + + ABSTRACT + abstract_title_1 + 418 + Methods + + + ABSTRACT + abstract + 426 + Study boxes containing vials of diazepam, lorazepam, and midazolam were distributed to 4 active EMS units in each of 2 EMS systems in the southwestern United States during May-August 2011. The boxes logged temperature every minute and were stored in EMS units per local agency policy. Two vials of each drug were removed from each box at 30-day intervals and underwent high-performance liquid chromatography to determine drug concentration. Concentration was analyzed as mean (and 95%CI) percent of initial labeled concentration as a function of time and mean kinetic temperature (MKT). + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + midazolam + + + + ABSTRACT + abstract_title_1 + 1015 + Results + + + ABSTRACT + abstract + 1023 + 192 samples were collected (2 samples of each drug from each of 4 units per city at 4 time-points). After 120 days, the mean relative concentration (95%CI) of diazepam was 97.0% (95.7-98.2%) and of midazolam was 99.0% (97.7-100.2%). Lorazepam experienced modest degradation by 60 days (95.6% [91.6-99.5%]) and substantial degradation at 90 days (90.3% [85.2-95.4%]) and 120 days (86.5% [80.7-92.3%]). Mean MKT was 31.6 C (95%CI 27.1-36.1). Increasing MKT was associated with greater degradation of lorazepam, but not midazolam or diazepam. + + MESH:D003975 + Chemical + + diazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D008874 + Chemical + + midazolam + + + + ABSTRACT + abstract_title_1 + 1574 + Conclusions + + + ABSTRACT + abstract + 1586 + Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. Lorazepam experienced significant degradation over 120 days and appeared especially sensitive to higher MKT exposure. + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + Midazolam + + + + INTRO + title_1 + 1824 + Introduction + + + INTRO + paragraph + 1837 + Emergency medical services (EMS) treatment of status epilepticus with benzodiazepines improves outcomes. Recent evidence suggests that, among the commonly used benzodiazepines, midazolam may be the most effective in achieving seizure cessation prior to hospital arrival. EMS medications are frequently stored without temperature-control procedures, which may negatively impact the medication through degradation, and heat stability is an important factor in determining which benzodiazepine to deploy in an EMS system. Diazepam and lorazepam experience some heat-dependent degradation while midazolam is heat-stable for at least 60 days. The effect of longer storage, especially in extreme heat conditions, is unknown. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D013226 + Disease + + status epilepticus + + + MESH:D001569 + Chemical + + benzodiazepine + + + MESH:D003975 + Chemical + + Diazepam + + + MESH:D001569 + Chemical + + benzodiazepines + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D012640 + Disease + + seizure + + + MESH:D001569 + Chemical + + benzodiazepines + + + + INTRO + paragraph + 2556 + We sought to expand on our previous work by describing the degradation of diazepam, lorazepam, and midazolam as a function of temperature exposure and time over a longer, 120-day storage period on active EMS units during the summer months in the southwestern United States. + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + + METHODS + title_1 + 2830 + Methods + + + METHODS + title_2 + 2838 + Study Design and Setting + + + METHODS + paragraph + 2863 + This experimental pharmaco-stability study of medications stored in active EMS units was designed as an independent extension of our previous work that demonstrated heat-dependant degradation of lorazepam over 60 days. The present study was conducted during the summer of 2011 (May through August) and focused on two EMS agencies in the southwestern United States with historically high ambient temperatures. We extended the period of observation to 120 days and added diazepam as a comparator to give insight on the behavior of all benzodiazepines currently available for prehospital use. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D001569 + Chemical + + benzodiazepines + + + MESH:D003975 + Chemical + + diazepam + + + + METHODS + paragraph + 3453 + Vials of diazepam, lorazepam, and midazolam were distributed to 4 active EMS units in each of the two EMS systems. Instrumented boxes logged temperature every minute and were stored in EMS units per local agency policy alongside other routine medications. Use of temperature-control systems beyond normal vehicle air conditioning or garaging practices were not specified in the study protocol. Mirroring routine EMS practices, some vehicles were kept in station garages unless responding to an emergency call, while others were constantly exposed to ambient temperatures during work shifts. Two samples of each benzodiazepine were removed from each box after 30, 60, 90, and 120 days of deployment. + + 32644 + Species + + not specified + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D001569 + Chemical + + benzodiazepine + + + + METHODS + paragraph + 4152 + The methods of measurement, data collection, and data processing were identical to our previous study. Briefly, the instrumented study boxes measured and recorded temperature every minute. Temperatures were analyzed and summarized by determination of the mean kinetic temperature (MKT), which is commonly used in the pharmaceutical industry to describe the overall effects of temperature changes on heat-sensitive materials. MKT expresses the cumulative heat stress to which a medication has been exposed over time and is not a simple average of ambient temperatures. + + + METHODS + paragraph + 4720 + Samples were analyzed in a commercial laboratory (DynaLabs, St. Louis, MO) by high-performance liquid chromatography (HPLC) to determine the concentration of the active drug. Samples were refrigerated after removal from the field, including during shipping, to minimize further heat-related degradation. + + + METHODS + paragraph + 5024 + Data were managed within Microsoft Excel (Microsoft, Redmond, WA) and analyzed using SPSS version 19 (International Business Machines, Armonk, NY). + + + METHODS + title_2 + 5172 + Outcome Measures + + + METHODS + paragraph + 5189 + The primary outcome was the relative reduction in medication concentration from labeled concentration after 30-120 days of exposure. + + + METHODS + title_2 + 5324 + Primary Data Analysis + + + METHODS + paragraph + 5346 + Concentration was analyzed as a function of time and MKT. For each benzodiazepine, the mean relative concentration at 60, 90, and 120 days was compared to the 30-day measurement using the Student t-test. The influence of MKT and time on each medication's degradation was determined with linear regression and oneway analysis of covariance (ANCOVA), respectively. + + MESH:D001569 + Chemical + + benzodiazepine + + + + METHODS + title_2 + 5709 + Sample Size Determination + + + METHODS + paragraph + 5735 + Sample size was estimated to provide a significance of 0.05 and a power of 0.8, assuming a mean difference of 7.5% between the 120-day relative concentrations of lorazepam compared to midazolam or diazepam. The assumed within-group sample variability (standard deviation) was 5%. + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + + RESULTS + title_1 + 6015 + Results + + + RESULTS + paragraph + 6023 + A total of 192 samples were collected (2 samples in each of 4 units per city at 4 timepoints for each drug) and underwent HPLC. The cumulative mean MKT over the 120-day period was 31.6 C (95%CI 27.1-36.1 C) (Table 1). + + + RESULTS + paragraph + 6245 + Benzodiazepine concentration over time is shown in Table 1 and the impact of time on degradation was significantly different among the benzodiazepines (ANCOVA p < 0.01). Diazepam and midazolam experienced minimal degradation at each time point. At 120 days, the mean relative concentration (95%CI) of diazepam was 97.0% (95.7-98.2%) and of midazolam was 99.0% (97.7-100.2%). Lorazepam experienced significant degradation by 60 days (95.6% [91.6-99.5]) with the concentration in half of all samples being less than 95% of labeled concentration. Relative concentration of lorazepam was 90.3% (85.2-95.4) at 90 days and 86.5% (80.7-92.3) at 120 days. + + MESH:D008874 + Chemical + + midazolam + + + MESH:D003975 + Chemical + + Diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D001569 + Chemical + + benzodiazepines + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D001569 + Chemical + + Benzodiazepine + + + + RESULTS + paragraph + 6903 + Midazolam and diazepam were stable across the range of mean kinetic temperatures, whereas increasing MKT was associated with greater degradation of lorazepam after 120 days (Figure 1; lorazepam R2 = 0.98). The mean (95%CI) daily ambient temperature for each site was not significantly different (30.6 C [23.3-37.2 C] vs. 33.3 C [26.7-40.0 C]; p = 0.83). However, there was a greater than expected observed difference of MKT between sites over the 120 days (27.0 C [22.9-31.1 C] vs. 35.0 C [30.3-39.8 C]; p = 0.009). There was no significant intersite difference in the relative concentrations of midazolam or diazepam at any timepoint. Lorazepam experienced statistically significant degradation at 60, 90, and 120 days (p = 0.009) at each site, and the magnitude of the temperature effect was different between the two sites (ANCOVA p = 0.001) (Figure 2). + + MESH:D003975 + Chemical + + diazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + Midazolam + + + + DISCUSS + title_1 + 7776 + DIscussion + + + DISCUSS + paragraph + 7787 + In this study, we found that midazolam and diazepam experienced minimal degradation during 120 days of EMS deployment in high-heat environments. Lorazepam maintained acceptable concentrations of active drug for at least 30 days. However, when exposed to high heat stress, many samples experienced significant and progressive degradation by 60 days. + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + Lorazepam + + + + DISCUSS + paragraph + 8136 + We have previously evaluated rates of degradation for lorazepam and midazolam over 60 days of EMS field deployment at multiple sites during the conduct of a multicenter clinical trial. In that study we found that midazolam remained stable at 60 days, but that lorazepam showed slight time- and temperature-dependent degradation. The current study builds upon this work by extending the period of observation to 120 days, by focusing on EMS systems with very high heat stress, and by including diazepam, the most common benzodiazepine in EMS use. The current study confirms the stability of midazolam for at least 120 days and that lorazepam is time and heat sensitive. + + MESH:D008874 + Chemical + + midazolam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D001569 + Chemical + + benzodiazepine + + + + DISCUSS + paragraph + 8805 + Gottwald et al. previously reported some experience with the degradation of diazepam and lorazepam deployed on two ambulances in San Francisco. Interestingly, despite higher ambient temperatures, we found that diazepam was more resilient than first reported, with no diazepam samples determined to have <90% of labeled concentration. Lorazepam's degradation was greater at 60 days and beyond, as well, reaffirming the relationship between heat stress and medication decomposition. + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D003975 + Chemical + + diazepam + + + + DISCUSS + paragraph + 9286 + Ambient temperatures were similar between the two sites. However, there was an unexpected difference in the MKTs encountered at each site (27.0 vs. 35.0 C). Although unplanned, this difference allows additional insight into lorazepam's instability in the prehospital setting. Post hoc evaluation of this difference found that the agency with the lower MKT frequently keeps EMS units running with the vehicle's air conditioning system engaged, while the other agency routinely turns off the EMS units and parks them in station garages between calls. It is probable that a combination of operational and environmental factors contributed to the higher degradation rate at one site. However, it appears that altering the deployment and storage procedures for vehicles does not prevent lorazepam degradation in hot environments and further temperature-control methods, such as on-board refrigeration units, may be needed to extend useful shelf life. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008140 + Chemical + + lorazepam + + + + DISCUSS + paragraph + 10233 + It is notable that ambient temperature cannot be used as a surrogate for MKT when assessing whether, or for how long, lorazepam can be stored in an EMS unit. MKT is a dynamic variable that accounts for the potential stress caused by changing temperatures, which is one reason that the pharmacology literature supports the use of MKT, rather than simple temperature means, for evaluating heat stability of drugs. These data support the notion that EMS agencies should take multiple variables into consideration, including temperature exposure and length of field deployment, when determining medication storage and restocking policies. + + - + Chemical + + MKT + + + - + Chemical + + MKT + + + MESH:D008140 + Chemical + + lorazepam + + + + DISCUSS + paragraph + 10868 + Based on our findings here and previously, EMS systems choosing to deploy lorazepam should employ lorazepam storage methods to limit high heat exposures and maintain controlled room temperature (MKT < 25 C) environments. Otherwise, it may be prudent to limit field deployment time of lorazepam to 30 days to minimize degradation risks. In some systems, preferential use of midazolam or diazepam may be warranted. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + + DISCUSS + title_1 + 11282 + Limitations And Future Research + + + DISCUSS + paragraph + 11314 + This study has limitations. First, baseline (day 0) samples were not obtained because our previous work demonstrated consistent baseline concentrations for midazolam and lorazepam. Furthermore, the differences in the pair of samples taken from each EMS unit for testing were insignificant. This is consistent with the quality controls and USP specifications expected in these commercial pharmaceuticals. + + MESH:D008874 + Chemical + + midazolam + + + MESH:D008140 + Chemical + + lorazepam + + + + DISCUSS + paragraph + 11718 + Second, we did not perform duplicate measures from each sample; instead, we performed redundant single measurements from independent but identically stored samples. Previous work has demonstrated the reliability of the HPLC testing, and our duplicate sample testing reduces the bias that an outlier may cause. + + + DISCUSS + paragraph + 12028 + Finally, this study was not designed to evaluate the impact of different vehicle deployment or medication storage practices on drug degradation. The impact of medication refrigerators and other techniques to control heat exposure and MKT may be more important for lorazepam than the other benzodiazepines and is a topic suitable for further study. + + MESH:D001569 + Chemical + + benzodiazepines + + + MESH:D008140 + Chemical + + lorazepam + + + + DISCUSS + paragraph + 12376 + Evaluating the impact of heat exposure on other medications commonly used by EMS should be a priority. + + + CONCL + title_1 + 12479 + Conclusion + + + CONCL + paragraph + 12490 + Midazolam and diazepam experienced minimal degradation throughout 120 days of EMS deployment in high-heat environments. In contrast, lorazepam degraded significantly over this time and appeared especially sensitive to higher mean kinetic temperatures. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + Midazolam + + + MESH:D003975 + Chemical + + diazepam + + + + CONCL + footnote + 12742 + Author contributions: JM and RS conceived the study and designed the trial. All authors supervised the conduct of the trial and data collection. JM and RS analyzed the data. JM drafted the manuscript, and all authors contributed substantially to its revision. JM takes responsibility for the paper as a whole. + + + CONCL + footnote + 13052 + + + + CONCL + footnote + 13069 + The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper. + + + CONCL + footnote + 13190 + + + + CONCL + footnote + 13207 + This work was presented in oral and poster format at the Society for Academic Emergency Medicine annual meeting (May 2013, Atlanta). + + + CONCL + footnote + 13340 + + + + METHODS + title + 13357 + The Neurological Emergencies Treatment Trials investigators + + + METHODS + paragraph + 13417 + Clinical Coordinating Center + + + METHODS + paragraph + 13446 + Robert Silbergleit, MD, Daniel Lowenstein, MD, William Barsan, MD, Arthur Pancioli, MD, Valerie Stevenson, BAS, RRT, CCRP, Erin Zaleski, MA, Deneil + + + METHODS + paragraph + 13594 + Harney, MPH, MSW, Donna Harsh, MS, Joy Pinkerton, BSN, RN, MS, Allison Kade, BA, Nicholas Siewert, BA, Ashley Pinawin, BS, Catherin Ring, Phebe Brenne + + + METHODS + paragraph + 13745 + National EMS Coordinator + + + METHODS + paragraph + 13770 + Kay Vonderschmidt, MPA, MS-EM, NREMT-P + + + METHODS + paragraph + 13809 + Statistical Data Management Center + + + METHODS + paragraph + 13844 + Valerie Durkalski, PhD, Yuko Palesch, PhD, Catherine Dillon, Keith Pauls, Qi Wu, Wenle Zhao, PhD + + + METHODS + paragraph + 13941 + National Institutes of Health + + + METHODS + paragraph + 13971 + Robin Conwit, MD, Scott Janis, PhD, David Jett, PhD, Brandy Fureman, PhD + + 9188 + Species + + Robin + + + + METHODS + paragraph + 14044 + Hubs (ordered by number of subjects enrolled) + + + METHODS + paragraph + 14090 + Wayne State University (178) + + + METHODS + paragraph + 14119 + Hub Principal Investigator: Robert D. Welch, MD, MS Primary Study Coordinators: Lynnmarie Mango, MPH, Valerie H. Mika, MS EMS Director(s)/Coordinator: Jenny Atas, MD Other Site Investigators: Robert Dunne, MD, Douglas Wheaton, MD, Phillip Levy, MD, MPH, Marc-Anthony Velilla, MD, Robert Sherwin, MD, Brian O'Neil, MD, Angela Groves, MD, Marc Rosenthal, DO, PhD Participating EMS Service: Detroit EMS + + 29780 + Species + + Mango + + + 301693 + Species + + Atas + + + MESH:D004881 + Disease + + Marc-Anthony Velilla + + + + METHODS + paragraph + 14519 + University of Cincinnati (133) + + + METHODS + paragraph + 14550 + Hub Principal Investigator: Arthur Pancioli, MD Primary Study Coordinators: Irene Ewing, RN, BSN, Peggy Waymeyer, RN EMS Director(s)/Coordinator: M. Kay Vonderschmidt, MPA, MS-EM, NREMT-P, Jason McMullan, MD Other Site Investigators: Hamilton Schwartz, MD, Brian Stettler, MD, William Knight, MD, Opeolu Adeoye, MD, Rhonda Cadena, MD, Jordan Bonomo, MD, Erin Grise, MD, Laura Heitsch, MD, George Shaw, MD, Nick Gagai, CCRP, Pamela Schmit, RN BSN, Sara Stark, Med, Traci Doellman, RN Participating EMS Services: Cincinnati Fire Department, BlueAsh Fire Department, Forest Park Fire Department, Green Township Fire Department, Florence Fire Department, Independence Fire Department + + CellLine + + Laura Heitsch + + + + METHODS + paragraph + 15230 + University of California San Francisco (121) + + + METHODS + paragraph + 15275 + Hub Principal Investigator: J. Claude Hemphill, III, MD, MAS Primary Study Coordinators: Michele Meeker, RN, BSN, Kelley Rosborough, BA EMS Director(s)/Coordinator: Jeany Duncan EMT-P Other Site Investigators: Karl Sporer, MD, FACEP, FACP, Alan Gelb, MD; Wade Smith, MD, PhD, Prasanthi Ramanujam, MD, Kazuma Nakagawa, MD, Asma Moheet, MD, Hooman Kamel, MD, Bharath Naravetla, MD, Mary Mercer, MD, Christine Wong, MD Participating EMS Services: San Francisco Fire Department, EMS Division + + MESH:D020526 + Disease + + Claude Hemphill + + + + METHODS + paragraph + 15763 + University of Texas - Houston (81) + + + METHODS + paragraph + 15800 + Hub Principal Investigator: Elizabeth Jones, MD Trial Principal Investigator: Truman J. Milling, MD Primary Study Coordinators: Misty Ottman, RN, BSN, Ben King, Laura LaChance EMS Directors/Coordinators: Jeff Brockman, RN, Pete Didonato, EMT-P Other Site Investigator: Paul Hinchey, MD Participating EMS Service: Austin-Travis County EMS + + CVCL:0030 + CellLine + + Laura La + + + + METHODS + paragraph + 16138 + Emory University (75) + + + METHODS + paragraph + 16160 + Hub Principal Investigator: David W. Wright, MD Trial Principal Investigators: Matthew D. Bitner, MD, Gerald W. Beltran, DO Primary Study Coordinator: Harriet Nevarez, RN, CCRC EMS Director/Coordinator: Rachel Barnhard, Andrea G. McDougal Other Site Investigators: Jeffrey F. Linzer Sr, MD, Lisa H. Merck, MD MPH, Tamara Espinoza, MD Participating EMS Service: Grady EMS + + + METHODS + paragraph + 16531 + Henry Ford Health System (64) + + + METHODS + paragraph + 16561 + Hub Principal Investigator: Christopher A. Lewandowski, MD Trial Principal Investigator: Taher T. Vohra, MD Primary Study Coordinators: Paula L. Crouse, RN, BSN., MA., Anna E. Baker, RN, BSN EMS Director/Coordinator: Dean R. Creech EMT-P, I/C Other Site Investigator: Andrew N. Russman, DO, Joseph B. Miller, MD, Jumana Nagarwala, MD, Daniel J. Miller, MD, Raymond Fowkes, MD, Anne Marie Lundell, RN, BSN Participating EMS Services: Detroit EMS, West Bloom-field Fire and EMS Services + + + METHODS + paragraph + 17046 + Stanford University (62) + + + METHODS + paragraph + 17071 + Hub Principal Investigator: James V. Quinn, MD, MS Primary Study Coordinators: Stephanie Casal. RN, CNS, Anke Hebig, Mark Liao EMS Director/Coordinator: Peter D'souza, MD Participating EMS Services: Palo Alto Fire Department, San Jose Fire Department, Redwood City Fire Department, San Mateo Fire Department + + 28980 + Species + + Redwood + + + + METHODS + paragraph + 17379 + University of Arizona (58) + + + METHODS + paragraph + 17406 + Hub Principal Investigator: Kurt R. Denninghoff, MD Trial Principal Investigator: Daniel W. Spaite, MD Primary Study Coordinator: Bruce Barnhart, RN, CEP EMS Director(s)/Coordinator: Willie Haro, CEP Other Site Investigator: Bentley J. Bobrow, MD Participating EMS Service: Glendale Fire Department + + + METHODS + paragraph + 17705 + Virginia Commonwealth University (55) + + + METHODS + paragraph + 17743 + Hub Principal Investigator: Joseph P. Ornato, MD Primary Study Coordinator: Sallie L. Noe, RN + + + METHODS + paragraph + 17837 + EMS Director/Coordinator: Alan D. Payne, CCEMTP Other Site Investigators: Alan R. Towne, MD, Michael C. Kurz, MD, John T. Carmack, MD Participating EMS Service: Richmond Ambulance Authority + + + METHODS + paragraph + 18027 + University of Minnesota (47) + + + METHODS + paragraph + 18056 + Hub Principal Investigator: Michelle Biros, MD Trial Principal Investigator: Brian Mahoney, MD Primary Study Coordinators: Corey Sargent, Kathleen Miller, BSN, CCRC Other Site Investigators: David Hildebrandt, Chris Kummer, Doug Gesme Participating EMS Services: Hennepin County EMS + + + METHODS + paragraph + 18339 + Medical College of Wisconsin (47) + + + METHODS + paragraph + 18373 + Hub Principal Investigator: Tom P. Aufderheide, MD Primary Study Coordinator: Joseph T. Brandt Jr., BS, EMT-P EMS Director/Coordinator: M. Riccardo Colella, DO Other Site Investigators: Ron Pirrallo, MD, MHSA, Walter Bialkowski, MS, Benjamin Hermanson, BS, Christopher Sandoval, BS, EMT-P, Kevin Morrow, MFA, Kelly McCormick, BS, MBA, Katherine Burpee, BA, Geri Price, BS, Dawn Kawa, BA Participating EMS Services: Milwaukee County EMS, Milwaukee Fire Department, Franklin Fire Department, Greenfield Fire Department, North Shore Fire Department, Oak Creek Fire Department, South Milwaukee Fire Department, Wauwatosa Fire Department, West Allis Fire Department + + Disease + + Riccardo Colella + + + + METHODS + paragraph + 19034 + University of Kentucky (31) + + + METHODS + paragraph + 19062 + Hub Principal Investigator: Roger L. Humphries, MD Primary Study Coordinator: Linda Dechtenberg, RN, BSN, CCRC EMS Director/Coordinator: Christofer Sweat Other Site Investigator: L. Creed Pettigrew, MD,MPH Participating EMS Service: Lexington-Fayette Urban County Government Division of Fire & Emergency Services + + + METHODS + paragraph + 19375 + University of Pennsylvania (26) + + + METHODS + paragraph + 19407 + Hub Principal Investigator: Jill M. Baren, MD, MBE Trial Principal Investigator: R. Daniel Bledsoe, MD Primary Study Coordinator: Barbie Stahlman, MS, Katherine Lamond, BA, Pamela G. Nathanson, MBE Other Site Investigator: Scott E. Kasner, MD, MSCE, Peter D. Le Roux, MD Participating EMS Services: York Hospital Medic 97, White Rose Ambulance, Grantley Fire Company, Jacobus Lions Ambulance Club, West York Ambulance + + 9689 + Species + + Lions + + + + METHODS + paragraph + 19825 + Oregon Health & Science University (24) + + + METHODS + paragraph + 19865 + Hub Principal Investigators: Craig R. Warden, MD, MPH, Robert A. Lowe, MD, MPH Primary Study Coordinator: Rachel N. Stone, CCRP Participating EMS Service: Clackamas Fire District #1 + + + METHODS + paragraph + 20047 + New York Presbyterian Hospital (10) + + + METHODS + paragraph + 20083 + Hub Principal Investigator: Stephan Mayer, MD, FCCM Trial Principal Investigator: Neal Flomenbaum, MD Primary Study Coordinators: M. Cristina Falo, PhD, Lisa-Vanessa Magitbay, RN, Chirag Surti EMS Directors/Coordinators: Heidi Cordi, MD, Daniel Ribaudo Other Site Investigators: Axel Rosengart, MD, PhD, Matthew Vibbert, MD, Santiago Ortega-Gutierrez, MD, H. Alex Choi, MD, Emily Gilmore, MD, Rishi Malhotra, MD, Lawrence Berger Participating EMS Services: New York Presbyterian + + + METHODS + paragraph + 20562 + Temple University(8) + + + METHODS + paragraph + 20583 + Hub Principal Investigator: Nina T. Gentile, MD Trial Principal Investigators: Alvin Wang, DO, Christopher Vates, MD, Ben Usatch, MD Primary Study Coordinators: Brent B. Freeman, Stacey L. Cleary Participating EMS Services: Volunteer Medical Services Corps of Lower Merion and Narberth (Narberth Ambulance), Life Lion EMS + + 9689 + Species + + Lion + + + + METHODS + paragraph + 20905 + University of Maryland (3) + + + METHODS + paragraph + 20932 + Hub Principal Investigator: Barney Stern, MD Trial Principal Investigators: Tricia Ting, MD, Gregory Krauss, MD Primary Study Coordinators: Virginia Ganley, RN, Susan Rice, RN, Jennifer Ronald EMS Director/Coordinator: Michelle Stevens, RN Other Site Investigators: Brian Browne, MD, Robert Rosenthal, MD, Peter Hill, MD Participating EMS Services: Maryland Institute for Emergency Medical Services Systems (MIEMSS), Baltimore City EMS + + 4530 + Species + + Rice + + + + REF + title + 21368 + References + + + surname:Corry;given-names:MD + surname:Isaacs;given-names:SM + surname:Gelb;given-names:AM + surname:Alldredge;given-names:BK + 631 + 11547716 + 2001 + N Engl J Med + ref + 345 + surname:Lowenstein;given-names:DH + REF + 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surname:Polykratis;given-names:IA + surname:de Guzman;given-names:R + 394 + 23734991 + 2013 + Prehosp Emerg Care + ref + 17 + REF + 400 + surname:Dubick;given-names:MA + surname:Cap;given-names:AP + 21878 + Stability of tranexamic acid after 12-week storage at temperatures from -20 degrees C to 50 degrees C. + + MESH:D014148 + Chemical + + tranexamic acid + + + + surname:Payton;given-names:TF + 16 + surname:Green;given-names:P + surname:Shayhorn;given-names:MA + REF + surname:Kupas;given-names:DF + 67 + 22035224 + 2012 + Prehosp Emerg Care + ref + 75 + 21983 + Structured inspection of medications carried and stored by emergency medical services agencies identifies practices that may lead to medication errors. + + + surname:Orsulak;given-names:PJ + surname:Liu;given-names:PK + surname:Akers;given-names:LC + surname:Gottwald;given-names:MD + 333 + 10452426 + 1999 + Am J Emerg Med + ref + 17 + REF + surname:Alldredge;given-names:BK + surname:Lowenstein;given-names:DH + 7 + surname:Fields;given-names:SM + surname:Bacchetti;given-names:P + surname:Corry;given-names:MD + 22135 + Prehospi tal stability of diazepam and lorazepam. + + MESH:D003975 + Chemical + + diazepam + + + MESH:D008140 + Chemical + + lorazepam + + + + surname:Denninghoff;given-names:K + surname:Jones;given-names:E + surname:Pinnawin;given-names:A + surname:McMullan;given-names:JT + 1 + 23148574 + 2013 + Prehosp Emerg Care + ref + 17 + REF + surname:Silbergleit;given-names:R + 7 + surname:Zaleski;given-names:E + surname:Spaite;given-names:DW + surname:Siewart;given-names:N + 22185 + The 60-day temperature-dependent degradation of midazolam and lorazepam in the prehospital environment. + + MESH:D008140 + Chemical + + lorazepam + + + MESH:D008874 + Chemical + + midazolam + + + + surname:Grady;given-names:LT + 57 + surname:Medwick;given-names:T + surname:Bailey;given-names:L + REF + surname:Okeke;given-names:CC + 1441 + 10938985 + 2000 + Am J Health Syst Pharm + ref + 5 + 22289 + Revised USP standards for product dating, packaging, and temperature monitoring. + + + FIG + nihms-644369-f0001.jpg + F1 + fig_caption + 22370 + Relative concentrations of benzodiazepines at 120 days as a function of cumulative mean kinetic temperatures (MKT). + + MESH:D001569 + Chemical + + benzodiazepines + + + + FIG + nihms-644369-f0002.jpg + F2 + fig_caption + 22486 + Comparison by site of relative concentration of lorazepam and cumulative mean kinetic temperature (MKT) at 30, 60, 90, and 120 days. + + MESH:D008140 + Chemical + + lorazepam + + + + TABLE + table_footnote + 22619 + p = 0.014 + + + TABLE + table_footnote + 22629 + + + + TABLE + table_footnote + 22661 + p < 0.001 when compared to 30-day concentration. + + + TABLE + table_footnote + 22710 + + + + TABLE + table + 22742 + + + + TABLE + T1.xml + T1 + table_caption + 22768 + Average mean kinetic temperature (MKT) and relative concentration of benzodiazepines compared to label at each measured timepoint. + + MESH:D001569 + Chemical + + benzodiazepines + + + + TABLE + T1 + table + T1.xml + 22899 + Concentration, mean % (95% CI) 30-day 60-day 90-day 120-day Diazepam 97.0 (96.3-97.6) 97.1 (96.6-97.6) 97.4 (96.6-98.3) 97.0 (95.7-98.2) Lorazepam 101.0 (99.0-102.9) 95.6* (91.6-99.5) 90.3** (85.2-95.4) 86.5** (80.7-92.3) Midazolam 101.0 (99.8-101.4) 100.6 (99.8-101.4) 99.0 (98.0-99.9) 99.0 (98.1-100.2) MKT 28.1 (24.9-31.2) 30.4 (26.0-34.7) 31.0 (26.8-35.1) 31.6 (27.1-36.1) + + MESH:D008874 + Chemical + + Midazolam + + + MESH:D008140 + Chemical + + Lorazepam + + + MESH:D003975 + Chemical + + Diazepam + + + + + 4272821 + + surname:Braun;given-names:Thomas + surname:Nolte;given-names:Hendrik + surname:Wust;given-names:Stas + 25530839 + surname:Boettger;given-names:Thomas + 2014 + 4272821 + 23 + 10.1186/s13395-014-0023-5 + Keywords + front + 23 + 4 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + Skelet Muscle; 2014; 4(1) 23. doi:10.1186/s13395-014-0023-5 + miR-206 miR-133b miR-1 MDX Muscle regeneration Pax7 + Boettger T, Wüst S, Nolte H, Braun T + 0 + The miR-206/133b cluster is dispensable for development, survival and regeneration of skeletal muscle + + 387202 + Gene + + miR-206 + + + + ABSTRACT + abstract_title_1 + 102 + Background + + + ABSTRACT + abstract + 113 + Three different gene clusters code for the muscle-specific miRNAs miR-206, miR-1 and miR-133a/b. The two miR-1/133a clusters generate identical mature miR-1 and miR-133a miRNAs in heart and skeletal muscle, while the cognate miR-206/133b cluster is exclusively expressed in skeletal muscle. Since sequences of the miRNAs miR-133a and miR-133b are almost identical, it seems likely that they share potential targets. Similarly, miR-1 and miR-206 are structurally related and contain identical seed sequences important for miRNA-target recognition. In the past, different functions of these miRNAs were suggested for development, function and regeneration of skeletal muscle using different in vivo and in vitro models; however, mutants lacking the complete miR-206/133b cluster, which generates a single pri-miRNA constituting a functional unit, have not been analyzed. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + ABSTRACT + abstract_title_1 + 982 + Methods + + + ABSTRACT + abstract + 990 + We generated miR-206/133b knock-out mice and analyzed these mice morphologically; at the transcriptome and proteome level to elucidate the contribution of this miRNA cluster for skeletal muscle development, differentiation, regeneration in vivo; and by systematic analysis. In addition, we studied the consequences of a genetic loss of miR-206/133b for expression of Pax7 and satellite cell differentiation in vitro. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + ABSTRACT + abstract_title_1 + 1407 + Results + + + ABSTRACT + abstract + 1415 + Deletion of the miR-206/133b cluster did not reveal any obvious essential function of the miRNA-cluster for development and differentiation of skeletal muscle. Careful examination of skeletal muscles of miR-206/133b mutants revealed no structural alterations or molecular changes at the transcriptome and proteome level. In contrast to previous studies, deletion of the miR-206/133b cluster did not impair regeneration of skeletal muscle in mdx mice. Likewise, differentiation of miR-206/133b deficient satellite cells in vitro was unaffected and no change in Pax7 protein concentration was apparent. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + + ABSTRACT + abstract_title_1 + 2016 + Conclusions + + + ABSTRACT + abstract + 2028 + We conclude that the miR-206/133b cluster is dispensable for development, function and regeneration of skeletal muscle, probably due to overlapping functions of the related miR-1/133a clusters, which are strongly expressed in skeletal muscle. We reason that the miR-206/133b cluster alone is not an essential regulator of skeletal muscle regeneration, although more subtle functions might exist that are not apparent under laboratory conditions. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + + INTRO + title_1 + 2474 + Background + + + INTRO + paragraph + 2485 + miRNAs regulate protein expression at the post-transcriptional level by decreasing transcript abundance or inhibiting protein translation. The miRNAs miR-1, miR-206 and miR-133a/b are specifically expressed in striated muscle. The mouse genome contains two miR-1/133a gene clusters located on chromosome 2 and 18, giving rise to identical mature miR-1 and miR-133a miRNAs while the structurally related miR-206/133b cluster is located on mouse chromosome 1. The mature miR-133b differs in only one nucleotide from miR-133a and the primary sequence of miR-206 is highly related to miR-1. Importantly, miR-1 and miR-206 do not differ in the seed sequence that is assumed to determine target specificity of miRNAs. Deletion of both miR-1/133a clusters, which are expressed in heart and all skeletal muscles, results in early embryonic lethality due to defects in heart development caused by the failure to restrict myocardin expression and concomitant upregulation of smooth muscle genes. Deletion of both miR-1 copies leads to cardiac defects with partially penetrant neonatal lethality but did not cause a major skeletal muscle phenotype, which was attributed to the remaining expression of miR-206 in skeletal muscle. Deletion of both miR-133a copies affects cardiomyocyte proliferation and heart physiology and results in a centronuclear skeletal myopathy and a shift of muscle fiber identity from glycolytic to oxidative muscle fibers in the soleus muscle. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 214384 + Gene + 17043 + + myocardin + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + MESH:D009135 + Disease + + myopathy + + + MESH:D020964 + Disease + + embryonic lethality + + + MESH:D006331 + Disease + + cardiac defects + + + + INTRO + paragraph + 3944 + The miR-206/133b cluster is not expressed in the heart, but its expression is confined to developing skeletal muscle. In adult muscles miR-206/133b are preferentially found in slow myofibers. Expression of miR-206/133b is controlled by a network of myogenic regulatory genes, including MyoD, which binds to the miR-206/133b locus in C2C12 cells. The miR-206/133b locus not only encodes the miRNAs miR-206 and miR-133b, but also the long non-coding RNA linc-MD1, which is expressed during muscle development similar to miR-206 and miR-133b and assumed to act as a competing endogenous RNA or miRNA decoy. Genomic deletion of miR-206 did not cause an obvious clinical phenotype in mice, although it was determined that miR-206 is required for re-innervation of muscle tissue in mice with mutant Sod1. It was proposed that miR-1/206 repress Hdac4, which in turn might regulate genes involved in controlling muscle-derived signals that enhance synapse formation under pathological conditions. More recently, miR-206 was claimed to act as a modifier of muscular dystrophy. Deletion of miR-206 in the mdx mouse model of Duchenne muscular dystrophy led to increased lethality in compound mutants and accumulation of degenerated muscle fibers. Surprisingly, no defects in muscle innervation were observed after the loss of miR-206, although muscle fibers are continuously lost and replaced in mdx mice. The dramatic enhancement of muscle dystrophy in miR-206/mdx compound mutants was attributed to impaired differentiation of myofibers due to the up-regulation of Pax7, which contains identical binding sites for miR-206 and miR-1 in its 3' untranslated region (UTR). The presence of miR-206 target sites in Pax7 did also motivate other researchers to investigate a regulation of Pax7 by miR-206, revealing that overexpression or knock-down of miR-206 can affect Pax7 levels and thereby muscle differentiation in vitro. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 17927 + Gene + 7857 + + MyoD + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 105661730 + Gene + + linc-MD1 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 20655 + Gene + 392 + + Sod1 + + + 208727 + Gene + 55946 + + Hdac4 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + CVCL:0188 + CellLine + + C2C12 + + + MESH:D009136 + Disease + + muscle dystrophy + + + MESH:D009136 + Disease + + muscular dystrophy + + + MESH:D020388 + Disease + + Duchenne muscular dystrophy + + + + INTRO + paragraph + 5858 + Here, we analyzed mice with a targeted deletion of the miR-206/133b cluster. miR-206/133b are processed from a common precursor and thus might be regarded as a functional unit similar to the miR-1/133a clusters. Concomitant loss of miR-206 and miR-133b neither leads to an obvious clinical phenotype or causes detectable molecular changes in skeletal muscles nor impairs muscle regeneration in the MDX mouse model of muscular dystrophy. Surprisingly, lack of miR-206/133b and the miR-133 decoy, contained in the third exon of linc-MD1, did not have obvious effects on satellite cell proliferation and differentiation. Our findings differ from a previous analysis of miR-206 mutants, which might suggest that other products processed from the primary miR-206/133b transcript balance effects of miR-206 in vivo. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 105661730 + Gene + + linc-MD1 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + MDX + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + MESH:D009136 + Disease + + muscular dystrophy + + + + METHODS + title_1 + 6668 + Methods + + + METHODS + title_2 + 6676 + Ethics statement + + + METHODS + paragraph + 6693 + Animal maintenance and animal experiments were in accordance with German animal protection laws and were approved by the local governmental animal protection committee (Regierungspraesidium Darmstadt, Federal State of Hessen). + + + METHODS + title_2 + 6920 + Deletion of miR-206/133b + + 387202 + Gene + + miR-206 + + + + METHODS + paragraph + 6945 + The miR-206/133b coding region was deleted from the mouse genome using a vector containing genomic sequences flanking the miRNA miR-206/133b gene and a PGK-neomycin selection cassette. The short arm (2.3 kb) used for recombination encompassed the miR-206 5' flanking region from TTAGGCATATAAAGTTTGCACGACC to GATATAAAGAAGCATGTGGCCTGGG). The long arm of the targeting vector consisted of a 5.9 kb NcoI fragment directly downstream of miR-133b. A DTA selection cassette was used for negative selection against nonhomologous insertion of the vector into the genome. Homologous recombination in mouse embryonic stem (ES) cells was ascertained by Southern blot analysis using a SphI digest and a 453 bp 5' probe amplified by the oligonucleotides (TAAGTCCTGATGCTTCTCAATACCC; GTTGATAAAGAAACTGTGTGTTACG), resulting in an 8.4-kb WT and an 11.8-kb knock-out signal. Although the inserted beta-galactosidase coding (lacZ) cassette contained an IRES sequence, no beta-galactosidase was expressed from the engineered locus. Genotyping of mdx mice followed the protocol described by Shin et al.. Primary sequences of mature miRNAs were derived from mirBase20 (MIMAT0000239, MIMAT0000123, MIMAT0000769 and MIMAT0000145). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 12091 + Gene + 47922 + + beta-galactosidase + + + 12091 + Gene + 47922 + + beta-galactosidase + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 13405 + Gene + 20856 + + mdx + + + 387202 + Gene + + miR-206 + + + Disease + + MIMAT0000239, MIMAT0000123, MIMAT0000769 + + + MESH:D009355 + Chemical + + neomycin + + + + METHODS + title_2 + 8153 + RNAse protection assay and northern blot + + + METHODS + paragraph + 8194 + Total RNA was isolated from M. soleus of PBS perfused animals using the TRIZOL method (Invitrogen, Thermo Fisher Scientific, Darmstadt, Germany). The RNase protection assay was performed using the mirVana Probe construction Kit (Ambion, Thermo Fisher Scientific, Darmstadt, Germany; Cat#1550) using oligos (miR-206: TGGAATGTAAGGAAGTGTGTGG, miR-145: GTCCAGTTTTCCCAGGAATCCCTTTTTTTT) together with the mirVana miRNA Detection Kit (Ambion#1552) following the manufacturer's instructions. A polyT at the control oligo was added to allow size resolution of the control oligo. The sequence CCTGTCTC was added to the 3' end of the oligos to allow binding of the Ambion T7-Promoter Primer. Amersham alpha-32P UTP (GE Healthcare, Freiburg, Germany; PB20383, 800 Ci/mmol) was used for labeling. Probes were purified using a 15% Invitrogen TBE-Urea Gel (EC68855BOX), the gel was exposed to a Kodak X-ray film, labeled probes were excised und eluted 30 min in elution buffer. 5 mug of total RNA isolated from WT, heterozygous or KO-muscle or yeast-RNA (Roche, Mannheim, Germany), respectively was hybridized at 42 C overnight with approximately 104 cpm of probe in hybridization buffer in a total volume of 20 mul. Samples were analyzed using 15% TBE-Urea gels after RNase A/T1 digestion. Signals were detected using Kodak X-Ray film. Northern blot analysis was performed as previously described. Antisense oligonucleotides specific for miR-206 (CCACACACTTCCTTACATTCCA), miR-1 (ATACATACTTCTTTACATTCCA) and U6 (ATATGGAACGCTTCACGAATT) were used. + + 387202 + Gene + + miR-206 + + + 387163 + Gene + + miR-145 + + + 387202 + Gene + + miR-206 + + + 4932 + Species + + yeast + + + 735281 + Gene + + miR + + + MESH:D009841 + Chemical + + oligonucleotides + + + - + Chemical + + TBE + + + MESH:D014508 + Chemical + + Urea + + + MESH:D014508 + Chemical + + Urea + + + - + Chemical + + alpha-32P UTP + + + - + Chemical + + TBE + + + MESH:D007854 + Chemical + + PBS + + + c.1A>T + DNAMutation + + A/T1 + + + MESH:C411644 + Chemical + + TRIZOL + + + + METHODS + title_2 + 9732 + Microarray analysis, quantitative reverse transcription PCR and small RNA sequencing + + + METHODS + paragraph + 9817 + RNA quality was analyzed with the Agilent Bioanalyzer and the RNA 6000 Nano Kit (Agilent Technologies, Santa Clara, CA). For RNA expression analysis, the Affymetrix GeneChip MouseGenome 430 2.0 Array was used, employing the one-cycle target labeling protocol. Data were analyzed using the Affymetrix expression console and the RMA algorithm. Microarray data have been submitted to Arrayexpress (Acc#E-MTAB-2439). TaqMan MicroRNA Expression Assays and the Applied Biosystems StepOnePlus system were used to quantify miR-206 and miR-1 expression. The RT reaction was done using the miR-206 and the U6 specific primer with the Taqman MicroRNA Reverse Transcription Kit (#4366596). qPCR assays were performed with three independent samples each with Taqman Universal PCR Master Mix (#4324018), FAM labeled miR-206 or miR-1 assay (#000510; #002222) and VIC labeled U6 assay (#001973) for normalization. Relative expression was calculated using the Ct method. RNA isolated from a pool of M. soleus of five male C57Bl6 mice was sequenced using IonTorrent sequencing and analyzed as described. Data were submitted to the NCBI's Gene Expression Omnibus (Acc#GSE63342). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13615 + Gene + 1477 + + VIC + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + METHODS + title_2 + 10985 + Whole-mount in situ hybridization + + + METHODS + paragraph + 11019 + For whole mount in-situ hybridization, a myogenin cDNA corresponding to 307 to 1393 bp of ENSMUST00000027730 was used to synthesize a DIG-labeled antisense probe by T7-RNA-polymerase. E10.5 embryos were isolated and genotyped after mating of heterozygous parents. + + 17928 + Gene + 1854 + + myogenin + + + - + Chemical + + DIG + + + + METHODS + title_2 + 11284 + Fiber type determination, histology + + + METHODS + paragraph + 11320 + For histological analysis, muscle tissue was perfused in situ with 4% PFA, dissected, washed in PBS, dehydrated and embedded in paraffin, followed by preparation of 10 mum sections, deparaffinization and hydration in distilled water. Von Kossa staining was performed by incubating slides with 1% silver nitrate solution and exposed to light for 30 min. Slides were washed two times for 3 min in H2O and incubated in 5% sodium thiosulfate solution for 5 min. Thereafter, slides were washed two times in H2O, followed by counterstaining for 7 min in 0.1% EosinG (Merck, Darmstadt, Germany) with 0.05% acetic acid. Slides were dehydrated and mounted using Entellan. For Sirius red staining, slides were stained in Weigerts Iron-Hematoxylin solution (Sigma, Munich, Germany; Cat#HT1079) for 8 min, dipped twice in distilled water and stained one hour in 0.1% Direct Red 80 (Sigma#365548)/saturated picric acid. Subsequently, slides were washed using 0.5% acetic acid, dipped in distilled water, dehydrated and mounted using Entellan. + + MESH:D014867 + Chemical + + H2O + + + MESH:D014867 + Chemical + + water + + + MESH:C009798 + Chemical + + Direct Red 80 + + + MESH:D007854 + Chemical + + PBS + + + MESH:D017245 + Chemical + + PFA + + + MESH:D012835 + Chemical + + silver nitrate + + + CVCL:5008 + CellLine + + HT1079 + + + MESH:D014867 + Chemical + + water + + + MESH:C017717 + Chemical + + sodium thiosulfate + + + MESH:D019342 + Chemical + + acetic acid + + + MESH:C005858 + Chemical + + picric acid + + + MESH:D014867 + Chemical + + water + + + - + Chemical + + Sirius red + + + MESH:D010232 + Chemical + + paraffin + + + MESH:D014867 + Chemical + + H2O + + + MESH:D019342 + Chemical + + acetic acid + + + - + Chemical + + Iron-Hematoxylin + + + + METHODS + paragraph + 12356 + For determination and quantification of fiber types in the muscle tissue, tissue was perfused in situ with 4% PFA, isolated and incubated in 15% and 30% sucrose/PBS for 2 hours and overnight, respectively. Tissue was frozen on dry ice and cryotome-sectioned. Sections were mounted on Superfrost slides. Tissue was dried and subsequently treated with 4% PFA/0.1% sodium desoxycholate/0.02% NP-40 for 5 min, washed 3 times with PBS for 5 min, blocked in 2% FCS, 0.5% NP-40/PBS for 1 h and then incubated with monoclonal anti-myosin (skeletal, slow; Sigma, M8421) in 2% FCS/0.5% NP-40/PBS overnight at 4 C. Slides were washed three times with PBS for 5 min and then incubated with biotinylated secondary antibody (Vector Labs, Burlingame, CA; BA-1400) for 2 h at RT and further processed according to the Vectastain Elite ABC Kit. + + MESH:D007854 + Chemical + + PBS + + + MESH:D013395 + Chemical + + sucrose + + + MESH:D017245 + Chemical + + PFA + + + + METHODS + title_2 + 13191 + Satellite cell isolation and culture + + + METHODS + paragraph + 13228 + Satellite cells were isolated from the hind leg muscles of approximately 20 to 25-week-old wildtype and miR-206/133b knock-out animals by using the skeletal muscle dissociation kit (Miltenyi Biotech, Bergisch Gladbach, Germany, 130-098-305) and enriched by the satellite cell isolation Kit, mouse (Miltenyi Biotech, 130-104-268) according to the manufacturer's instructions. Isolated cells were plated on gelatin-coated muclear 96-well plates (Sigma#M0562). Satellite cells were grown in proliferation medium (40% DMEM, 40% Ham F-10; 20% FCS; Pen/Strep; 2.5 ng/ml human FGF-2, Miltenyi Biotech#130-093-840) for 3 days, followed by switch to differentiation medium (DMEM, 2% horse serum, and Pen/Strep). Cells were incubated for 5 min in fixative (4% PFA/PBS, 0.1% sodium desoxycholate, 0.2% NP-40), washed 3 times in PBS, blocked in carrier (PBS, 5% BSA, 0.5% NP-40) for 1 h and then incubated with MF 20 supernatant in carrier (1:100, DSHB, Iowa City, Iowa). Secondary antibody was goat anti-mouse IgG1 Alexa 488 (1:2000, Life technologies). + + 27344 + Gene + 8315 + + Pen + + + 2247 + Gene + 1521 + + FGF-2 + + + 27344 + Gene + 8315 + + Pen + + + 16017 + Gene + + IgG1 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 9796 + Species + + horse + + + 9925 + Species + + goat + + + 10090 + Species + + mouse + + + MESH:D007854 + Chemical + + PBS + + + - + Chemical + + DMEM + + + MESH:D003840 + Chemical + + sodium desoxycholate + + + MESH:D007854 + Chemical + + PBS + + + MESH:D007854 + Chemical + + PBS + + + - + Chemical + + Alexa 488 + + + MESH:D017245 + Chemical + + PFA + + + - + Chemical + + DMEM + + + + METHODS + title_2 + 14277 + Quantitative proteomics + + + METHODS + paragraph + 14301 + Soleus muscle was dissected from adult 13C6Lys-labeled (SILAC) WT and from nonlabeled WT and miR-206/133b mutant littermates. Protein extracts were processed and mass spectrometry was performed as described previously. Briefly, 30 mug protein extracts of soleus muscle of labeled and the respective unlabeled muscle were combined, size-fractionated into 10 fractions using SDS-PAGE, and the proteins were digested in-gel using the endoproteinase LysC. After digestion peptides were eluted from gel-fractions using an acetonitrile gradient. Peptides were desalted prior to LC-MS/MS analysis and loaded onto a 15 cm in-house packed column (75 mum ID, C18 Beads 3 mum diameter, Dr. Maisch, Germany) and eluted by an linear increase of buffer B within a binary buffer system: A) 0.5% acetic acid and B) 80% ACN, 0.5% acetic acid. The flow-rate was set to 200 nL/min and gradient time to 150 min. The Agilent 1200 nanoflow HPLC system was coupled to an LQT-Orbitrap XL (Thermo Scientific) via an electrospray ionization source (Thermo Fisher Scientific). The mass spectrometer was operated in the data-dependent mode to automatically measure full MS scans and MS/MS spectra. In detail, survey scans were measured after accumulation of 1E6 ions at a resolution of 60,000 and subsequently isolation and fragmentation (CID, 35% norm. collision energy, target value: 5,000) of the top five intense peaks was performed in the linear ion trap. Peptides were identified by correlation of acquired MS/MS spectra against the mouse IPI (ftp://ftp.ebi.ac.uk/pub/databases/IPI) using MaxQuant software. A FDR cutoff of 1% was set on the protein and peptide level. Two mis-cleavages were allowed. Amidomethylation at cysteine residues was set as fixed modification, while oxidation on methione residues and acetylation at the N-term of proteins were defined as variable modifications. Quantification of proteins was performed by MaxQuant software. Only peptides with a minimal length of six amino acids and at least two ratio counts for SILAC pairs were used for quantification. Using SILAC as an internal standard, ratios miR-206/133b KO/13C6Lys-WT and 13C6Lys-WT/WT were obtained and were used for calculation of the direct KO/WT ratios for relative protein abundance. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mouse + + + - + Chemical + + 13C6Lys + + + Disease + + CID + + + MESH:D019342 + Chemical + + acetic acid + + + MESH:D003545 + Chemical + + cysteine + + + MESH:D019342 + Chemical + + acetic acid + + + MESH:C084683 + Chemical + + ACN + + + - + Chemical + + 13C6Lys + + + MESH:D012967 + Chemical + + SDS + + + MESH:C032159 + Chemical + + acetonitrile + + + - + Chemical + + methione + + + - + Chemical + + 13C6Lys + + + MESH:D004194 + Disease + + ionization + + + + METHODS + title_2 + 16560 + Western blot + + + METHODS + paragraph + 16573 + For western-blot analysis of embryonic tissue, the trunk of E10.5 embryos posterior of the otic placode was used. A total of 10 mug of protein extract were loaded onto a 9% SDS-PAGE and blotted onto nitrocellulose membranes. The following antibodies were used: anti-Pax7 (supernatant of DSHB#P3U1), anti-pan-actin (1:1000, CST, Frankfurt, Germany; Cat#4968), anti-GAPDH (1:1000, CST; Cat#2118), secondary antibodies were anti-mouse-HRP (1:1000, eBioscience, Affymetrix, Frankfurt Germany, Cat#18-8816-31) and anti-rabbit-HRP (1:5000, Pierce, Thermo Fisher Scientific, Darmstadt, Germany, Cat#1858415). + + 18509 + Gene + 55665 + + Pax7 + + + 12854 + Gene + 997 + + CST + + + 12854 + Gene + 997 + + CST + + + 10090 + Species + + mouse + + + 9986 + Species + + rabbit + + + MESH:D012967 + Chemical + + SDS + + + + METHODS + title_2 + 17176 + Statistics + + + METHODS + paragraph + 17187 + Data are reported as mean +- SEM. Differences between groups were tested for statistical significance using the unpaired Student's. For microarray experiments the median of groups was used and statistical analysis using an unpaired t-test was performed using DNAStar Arraystar 11 software (annotation na33). The Benjamini-Hochberg FDR correction was used to control the multiple hypothesis testing in case of the microarray experiments. + + + RESULTS + title_1 + 17630 + Results + + + RESULTS + title_2 + 17638 + Deletion of the miR-206/133b cluster does not alter skeletal muscle morphology + + 387202 + Gene + + miR-206 + + + + FIG + 13395_2014_23_Fig1_HTML.jpg + Fig1 + fig_caption + 17717 + Generation of miR-206/133b knock-out mice. The miRNAs miR-206, miR-1 and 133a/b are encoded in three clusters on mouse chromosome 1, 2 and 18. The primary sequence (A) of mature miR-1/133a encoded on chromosome 2 and 18 is identical, miR-206 differs in four bases from miR-1, and miR-133b differs in one base from miR-133a (red). The seed sequence, which is instrumental for target binding of miRNAs, is identical between the corresponding miRNAs (blue). Schematic representation of the miR-206/133b genomic region (B) with deleted parts indicated (red). Southern blot analysis (C) of genomic DNA isolated from wildtype (WT), heterozygous, and homozygous mutant animals proves deletion of the miR-206/133b locus. An SphI digest of genomic DNA and a probe hybridizing 5' prime of the targeting vector were used to monitor targeting of the miR-206/133b locus by homologous recombination. RNase protection assay (D) and northern blot (E) analysis demonstrates the loss of miR-206 in the soleus muscle of knock-out animals. Expression of miR-1 is not changed in soleus muscle of miR-206/133b knock-out mice. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + TABLE + table + 18824 + Deletion of the miR-206/133b cluster does not lead to abnormalities in the Mendelian distribution of mutant animals (a) at the time of weaning. Additional mdx mutation of the dystrophin gene does not lead to further lethality in double mutant animals (b). Distribution of male offspring after mating of male miR-206/133b+/- mdxy/+ or miR-206/133b+/- mdx+/+ to female miR-206/133b+/- mdx+/- is presented. + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + dystrophin + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + + TABLE + Tab1.xml + Tab1 + table_caption + 19236 + Normal Mendelian distribution of offspring at the time of weaning + + + TABLE + Tab1 + table + Tab1.xml + 19304 + Genotype N % observed Expected miR-206/133b+/+ mdxy/+ 29 13.0% 12.5% miR-206/133b+/+ mdxy/- 25 11.2% 12.5% miR-206/133b+/- mdxy/+ 57 25.6% 25.0% miR-206/133b+/- mdxy/- 41 18.4% 25.0% miR-206/133b-/- mdxy/+ (a) 35 15.7% 12.5% miR-206/133b-/- mdxy/- (b) 36 16.1% 12.5% Total 223 100.0% 100.0% + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + + FIG + 13395_2014_23_Fig2_HTML.jpg + Fig2 + fig_caption + 19630 + Deletion of miR-206/133b does not impair myogenesis during somite development. miR-206/133b knock-out animals are born with the expected Mendelian distribution (see text), whole mount in situ hybridization indicates normal myogenin expression in wildtype (WT; A, C) and knock-out embryos (KO; B, D) at embryonic day 10.5. Sections (C, D) are at the level of the forelimbs. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 17928 + Gene + 1854 + + myogenin + + + + RESULTS + paragraph + 20004 + To analyze the function of the miR-206/133b miRNA cluster we deleted the coding region of the miR-206/133b gene on mouse chromosome 1 by homologous recombination (Figure 1A, B). The targeting strategy also removed the last exon of the ncRNA linc-MD1 including the putative miR-133a/b binding site in linc-MD1, which is identical to the miR-133b* sequence. Neither the putative miR-135 binding sites in the second exon of linc-MD1, nor the described distal or proximal regulatory elements, identified by myogenin or myoD ChIP-seq approaches in C2C12 myocytes, were affected by the targeting strategy. Targeted deletion of the miR-206/133b coding region was validated by southern blot analysis (Figure 1C). Viable knock-out mice were obtained by breeding of heterozygous animals. Deletion of the miR-206/133b cluster did not impair embryonic or early postnatal survival of homozygous animals (Table 1; mdxY/+ mice). To exclude a function of miR-206/133b in early development that might be compensated during later stages we analyzed the spatial distribution of myogenin transcripts at the embryonic stage E10.5. No differences in somite morphology and myogenin expression were detected (Figure 2). Development of body weights and survival rate of miR-206/133b knock-out mice was indistinguishable from WT animals. RNase protection analysis as well as northern blot analysis using RNA isolated from hind limb muscle confirmed the absence of miR-206 in homozygous mutant animals (Figure 1D, E). Due to the presence of miR-133a in skeletal muscles, which differs by only one nucleotide from miR-133b, it was not possible to demonstrate the absence of mature miR-133b in homozygous mutant animals. + + 387202 + Gene + + miR-206 + + + 387202;723817 + Gene + + miR-206/133b + + + 105661730 + Gene + + linc-MD1 + + + 723817 + Gene + + miR-133b + + + 105661730 + Gene + + linc-MD1 + + + 17928 + Gene + 1854 + + myogenin + + + 17927 + Gene + 7857 + + myoD + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 17928 + Gene + 1854 + + myogenin + + + 17928 + Gene + 1854 + + myogenin + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 723817 + Gene + + miR-133b + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 105661730 + Gene + + linc-MD1 + + + CVCL:0188 + CellLine + + C2C12 + + + + FIG + 13395_2014_23_Fig3_HTML.jpg + Fig3 + fig_caption + 21701 + Expression of miR-206 is confined to skeletal muscle. Expression of miR-206 (A) is detected by quantitative reverse transcription PCR (qRT-PCR) in tissues isolated from wildtype (WT) (open bars), miR-206/133b knock-out and mdxy/- (gray bars) mice. Expression of miR-206 is detected in skeletal muscle of the WT mice. miR-206 is not detected in muscles isolated from knock-out mice, confirming the loss of miR-206 in the knock-out tissue and specificity of the Taqman assay. miR-206 was at the qRT-PCR detection limit in heart and brain of WT mice. In WT mice, miR-206 was strongly expressed in the soleus muscle, which has a high content of type 1/oxidative skeletal muscle fibers. A lower amount of miR-206 was detected in the diaphragm, as well as in other skeletal muscles. In mdx mice, the expression of miR-206 was not increased in soleus muscle. Increased miR-206 expression was detected in the diaphragm and other skeletal muscles. Expression of miR-1 (B) was also analyzed in the respective muscles and appeared unchanged in the soleus muscle of miR-206/133b knock-out mice (black bar). Mdx mutation did not cause significant changes in the expression of miR-1. Samples were isolated from >=3 different animals each, and qRT-PCR was performed in triplicate for each of the samples. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 100316783 + Gene + + miR-1 (B) + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + Mdx + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + + FIG + 13395_2014_23_Fig4_HTML.jpg + Fig4 + fig_caption + 22993 + No alterations in fiber type identity and fibers size in miR-206/133b knock-out mice. Sections of the soleus muscle isolated from adult wildtype (WT) (A) and mutant mice (B) were stained for markers of slow myofibers. No changes in fiber size of slow and fast muscle fibers were detected (C; n >=8 per group, 30 fibers per animals). The ratio of slow/fast muscle fibers was unchanged in the soleus muscle of knock-out compared to WT mice (D). The scale bar in A corresponds to 100 mum in A and B. + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + RESULTS + paragraph + 23493 + The miR-206/133b cluster is predominantly expressed in muscles with a high content of oxidative type I myofibers such as the soleus muscle and less abundant in muscles with only few oxidative fibers (Figure 3A). Analysis of published small RNA sequencing results reveals that miR-206 is much less abundant in skeletal muscle than the related miR-1 (GSM539875; 20124 miR-206 versus 745064 miR-1 tags per million miRNA tags). Small RNA sequencing of mouse soleus muscle revealed that even in the muscle with highest expression of miR-206 miR-1 is by far the most abundant miRNA (GSE63342; 30812 miR-206 versus 281944 miR-1 tags per million miRNA tags). The expression of miR-1 was not significantly altered after loss of miR-206 (Figure 3B, M. soleus samples). Histological examination of M. soleus did not reveal obvious morphological alterations. We detected no changes in fiber size, fiber type distribution (Figure 4) or localization of myofiber nuclei. Similar results were obtained for other skeletal muscles (data not shown). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mouse + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + RESULTS + title_2 + 24527 + miR-206/133b mutant skeletal muscles do not show significant molecular abnormalities + + 387202 + Gene + + miR-206 + + + MESH:C567116 + Disease + + molecular abnormalities + + + + FIG + 13395_2014_23_Fig5_HTML.jpg + Fig5 + fig_caption + 24612 + No transcriptional changes in the soleus muscle of miR-206/133b knock-out (KO) mice. The scatter blot of the median of the log2 signals of wildtype (WT) (n = 9) versus KO (n = 8) indicates only few genes with expression changes more than twofold (green lines). The probe set 1446563_at detects RNA originating from the genome between miR-206 and miR-133b most likely the pri-miRNA of these miRNAs. The weak signal for this probe set in WT samples is reduced to the detection limit in the KO samples. Only a few of the detected changes in gene expression are significant after statistical analysis using Student's t-test. None of these changes could be verified by quantitative reverse transcription PCR (qRT-PCR) or analysis of protein expression. After false discovery rate correction by the Benjamini-Hochberg algorithm, only one gene directly downstream of the manipulated genomic locus significantly changed expression. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 10090 + Species + + mice + + + + RESULTS + paragraph + 25547 + To investigate whether the loss of expression of miR-206/133b leads to changes of gene expression levels we performed Affymetrix GeneChip analysis using RNA isolated from M. soleus and mouse genome 430 2.0 arrays (n = 9 WT, n = 8 KO). Only few and subtle changes were detected by this type of analysis (Figure 5). The strongly and significantly downregulated probe set 1446563_at detects RNA transcribed from the miR-206/133b locus with the probes binding between the both miRNAs but not within the known linc-MD1 exons. This finding additionally confirms the successful knock-out strategy and suggests that this probe set detects the common miR-206/133b precursor RNA. The only gene that was significantly (2.5-fold) upregulated after false discovery rate correction was IL-17a, which is located downstream of the miR-206/133b locus, suggesting a cis-regulatory mechanism caused by manipulation of the neighboring miRNA locus. No other gene was significantly deregulated in soleus muscle. In particular, we did not observe up-regulation of predicted miR-206 or miR-133b target genes. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 105661730 + Gene + + linc-MD1 + + + 16171 + Gene + 1651 + + IL-17a + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mouse + + + + FIG + 13395_2014_23_Fig6_HTML.jpg + Fig6 + fig_caption + 26641 + miR-206/133b does not modulate abundance of PAX7 in E10.5 embryos. SILAC-based screening of changes in protein abundance based on two independent pairs of miR-206/133b knock-out / wildtype (WT) samples revealed few changes that were greater than 1.5 fold up or down in miR-206/133b knock-out versus WT M. soleus (A). Not one of the proteins expected to be potentially upregulated was a predicted target of miR-206 or miR-133b (microRNA.org). In accordance with transcriptome data, no change in protein abundance could be confirmed in western blot analysis. Loss of miR-206/133b does not change the abundance of PAX7 protein in the trunk of E10.5 embryos (B, C). Analysis of the 3' untranslated region (UTR) of Pax7 using miRanda indicates identical binding sites for miR-206 and miR-1. Differences in the primary sequence of miR-206 and miR-1 do not lead to substantial changes in the predicted miRNA-Pax7 UTR interaction (D). + + 387202;723817 + Gene + + miR-206/133b + + + 18509 + Gene + 55665 + + PAX7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + PAX7 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + RESULTS + paragraph + 27571 + Since microRNAs regulate expression on the transcript or protein level, we performed an unbiased SILAC based proteomics screen to detect potential miRNA targets that are regulated at the translational level. Proteins were isolated from M. soleus of WT and miR-206/133b mutant mice and combined with proteins obtained from soleus muscles of 13C6Lys-labled mice, which serve as a reference for relative quantification. In total, we identified 1,500 proteins and 805 were quantified in two independent experiments. SILAC ratios of WT/13C6-Lys-WT and in knock-out/13C6-Lys-WT were used to calculate the direct KO/WT ratios for each of the experiments. In accordance with our transcriptome analysis, only few changes in protein concentrations between knock-out and WT soleus muscle were observed (Figure 6A). Although we quantified 94 potential miR-206 targets that displayed a good score according to the miRanda based microRNA.org target prediction database (August 2010 release), none of the potential target protein was upregulated more than 1.5 fold in the knock-out tissue. Likewise, none of the 57 proteins found in our SILAC data set that were predicted as miR-133b targets according to the microRNA.org database were upregulated. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + - + Chemical + + 13C6 + + + MESH:D008239 + Chemical + + Lys + + + MESH:D008239 + Chemical + + Lys + + + - + Chemical + + 13C6 + + + - + Chemical + + 13C6Lys + + + + RESULTS + paragraph + 28806 + Although no alterations in skeletal muscle development of miR-206/133b mutants were apparent, we investigated the potential regulatory interaction between miR-206 and Pax7 in somites of E10.5 embryos. No changes in Pax7 protein expression were detected in mutant compared to WT embryos indicating that miR-206 is not required to suppress expression of Pax7 in somites (Figure 6B, C), probably due to the presence of miR-1, which shows a similar expression pattern as miR-206 during skeletal muscle development but is more widely expressed compared to miR-206 in adult muscles. In fact, the seed sequences of miR-1 and miR-206 are identical, which predicts a comparable repression of Pax7 by either miR-206 or miR-1 (Figure 6D) according to target algorithms like miRanda. Taken together, neither transcriptome nor proteome analysis revealed significant changes in gene expression between WT and KO mice, which is in line with the lack of a muscle specific phenotype in miR-206/133b knock-out mice. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + RESULTS + title_2 + 29806 + Loss of the miR-206/133 cluster does not affect skeletal muscle regeneration in Duchenne muscular dystrophy model mice + + 387202 + Gene + + miR-206 + + + MESH:D020388 + Disease + + Duchenne muscular dystrophy + + + + FIG + 13395_2014_23_Fig7_HTML.jpg + Fig7 + fig_caption + 29925 + No difference in body weight and survival of miR-206/133b and mdx mutant mice. (A) The body weight of male animals is not different at 6 weeks of age (n >=11/group); however, at later ages (B), the mdx mutation leads to an increase in body weight that is not further modified by mutation of the miRNA cluster (wildtype (WT): miR-206/133b+/+, knock-out (KO): miR-206/133b-/-; MDX: mdxY/-; n >=5/group). (C) Survival of miR-206/133b mutants is not impaired. The mdx mutation does not affect survival of miR-206/133b knock-out mice. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + MDX + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + FIG + 13395_2014_23_Fig8_HTML.jpg + Fig8 + fig_caption + 30463 + Loss of miR-206/133b does not change serum creatine kinase activity. Creatine kinase activity in the serum in miR-206/133b mice knock-out mice is not altered. The mdx mutation leads to strong increase of creatine kinase activity. No differences were observed in mdx versus miR-206/133b KO / MDX mice (n >=6/group). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + MDX + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + FIG + 13395_2014_23_Fig9_HTML.jpg + Fig9 + fig_caption + 30780 + miR-206/133b mutation does not modulate muscle loss or regeneration in Mdx mice. No histological changes are observed in diaphragm or M. quadriceps after loss of miR-206/133b. Mutation of the mouse dystrophin gene (MDX) leads to increased muscle mass, calcifications (von Kossa staining) and increased fibrosis (red signal in Sirius red staining). Additional loss of miR-206/133b does not lead to changes of the mdx phenotype. Sections obtained from 12 weeks old animals are shown, similar results are obtained using tissue from 6-week- and 9-month-old mice. The scale bar corresponds to 200 mum in the upper three rows and to 50 mum in the lower two rows. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + dystrophin + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + MDX + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + Mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 10090 + Species + + mice + + + MESH:D005355 + Disease + + fibrosis + + + MESH:D009133 + Disease + + muscle loss + + + MESH:D014842 + Disease + + von + + + + RESULTS + paragraph + 31441 + Previously, miR-206 has been implicated in muscle fiber stability and regeneration of skeletal muscles, although the miR-206 seems dispensable for specification, proliferation and differentiation of skeletal muscle. To analyze the potential function of miR-206/133b miRNAs and their targets in maintenance and regeneration of muscle fibers we generated miR-206/133b-/-//mdxY/- compound mutant mice. Mdx mice carry a point mutation in the dystrophin gene on the X-chromosome leading to constant damage and loss of muscle fibers, which is compensated by increased regeneration. miR-206/133b-/-/mdx mutant mice were born at the expected Mendelian ratio (Table 1) and developed normally, which is in contrast to the previously described miR-206-/-/mdx mutant mice. Body weight development (Figure 7A, B) and survival (Figure 7C) of compound mutant animals was indistinguishable from mdx mice. Regeneration of skeletal muscles in mdx mice leads to increased expression of the miR-206/133b cluster in muscles with a low content of type I muscle fibers that express only low levels of miR-206/133b under normal conditions (Figure 3A,). In contrast we observed a moderate, not significant reduction of miR-1 in some mdx muscles (Figure 3B), possibly related to the emergence of other non-muscle cell types in the dystrophic muscles. Hence, it seems possible that such muscle might be affected by inactivation of miR-206/133b in conditions of constant regeneration. Loss of dystrophin expression results in the release of muscle creatine kinase (MCK) from muscle fibers into the blood, which serves as an indicator of muscle injury or muscle degenerative processes. To access the degree of muscle damage in miR-206/133b-/-/mdx mutant mice we determined MCK activity in serum of adult mice. Only very low MCK activity was detected in WT and miR-206/133b mice, which did not differ significantly between WT (297 +- 85 U/l, n = 6) and mutants (152 +- 62 U/l, n = 11). Importantly, we observed much higher MCK activities in mdx mice (8590 +- 1507 U/l, n = 6), which were not increased any further in miR-206/133b/mdx mutants (8024 +- 2464 U/l, n = 6; Figure 8) indicating that compound mutants do not suffer from increased muscle fiber damage compared to mdx mice. Next, we performed a comprehensive histological analysis of different muscles of the hind limb, as well as of the diaphragm from WT, miR-206/133b-/-, mdxY/- and miR-206/133b-/-//mdxY/- mice. Trichrome- and Sirius Red-, and von Kossa staining were used to determine myofiber organization, fibrosis, and intramuscular calcifications often present in mdx mice (Figure 9). No differences were found between WT and miR-206/133b skeletal muscle based on Trichrome- (not shown), von Kossa or Sirius red staining including absence of fibrosis and calcifications. Mutation of dystrophin caused the typical signs of muscle dystrophy, namely massive thickening of the diaphragm, enhanced fibrosis and patches of calcified tissue within the diaphragm and the M. quadriceps (Figure 9). No calcified fibers were observed in the soleus muscle. Deletion of miR-206/133b in mdx mice did not cause a more severe phenotype in the soleus, quadriceps or diaphragm muscles compared to single mutant mdx mice. Together these data indicate that loss of miR-206/133b cluster does not increase muscle damage in mdx mice or compromise skeletal muscle regeneration in constantly renewing muscle tissue. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + dystrophin + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + dystrophin + + + 12715 + Gene + 20432 + + muscle creatine kinase + + + 12715 + Gene + 20432 + + MCK + + + 12715 + Gene + 20432 + + MCK + + + 12715 + Gene + 20432 + + MCK + + + 387202 + Gene + + miR-206 + + + 12715 + Gene + 20432 + + MCK + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + dystrophin + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + mdx + + + 13405 + Gene + 20856 + + Mdx + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + MESH:D009136 + Disease + + dystrophic muscles + + + MESH:D009136 + Disease + + muscle dystrophy + + + MESH:D005355 + Disease + + fibrosis + + + MESH:D014842 + Disease + + von + + + MESH:D009135 + Disease + + muscle injury + + + - + Chemical + + Sirius red + + + MESH:D005355 + Disease + + fibrosis + + + MESH:D005355 + Disease + + fibrosis + + + MESH:D009135 + Disease + + muscle damage + + + MESH:D009135 + Disease + + muscle damage + + + MESH:D014842 + Disease + + von Kossa + + + + FIG + 13395_2014_23_Fig10_HTML.jpg + Fig10 + fig_caption + 34932 + No differences in differentiation of muscle progenitor cells in vitro . Proliferating muscle stem cells isolated from hind limb muscles of wildtype (WT) and miR-206/133b knock-out mice (KO) were induced to differentiate and stained after 3 or 5 days in differentiation medium (DM) for expression of myosin (green: MF20 staining, blue: nuclear staining; the scale bar corresponds to 50 mum). No differences in cell morphology or differences in the expression of myosin were detected between WT and mutant cells (A). Formation of fused myotubes from differentiating myoblasts was not impaired by the loss of the miR-206/133b cluster (B). Pax7 expression in differentiating myogenic progenitor cells was detected by western blot. No differences in Pax7 expression were detected after loss of miR-206/133b (C). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mice + + + MESH:D009223 + Disease + + DM + + + + RESULTS + paragraph + 35742 + To further analyze a possible function of the miR-206/133b cluster in the differentiation of muscle stem cells, we isolated muscle satellite cells from WT and miR-206/133b mutant mice. Loss of the miR-206/133b cluster did not impair the ability of muscle stem cells to differentiate to myotubes in vitro (Figure 10A). We did not observe reduced differentiation as accessed by the number of MF20 positive cells nor changes in the fusion of myocytes as accessed by the number of myotubes with more than one nucleus (Figure 10B). Moreover, we found that the lack of miR-206/133b did not increase the concentration of the potential miR-206 and miR-1 target Pax7 in differentiating satellite cells on the 5th day after induction of differentiation (Figure 10C). + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + + DISCUSS + title_1 + 36502 + Discussion + + + DISCUSS + paragraph + 36513 + Physiological properties of skeletal muscle cells including specification, differentiation, function and regeneration are controlled by an extensive network of regulatory molecules. Many key genes have been identified that are essential for specification and differentiation of muscle precursor cells during development. Commitment of muscle cells is controlled by a core program of myogenic regulatory factors of the MyoD family, which also control transcription of the miR-1/206/133 miRNA clusters resulting in a prominent expression of these miRNAs in muscle cells. Yet, important differences in the expression pattern of miR-1/206/133 miRNA clusters exist: the miR-1/133a clusters are expressed in heart and all skeletal muscles, while expression of miR-206/133b becomes restricted to a subset of oxidative muscle fibers by hitherto unknown molecular mechanisms. Since the two miR-1/133a clusters display virtually identical expression patterns, partially overlapping or redundant functions of these miRNAs seem likely. In contrast, the miR-206/133b cluster shows a more restricted expression, which might argue for a distinct function of this cluster albeit muscle cells expressing miR-206/133b also transcribe miR-1/133a. In addition, the primary sequence of miR-206 differs from miR-1 while miR-133b is very similar to miR-133a. However, the seed sequences of miR-206 and miR-1 important for target recognition are identical, indicating only limited differences in the target specificity of miR-206 and miR-1. The miR-206/133b locus also directs expression of the long non-coding RNA linc-MD1, which was postulated to act as a competing endogenous RNA or miRNA decoy adding a further potential function to the miR-206/133b locus. + + 17927 + Gene + 7857 + + MyoD + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 105661730 + Gene + + linc-MD1 + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + DISCUSS + paragraph + 38250 + We have generated a miR-206/133b knock-out allele by deletion of the genomic region spanning from miR-206 to miR-133b and resulting in removal of the third exon of the linc-MD1 that might act as a miR-133 sponge. Surprisingly, homozygous deletion of the miR-206/133b locus did not cause an apparent phenotype in adult mice. No changes in the morphology and distribution of muscle fiber types were apparent and the transcriptional and proteomics signatures of miR-206/133b mutant muscles were virtually unchanged, ruling out that miR-206 and/or 133b alone play an essential role in muscle physiology. These observations are consistent with to observations made in miR-206 knock-out mice, which also show no obvious phenotype under baseline conditions. Apparently, the concomitant loss of miR-206, miR-133b and the potential miR-133 sponge in linc-MD1 is compatible with normal muscle development and functions, which might be explained by expression of the miR-1/133a clusters in type I myofibers compensating for deletion of miR-206/133b. Theoretically, the inactivation of miR-133b might also be counteracted by the loss of the miR-133 sponge function, although the physiological relevance of endogenous competing RNAs has been questioned making this explanation less likely. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 105661730 + Gene + + linc-MD1 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 105661730 + Gene + + linc-MD1 + + + 387202 + Gene + + miR-206 + + + 723817 + Gene + + miR-133b + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + DISCUSS + paragraph + 39527 + We found a more widespread and abundant expression of the miR-206/133b cluster in regenerating muscles, which confirms previous findings. It seems possible that the expression of miR-206 in regenerating muscle reflects a specific function of the miR-206/133b in the regenerative response. However, it is important to point out that miR-206/133b is broadly expressed during development in immature muscle cells. Hence, it is difficult to exclude that the increased expression of miR-206/133b during skeletal muscle regeneration simply mirrors the immature state of newly formed myofibers that have not yet terminated expression of the miR-206/133b cluster in the majority of mature type II fibers. The lack of a more severe phenotype in miR-206/133b-/-//mdxY/- compound mutants compared to mdxY/- mice clearly argues against an important role of the miR-206/133b cluster for skeletal muscle regeneration. Moreover, we did not see an increase in lethality or an increase in serum creatine kinase levels in compound mutant mice, which also excludes an important role of the miR-206/133b cluster in restricting myofiber damage. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + DISCUSS + paragraph + 40655 + miR-206 has been claimed to suppress Pax7 expression thereby regulating muscle stem cell renewal and differentiation. The normal expression of Pax7 in isolated miR-206/133b mutant satellite cells and the normal differentiation of mutant satellite cells to myotubes in vitro do not argue for an important role of miR-206 in Pax7 expression. These results are in contrast to previous publications mostly relying on the inhibition of miR-206 by anti-miRs, overexpression of miR-206, or by correlation of expression profiles. It seems possible that previous anti-miR approaches were biased by off-target effects or affected expression of miR-1, which can also suppress Pax7. Generation of skeletal muscle specific miR-1/miR133a//miR-206/133b triple mutants will probably solve this controversy in the future. We do not have a convincing explanation for the attenuation of Pax7 down-regulation in differentiating miR-206-deficient satellite cells reported by Liu et al.. The same authors also described that miR-206/mdx mutant mice suffer from increased muscle degeneration, impaired regeneration and premature death, which was all not seen in our miR-206/133b mutants. In principle, it is feasible that the concomitant loss of mir-206 and miR-133b in our model accounts for these differences, which would strengthen the notion that the miR-206/133b cluster acts as a functional unit targeting similar biological processes sometimes in parallel and sometimes in opposing directions. Alternatively, subtle differences in the genetic background and/or strain-depended differences in the expression of miR-1/133a compensating for the loss of miR-206/133b might differentially affect the course of muscular dystrophy in miR-206/133b-/-//mdxy/- and miR-206-/-//mdxy/- compound mutants. + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 18509 + Gene + 55665 + + Pax7 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + mir-206 + + + 723817 + Gene + + miR-133b + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 735281 + Gene + + miR + + + 13405 + Gene + 20856 + + mdx + + + 10090 + Species + + mice + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + MESH:D009422 + Disease + + impaired regeneration and premature death + + + MESH:D009135 + Disease + + muscle degeneration + + + MESH:D009136 + Disease + + muscular dystrophy + + + + CONCL + title_1 + 42439 + Conclusion + + + CONCL + paragraph + 42450 + Taken together, our results question a major role of the miR-206/133b cluster in development, function and regeneration of skeletal muscle in mouse and argue for overlapping functions of the miR-206/133b and miR-1/133a gene clusters during myogenesis. Our findings are in line with the hypothesis that individual miRNAs often fine-tune biological processes and play modulatory roles. Further genetic studies targeting the miR-1/133a together with the miR-206/133b gene clusters specifically in skeletal muscle will probably reveal the prevalent physiological function of miR-1/206/133 in mammals. + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 387202 + Gene + + miR-206 + + + 10090 + Species + + mouse + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + 735281 + Gene + + miR + + + + ABBR + title + 43047 + Abbreviations + + + ABBR + paragraph + 43061 + BSA + + + ABBR + paragraph + 43065 + bovine serum albumin + + 9913 + Species + + bovine + + + + ABBR + paragraph + 43086 + ChIP + + + ABBR + paragraph + 43091 + chromatin immunoprecipitation + + + ABBR + paragraph + 43121 + DM + + MESH:D009223 + Disease + + DM + + + + ABBR + paragraph + 43124 + differentiation medium + + + ABBR + paragraph + 43147 + DSHB + + - + Chemical + + DSHB + + + + ABBR + paragraph + 43152 + developmental studies hybridoma bank + + + ABBR + paragraph + 43189 + DTA + + MESH:C042899 + Chemical + + DTA + + + + ABBR + paragraph + 43193 + diphtheria toxin A + + + ABBR + paragraph + 43212 + ES cell + + + ABBR + paragraph + 43220 + embryonic stem cell + + + ABBR + paragraph + 43240 + FCS + + + ABBR + paragraph + 43244 + fetal calf serum + + 9913 + Species + + calf + + + + ABBR + paragraph + 43261 + FDR + + + ABBR + paragraph + 43265 + false discovery rate + + + ABBR + paragraph + 43286 + HRP + + + ABBR + paragraph + 43290 + horseradish peroxidase + + 3704 + Species + + horseradish + + + + ABBR + paragraph + 43313 + IRES + + + ABBR + paragraph + 43318 + internal ribosome entry site + + + ABBR + paragraph + 43347 + KO + + + ABBR + paragraph + 43350 + knock-out + + + ABBR + paragraph + 43360 + lacZ + + + ABBR + paragraph + 43365 + beta-galactosidase coding + + 12091 + Gene + 47922 + + beta-galactosidase + + + + ABBR + paragraph + 43391 + MCK + + + ABBR + paragraph + 43395 + creatine kinase, muscle + + + ABBR + paragraph + 43419 + MDX + + + ABBR + paragraph + 43423 + mouse model of Duchenne Muscular Dystrophy, X-linked + + 10090 + Species + + mouse + + + MESH:D020388 + Disease + + Duchenne Muscular Dystrophy + + + + ABBR + paragraph + 43476 + miR + + + ABBR + paragraph + 43480 + miRNA + + + ABBR + paragraph + 43486 + PBS + + MESH:D007854 + Chemical + + PBS + + + + ABBR + paragraph + 43490 + phosphate buffered saline + + - + Chemical + + phosphate buffered saline + + + + ABBR + paragraph + 43516 + PFA + + MESH:D017245 + Chemical + + PFA + + + + ABBR + paragraph + 43520 + paraformaldehyde + + MESH:C003043 + Chemical + + paraformaldehyde + + + + ABBR + paragraph + 43537 + qRT-PCR + + + ABBR + paragraph + 43545 + quantitative reverse transcription polymerase chain reaction + + + ABBR + paragraph + 43606 + SDS-PAGE + + MESH:D012967 + Chemical + + SDS + + + + ABBR + paragraph + 43615 + sodium dodecyl sulfate polyacrylamide gel electrophoresis + + MESH:C016679 + Chemical + + polyacrylamide + + + MESH:D012967 + Chemical + + sodium dodecyl sulfate + + + + ABBR + paragraph + 43673 + SEM + + + ABBR + paragraph + 43677 + standard error of the mean + + + ABBR + paragraph + 43704 + SILAC + + + ABBR + paragraph + 43710 + stable isotope labeling by amino acids in cell culture + + + ABBR + paragraph + 43765 + UTR + + + ABBR + paragraph + 43769 + untranslated region + + + ABBR + paragraph + 43789 + WT + + + ABBR + paragraph + 43792 + wildtype + + + ABBR + footnote + 43801 + Competing interests + + + ABBR + footnote + 43821 + The authors declare that they have no competing interests. + + + ABBR + footnote + 43880 + Authors' contributions + + + ABBR + footnote + 43905 + TBo, SW, and HN conducted the experiments. TBo and TBr designed and interpreted the experiments and wrote the manuscript. 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+ surname:Li;given-names:Hegang + 25511509 + surname:Liu;given-names:Nan + 2014 + 4272822 + 144 + 10.1186/s12863-014-0144-1 + Keywords + front + 144 + 15 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + TITLE + BMC Genet.; 2014 Dec 16; 15144. doi:10.1186/s12863-014-0144-1 + Microarray Proteomic technology Wool growth Differential expression + surname:Zhao;given-names:Jinshan + surname:He;given-names:Guangling + surname:Liu;given-names:Jifeng + surname:De;given-names:Wei + surname:Cheng;given-names:Ming + surname:Bu;given-names:Ran + Liu N, Li H, Liu K, Yu J, Bu R, Cheng M, De W, Liu J, He G, Zhao J + 0 + Identification of skin-expressed genes possibly associated with wool growth regulation of Aohan fine wool sheep + + + ABSTRACT + abstract_title_1 + 112 + Background + + + ABSTRACT + abstract + 123 + Sheep are valuable resources for the animal fibre industry. Therefore, identifying genes which regulate wool growth would offer strategies for improving the quality of fine wool. In this study, we employed Agilent sheep gene expression microarray and proteomic technology to compare the gene expression patterns of the body side (hair-rich) and groin (hairless) skins of Aohan fine wool sheep (a Chinese indigenous breed). + + 9940 + Species + + Sheep + + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + + ABSTRACT + abstract_title_1 + 546 + Results + + + ABSTRACT + abstract + 554 + Comparing the body side to the groin skins (S/G) of Aohan fine wool sheep, the microarray study revealed that 1494 probes were differentially expressed, including 602 more highly expressed and 892 less highly expressed probes. The microarray results were verified by means of quantitative PCR. Cluster analysis could distinguish the body side skin and the groin skin. Based on the Database for Annotation, Visualization and Integrated Discovery (DAVID), 38 of the differentially expressed genes were classified into four categories, namely regulation of receptor binding, multicellular organismal process, protein binding and macromolecular complex. Proteomic study revealed that 187 protein spots showed significant (p < 0.05) differences in their respective expression levels. Among them, 46 protein entries were further identified by MALDI-TOF/MS analyses. + + 9940 + Species + + sheep + + + + ABSTRACT + abstract_title_1 + 1418 + Conclusions + + + ABSTRACT + abstract + 1430 + Microarray analysis revealed thousands of differentially expressed genes, many of which were possibly associated with wool growth. Several potential gene families might participate in hair growth regulation. Proteomic analysis also indentified hundreds of differentially expressed proteins. + + + ABSTRACT + abstract_title_1 + 1721 + Electronic supplementary material + + + ABSTRACT + abstract + 1755 + The online version of this article (doi:10.1186/s12863-014-0144-1) contains supplementary material, which is available to authorized users. + + + INTRO + title_1 + 1895 + Background + + + INTRO + paragraph + 1906 + Sheep are valuable resources for the animal fibre industry. Identifying genes which regulate wool growth offers the opportunity to improve the wool production efficiency, product quality and diversity in breeding programs. It can also offer the opportunity to develop transgenic lines and to develop therapeutic agents that can be used to tailor for desirable fibre attributes by altering gene expression. The genetic polymorphisms and their mechanisms of wool and cashmere growth and regulation have been thoroughly studied. In mammals, several gene families, such as WNTs, tumor necrosis factors (TNFs), fibroblast growth factors (FGFs) and transforming growth factor(TGFs), have been implicated in hair follicle initiation, morphogenesis and cycling. Recently, the molecular characteristics of primary wool follicle initiation in Merino sheep have been reported recently. + + 9940 + Species + + Sheep + + + 9940 + Species + + sheep + + + MESH:D009336 + Disease + + tumor necrosis + + + + INTRO + paragraph + 2781 + Transcriptomic research such as microarray analysis, has been successfully applied to investigate the characteristics of hair follicle stem cells in mice. Microarray studies have been reported for different traits in sheep and goats, such as resistance to parasites, mammary development and milk quality, wool follicle development, natural fleece rot resistance and pigmentation traits of skin and wool. A subset of skin-expressed microRNAs with possible roles in goat and sheep hair growth has also been reported. Several studies have demonstrated the usefulness of cDNA microarray for expression profiling of wool follicle growth cycles in whole skin. More recently the RNA-seq method was also used to determine the genes differentially expressed among various tissues (including whole skin) of sheep. + + 10090 + Species + + mice + + + 9940 + Species + + sheep + + + 9925 + Species + + goats + + + 9925 + Species + + goat + + + 9940 + Species + + sheep + + + MESH:D012757 + Disease + + sheep hair growth + + + + INTRO + paragraph + 3585 + The Aohan fine wool sheep, bred in Inner Mongolia, is an outstanding breed, providing both wool and meat. The major characteristics of this breed are high quality wool,high disease resistance, and high adaptability. According to figures, Aohan fine wool sheep can provide up to 9 kg of quality wool per year (fiber length up to 10.5 cm, fibre diameter less than 22 mum). Therefore, Aohan fine wool sheep are considered as a valuable genetic resource for fine wool production. Wang et al. demonstrated that seasonal factors significantly influenced the wool growth of Aohan fine wool sheep. The peak of the growth rate occurs in summer and the low in winter. The expression profiling of immune genes and type I inner root sheath (IRS) keratin genes in the whole skin of Aohan fine wool sheep has previously been reported by our laboratory team. + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + 1214577 + Species + + to 9 + + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + + INTRO + paragraph + 4432 + However, to our knowledge, as of yet no microarray or proteomic study at a genome-wide level has been reported on protein-coding genes which are possibly responsible for regulating hair growth of adult sheep so far. The aims of the present work are to investigate and compare the gene expression level of the body side skin and groin skin using microarray and proteomic technology, and to identify the possible genes and proteins responsible for the wool growth regulation of Aohan fine wool sheep. + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + + RESULTS + title_1 + 4931 + Results + + + RESULTS + title_2 + 4939 + Summary of microarray analysis + + + RESULTS + paragraph + 4970 + A total of 1494 probes were differentially expressed comparing the body side to groin skins (S/G) in Aohan fine wool sheep, including 602 up-regulated and 892 down-regulated probes, as shown in Additional file 1: Table S1. Most probes (1110) were not assigned to unique transcripts, due to the lack of information. The number of distinct genes/transcripts (annotated) was 331, of which 112 were up-regulated and 219 were down-regulated. In S/G, 7 genes (CYP1A1, LOC100137068, LOC443300, LOC101106865, Connexin 43, SCD and LOC101122398) were down-regulated by more than 10-fold. + + 100170113 + Gene + + CYP1A1 + + + 443455 + Gene + + Connexin 43 + + + 443185 + Gene + + SCD + + + 101122398 + Gene + + LOC101122398 + + + 9940 + Species + + sheep + + + + RESULTS + paragraph + 5548 + Furthermore, many gene families which regulate different aspects of hair follicle growth showed differential expression in S/G (see Additional file 2: Table S3), such as growth factors, immune cytokines, Keratins (KRT) and Keratin-associated proteins (KAP), and so on. + + + RESULTS + title_2 + 5817 + Selective verification for microarray data by QPCR + + + FIG + 12863_2014_144_Fig1_HTML.jpg + Fig1 + fig_caption + 5868 + Q-PCR validation of the microarray data. P values (T-test) of the Q-PCR data are 0.036 (FGF10), 0.019 (LOC443300), 0.028 (FGF18), 0.018 (Connexin 43), 0.044 (SCD) and 0.019 (ZO1), respectively. S/G represent body side skin group/groin skin group, respectively. + + 443074 + Gene + + FGF10 + + + 780460 + Gene + + FGF18 + + + 443455 + Gene + + Connexin 43 + + + 443185 + Gene + + SCD + + + 443200 + Gene + + ZO1 + + + + RESULTS + paragraph + 6130 + In order to verify the microarray results, we selected 10 genes, namely FGF10, LOC443300, FGF18, Connexin43, SCD, ZO1, MMP2, ITGB1, PAG11 and CRYAB, to comparatively analyze their expression patterns by qPCR. As shown in Figure 1, the qPCR results for the selected seven genes were consistent with the microarray results, except for FGF10, ZO1 and CRYAB, thus reflecting the reliability of our microarray data. + + 443074 + Gene + + FGF10 + + + 780460 + Gene + + FGF18 + + + 443455 + Gene + + Connexin43 + + + 443185 + Gene + + SCD + + + 443200 + Gene + + ZO1 + + + 443115 + Gene + + MMP2 + + + 443141 + Gene + + ITGB1 + + + 100034671 + Gene + + PAG11 + + + 497274 + Gene + + CRYAB + + + 443074 + Gene + + FGF10 + + + 443200 + Gene + + ZO1 + + + 497274 + Gene + + CRYAB + + + + RESULTS + title_2 + 6542 + Hierarchical cluster analysis, biological process gene ontology (GO) analyses and putative gene networks + + + FIG + 12863_2014_144_Fig2_HTML.jpg + Fig2 + fig_caption + 6647 + Hierarchical cluster analysis of data between different skin areas of the Aohan fine wool sheep in anagen phase. Each column represents one sheep, and each horizontal line refers to a gene. Color legend is on the top of the figure. Red indicates genes with a greater expression relative to the geometrical means, green indicates genes with a lower expression relative to the geometrical means. S1, S2 and S3 represent 3 repeats of body side skin group, and G1, G2, G3 represent 3 repeats of groin skin group. + + 9940 + Species + + sheep + + + 9940 + Species + + sheep + + + + RESULTS + paragraph + 7157 + To further investigate the similarity in the expression patterns of protein-coding genes between the two skin areas, we performed cluster analysis using the Cluster 3.0 tool. As shown in Figure 2, cluster analysis could make a distinction between the body side skin and the groin skin. + + + RESULTS + paragraph + 7444 + Based on the Database for Annotation, Visualization and Integrated Discovery (DAVID), 38 of the differentially expressed genes were classified into four categories, many of which shared the same genes, according to their functional correlation (Additional file 3: Table S2). The majority of the genes possibly related to the wool growth control could be assigned into four categories including regulation of receptor binding, multicellular organismal process, protein binding and macromolecular complex. + + + FIG + 12863_2014_144_Fig3_HTML.jpg + Fig3 + fig_caption + 7948 + Biological pathways having more differentially expressed genes. A: PI3K-AKT Pathway; B: JAK-STAT Pathway. Gene name in red in the gene box indicates higher gene expression in S/G, green indicates lower gene expression, and black indicates no change of the gene expression. + + + RESULTS + paragraph + 8222 + Two singnaling pathways, PI3K-AKT pathway and JAK-STAT pathway, were identified as biological pathways having more differentially expressed genes. Figure 3 displayed the putative interactions related to the differentially expressed genes of the two pathways. These interactions were intensively involved in cell cycle and apoptosis processes. + + + FIG + 12863_2014_144_Fig4_HTML.jpg + Fig4 + fig_caption + 8566 + Selected factors with possible hair growth regulatory roles . In the hair growth cycle, some factors play roles in anagen promotion or maintenance (left), some factors play roles in catagen-telogen promotion or maintenance (middle), and others have unkown funtions (right). The differentially expressed genes in this study was highlighted in colour (green or red). Genes in green represent that their regulation direction were in agreement with their putative function, while genes in red represent the opposite situation. + + + RESULTS + paragraph + 9092 + We speculate that wool growth regulation shared similar gene networks with hair. So particular differentially expressed genes, which also appeared in the conclusion of factors with known hair growth regulatory roles, were included in the putative gene lists of wool growth regulation (Figure 4). In the networks, 3 genes (FGF7, IGFBP3 and PRL) were contained in the category of anagen promotion and maintenance, 3 genes (IL1A, IL6 and TAC1) were contained in the category of catagen-telogen promotion and maintenance, while 2 genes (CDKN1B and FGF18) were contained in the category of function unknown. + + 443095 + Gene + + FGF7 + + + 100286737 + Gene + + IGFBP3 + + + 443317 + Gene + + PRL + + + 443404 + Gene + + IL1A + + + 443406 + Gene + + IL6 + + + 100034669 + Gene + + TAC1 + + + 100127219 + Gene + + CDKN1B + + + 780460 + Gene + + FGF18 + + + + RESULTS + title_2 + 9696 + Quantitative comparison and identification of protein spots on 2-DE gels + + + FIG + 12863_2014_144_Fig5_HTML.jpg + Fig5 + fig_caption + 9769 + Representative image of 2-DE silver stained polyacrylamide gel. + + MESH:C016679 + Chemical + + polyacrylamide + + + + RESULTS + paragraph + 9835 + To detect differential protein expression between the two skin areas, we constructed triplicate 2D maps of protein samples for each group were created. Figure 5 shows two representative 2-DE gel images of the protein expression patterns of the two groups. One hundred and eighty-seven protein spots showed significant (p < 0.05, t-test) differences in expression levels between the body side skin group and the control. Of the 187, 85 protein spots were more highly expressed and 102 were less highly expressed in body side skin as compared to the groin skin. Some of them could not be identified because of incomplete polypeptide fragments or low abundance (beyond the identification limit). In total, 46 protein entries were identified by MALDI-TOF/MS analyses. An overview of these proteins is presented in Additional file 4: Table S4. + + + DISCUSS + title_1 + 10679 + Discussion + + + DISCUSS + title_2 + 10690 + Methodologies + + + DISCUSS + paragraph + 10704 + This study is a pilot to establish microarray methodologies in the wool growth field. Our results contrasted the gene expressed in the skin cells from hair-rich and hairless regions at transcriptional and translational levels. The number and fold-change of DE genes in December were much lower than that in August (5605 probes were differentially expressed in August; data not shown). That was compatible with reduced follicle activities in winter. And most of DE genes in December were also differentially expressed in August (data not shown). + + + DISCUSS + paragraph + 11249 + The advantage of this study is that samples could be obtained easily while the weakness of this study is the low specificity in the samples because the skin samples compose not only wool follicle, but also other parts of the skin. + + + DISCUSS + title_2 + 11480 + The limitations encountered during the study and how we determine the results + + + DISCUSS + paragraph + 11558 + The small sample size is another weakness of this study. These three animals used in the study were half sibs (sharing the same father). The number of our resource population was not very large, so it was difficult to select more than three even-aged half sibs. It is unusual using only three animals, but this shortcoming could be partly made up for through using genetically related animals. Additionally, to confirm the reliability of the results from the microarray experiments, we conduct the qPCR validation for the expression levels of selected 10 genes, whose fold-changes between the two skin areas ranged from 2.15 to 19.42. Seven of these 10 genes displayed the same regulation directions compared to the microarray results, indicating our differential expression data had relatively reliable value in the field. + + + DISCUSS + paragraph + 12382 + The technical approach (microarrays) is another limitation of the study. First, the number of the probes (15208) designed by Agilent is limited compared to the RNA-seq method. Most of the probes were inadequately annotated. The full utility of the microarray data depends on the complete annotation of the whole sheep genome. Second, the specificity of the probes is relatively lower than RNA-sequencing, although using 60-mers long probes in our study. False positive and false negative phenomena exsit unavoidably in the microarray data. + + 9940 + Species + + sheep + + + 1335626 + Species + + mers + + + + DISCUSS + title_2 + 12922 + Functional implication of differentially expressed genes in the microarray study + + + DISCUSS + paragraph + 13003 + In the present study, we investigated the molecular events possibly related to sheep hair growth control using microarray and proteomic technologies. Transcriptomic analysis identified hundreds of differentially expressed genes displaying over 2-fold difference between the two sampled skin regions of Aohan fine wool sheep at December time point, and the number of less highly expressed transcripts (892) in body side skin was greater than that of the more highly expressed ones (602). The qPCR results validated the reliability of our microarray data. + + 9940 + Species + + sheep + + + MESH:D012757 + Disease + + sheep hair + + + + DISCUSS + paragraph + 13557 + IL-1A and IL-1B inhibit hair growth in vitro as reported previously, but IL-1A has shown downregulation in body side skin in our study. One possibility is in vivo IL-1A controls wool growth in a time dependent or a negative feedback manner. + + 443404 + Gene + + IL-1A + + + 443539 + Gene + + IL-1B + + + 443404 + Gene + + IL-1A + + + 443404 + Gene + + IL-1A + + + + DISCUSS + paragraph + 13798 + The skin of skin-specific SCD1 knock-out (SKO) mice exhibited variable orthokeratotic hyperkeratosis and parakeratotic hyperkeratosis, occasional mal-aligned hair follicles, and instances of protrusion of hair shafts out of the hair follicle and into the surrounding connective tissue, with an infiltration of inflammatory cells surrounding the exposed hair shaft and keratin material. In our study, SCD was less highly expressed at 11.91 fold change in body side skin. This maybe indicate that the SCD could play an important role in groin skin. + + 20249 + Gene + 74538 + + SCD1 + + + 443185 + Gene + + SCD + + + 443185 + Gene + + SCD + + + 10090 + Species + + mice + + + MESH:D017488 + Disease + + orthokeratotic hyperkeratosis and parakeratotic hyperkeratosis + + + + DISCUSS + paragraph + 14345 + MMP2 activity is associated with the disappearance of collagen VII during the invasion of epithelial cords of hair follicles and sweat glands in human skin. MMP2 plays a role in hair growth-associated extracellular matrix remodeling and cell migration, and may be a downstream effector through which thymosin ss4 exerts its effect on hair growth. However, MMP2 showed significant lower expression in body side skin. This may suggest sheep has a different hair growth regulatory mechanism than human. + + 4313 + Gene + 3329 + + MMP2 + + + 443115 + Gene + + MMP2 + + + 443115 + Gene + + MMP2 + + + 9606 + Species + + human + + + 9940 + Species + + sheep + + + 9606 + Species + + human + + + + DISCUSS + paragraph + 14845 + There is a log linear relationship between the relative level of beta 1 integrins (ITGB1) on the cell surface and proliferative capacity in keratinocytes. ITGB1-mediated signalling is also important in human hair growth control. Skin and hair follicle integrity is crucially dependent on ITGB1 expression in keratinocytes. In our study, however,ITGB1 expression in the body side skin was less highly expressed. The mechanism how it happens deserves further investigation. + + 443141 + Gene + + ITGB1 + + + 3688 + Gene + 22999 + + ITGB1 + + + 443141 + Gene + + ITGB1 + + + 443141 + Gene + + ITGB1 + + + 9606 + Species + + human + + + + DISCUSS + paragraph + 15317 + Rowe et al. confirmed that predominantly CYP1A1 was located to the sebaceous gland surrounding the hair shaft. It is not known why CYP1A1 expression was less highly expressed more than 10 folds in the body side skin from our data. This interesting phenomenon is worth pursuing in future studies. + + 100170113 + Gene + + CYP1A1 + + + 100170113 + Gene + + CYP1A1 + + + + DISCUSS + paragraph + 15613 + As development proceeds, GluD1 expression becomes restricted to the hippocampus, cochlear and vestibular hair cells, and spiral ganglion cells. Hair follicles are also derived from ectoderm. So why GLUD1 was less highly expressed in body side skin really deserves further investigation. + + 443239 + Gene + + GluD1 + + + 443239 + Gene + + GLUD1 + + + MESH:D015837 + Disease + + vestibular hair cells + + + + DISCUSS + paragraph + 15900 + Connexin 43 (Cx43) is a gap junction protein expressed in the follicular dermal papilla (DP). A recent mouse model research suggested an important role for Cx43 in hair regeneration, growth, and cuticle formation. However, Connexin 43 was less highly expressed in body side skin in the microarray analysis of this study, and it was also confirmed by qPCR. + + 443455 + Gene + + Connexin 43 + + + 443455 + Gene + + Cx43 + + + 14611 + Gene + 9670 + + Cx43 + + + 443455 + Gene + + Connexin 43 + + + 10090 + Species + + mouse + + + MESH:D010211 + Disease + + follicular dermal papilla + + + + DISCUSS + paragraph + 16256 + Several papers indicated that prostaglandin induced hair growth, but in our results, one prostaglandin synthase (PGFS) were less highly expressed in body side skin. This interesting contradiction remains to be further elucidated. + + 443108 + Gene + + PGFS + + + MESH:D011453 + Chemical + + prostaglandin + + + + DISCUSS + paragraph + 16486 + We have indeed tried our best to find out differentially expressed genes associated with LCE and MOGAT as reported in a Science paper. However, we could find only a few genes (DGAT2, LPL, ZO1, GJB2 and ITGB1) that have the possibility to play roles in epidermal development complex (EDC) or skin lipid metabolism. One reason for limited DE genes is that what we compared were different regions of sheep skin tissue (which might share similar principles in EDC regulation and lipid synthesis), not distinct tissues such as rumen and skin. That might also be caused by inadequate annotation of the probes or less gene coverage redundancy of the Agilent microarray. + + 101110535 + Gene + + DGAT2 + + + 443408 + Gene + + LPL + + + 443200 + Gene + + ZO1 + + + 443345 + Gene + + GJB2 + + + 443141 + Gene + + ITGB1 + + + 9940 + Species + + sheep + + + MESH:D008055 + Chemical + + lipid + + + MESH:D008055 + Chemical + + lipid + + + + DISCUSS + title_2 + 17149 + Functional implication of differentially expressed proteins in the proteomic study + + + DISCUSS + paragraph + 17232 + Most of the differentially expressed proteins were not listed above in the Discussion. + + + DISCUSS + paragraph + 17319 + Cryab protein was detected in 2DE in groin skin but not in body side skin. But the Cryab transcripts was more highly expressed in body side skin by the microarray study, though the differential expression was not validated by qPCR. This gene, downregulated in the transcriptional level during the depilation-induced hair cycle, was considered to play a role in regulation of apoptosis. + + 497274 + Gene + + Cryab + + + 497274 + Gene + + Cryab + + + + DISCUSS + paragraph + 17705 + FGF18 protein was less highly expressed in body side skin compared to the control. But the FGF18 transcripts was more highly expressed by the microarray study. Consistent to our microarray results, FGF18 is highly expressed in hair follicles and capable of inducing anagen from telogen stage hair follicles. + + 780460 + Gene + + FGF18 + + + 780460 + Gene + + FGF18 + + + 780460 + Gene + + FGF18 + + + + DISCUSS + paragraph + 18013 + KRT14 and KRT2.11 proteins were both less highly expressed in body side skin compared to the control. But these proteins were expected to play roles in wool follicles. + + 100303604 + Gene + + KRT14 + + + 443415 + Gene + + KRT2.11 + + + + DISCUSS + paragraph + 18181 + PADI3 played roles in the assembly of a globular S100A3 homotetramer,a putative Ca2+ modulator maturing human hair cuticle. But the PADI3 protein was less highly expressed in body side skin compared to the control in our study. This interesting contradiction remains to be further elucidated. + + 51702 + Gene + 7882 + + PADI3 + + + 443369 + Gene + + PADI3 + + + 6274 + Gene + 2223 + + S100A3 + + + 9606 + Species + + human + + + MESH:D000069285 + Chemical + + Ca2+ + + + + DISCUSS + paragraph + 18474 + FGFR3 RNA was detected in precuticle cells in the periphery of the hair bulb. But FGFR3 protein was less highly expressed in body side skin compared to the control in our study. + + 554324 + Gene + + FGFR3 + + + 554324 + Gene + + FGFR3 + + + + DISCUSS + title_2 + 18652 + Concordance between the results of transcriptomic and proteomic levels + + + DISCUSS + paragraph + 18723 + In the results of transcriptomic level (microarray), the number of more highly expressed genes was less than that of less highly expressed ones (602 versus 892). Consistently, more highly expressed protein spots were less than more lowly expressed ones (85 versus 102). + + + DISCUSS + paragraph + 18993 + The concordance between these two results from microarray and proteomic experiments was shown in Additional file 4: Table S4. Of the 46 identified proteins, 13 corresponding transcripts also shown differential expression. Among these 13 transcripts, the regulation trends of only 6 (in green) were consistent with their protein counterparts. The inconsistency of the two levels of other 7 DE genes (in red) might be produced by post transcriptional regulation or for unknown reasons. + + + DISCUSS + title_2 + 19477 + The confidence of the differentially expressed transcripts + + + DISCUSS + paragraph + 19536 + For the Agilent microarray, the probe number of each gene is indefinite, from 1 to 5. There exists alternative splicing after gene transcription, so the inconsistency between different probes for the same gene is sometimes reasonable. Certainly the confidence is higher if all probes for the same gene were differentially expressed. For example, there are 4 probes for FGF7 and LOC443300(YWHAE) separately, of which 2 were both less highly expressed in S/G. So we speculated that the reliabilities of differential expression of FGF7 and LOC443300 were close to each other. While MMP2 differential expression was only detected in one of three probes, then its confidence might be lower than the former two genes (FGF7 and LOC443300). Nevertheless, because of the inadequate annotation of most probes, we found it difficult to calculate probe number of all the differentially expressed genes. + + 443095 + Gene + + FGF7 + + + 443300 + Gene + + YWHAE + + + 443095 + Gene + + FGF7 + + + 443115 + Gene + + MMP2 + + + 443095 + Gene + + FGF7 + + + + DISCUSS + title_2 + 20427 + Gene networks + + + DISCUSS + paragraph + 20441 + Figure 5 listed some of the factors which possibly play roles in wool growth regulation. We speculate that lower temperature and shorter day photoperiod were key causes of the downregulation of wool growth rate. So we could speculate that those 8 genes (IL1A,FGF7,TAC1,IGFBP3,CDKN1B,IL6,FGF18 and PRL) played roles in wool growth regulation during winter. + + 443404 + Gene + + IL1A + + + 443095 + Gene + + FGF7 + + + 100034669 + Gene + + TAC1 + + + 100286737 + Gene + + IGFBP3 + + + 100127219 + Gene + + CDKN1B + + + 443406 + Gene + + IL6 + + + 780460 + Gene + + FGF18 + + + 443317 + Gene + + PRL + + + + CONCL + title_1 + 20798 + Conclusions + + + CONCL + paragraph + 20810 + In summary, the data presented in this study suggested that the body side skin displays a differentially expressed pattern in comparison with the groin skin at December time point. The majority of these genes possibly related to the wool growth control, and they could be assigned into the categories including regulation of receptor binding, multicellular organismal process, protein binding and macromolecular complex. Several potential gene families might participate in hair growth regulation, including fibroblast growth factors, transforming growth factor-beta, insulin-like growth factor, and so on. Proteomic analysis also identified hundreds of differentially expressed proteins. This systematic analysis could lead to a better understanding of the wool growth control mechanism in Aohan fine wool sheep. + + 9940 + Species + + sheep + + + + METHODS + title_1 + 21624 + Methods + + + METHODS + title_2 + 21632 + Animals and sample preparation + + + METHODS + paragraph + 21663 + All animals were treated in accordance with the animal protocols defined by national and local animal welfare bodies, and all animal work was approved by the Shandong Province Biological Studies Animal Care and Use Committee. + + + METHODS + paragraph + 21889 + Sampling methods were described previously. One ram and two ewes of 16-month-old Aohan fine wool sheep were used in the microarray study. These animals were half sibs (sharing the same father). In December 2010, two areas of full-thickness skin were sampled from the same animal under local anaesthesia: body side skin (wool bearing) and groin skin (non-wool bearing) for microarray and proteomic experiments. The area of each sample was about 1 cm2. All samples were immediately put into collection tubes and stored in liquid nitrogen for RNA and protein extraction. A total of 15, 208 probes were spotted on this Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA). + + 9940 + Species + + sheep + + + 9940 + Species + + Sheep + + + MESH:D009584 + Chemical + + nitrogen + + + + METHODS + title_2 + 22570 + RNA extraction and microarray hybridization + + + METHODS + paragraph + 22614 + TRIzol (Invitrogen) was used for total RNA extractions according to the manufacturer's protocol. RNA quantity and quality were measured by NanoDrop ND-1000. Its OD260/OD280 ratio was confirmed to be higher than 1.8. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. The RNA samples were sent to Kangchen Biotechnology Limited Company (Shanghai, China) for hybridization to the Agilent Sheep Gene Expression Microarray (Santa Clara, CA, USA). Each RNA sample was hybridized to one microarray slide. 1 mug of total RNA from each sample was amplified and transcribed into fluorescent cRNA with using the manufacturer's Agilent's Quick Amp Labeling protocol (version 5.7, Agilent Technologies). The labeled cRNAs were hybridized onto the Whole Genome Oligo Array (4x44K, Agilent Technologies). + + 9940 + Species + + Sheep + + + MESH:D012685 + Chemical + + agarose + + + MESH:D000249 + Chemical + + Amp + + + MESH:C411644 + Chemical + + TRIzol + + + + METHODS + title_2 + 23440 + Microarrays data analysis + + + METHODS + paragraph + 23466 + After hybridization and washing, the microarray slides were scanned with the Agilent Scanner G2505B. The resulting text files extracted from Agilent Feature Extraction Software (version 10.5.1.1) were imported into the Agilent GeneSpring GX software (version 11.0) for further analysis. Quantile normalization, probe annotation and subsequent data processing were performed using the GeneSpring GX v11.0 software package (Agilent Technologies). After Quantile normalization of the raw data, genes that at least 1 out of 6 samples have flags in Present ("All Targets Value") were chosen for differentially expressed genes screening. Gene expression levels were quantified relative to the expression of GAPDH. Differentially expressed genes were identified through fold-change screening. The fold-change of 2.0 and a false discovery rate of approximately 5% were set as the threshold. All data have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE62552 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc). Clustering analysis of all differentially expressed genes was performed using Cluster 3.0 to analyze the similarity in the expression patterns among different skin sites. The functional annotation of differentially expressed genes was performed by the DAVID (The Database for Annotation, Visualization and Integrated Discovery) gene annotation tool (http://david.abcc.ncifcrf.gov/). The KEGG pathway analysis was done manually (http://www.genome.jp/kegg/). + + 443005 + Gene + + GAPDH + + + 34205 + Species + + flags + + + + METHODS + paragraph + 24992 + Particular differentially expressed genes, which also appeared in the Figure 1 of reference No. 29 summarizing selective factors with known hair growth regulatory roles, were included in the putative gene networks of wool growth regulation. The networks contained genes possibly playing roles in anagen promotion and maintenance, in catagen-telogen promotion and maintenance, or with function unknown. + + + METHODS + title_2 + 25395 + QPCR + + + TABLE + table + 25400 + aThe annealing temperature represents the optimal temperature during quantitative PCR. + + + TABLE + table + 25488 + bRNA levels of GAPDH was assayed for normalization during quantitative PCR. + + 443005 + Gene + + GAPDH + + + + TABLE + Tab1.xml + Tab1 + table_caption + 25565 + Primers used for Q-PCR validation + + + TABLE + Tab1 + table + Tab1.xml + 25601 + Gene Primer sequence (5'-3') Tm ( C) a Target size (bp) GAPDHb Forward: ATGCCTCCTGCACCACCA 60 76 Reverse: AGTCCCTCCACGATGCCAA FGF10 Forward: GATCCGAGAAAGGAGCGAGG 60 554 Reverse: TCCAGGATACTGTACGGGCA LOC443300 Forward: ACCAACACATCCCATTCGCT 60 140 Reverse: CACTCAGCGTGTCCAGTTCT FGF18 Forward: AAGTCCGGATCAAGGGCAAG 60 98 Reverse: CACACACTCTTTGCTGGTGC Connexin43 Forward: GTCGTGTCGTTGGTGTCTCT 60 291 Reverse: CACTCAGCGTGTCCAGTTCT SCD Forward: AAGAGTGGCTGAGTTTCTGGTC 60 277 Reverse: GAAAGGAAGGTGATAGGGACAA Zo1 Forward: AGATAGCCCTGCAGCCAAAG 60 117 Reverse: GGGAGGTCAAGCAGGAAGAG MMP2 Forward: AACGCCATCCCTGATAACCT 60 126 Reverse: GCTTCCGAACTTCACGCTC ITGB1 Forward: AGCACGGATGAGGTGAACAG 60 407 Reverse: CCAAGGCAGGTCTGACACAT PAG11 Forward: AGCGTCGCCTACGAATCTG 60 120 Reverse: CTCAAACCCATATTCCGTCACA CRYAB Forward: CACCCAGCTGGATTGACACT 60 147 Reverse: CCTCATGTTTGCCATGCACC + + 443074 + Gene + + FGF10 + + + 780460 + Gene + + FGF18 + + + 443455 + Gene + + Connexin43 + + + 443115 + Gene + + MMP2 + + + 443141 + Gene + + ITGB1 + + + 100034671 + Gene + + PAG11 + + + 497274 + Gene + + CRYAB + + + 443200 + Gene + + Zo1 + + + 443185 + Gene + + SCD + + + + METHODS + paragraph + 26479 + The total RNA samples prepared for microarray analysis were also used for qPCR analysis. Reverse transcriptions were performed using RevertAid First Strand cDNA Synthesis Kit (MBI Fermentas, Vilnius, Lithuania) according to the manufacturer's protocols. The primers were designed with the Oligo 6.0 program. The primer sequences, melting temperatures and product sizes are shown in Table 1. Gene expression levels were quantified relative to the expression of GAPDH. + + 443005 + Gene + + GAPDH + + + + METHODS + title_2 + 26949 + Tissue protein extraction + + + METHODS + paragraph + 26975 + Lysis buffer preparation: 42% Urea, 15.2% Thiourea, 4% CHAPS, 1% DTT. Sampled tissues were homogenziated in lysis buffer (containing 1% cocktail and 2% IPG-buffer, added right before use) at the ratio of 1:7 (weight/volume). The tissues were cut into small pieces by ophthalmic scissors, and left at 4 C for 1 h, vortexed it every 15 min. Then, the tissue homogenate was centrifuge at 40,000 g for 30 min. Supernatants were collected and stored at -80 C. Protein concerntrations were determined by Bradford method. + + MESH:C028213 + Chemical + + CHAPS + + + MESH:D004229 + Chemical + + DTT + + + MESH:D013890 + Chemical + + Thiourea + + + MESH:D014508 + Chemical + + Urea + + + - + Chemical + + IPG + + + + METHODS + title_2 + 27498 + One-dimensional electrophoresis + + + METHODS + paragraph + 27530 + 0.5% IPG-buffer was added into each 150 mug protein sample (in a final volume of 400-600 muL), and was loaded in the One-dimensional electrophoresis instrument. The progamme is as follows: Step-n-hold (S1, 30 V for 6 h; S2, 60 V for 6 h); Gradient (S3, 500 V for 1 h;S4,1000 V for 1 h; S5, 3000 V for 3 h; S6, 8000 V for 3 h); Step-n-hold (S7, 8000 V for 20 h). + + - + Chemical + + IPG + + + + METHODS + title_2 + 27908 + 2-dimensional (2-D) SDS-PAGE preparation + + MESH:D012967 + Chemical + + SDS + + + + METHODS + paragraph + 27949 + Tris-HCl (PH = 8.8), Monomer storage (30% Acrylamid and 0.8% NN'-methy lenebisacry lamid), 10 x electrophoresis buffer (3.03% Tris-Base, 14.4% Glycine, 1% SDS), balanced solution (36.05% Urea, 5% Tris-HCl, 2% SDS, 34.5% Glycerine). + + MESH:D005990 + Chemical + + Glycerine + + + MESH:D005998 + Chemical + + Glycine + + + MESH:D014325 + Chemical + + Tris-Base + + + - + Chemical + + Tris-HCl + + + MESH:D012967 + Chemical + + SDS + + + - + Chemical + + NN'-methy lenebisacry lamid + + + MESH:D012967 + Chemical + + SDS + + + MESH:D020106 + Chemical + + Acrylamid + + + - + Chemical + + Tris-HCl + + + MESH:D014508 + Chemical + + Urea + + + + METHODS + title_2 + 28196 + The second dimensional SDS-PAGE + + MESH:D012967 + Chemical + + SDS + + + + METHODS + paragraph + 28228 + Transfer: Voltage 300v, Current 50 mA, Time 1 h. + + + METHODS + paragraph + 28279 + Separation: Voltage 300v, Current 200 ~ 250 mA, Time 4 ~ 5 h. + + + METHODS + paragraph + 28351 + Fixative preparation: 40% Ethanol and 10% Acetic acid. + + MESH:D019342 + Chemical + + Acetic acid + + + MESH:D000431 + Chemical + + Ethanol + + + + METHODS + paragraph + 28406 + Electrophoresis was carried out until the blue dye front had just disappeared from the bottom of the gel. + + + METHODS + paragraph + 28512 + Fixation: take out the rubber strip and put it into Fixative for 1 h. + + + METHODS + paragraph + 28583 + The electrophoresis programme is as follows: + + + METHODS + title_2 + 28628 + Staining and visualization + + + METHODS + paragraph + 28655 + Sensitizing solution preparation: 30% Ethanol, 0.314% Na2S2O3, 6.8%NaAc. + + - + Chemical + + Na2S2O3 + + + MESH:D000431 + Chemical + + Ethanol + + + - + Chemical + + NaAc + + + + METHODS + paragraph + 28728 + Sensitizing: the gels were sensitized for 30 min. + + + METHODS + paragraph + 28779 + Washing: the gels were washed for three times using ddH2O. Ten minutes each time. + + - + Chemical + + ddH2O + + + + METHODS + paragraph + 28861 + Ilver staining: Silver staining solution was prepared (1.25 AgNO3 and 200 muL Formaldehyde in 500 mL ddH2O). The gels were stained by the solution for 20 min. + + MESH:D012835 + Chemical + + AgNO3 + + + MESH:D005557 + Chemical + + Formaldehyde + + + - + Chemical + + ddH2O + + + + METHODS + paragraph + 29023 + Washing: the gels were washed for 2 min using ddH2O. + + - + Chemical + + ddH2O + + + + METHODS + paragraph + 29077 + Visualizing solution preparation: 12.5 g Na2CO3 and 100 muL Formaldehyde in 500 mL ddH2O. + + MESH:C005686 + Chemical + + Na2CO3 + + + - + Chemical + + ddH2O + + + MESH:D005557 + Chemical + + Formaldehyde + + + + METHODS + paragraph + 29170 + Termination solution preparation: 2 g Glycine in 50 ml ddH2O. + + - + Chemical + + ddH2O + + + MESH:D005998 + Chemical + + Glycine + + + + METHODS + paragraph + 29234 + Visualization until the solution became muddy, then terminating for 30 min. + + + METHODS + title_2 + 29311 + Determination of relative protein expression + + + METHODS + paragraph + 29356 + Gels were then scanned and analyzed using ImageMaster TM 2D platinum software (Version 5.0, GE Healthcare, San Francisco, CA, USA). The expression level was determined by the relative volume of each spot in the gel and expressed as %Vol (%Vol = [spot volume/Sigmavolumes of all spots resolved in the gel]). The means and standard deviations of both sample groups were calculated. Statistical significance with Student's t-tests using ImageMaster TM 2D platinum software. P values <0.05 were considered statistically significant. + + + METHODS + title_2 + 29888 + Identification of differentially expressed proteins by mass spectrometry (MS) + + + METHODS + paragraph + 29966 + Protein spots with significant differences between the two groups were excised, dehydrated in acetonitrile, and dried at room temperature. Gel pieces were denatured, alkylated, trypsin digested and analyzed by an Ultraflex II MALDI-TOF-TOF mass spectrometer (Bruker Daltonics GmbH, Bremen, Germany) under the control of FlexControl TM 2.4 software (Bruker Daltonics GmbH). Acquired peptide mass fingerprint (PMF) were processed using the software FlexAnalysis 3.0 (Bruker Daltonics, Bremen, Germany). The peak detection algorithm was: SNAP (Sort Neaten Assign and Place); S/N threshold: 1.5; Quality Factor Threshold: 50. The tryptic auto-digestion ion picks (trypsin [108-115] 842.5094 Da, trypsin [58-77] 2211.104 Da) were used as internal standards. The resulting peptide mass lists were used to search the Matrixscience database (http://www.matrixscience.com). The following search parameter criteria were used: mass tolerance 100 ppm, miss cleavage 1, modification comprises carbamidomethyl and methionine oxidation. Matched peptides number between experimental PMF and theoretical PMF 5. All MS data have been deposited in PeptideAtlas and are accessible through Dataset Identifier PASS00597 (http://www.peptideatlas.org/PASS/PASS00597). + + - + Chemical + + carbamidomethyl + + + MESH:C032159 + Chemical + + acetonitrile + + + MESH:D008715 + Chemical + + methionine + + + + SUPPL + title_1 + 31224 + Availability of supporting data + + + SUPPL + paragraph + 31256 + All cDNA microarray data have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE62552 (http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE62552). + + + SUPPL + paragraph + 31463 + Proteomic data have been deposited in PeptideAtlas and are accessible through Dataset Identifier PASS00597 (http://www.peptideatlas.org/PASS/PASS00597). + + + SUPPL + title_1 + 31616 + Additional files + + + SUPPL + footnote + 31633 + Nan Liu and Hegang Li contributed equally to this work. + + + SUPPL + footnote + 31689 + Competing interests + + + SUPPL + footnote + 31709 + The authors declare that they have no competing interests. + + + SUPPL + footnote + 31768 + Authors' contributions + + + SUPPL + footnote + 31793 + JZ and HL carried out data collection and analysis and manuscript preparation. KL carried out data analysis. JY participated in data analysis. MC, participated in data collection. WD participated in data discussion and manuscript preparation. RB participated in manuscript preparation. JL and GH participated in discussion of the study. NL participated in the design of the study, coordination, data analysis and manuscript preparation. 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surname:Gao;given-names:W + 37385 + DAVID: database for annotation, visualization, and integrated discovery + + + 20 + surname:Langen;given-names:H + surname:Hobohm;given-names:U + REF + surname:Berndt;given-names:P + 3521 + 10.1002/(SICI)1522-2683(19991201)20:18<3521::AID-ELPS3521>3.0.CO;2-8 + 10612278 + 1999 + Electrophoresis + ref + 3526 + 37457 + Reliable automatic protein identification from matrix-assisted laser desorption/ionization mass spectrometric peptide fingerprints + + + + 4272823 + + surname:Bhagat;given-names:Aditi + surname:Porter;given-names:Gerald + surname:Chen;given-names:Leonard N + 25497602 + surname:Joh;given-names:Daniel Y + 2014 + 4272823 + 277 + 10.1186/s13014-014-0277-4 + Keywords + front + TITLE + Radiat Oncol; 2014 Dec 12; 9277. doi:10.1186/s13014-014-0277-4 + 277 + 9 + This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. + surname:Ju;given-names:Andrew W + surname:Lynch;given-names:John H + surname:Dritschilo;given-names:Anatoly + surname:Suy;given-names:Simeng + surname:Carroll;given-names:John + Prostate cancer SBRT Rectal endoscopy Telangiectasias CyberKnife Expanded prostate index composite Bother Proctitis Rectal bleeding Vienna rectoscopy score + surname:Collins;given-names:Sean P + surname:Collins;given-names:Brian T + surname:Lei;given-names:Siyuan + surname:Yung;given-names:Thomas + surname:Moures;given-names:Rudy + surname:Kim;given-names:Joy S + surname:Sood;given-names:Sumit + Joh DY, Chen LN, Porter G, Bhagat A, Sood S, Kim JS, Moures R, Yung T, Lei S, Collins BT, Ju AW, Suy S, Carroll J, Lynch JH, Dritschilo A, Collins SP + 0 + Proctitis following stereotactic body radiation therapy for prostate cancer + + MESH:D011349 + Disease + + Proctitis + + + MESH:D011471 + Disease + + prostate cancer + + + + ABSTRACT + abstract_title_1 + 76 + Background + + + ABSTRACT + abstract + 87 + Proctitis after radiation therapy for prostate cancer remains an ongoing clinical challenge and critical quality of life issue. SBRT could minimize rectal toxicity by reducing the volume of rectum receiving high radiation doses and offers the potential radiobiologic benefits of hypofractionation. This study sought to evaluate the incidence and severity of proctitis following SBRT for prostate cancer. + + MESH:D011349 + Disease + + proctitis + + + MESH:D064420 + Disease + + toxicity + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011349 + Disease + + Proctitis + + + + ABSTRACT + abstract_title_1 + 491 + Methods + + + ABSTRACT + abstract + 499 + Between February 2008 and July 2011, 269 men with clinically localized prostate cancer were treated definitively with SBRT monotherapy at Georgetown University Hospital. All patients were treated to 35-36.25Gy in 5 fractions delivered with the CyberKnife Radiosurgical System (Accuray). Rectal bleeding was recorded and scored using the CTCAE v.4. Telangiectasias were graded using the Vienna Rectoscopy Score (VRS). Proctitis was assessed via the Bowel domain of the Expanded Prostate Index Composite (EPIC)-26 at baseline and at 1, 3, 6, 9, 12, 18 and 24 months post-SBRT. + + 9606 + Species + + men + + + 9606 + Species + + patients + + + MESH:D011471 + Disease + + localized prostate cancer + + + MESH:D011349 + Disease + + Proctitis + + + MESH:D006470 + Disease + + Rectal bleeding + + + MESH:D013684 + Disease + + Telangiectasias + + + - + Chemical + + SBRT + + + + ABSTRACT + abstract_title_1 + 1075 + Results + + + ABSTRACT + abstract + 1083 + The median age was 69 years with a median prostate volume of 39 cc. The median follow-up was 3.9 years with a minimum follow-up of two years. The 2-year actuarial incidence of late rectal bleeding >= grade 2 was 1.5%. Endoscopy revealed VRS Grade 2 rectal telangiectasias in 11% of patients. All proctitis symptoms increased at one month post-SBRT but returned to near-baseline with longer follow-up. The most bothersome symptoms were bowel urgency and frequency. At one month post-SBRT, 11.2% and 8.5% of patients reported a moderate to big problem with bowel urgency and frequency, respectively. The EPIC bowel summary scores declined transiently at 1 month and experienced a second, more protracted decline between 6 months and 18 months before returning to near-baseline at two years post-SBRT. Prior to treatment, 4.1% of men felt their bowel function was a moderate to big problem which increased to 11.5% one month post-SBRT but returned to near-baseline at two years post-SBRT. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + men + + + MESH:D011349 + Disease + + proctitis + + + MESH:D006470 + Disease + + late rectal bleeding + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D013684 + Disease + + telangiectasias + + + MESH:D015212 + Disease + + bowel function + + + + ABSTRACT + abstract_title_1 + 2070 + Conclusions + + + ABSTRACT + abstract + 2082 + In this single institution cohort, the rate and severity of proctitis observed following SBRT is low. QOL decreased on follow-up; however, our results compare favorably to those reported for patients treated with alternative radiation modalities. Future prospective randomized studies are needed to confirm these observations. + + 9606 + Species + + patients + + + MESH:D011349 + Disease + + proctitis + + + + INTRO + title_1 + 2409 + Background + + + INTRO + paragraph + 2420 + Post-treatment bowel function is a primary determinant of quality of life following radiation therapy for prostate cancer. Late proctitis occurs at a frequency of 5-20% with conventionally fractionated treatment. The incidence of proctitis is dependent on the assessment method and whether it is patient or physician reported. Patients with radiation-induced proctitis report increased bowel frequency/urgency, incontinence, bleeding and pain. These symptoms occur months to years after treatment with the large majority of patients reporting symptoms within two years following radiation therapy. Patient characteristics such as age, comorbidities, hemorrhoids, inflammatory bowel disease and/or anticoagulation may increase an individual's risk of radiation-induced proctitis. Unfortunately, treatment options for radiation proctitis are limited and of unclear clinical benefit. + + 9606 + Species + + patient + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + 9606 + Species + + Patient + + + MESH:D015212 + Disease + + inflammatory bowel disease + + + MESH:D006484 + Disease + + hemorrhoids + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D006470 + Disease + + bleeding + + + MESH:D011349 + Disease + + proctitis + + + MESH:D015212 + Disease + + bowel function + + + MESH:D011349 + Disease + + proctitis + + + MESH:D014549 + Disease + + incontinence + + + MESH:D010146 + Disease + + pain + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis + + + + INTRO + paragraph + 3305 + Proctitis is the principle dose-limiting toxicity of prostate radiotherapy. Acute proctitis has been attributed to radiation-induced injury to the epithelial lining of the rectal mucosa, leading to mucosal sloughing, acute inflammatory infiltrates, and vascular permeability leading to edema. These early changes are associated with bowel frequency, bowel urgency and rectal pain. Chronic proctitis results from epithelial atrophy and obliterative endarteritis, resulting in tissue ischemia, rectal fibrosis, stricture formation, and neovascularization; this presents clinically with bowel frequency, urgency and rectal bleeding. Historically, endoscopic findings in patients with proctitis include telangiectasia, congested mucosa, and ulcers. Telangiectasias are observed in 20-80% of patients receiving conventionally fractionated radiation therapy. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + MESH:D058186 + Disease + + injury + + + MESH:D013684 + Disease + + telangiectasia + + + MESH:D011349 + Disease + + proctitis + + + MESH:D005355 + Disease + + fibrosis + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D064420 + Disease + + toxicity + + + MESH:D007511 + Disease + + ischemia + + + MESH:D006470 + Disease + + bleeding + + + MESH:D002277 + Disease + + epithelial atrophy + + + MESH:D004487 + Disease + + edema + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + Chronic proctitis + + + MESH:D014456 + Disease + + ulcers + + + MESH:D011349 + Disease + + Proctitis + + + MESH:D004692 + Disease + + endarteritis + + + MESH:D013684 + Disease + + Telangiectasias + + + + INTRO + paragraph + 4158 + The risk of proctitis appears to be dependent upon both the total radiation dose and the volume of the rectum in the high dose area. Technical factors such as treatment of the proximal seminal vesicles and expansion of planning target volume (PTV) to compensate for intra-fraction prostate motion may contribute to the severity of rectal toxicities. To minimize bowel toxicity, it is currently recommended that the volume of the rectum receiving > 75 Gray (Gy) be limited to < 10% with conventionally fractionated radiation therapy. Studies have shown that advanced radiation technologies such as intensity modulated radiation therapy (IMRT) may decrease the dose to the rectum and potentially decrease proctitis. + + MESH:D012002 + Disease + + rectal toxicities + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis + + + MESH:D015212 + Disease + + bowel toxicity + + + + INTRO + paragraph + 4880 + Recent data suggest that large radiation fraction sizes are radiobiologically favorable over lower fraction sizes in prostate cancer. The alpha/beta for prostate cancer may be as low as 1.5 Gy. If the alpha/beta for prostate cancer is less than 3-5 Gy, which is generally the value accepted for late rectal complications, the linear-quadratic model predicts that delivering large radiation fraction sizes will result in improved local control with a similar rate of bowel complications. Therefore, using large fraction sizes in SBRT provides an attractive modality to leverage the potential radiobiologic benefits of hypofractionation with the minimal invasiveness of an external-beam treatment modality. Furthermore, by reducing the number of treatment visits, hypofractionated therapy can also be logistically favorable for patients and reduce financial burden, ultimately decreasing barriers to care. + + 9606 + Species + + patients + + + MESH:D015212 + Disease + + bowel complications + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011471 + Disease + + prostate cancer + + + + INTRO + paragraph + 5784 + The safety and efficacy of SBRT in the treatment of clinically localized prostate cancer has been established in a number of studies. The use of large fraction sizes in SBRT offers the potential radiobiological benefits of hypofractionation and potentially may minimize radiation proctitis by reducing the volume of the rectum receiving high radiation doses. The CyberKnife robotic radiosurgical system uses image guidance to track implanted fiducials to account for intrafraction prostatic motion. This reduces the uncertainty of the location of the prostate and allows treatment to be delivered with a smaller PTV expansion, which may reduce the doses delivered to the anterior rectal wall. The goal of this study is to report the bowel outcomes following SBRT for clinically localized prostate cancer. + + MESH:D011349 + Disease + + proctitis + + + MESH:D011471 + Disease + + localized prostate cancer + + + MESH:D011471 + Disease + + localized prostate cancer + + + + METHODS + title_1 + 6589 + Methods + + + METHODS + title_2 + 6597 + Patient selection + + 9606 + Species + + Patient + + + + METHODS + paragraph + 6615 + Patients eligible for study inclusion had histologically-confirmed prostate cancer treated per our institutional protocol. Clinical stage was defined according to the 6th edition of the American Joint Committee on Cancer criteria. Risk groups were defined using the D'Amico criteria. Internal Review Board (IRB)-approval was obtained for retrospective review of data that was prospectively collected in our institutional database. + + 9606 + Species + + Patients + + + MESH:D011471 + Disease + + prostate cancer + + + + METHODS + title_2 + 7048 + SBRT treatment planning and delivery + + + METHODS + paragraph + 7085 + SBRT was delivered using the CyberKnife robotic radiosurgical system (Accuray Inc., Sunnyvale, CA). The fiducial placement and computed tomography (CT)/magnetic resonance (MR) simulation procedures have been previously described in Lei et al.. The clinical target volume (CTV) was defined as the prostatic capsule and proximal seminal vesicles (to the point where the seminal vesicles separate). To cover areas of potential ECE, the expansion from the CTV to the PTV was 5 mm in all directions except 3 mm posteriorly into the rectum. There was no difference in the target volume delineation for different risk groups. Fiducial-based tracking was used to account for interfraction and intrafraction prostate motion. Treatment planning was performed using Multiplan (Accuray Inc., Sunnyvale, CA). Patients were treated with 35 or 36.25 Gy of radiotherapy delivered in 5 fractions of 7-7.25 Gy each to the PTV, administered over a timespan of 1-2 weeks. This corresponds to a tumor equivalent dose in 2-Gy fractions (EQD2) of approximately 85-90 Gy assuming an alpha/beta ratio of 1.5. The rectum was contoured as a solid structure from the anus (at the level of the ischial tuberosities) to the rectosigmoid flexure and evaluated with dose-volume histogram analysis during treatment planning using Multiplan (Accuray Inc., Sunnyvale, CA) inverse treatment planning. Rectal volume receiving 36 Gy was limited to <= 1 cc. Assuming an alpha/beta of 3 Gy for late bowel complications, this is biologically equivalent to approximately 74 Gy administered in 2 Gy fractions. The rectal dose-volume histogram (DVH) goals were <50% rectal volume receiving 50% of the prescribed dose, <20% receiving 80% of the dose, <10% receiving 90% of the dose, and < 5% receiving 100% of the dose. Typical dose distributions have been previously described. Patients were placed on a low-residual diet and given enemas prior to simulation and treatment delivery to maximize the potential distance between the prostate and the rectal wall and minimize intrafraction prostate motion. + + 1889 + Gene + 1068 + + ECE + + + 9606 + Species + + Patients + + + 9606 + Species + + Patients + + + MESH:D015212 + Disease + + late bowel complications + + + - + Chemical + + PTV + + + MESH:D009369 + Disease + + tumor + + + MESH:D014402 + Disease + + ischial tuberosities + + + + METHODS + title_2 + 9154 + Follow-up and statistical analysis + + + METHODS + paragraph + 9189 + Rectal bleeding was prospectively documented at follow-up visits using the National Cancer Institute (NCI) Common Toxicity Criteria (CTC) version 4.0. Acute rectal bleeding was defined as experiencing toxicity within 6 months of radiation therapy. Late rectal bleeding was defined as occurring at least 6 months after delivery of radiation therapy. Grade 1 represents minimal bleeding not requiring medications for symptom control. Grade 2 indicates rectal bleeding requiring new medication (i.e. steroid suppository) or minor laser coagulation. Telangiectasia were graded using the Vienna Rectoscopy Score (VRS): Grade 1 (a single telangiectasia), Grade 2 (multiple non-confluent telangiectasia) and Grade 3 (multiple confluent telangiectasia). + + MESH:D064420 + Disease + + toxicity + + + MESH:D006470 + Disease + + bleeding + + + MESH:D013684 + Disease + + telangiectasia + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D013684 + Disease + + telangiectasia + + + MESH:D064420 + Disease + + Toxicity Criteria + + + MESH:D006470 + Disease + + bleeding + + + MESH:D013684 + Disease + + Telangiectasia + + + MESH:D013256 + Chemical + + steroid + + + MESH:D006470 + Disease + + Rectal bleeding + + + MESH:D013684 + Disease + + telangiectasia + + + + METHODS + paragraph + 9937 + Proctitis was assessed via the bowel domain of the Expanded Prostate Index Composite (EPIC)-26 at baseline and at 1, 3, 6, 9, 12, 18 and 24 months. The timing of the quality of life (QOL) assessments was relative to the last day of SBRT treatment. The EPIC-26 bowel domain includes five questions related to individual symptoms (questions 6a-e: urgency, frequency, pain, bloody stool, incontinence) and one question (question 7) related to overall bother (degree of interference or annoyance caused by bowel symptoms. + + MESH:D011349 + Disease + + Proctitis + + + + METHODS + paragraph + 10456 + Wilcoxon signed-rank test and chi-square analysis were used to assess differences in ongoing quality of life scores in comparison to baseline. For each EPIC question, the responses were grouped into three clinically relevant categories (no problem, very small-small problem and moderate to big problem). To statistically compare changes between time points, the levels of responses were assigned a score and the significance of the mean changes in the scores was assessed by Wilcoxon rank test. EPIC scores for the bowel domain and its individual questions range from 0 - 100 with lower values representing worsening bowel symptoms. The minimally important difference (MID) in EPIC score was defined as a change of one-half standard deviation (SD) from the baseline. To limit the effect of attrition bias, statistical analysis was limited to time points in which >= 80% of the patient data were available. + + 9606 + Species + + patient + + + Disease + + MID + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D019217 + Disease + + attrition bias + + + MESH:D029461 + Disease + + SD + + + + RESULTS + title_1 + 11367 + Results + + + TABLE + Tab1.xml + Tab1 + table_caption + 11375 + Baseline patient characteristics and treatment + + 9606 + Species + + patient + + + + TABLE + Tab1 + table + Tab1.xml + 11424 + % Patients (N = 269) Age (y/o) Median 69 (44-90) <60 8.20% 60-69 42.40% 70-79 41.30% >=80 8.20% Race White 55.80% Black 37.20% Other 7.10% Prostate volume (cc) Median 39.04 (11.56-138.69) Charlson comorbidity index (CCI) 0 66.9% 1 22.7% 2 6.7% >=3 3.7% Body mass index BMI <25 21.9% 25 <= BMI < 30 47.2% 30 <= BMI < 35 21.9% BMI >=35 8.9% Use of anticoagulants Yes 35.3% No 64.7% Risk group Low 36.80% Intermediate 53.20% High 10.00% Hormonal therapy Yes 16.40% No 83.60% Dose 36.25 Gy 83.30% 35 Gy 16.40% Other 0.40% + + 9606 + Species + + Patients + + + + RESULTS + paragraph + 12029 + From February 2008 to July 2011, 269 prostate cancer patients were treated per our institutional SBRT monotherapy protocol (Table 1). The median follow-up was 3.9 years. The median age was 69 years and 44.2% were of non-Caucasian ancestry. Comorbidities were common with 35% were taking anticoagulants (including aspirin) prior to treatment. The median prostate volume was 39 cc. By D'Amico classification, 99 patients were low-, 143 intermediate-, and 27 high-risk. 16.4% also received androgen deprivation therapy (ADT) at the discretion of the treating Urologist. 83.3% of the patients were treated with 36.25 Gy in five 7.25 Gy fractions. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + - + Chemical + + ADT + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D001241 + Chemical + + aspirin + + + + RESULTS + paragraph + 12680 + A total of 61 (22.7%) patients reported rectal bleeding after SBRT, with 28 (10.4%) patients reporting acute bleeding and 38 patients (14.1%) reporting late bleeding. Overall, Grade 2 acute and late rectal bleeding were observed in 0 (0%) and 4 (1.5%) of patients, respectively. There was no Grade 3 or higher rectal bleeds. The majority of acute rectal bleeding occurred within 1-month post-SBRT treatment (79%). The bleeding completely resolved in the majority of the patients by the subsequent follow-up visit. Likewise, the majority of late rectal bleeding was observed at one specific follow-up appointment and did not persist on subsequent follow-ups. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + + RESULTS + paragraph + 13338 + Seventy-three patients had one or more rectal endoscopies during the follow-up period. Endoscopy was performed as a part of normal colorectal cancer screening or if patient had clinically significant rectal bleeding. The median interval from completion of SBRT to endoscopy was 24 months (3 months to 53 months). On endoscopy, rectal telangiectasias were found in 8 (11.0%) patients. All 8 patients were observed to have multiple non-confluent telangiectasias (VRS Grade 2). No patient had confluent or circumferential telangiectasia. Minor laser coagulation was performed in three patients. No rectal ulcerations, strictures, or fistulas were observed. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D013684 + Disease + + telangiectasia + + + MESH:D013684 + Disease + + telangiectasias + + + MESH:D015179 + Disease + + colorectal cancer + + + MESH:D005402 + Disease + + fistulas + + + MESH:D006470 + Disease + + bleeding + + + MESH:D013684 + Disease + + telangiectasias + + + + TABLE + table + 13995 + Patient-reported responses to EPIC-26 questions 6A (Urgency to have a bowel movement), 6B (Frequency of bowel movements), 6C (Losing control of your stools), 6D (Bloody stools) and 6E (Abdominal, pelvic or rectal pain). + + 9606 + Species + + Patient + + + MESH:D009069 + Disease + + bowel movements + + + MESH:D009069 + Disease + + bowel movement + + + MESH:D010146 + Disease + + pain + + + + TABLE + Tab2.xml + Tab2 + table_caption + 14215 + Bowel symptoms following SBRT for prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + TABLE + Tab2 + table + Tab2.xml + 14267 + Start 1 3 6 9 12 18 24 Bowel urgency No problem 77.2% 49.8% 62.0% 60.2% 64.5% 62.3% 64.7% 66.8% Very small-small problem 21.0% 39.0% 33.7% 34.1% 29.8% 31.6% 29.5% 31.1% Moderate-big problem 1.9% 11.2% 4.3% 5.6% 5.6% 6.1% 5.8% 2.1% p value <0.0001 0.0002 <0.0001 0.0002 0.0001 0.0004 0.01568 Bowel frequency No problem 82.0% 57.0% 71.9% 71.5% 70.6% 72.5% 76.3% 77.4% Very small-small problem 15.7% 34.5% 25.0% 25.3% 23.4% 23.0% 18.8% 19.7% Moderate-big problem 2.2% 8.5% 3.1% 3.2% 6.0% 4.5% 4.9% 2.9% p value <0.0001 0.0079 0.0038 0.0031 0.0214 0.0169 0.2508 Incontinence No problem 94.8% 79.5% 85.2% 86.7% 88.3% 85.2% 85.7% 89.5% Very small-small problem 4.1% 17.0% 14.5% 11.2% 9.7% 12.3% 11.6% 8.8% Moderate-big problem 1.1% 3.5% 0.4% 2.0% 2.0% 2.5% 2.7% 1.7% p value <0.0001 0.0033 0.0021 0.03 0.0001 0.0001 0.0147 Bloody stools No problem 98.1% 90.8% 96.9% 95.6% 96.0% 96.3% 94.2% 94.2% Very small-small problem 1.9% 8.1% 2.3% 3.6% 2.4% 2.5% 5.4% 5.8% Moderate-big problem 0.0% 1.2% 0.8% 0.8% 1.6% 1.2% 0.4% 0.0% p value 0.0002 0.1677 0.0934 0.0574 0.0906 0.0066 0.0258 Pain (abdominal, pelvic or rectal) No problem 86.1% 76.8% 89.8% 86.7% 86.7% 88.5% 86.6% 90.3% Very small-small problem 12.4% 17.8% 7.8% 10.4% 9.7% 8.2% 12.1% 7.1% Moderate-big problem 1.5% 5.4% 2.3% 2.8% 3.6% 3.3% 1.3% 2.5% p value 0.0002 0.5516 0.6606 0.232 0.5713 0.8565 0.7498 N= 267 259 255 249 248 244 224 238 + + MESH:D010146 + Disease + + Pain + + + + TABLE + Tab3.xml + Tab3 + table_caption + 15733 + Changes in EPIC bowel summary and overall bowel bother scores following SBRT for prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + TABLE + Tab3 + table + Tab3.xml + 15832 + EPIC criteria Baseline 1 Month 6 Month 12 Month 18 Month 24 Month Change S.D. Change S.D. Change S.D. Change S.D. Change S.D. Bloody stool bother 99.5 -2.9 12.26 -1.1 8.54 -1.3 9.86 -2.1 10.93 -1.4 8.03 Bowel summary 94.8 -9.8 17.89 -3.2 12.21 -3.5 13.04 -3.3 13.40 -1.6 10.73 Bowel bother 90.7 -14.8 26.98 -2.6 20.82 -5.5 23.95 -3.5 20.91 -0.9 18.86 + + + FIG + 13014_2014_277_Fig1_HTML.jpg + Fig1 + fig_caption + 16256 + Individual EPIC-26 bowel symptoms (Questions 6a-e). Average individual symptom EPIC bother scores at baseline and following SBRT for prostate cancer: (a) urgency to have a bowel movement-Question 6a of the EPIC-26; (b) increase frequency of bowel movements- Question 6b of the EPIC-26; (c) losing control of stools- Question 6c of the EPIC-26; (d) bloody stools- Question 6d of the EPIC-26; (e) abdominal/pelvic/rectal pain- Question 6e of the EPIC-26. Thresholds for clinically significant changes in scores (1/2 standard deviation above and below the baseline) are marked with dashed lines. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. + + MESH:D009069 + Disease + + bowel movements + + + MESH:D010146 + Disease + + pain + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D009069 + Disease + + bowel movement + + + MESH:D051271 + Disease + + bowel symptoms + + + + RESULTS + paragraph + 16949 + At baseline, 1.9% of our cohort reported some level of annoyance due to bloody stools, however no patient felt it was a moderate to big problem (Table 2). The mean changes in EPIC bloody stool bother scores from baseline to 2 years of follow-up are shown in Table 3. The mean EPIC bloody stool bother score was 99.5 at baseline (Table 3). Bloody stool bother increased following treatment with the mean score decreasing to 96.6 at 1 month post-treatment (mean change, -2.9) (p = 0.0002) (Table 3, Figure 1d). However, only 1.2% of patients felt that that bloody stools were a moderate to big problem at 1 month following treatment (Table 2). Although bloody stool bother declined quickly, a second late increase in bloody stool bother was observed with the mean bloody stool bother score decreasing to 97.4 at 18 months (mean change from baseline, -2.1) (p = 0.0066) (Table 3, Figure 1d). Both declines met the threshold for clinically significant change (MID = 1.7). However, only 1.2% of patients felt that that bloody stools were a moderate to big problem at 12 months following treatment (Table 2). By two years following SBRT, bloody stool bother returned to near-baseline with bloody stool bother score of 98.1 (mean change from baseline, -1.4). By two years post-SBRT, no patient felt that blood stools was a moderate to big problem (Table 2). + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + Disease + + MID + + + + RESULTS + paragraph + 18327 + Baseline proctitis symptoms were uncommon in our patients (Table 2). All proctitis symptoms increased at one month post-SBRT. The most bothersome symptoms were bowel urgency and frequency. At one month post SBRT, 11.2% and 8.5% of patients reported moderate to big problems with bowel urgency and frequency, respectively (Table 2). 9.7% of patients required anti-diarrheals within the first month post-SBRT. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D011349 + Disease + + proctitis + + + MESH:D011349 + Disease + + proctitis symptoms + + + MESH:D015212 + Disease + + bowel urgency + + + + FIG + 13014_2014_277_Fig2_HTML.jpg + Fig2 + fig_caption + 18737 + Average EPIC bowel summary scores at baseline and following SBRT for prostate cancer. Thresholds for clinically significant changes in scores (1/2 standard deviation above and below the baseline) are marked with dashed lines. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. + + MESH:D011471 + Disease + + prostate cancer + + + + RESULTS + paragraph + 19063 + Likewise, the EPIC bowel summary score declined transiently at 1 month (mean change, -9.8) (Table 3). In addition, a second more protracted decline occurred between 6 and 18 months (mean change from baseline at 12 months, -3.5). Bowel declines at 1 month and 12 months were statistically significant (p < 0.0001); however, only the 1 month change met the threshold for clinically significant change (MID = 4.6) (Figure 2). The EPIC bowel summary score returned to near-baseline at two years post-SBRT (mean change from baseline, -1.6). Individual bowel symptoms changed with time in a similar manner (Figure 1). + + Disease + + MID + + + MESH:D051271 + Disease + + bowel symptoms + + + + TABLE + Tab4.xml + Tab4 + table_caption + 19692 + Overall bowel bother following SBRT for prostate cancer (Patient-reported responses to Question 7 of the EPIC-26) + + 9606 + Species + + Patient + + + MESH:D011471 + Disease + + prostate cancer + + + + TABLE + Tab4 + table + Tab4.xml + 19808 + Start 1 3 6 9 12 18 24 No problem 76.0% 44.6% 68.0% 69.1% 63.7% 63.5% 66.1% 70.4% Very small-small problem 19.9% 43.8% 28.9% 26.9% 30.2% 29.5% 29.5% 26.3% Moderate-big problem 4.1% 11.5% 3.1% 4.0% 6.0% 7.0% 4.5% 3.3% p value - <0.0001 0.1044 0.0828 0.0051 0.0005 0.0172 0.3499 N= 267 260 256 249 248 244 224 240 + + + FIG + 13014_2014_277_Fig3_HTML.jpg + Fig3 + fig_caption + 20163 + Overall bowel bother score (baseline and following SBRT; Question 7 of EPIC-26). Thresholds for clinically significant changes in scores (1/2 standard deviation above and below the baseline) are marked with dashed lines. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. + + + RESULTS + paragraph + 20484 + Overall bowel bother showed a similar pattern as the bowel summary score. At baseline, 24.0% of patients reported some level of annoyance due to bowel symptoms with 4.1% of patients feeling that bowel function was a moderate to big problem (Table 4). The mean EPIC bowel bother score was 90.7 at baseline (Table 3). Overall bowel bother worsened post-treatment and the mean score decreased to 75 at 1 month (mean change, -14.8) (Table 3, Figure 3). However, only 11.5% of patients felt that their bowel function was a moderate to big problem at 1 month following SBRT (Table 4). Bowel bother scores worsened over a second, more protracted time period (Figure 3). Once again, only the first decline met the threshold for clinically significant change (MID = 9.8). Only 7% of patients felt that bowel symptoms were a moderate to big problem at 12 month following treatment (Table 4). By two years following SBRT, overall bowel bother returned to near baseline with a bowel bother score of 89.8 (mean change from baseline, -0.9) (Table 3) and 3.3% of patients feeling bowel symptoms were a moderate to big problem (Table 4). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + Disease + + MID + + + MESH:D015212 + Disease + + bowel function + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D051271 + Disease + + feeling bowel symptoms + + + MESH:D015212 + Disease + + bowel function + + + + DISCUSS + title_1 + 21622 + Discussion + + + DISCUSS + paragraph + 21633 + Proctitis following prostate cancer radiotherapy is a critical quality of life issue and the principle dose-limiting toxicity. Currently, there is limited data on proctitis following SBRT for prostate cancer. A better understanding of bowel symptoms following SBRT would enable clinicians to provide more realistic expectations to patients as they weigh their treatment options. Our prior paper reported the prevalence, severity and overall incidence of bowel frequency/urgency, rectal pain and rectal bleeding following SBRT. However, it is reliant on physician reporting which may under report the incidence of bowel symptoms and provides no information on the associated bother. In this study, we utilized the EPIC-26 bowel domain to evaluate bowel symptoms but also assess related bother. + + 9606 + Species + + patients + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D010146 + Disease + + pain + + + MESH:D064420 + Disease + + toxicity + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011349 + Disease + + Proctitis following prostate cancer + + + MESH:D011349 + Disease + + proctitis + + + MESH:D051271 + Disease + + bowel symptoms + + + MESH:D006470 + Disease + + bleeding + + + + DISCUSS + paragraph + 22426 + Stereotactic body radiation therapy (SBRT) offers to minimize radiation-associated rectal toxicity by reducing the volume of rectum receiving high radiation doses. The overall incidence of post-SBRT rectal bleeding in this series was 23%. The low rate of late Grade >= 2 rectal bleeding (1.5%) seen in this study is consistent with the results from a prior review by our institution and results from other institutions which all report a rate of late Grade >= 2 rectal bleeding of < 5%. In fact, many of the bleeds were acute and transient suggestive of acute anal irritation or hemorrhoid exacerbation due to bowel frequency. Furthermore, the incidence of telangiectasia in patients who completed post-SBRT rectal endoscopy for the assessment of rectal bleeding or cancer screening was 11% which is significantly lower than the rates reported in prospective studies that looked at endoscopic outcomes after conventionally fractionated radiation therapy. + + 9606 + Species + + patients + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006484 + Disease + + hemorrhoid + + + MESH:D013684 + Disease + + telangiectasia + + + MESH:D009369 + Disease + + cancer + + + MESH:D006470 + Disease + + bleeding + + + MESH:D001523 + Disease + + irritation + + + MESH:D064420 + Disease + + toxicity + + + MESH:D006470 + Disease + + bleeding + + + + DISCUSS + paragraph + 23395 + These findings are particularly significant given that rectal bleeding is one of the principle dose-limiting toxicities, and is thus a potential barrier to administering radiation at appropriately therapeutic levels. In the work by Sanda et al, the proportion of patients reporting rectal bleeding as a moderate-big problem on the EPIC 26 survey progressively increased over time (reaching 5% at 12 and 24 months), ultimately manifesting as a late complication post-treatment. However, we observed that with SBRT rectal bleeding occurred at lower rates and presented as a relatively earlier complication: the incidence of patients reporting rectal bleeding as a moderate-big problem in this study reached a peak at 9 months at 1.6%, and by 24 months the incidence dropped to null. Additionally, in our series the rate of Grade 2-3 late rectal bleeding was 1.5%. This compares favorably to the 5%-15% seen with contemporary high-dose IMRT. We speculate that this reassuring profile with SBRT may be a result of the enhanced accuracy with fiducial tracking and narrowed target volumes, thus enhancing our efforts to spare normal tissue from inadvertent irradiation. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D064420 + Disease + + toxicities + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + late rectal bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + + FIG + 13014_2014_277_Fig4_HTML.jpg + Fig4 + fig_caption + 24562 + Radar plots showing the distribution of individual symptom bother following SBRT for prostate cancer. EPIC scores range from 0-100 with higher values representing a more favorable health-related QOL. Points further out from the center indicate higher levels of bother with a given symptom. + + MESH:D011471 + Disease + + prostate cancer + + + + DISCUSS + paragraph + 24855 + The pattern seen in bowel QOL after SBRT in our study is similar to the pattern seen after conventionally fractionated radiotherapy, proton therapy or brachytherapy. The bowel QOL score is at its lowest 1 or 2 months after treatment, but improves slowly thereafter to near baseline by 1-2 years after treatment. In our series, most moderate to big problems were seen at 1 month post-SBRT with approximately 10% of patients reporting moderate to big problems with bowel urgency and/or frequency (Figure 4). This compares favorably with 15-19% incidence of moderate to severe bowel urgency and frequency seen two months after conventionally fractionated external beam radiation therapy or brachytherapy. Unlike conventional modalities, this increase in bowel bother was transient and returned to near baseline by 3 months post-SBRT. A second transient increase in bowel bother occurred at 12 months with approximately 7% of patients reporting moderate to big problems. Unlike the pattern seen with conventional modalities, post-SBRT bowel bother returned to near baseline by two years. The mean bowel summary score change from baseline to 24 months in this study was -1.6. This change compares favorably to that seen at 24 months with conventionally fractionated IMRT and proton therapy -7.4 and -3.7, respectively. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel QOL + + + MESH:D015212 + Disease + + bowel urgency + + + MESH:D015212 + Disease + + bowel QOL + + + + DISCUSS + paragraph + 26176 + The present study has several identifiable limitations. The patient population was derived from a single high-volume institution cohort that can limit the translation of our work to the general population. Nevertheless, our work utilized a fairly large patient population with excellent follow-up rates, and was heterogeneous with respect to ethnicity and risk stratification, and considered key patient characteristics such as comorbidity, body mass index, and use of anticoagulants and hormonal therapy. It is conceivable, however, that institutions without the same experience may have a learning curve before they can achieve similar results. Also noteworthy is that our analysis was conducted without a concurrent comparator arm and thus must be carefully weighed against previously published work. Reassuringly, given that the observed toxicity rates in the acute phase are within range of previous studies, the superior toxicity profile at two years follow-up with SBRT compared to other modalities offers intriguing insight to guide subsequent trials for a more comprehensive assessments of SBRT-related proctitis in the future. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D064420 + Disease + + toxicity + + + MESH:D011349 + Disease + + proctitis + + + MESH:D064420 + Disease + + toxicity + + + + CONCL + title_1 + 27313 + Conclusions + + + CONCL + paragraph + 27325 + In this single institution cohort, the rate and severity of proctitis observed following SBRT is low. QOL tended to improve with longer follow-up and was near baseline at two years post-SBRT. Our results compare favorably to those reported for patients treated with alternative radiation modalities. Future prospective randomized studies are needed to confirm these observations. + + 9606 + Species + + patients + + + MESH:D011349 + Disease + + proctitis + + + + CONCL + title_2 + 27705 + Consent + + + CONCL + paragraph + 27713 + This study was a retrospective review of prospectively collected data that was approved by the Georgetown University Institutional Review Board. + + + ABBR + title + 27858 + Abbreviations + + + ABBR + paragraph + 27872 + ADT + + - + Chemical + + ADT + + + + ABBR + paragraph + 27876 + Androgen deprivation therapy + + + ABBR + paragraph + 27905 + CT + + + ABBR + paragraph + 27908 + Computed tomography + + + ABBR + paragraph + 27928 + CTC + + + ABBR + paragraph + 27932 + Common toxicity criteria + + MESH:D064420 + Disease + + toxicity + + + + ABBR + paragraph + 27957 + CTV + + + ABBR + paragraph + 27961 + Clinical target volume + + + ABBR + paragraph + 27984 + DVH + + - + Chemical + + DVH + + + + ABBR + paragraph + 27988 + Dose-volume histogram + + + ABBR + paragraph + 28010 + EQD2 + + + ABBR + paragraph + 28015 + Equivalent dose in 2-Gy fractions + + + ABBR + paragraph + 28049 + EPIC + + + ABBR + paragraph + 28054 + Expanded prostate index composite + + + ABBR + paragraph + 28088 + GTV + + - + Chemical + + GTV + + + + ABBR + paragraph + 28092 + Gross target volume + + + ABBR + paragraph + 28112 + Gy + + - + Chemical + + Gy + + + + ABBR + paragraph + 28115 + Gray + + + ABBR + paragraph + 28120 + IMRT + + + ABBR + paragraph + 28125 + Intensity modulated radiation therapy + + + ABBR + paragraph + 28163 + IRB + + + ABBR + paragraph + 28167 + Internal review board + + + ABBR + paragraph + 28189 + PTV + + - + Chemical + + PTV + + + + ABBR + paragraph + 28193 + Planning target volume + + + ABBR + paragraph + 28216 + QoL + + + ABBR + paragraph + 28220 + Quality of life + + + ABBR + paragraph + 28236 + MID + + Disease + + MID + + + + ABBR + paragraph + 28240 + Minimally important difference + + + ABBR + paragraph + 28271 + MR + + + ABBR + paragraph + 28274 + Magnetic resonance + + + ABBR + paragraph + 28293 + NCI + + + ABBR + paragraph + 28297 + National cancer institute + + MESH:D009369 + Disease + + cancer + + + + ABBR + paragraph + 28323 + SD + + MESH:D029461 + Disease + + SD + + + + ABBR + paragraph + 28326 + Standard deviation + + + ABBR + paragraph + 28345 + SBRT + + + ABBR + paragraph + 28350 + Stereotactic body radiation therapy + + + ABBR + paragraph + 28386 + VRS + + + ABBR + paragraph + 28390 + Vienna rectoscopy score + + + ABBR + footnote + 28414 + Daniel Y Joh and Leonard N Chen contributed equally to this work. + + MESH:C536108 + Disease + + Leonard N + + + + ABBR + footnote + 28480 + Competing interests + + + ABBR + footnote + 28500 + SP Collins and BT Collins serve as clinical consultants to Accuray Inc. The Department of Radiation Medicine at Georgetown University Hospital receives a grant from Accuray to support a research coordinator. The other authors declare that they have no competing interests. + + + ABBR + footnote + 28773 + Authors' contributions + + + ABBR + footnote + 28798 + DJ and LC are lead authors, who participated in data collection, data analysis, manuscript drafting, table/figure creation, and manuscript revision. GP participated in data analysis and manuscript drafting. AB, SS, RM, and JK aided in the quality of life data collection and maintaining the patient database. TY aided in clinical data collection. SL is the dosimetrist who developed the majority of patients' treatment plans, and contributed to the dosimetric data analysis and interpretation. BC and AJ participated in the design and coordination of the study. SS is a senior author who collected the dosimetric data, participated in its analysis, and helped draft the manuscript. JC, AD, and JL are senior authors who aided in drafting the manuscript. SC was the principal investigator who initially developed the concept of the study and the design, aided in data collection, drafted and revised the manuscript. 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Anderson randomized dose-escalation trial for prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Aird;given-names:EG + surname:Graham;given-names:JD + surname:Sydes;given-names:MR + surname:Dearnaley;given-names:DP + 10.1016/S1470-2045(07)70143-2 + 17482880 + 2007 + Lancet Oncol + ref + REF + 487 + 475 + 8 + surname:Moore;given-names:AR + surname:Scrase;given-names:CD + surname:Stephens;given-names:RJ + surname:Morgan;given-names:RC + surname:Russell;given-names:JM + surname:Syndikus;given-names:I + surname:Millar;given-names:J + surname:Parmar;given-names:MK + surname:Matthews;given-names:JH + surname:Jose;given-names:CC + surname:Huddart;given-names:RA + surname:Cowan;given-names:RA + surname:Bottomley;given-names:D + 31256 + Escalated-dose versus standard-dose conformal radiotherapy in prostate cancer: first results from the MRC RT01 randomised controlled trial + + CVCL:D923 + CellLine + + MRC RT01 + + + MESH:D011471 + Disease + + prostate cancer + + + + surname:van Leenders;given-names:GJ + surname:Heemsbergen;given-names:WD + surname:van Putten;given-names:WL + surname:Al-Mamgani;given-names:A + 980 + 10.1016/j.ijrobp.2008.02.073 + 18495377 + 2008 + Int J Radiat Oncol Biol Phys + ref + 72 + REF + surname:Lebesque;given-names:JV + 988 + surname:Incrocci;given-names:L + surname:Dielwart;given-names:MF + surname:Slot;given-names:A + 31395 + Update of Dutch multicenter dose-escalation trial of radiotherapy for localized prostate cancer + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Geinitz;given-names:H + surname:Becker;given-names:G + surname:Tomicek;given-names:B + surname:Goldner;given-names:G + 78 + 10.1016/j.ijrobp.2006.08.055 + 17189064 + 2007 + Int J Radiat Oncol Biol Phys + ref + 67 + surname:Feldmann;given-names:H + REF + surname:Potter;given-names:R + surname:Potzi;given-names:R + surname:Molls;given-names:M + surname:Bamberg;given-names:M + surname:Glocker;given-names:S + surname:Reibenwein;given-names:J + 83 + surname:Wachter-Gerstner;given-names:N + surname:Zimmermann;given-names:F + surname:Wachter;given-names:S + 31491 + Proctitis after external-beam radiotherapy for prostate cancer classified by Vienna Rectoscopy Score and correlated with EORTC/RTOG score for late rectal toxicity: results of a prospective multicenter study of 166 patients + + MESH:D011349 + Disease + + Proctitis + + + MESH:D064420 + Disease + + toxicity + + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Tucker;given-names:SL + surname:Jackson;given-names:A + surname:Gay;given-names:H + surname:Michalski;given-names:JM + S123 + 10.1016/j.ijrobp.2009.03.078 + 20171506 + 2010 + Int J Radiat Oncol Biol Phys + ref + 76 + REF + S129 + surname:Deasy;given-names:JO + 31714 + Radiation dose-volume effects in radiation-induced rectal injury + + MESH:D012002 + Disease + + rectal injury + + + + 19 + surname:Wylie;given-names:JP + REF + surname:Bayman;given-names:NA + 302 + 10.1016/j.clon.2007.03.005 + 17448647 + 2007 + Clin Oncol (R Coll Radiol) + ref + 307 + 31779 + When should the seminal vesicles be included in the target volume in prostate radiotherapy? + + + surname:Morton;given-names:G + surname:Franssen;given-names:E + surname:Mah;given-names:K + surname:Dawson;given-names:LA + 1155 + 10.1016/S0360-3016(98)00265-X + 9869243 + 1998 + Int J Radiat Oncol Biol Phys + ref + 42 + REF + 1161 + 31871 + Target position variability throughout prostate radiotherapy + + + surname:Bosch;given-names:WR + surname:Michalski;given-names:JM + surname:Dong;given-names:L + surname:Tucker;given-names:SL + 390 + 10.1016/j.ijrobp.2011.11.073 + 22342302 + 2012 + Int J Radiat Oncol Biol Phys + ref + 84 + REF + surname:Mohan;given-names:R + 395 + surname:Purdy;given-names:JA + surname:Cox;given-names:JD + surname:Winter;given-names:K + 31932 + Do intermediate radiation doses contribute to late rectal toxicity? 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surname:Chen;given-names:V + surname:Ju;given-names:AW + 32859 + Stereotactic body radiation therapy (SBRT) for clinically localized prostate cancer: the Georgetown University experience + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Ashley;given-names:R + 8 + surname:Diblasio;given-names:F + surname:Santoro;given-names:M + REF + surname:Katz;given-names:AJ + 118 + 10.1186/1748-717X-8-118 + 23668632 + 2013 + Radiat Oncol + ref + 32981 + Stereotactic body radiotherapy for localized prostate cancer: disease control and quality of life at 6 years + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Wang;given-names:P + surname:Fuller;given-names:D + surname:Collins;given-names:SP + surname:King;given-names:CR + 939 + 10.1016/j.ijrobp.2013.08.019 + 24119836 + 2013 + Int J Radiat Oncol Biol Phys + ref + 87 + REF + 945 + surname:Katz;given-names:A + surname:Steinberg;given-names:M + surname:Kupelian;given-names:P + 33091 + Health Related Quality of Life after Stereotactic Body Radiotherapy for Localized Prostate Cancer: Results from a Multi-Institutional Consortium of Prospective Trials + + MESH:D011471 + Disease + + Localized Prostate Cancer + + + + surname:Fuller;given-names:D + surname:Kaplan;given-names:I + surname:Freeman;given-names:D + surname:King;given-names:CR + 217 + 10.1016/j.radonc.2013.08.030 + 2013 + Radiat Ther Oncol + ref + 109 + surname:Katz;given-names:A + REF + surname:Steinberg;given-names:M + surname:Kupelian;given-names:P + surname:Wang;given-names:J + 221 + surname:Meier;given-names:R + surname:Collins;given-names:SP + surname:Bolzicco;given-names:G + 33258 + Stereotactic Body Radiotherapy for Localized Prostate Cancer: Pooled Analysis of Multi-Insitutional Prospective Trials + + MESH:D011471 + Disease + + Localized Prostate Cancer + + + + surname:Presti;given-names:JC;suffix:Jr + surname:Gill;given-names:H + surname:Brooks;given-names:JD + surname:King;given-names:CR + 877 + 10.1016/j.ijrobp.2010.11.054 + 21300474 + 2011 + Int J Radiat Oncol Biol Phys + ref + 82 + REF + 882 + 33377 + Long-term outcomes from a prospective trial of stereotactic body radiotherapy for low-risk prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Harkins;given-names:B + surname:Dombrowski;given-names:JJ + surname:Wong;given-names:DS + surname:McBride;given-names:SM + 3681 + 10.1002/cncr.26699 + 22170628 + 2012 + Cancer + ref + 118 + REF + surname:Kaplan;given-names:ID + 3690 + surname:Collins;given-names:SP + surname:Hanscom;given-names:HN + surname:Tapella;given-names:P + 33484 + Hypofractionated stereotactic body radiotherapy in low-risk prostate adenocarcinoma: Preliminary results of a multi-institutional phase 1 feasibility trial + + MESH:D011471 + Disease + + prostate adenocarcinoma + + + + surname:Wang;given-names:H + surname:Lei;given-names:S + surname:Suy;given-names:S + surname:Ju;given-names:AW + 30 + 10.1186/1748-717X-8-30 + 23369294 + 2013 + Radiat Oncol + ref + 8 + surname:Dritschilo;given-names:A + REF + surname:Collins;given-names:SP + surname:Lynch;given-names:JH + surname:Kim;given-names:JS + surname:Hanscom;given-names:HN + surname:Uhm;given-names:S + surname:Sherer;given-names:BA + surname:Oermann;given-names:EK + 33640 + Hypofractionated stereotactic body radiation therapy as monotherapy for intermediate-risk prostate cancer. Radiation Oncology + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Hossain;given-names:S + surname:King;given-names:CR + surname:Djajaputra;given-names:D + surname:Xie;given-names:Y + 236 + 10.1016/j.ijrobp.2008.04.051 + 18722274 + 2008 + Int J Radiat Oncol Biol Phys + ref + 72 + REF + 246 + surname:Xing;given-names:L + surname:Ma;given-names:L + 33766 + Intrafractional motion of the prostate during hypofractionated radiotherapy + + + surname:Chen;given-names:V + surname:Oermann;given-names:EK + surname:Piel;given-names:N + surname:Lei;given-names:S + 48 + 10.3389/fonc.2011.00048 + 22655248 + 2011 + Front Oncol + ref + 1 + surname:Collins;given-names:BT + REF + surname:Suy;given-names:S + surname:Collins;given-names:SP + surname:Jha;given-names:R + surname:Dritschilo;given-names:A + surname:Zhang;given-names:G + surname:Yu;given-names:X + surname:Kim;given-names:JS + surname:Hanscom;given-names:HN + surname:Dahal;given-names:KN + surname:Ju;given-names:AW + 33842 + Six-Dimensional Correction of Intra-Fractional Prostate Motion with CyberKnife Stereotactic Body Radiation Therapy + + + surname:Schultz;given-names:D + surname:Malkowicz;given-names:SB + surname:Whittington;given-names:R + surname:D'Amico;given-names:AV + 969 + 10.1001/jama.280.11.969 + 9749478 + 1998 + Jama + ref + 280 + surname:Wein;given-names:A + REF + surname:Beard;given-names:CJ + surname:Kaplan;given-names:I + surname:Renshaw;given-names:AA + 974 + surname:Tomaszewski;given-names:JE + surname:Broderick;given-names:GA + surname:Blank;given-names:K + 33957 + Biochemical outcome after radical prostatectomy, external beam radiation therapy, or interstitial radiation therapy for clinically localized prostate cancer + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Lei;given-names:S + surname:Wang;given-names:H + surname:Ju;given-names:AW + surname:Sood;given-names:S + 197 + 10.1186/1748-717X-8-197 + 23937800 + 2013 + Radiat Oncol + ref + 8 + surname:Collins;given-names:SP + REF + surname:Dritschilo;given-names:A + surname:Lynch;given-names:J + surname:Carroll;given-names:J + surname:Suy;given-names:S + surname:Zhang;given-names:G + surname:Uhm;given-names:S + 34114 + Rectal endoscopy findings following stereotactic body radiation therapy for clinically localized prostate cancer + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Potzi;given-names:R + surname:Goldner;given-names:G + surname:Gerstner;given-names:N + surname:Wachter;given-names:S + 11 + 10.1016/S0167-8140(99)00173-5 + 10719695 + 2000 + Radiother Oncol + ref + 54 + REF + 19 + surname:Potter;given-names:R + surname:Wambersie;given-names:A + 34227 + Endoscopic scoring of late rectal mucosal damage after conformal radiotherapy for prostatic carcinoma + + MESH:D011472 + Disease + + prostatic carcinoma + + + MESH:D009059 + Disease + + mucosal damage + + + + surname:Sandler;given-names:HM + surname:Litwin;given-names:MS + surname:Dunn;given-names:RL + surname:Wei;given-names:JT + 899 + 10.1016/S0090-4295(00)00858-X + 11113727 + 2000 + Urology + ref + 56 + REF + 905 + surname:Sanda;given-names:MG + 34329 + Development and validation of the expanded prostate cancer index composite (EPIC) for comprehensive assessment of health-related quality of life in men with prostate cancer + + 9606 + Species + + men + + + MESH:D011471 + Disease + + prostate cancer + + + MESH:D011471 + Disease + + prostate cancer + + + + 68 + surname:Willis;given-names:GB + surname:Potosky;given-names:AL + REF + surname:Reeve;given-names:BB + 599 + 10.1016/j.urology.2006.03.037 + 16979720 + 2006 + Urology + ref + 603 + 34502 + Should function and bother be measured and reported separately for prostate cancer quality-of-life domains? + + MESH:D011471 + Disease + + prostate cancer + + + + 41 + surname:Wyrwich;given-names:KW + surname:Sloan;given-names:JA + REF + surname:Norman;given-names:GR + 582 + 12719681 + 2003 + Med Care + ref + 592 + 34610 + Interpretation of changes in health-related quality of life: the remarkable universality of half a standard deviation + + + surname:Kwan;given-names:L + surname:Bergman;given-names:J + surname:Gollapudi;given-names:K + surname:Gore;given-names:JL + 1309 + 10.1016/j.juro.2010.06.012 + 20723914 + 2010 + J Urol + ref + 184 + REF + 1315 + surname:Litwin;given-names:MS + surname:Krupski;given-names:TL + 34728 + Correlates of bother following treatment for clinically localized prostate cancer + + MESH:D011471 + Disease + + localized prostate cancer + + + + surname:Brandeis;given-names:JM + surname:Kwan;given-names:L + surname:Gore;given-names:JL + surname:Litwin;given-names:MS + 2239 + 10.1002/cncr.22676 + 17455209 + 2007 + Cancer + ref + 109 + REF + 2247 + surname:Reiter;given-names:RE + surname:Withers;given-names:HR + surname:Lee;given-names:SP + 34810 + Quality of life after surgery, external beam irradiation, or brachytherapy for early-stage prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Diblasio;given-names:F + surname:Ashley;given-names:R + surname:Santoro;given-names:M + surname:Katz;given-names:AJ + 1 + 10.1186/1471-2490-10-1 + 20122161 + 2010 + BMC Urol + ref + 10 + REF + surname:Witten;given-names:M + 34917 + Stereotactic body radiotherapy for organ-confined prostate cancer + + MESH:D011471 + Disease + + prostate cancer + + + + surname:Deodato;given-names:F + surname:Digesu;given-names:C + surname:Massaccesi;given-names:M + surname:Ippolito;given-names:E + 10.1016/j.ijrobp.2011.12.046 + 22361084 + 2012 + Int J Radiat Oncol Biol Phys + ref + REF + e195 + e191 + 83 + surname:Mantini;given-names:G + surname:Cellini;given-names:N + surname:Ingrosso;given-names:M + surname:Pacelli;given-names:F + surname:Valentini;given-names:V + surname:Morganti;given-names:AG + surname:Mattiucci;given-names:GC + surname:Di Lallo;given-names:A + surname:Cuscuna;given-names:D + surname:Cilla;given-names:S + surname:Pirozzi;given-names:GA + surname:Macchia;given-names:G + 34983 + Early proctoscopy is a surrogate endpoint of late rectal toxicity in prostate cancer treated with radiotherapy + + MESH:D011471 + Disease + + toxicity in prostate cancer + + + + surname:de Jong;given-names:DJ + surname:Philippens;given-names:ME + surname:Kristinsson;given-names:J + surname:van Lin;given-names:EN + 799 + 10.1016/j.ijrobp.2006.09.034 + 17161552 + 2007 + Int J Radiat Oncol Biol Phys + ref + 67 + REF + surname:Visser;given-names:AG + 811 + surname:Leer;given-names:JW + surname:Kaanders;given-names:JH + surname:van der Vight;given-names:LP + 35094 + Reduced late rectal mucosal changes after prostate three-dimensional conformal radiotherapy with endorectal balloon as observed in repeated endoscopy + + MESH:D054549 + Disease + + endorectal balloon + + + + surname:Kollmeier;given-names:MA + surname:Yamada;given-names:J + surname:Pei;given-names:X + surname:Spratt;given-names:DE + 686 + 10.1016/j.ijrobp.2012.05.023 + 22795805 + 2013 + Int J Radiat Oncol Biol Phys + ref + 85 + REF + 692 + surname:Zelefsky;given-names:MJ + surname:Cox;given-names:B + 35244 + Long-term survival and toxicity in patients treated with high-dose intensity modulated radiation therapy for localized prostate cancer + + 9606 + Species + + patients + + + MESH:D064420 + Disease + + toxicity + + + MESH:D011471 + Disease + + localized prostate cancer + + + + + 4272824 + + surname:Banks;given-names:Matthew L. + surname:Negus;given-names:S. Stevens + 25439054 + surname:Hutsell;given-names:Blake A. + 52 + 2015 + 4272824 + 10.1016/j.drugalcdep.2014.11.003 + front + 0 + This file is available for text mining. It may also be used consistent with the principles of fair use under the copyright law. + TITLE + Drug Alcohol Depend; 2015 Jan 01; 14652-60. doi:10.1016/j.drugalcdep.2014.11.003 + Cocaine Choice Rhesus monkey Monoamine releaser Generalized matching law Sensitivity to reinforcement + 60 + NIHMS643973 + Hutsell BA, Negus SS, Banks ML + 0 + A generalized matching law analysis of cocaine vs. food choice in rhesus monkeys: Effects of candidate 'agonist-based' medications on sensitivity to reinforcement + + 9544 + Species + + rhesus monkeys + + + MESH:D003042 + Chemical + + cocaine + + + + ABSTRACT + abstract_title_1 + 170 + Background + + + ABSTRACT + abstract + 181 + We have previously demonstrated reductions in cocaine choice produced by either continuous 14-day phendimetrazine and D-amphetamine treatment or removing cocaine availability under a cocaine vs. food choice procedure in rhesus monkeys. The aim of the present investigation was to apply the concatenated generalized matching law (GML) to cocaine vs. food choice dose-effect functions incorporating sensitivity to both the relative magnitude and price of each reinforcer. Our goal was to determine potential behavioral mechanisms underlying pharmacological treatment efficacy to decrease cocaine choice. + + 9544 + Species + + rhesus monkeys + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:C100294 + Chemical + + phendimetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + + ABSTRACT + abstract_title_1 + 783 + Methods + + + ABSTRACT + abstract + 791 + A multi-model comparison approach was used to characterize dose- and time-course effects of both pharmacological and environmental manipulations on sensitivity to reinforcement. + + + ABSTRACT + abstract_title_1 + 969 + Results + + + ABSTRACT + abstract + 977 + GML models provided an excellent fit of the cocaine choice dose-effect functions in individual monkeys. Reductions in cocaine choice by both pharmacological and environmental manipulations were principally produced by systematic decreases in sensitivity to reinforcer price and non-systematic changes in sensitivity to reinforcer magnitude. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + + ABSTRACT + abstract_title_1 + 1318 + Conclusions + + + ABSTRACT + abstract + 1330 + The modeling approach used provides a theoretical link between the experimental analysis of choice and pharmacological treatments being evaluated as candidate 'agonist-based' medications for cocaine addiction. The analysis suggests that monoamine releaser treatment efficacy to decrease cocaine choice was mediated by selectively increasing the relative price of cocaine. Overall, the net behavioral effect of these pharmacological treatments was to increase substitutability of food pellets, a nondrug reinforcer, for cocaine. + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + monoamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D019970 + Disease + + cocaine addiction + + + MESH:D003042 + Chemical + + cocaine + + + + INTRO + title_1 + 1862 + 1. Introduction + + + INTRO + paragraph + 1878 + Preclinical self-administration models provide a measure of abuse-related reinforcing drug effects and have been the most reliable predictor of medication efficacy in clinical settings. In particular, drug self-administration procedures that involve the concurrent availability of an alternative nondrug reinforcer and determine treatment effects on behavior maintained by both drug and nondrug reinforcers may be especially predictive of medication effects. Important among these proposed advantages is the provision of a dependent variable, response allocation between two concurrently available reinforcers, which may be less sensitive to reinforcement-independent rate-altering effects and may more directly evince relative reinforcer value. + + + INTRO + paragraph + 2624 + Previous studies have examined the determinants of drug reinforcement in choice procedures in which subjects choose between drug (e.g., cocaine) and an alternative nondrug reinforcer (e.g., food; for review,). This literature body supports the general conclusion that choice between drug and food is sensitive to the magnitude, price, frequency, and delay dimensions of reinforcers. Although this research suggests that drug vs. food choice is sensitive to the relative value of drug reinforcement, relatively few studies have integrated these results with quantitative theories of operant choice. + + MESH:D003042 + Chemical + + cocaine + + + + INTRO + paragraph + 3222 + The generalized matching law (GML) is a quantitative framework, which predicts that behavior will be allocated among different reinforcers in proportion to their relative value. Relative reinforcer value has been quantified most effectively by the concatenated GML, which predicts that value is determined by a multiplicative combination of each reinforcer dimension listed above. A version of the GML suitable for drug vs. food choice may be written + + - + Chemical + + GML + + + + INTRO + paragraph + 3673 + where B represents behavior allocated to cocaine (Bc) or food (Bf), M represents the magnitude dimension of each reinforcer, and P represents the price dimension (fixed-ratio schedule) of each reinforcer. For cocaine, Mc equals the unit dose (mg/kg/injection) and for food Mf equals the value of food scaled in dose units of cocaine (Mf Mc). On the basis of preliminary model simulations, the proportionality constant was set to 0.032 in the present study; consequently, the magnitude of a 1-g food pellet was set equal to a 0.032-mg/kg-cocaine injection. Furthermore, we have previously demonstrated comparable reinforcing effects of a 0.032-mg/kg cocaine injection and a 1-g food pellet in both progressive-ratio and choice procedures. The present approach of fixing the scaling parameter at a single value is equivalent to previous applications of the GML using a bias parameter to account for choice between qualitatively different reinforcers (e.g.,). The free parameters sm (sensitivity to magnitude) and sp (sensitivity to price) capture the extent of changes in drug vs. food choice with changes in the relative magnitude and price of cocaine vs. food for an individual subject. For example, in the case of strict matching, sm = sp = 1, the proportion of responses allocated to the drug alternative is equal to the multiplicative combination of magnitude and price proportions. Fig. 1 shows cocaine choice dose-effect functions simulated by Eq. (1) with relative reinforcer magnitude and price equal to the values used in the present study. The main focus of these simulations is to demonstrate that the slope and horizontal position of the cocaine choice dose-effect functions are determined independently by sensitivity to reinforcer magnitude and reinforcer price, respectively. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + MESH:D008748 + Chemical + + Mc + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:C000604007 + Chemical + + sp + + + MESH:D003042 + Chemical + + cocaine + + + + INTRO + paragraph + 5469 + The purpose of the present investigation was to determine monoamine releaser treatment effects in a cocaine vs. food choice procedure on sensitivity parameters of the GML. We reanalyzed data from demonstrating reductions in cocaine vs. food choice produced by continuous 14-day D-amphetamine, phendimetrazine, and phenmetrazine treatment. A version of the concatenated GML incorporating the relative magnitude and price of cocaine and food reinforcers was fit to the cocaine choice dose-effect curves of individual monkeys. A multi-model comparison approach was used to evaluate competing hypotheses regarding dose- and time-course effects on sensitivity to reinforcement. For comparison, we also modeled 7-day effects of extinction on cocaine- or food-maintained responding during the choice procedure. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:C100294 + Chemical + + phendimetrazine + + + - + Chemical + + monoamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D010633 + Chemical + + phenmetrazine + + + - + Chemical + + GML + + + - + Chemical + + GML + + + + METHODS + title_1 + 6273 + 2. Methods + + + METHODS + title_2 + 6284 + 2.1. Study design and methods + + + METHODS + paragraph + 6314 + The experimental methods have been previously described in detail in). Briefly, studies were conducted in four-six adult male rhesus monkeys (Macaca mulatta) surgically implanted with a double-lumen catheter inserted in a major vein. The catheter was connected to a fluid swivel attached to the top of each subject's cage and was protected by a custom jacket and stainless steel tether. Animal research and maintenance were conducted according to the 8th edition of the Guide for the Care and Use of Laboratory Animals as adopted and promulgated by the National Institutes of Health (National Academies Press, 2011). Animal facilities were licensed by the United States Department of Agriculture and accredited by the Association for Assessment and Accreditation of Laboratory Animal Care. The Institutional Animal Care and Use Committee approved the research protocol. Monkeys had visual, auditory, and olfactory contact with other monkeys throughout the study. Operant procedures and foraging toys were provided for environmental manipulation and enrichment. Videos were played daily in animal housing rooms to provide additional environmental enrichment. + + 9544 + Species + + rhesus monkeys + + + 9544 + Species + + Macaca mulatta + + + + METHODS + paragraph + 7476 + Experimental sessions were conducted in each monkey's home cage. Monkeys responded in daily 2 h choice sessions (0900-1100 h) that consisted of a five-component concurrent schedule of food pellet and intravenous cocaine availability as described in detail previously. During each component, responses on the left key were reinforced with food (1-g banana-flavored pellets; Test Diets, Richmond, IN) according to a fixed-ratio (FR) 100 schedule, and responses on the right key were reinforced with intravenous cocaine (0-0.1 mg/kg/injection) according to an FR 10 schedule. A response on one key reset the ratio requirement on the alternative key. Each reinforcer delivery was followed by a 3-s timeout during which all stimulus lights were extinguished, and responding had no programmed consequences. During each component, the food key was transilluminated red. The stimulus lights for the cocaine key were flashed on and off in 3 s cycles, and longer flashes were associated with higher unit cocaine doses. Across components of the choice procedure, a different unit cocaine dose was available (0, 0.0032, 0.01, 0.032, and 0.1 mg/kg/injection during components 1-5, respectively) by manipulating the injection volume (0, 0.01, 0.03, 0.1, and 0.3 ml/injection, respectively) delivered via the 'cocaine' pump. Each component was in effect until 10 total reinforcers were earned or 20 min elapsed, whichever occurred first. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + METHODS + paragraph + 8911 + Once cocaine choice was stable, tests sessions were initiated. Subsequently, a 14-day treatment period was initiated during which a test solution was administered via the 'treatment' pump for 23 h/day. The treatment solutions and doses examined were D-amphetamine (0.032-0.1 mg/kg/h), (+)-phendimetrazine (0.32-1.0 mg/kg/h) or (+)-phenmetrazine (0.1-0.32 mg/kg/h). At the conclusion of each 14-day test period, a saline control treatment period was reinstated for at least 5 days and until cocaine choice returned to pretest levels. For the extinction studies, separate 7-day treatment periods were initiated where either cocaine was removed from the syringe or food pellets were removed from the dispenser. All other component-correlated stimuli were retained. + + MESH:D012965 + Chemical + + saline + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:D003913 + Chemical + + D-amphetamine + + + + METHODS + title_2 + 9683 + 2.2. Modeling approach and dose-effect models + + + METHODS + paragraph + 9729 + The dependent measure of interest was the proportion of cocaine choices (number of completed ratios on the cocaine-associated key/total completed ratios) for days 5-7 (7 days) and days 12-14 (14 days) of the continuous 14-day treatments or days 5-7 for the 7-day extinction condition. These data were then plotted as a function of unit cocaine dose. In the modeling presented below, the value of a single food pellet was held constant at 0.032 mg/kg/injection cocaine as in Fig. 1. Changes in response allocation produced by removal of cocaine or food availability were modeled using baseline reinforcer magnitudes. This approach allowed reinforcer sensitivity parameters to account for changes in response allocation and facilitated comparison with pharmacological manipulations. Parameters of Eq. (1) were estimated for individual monkeys by minimizing the residual sum of squares (RSS) using the Solver add-in of Microsoft Excel for Mac 2011. Model fitting was conducted separately for each manipulation. Data from a monkey were not included if (1) the monkey did not complete each dose of a treatment drug and (2) a choice proportion could not be calculated for two or more cocaine doses. The number of data points fit for individual monkeys are included in Table 1. Within a monkey and a treatment drug, a total RSS was computed by summing the RSS across each dose and treatment week. Therefore, if a monkey completed 14-day treatments for each test drug dose, the total RSS was computed from five cocaine choice dose-effect functions. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + RSS + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + RSS + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + RSS + + + + METHODS + paragraph + 11276 + Different versions of Eq. (1) were fit to the data for days 5-7 (7 days) and days 12-14 (14 days) of each continuous 14-day pharmacological treatment to test different hypotheses regarding treatment effects on sensitivity to reinforcement. Thus, different combinations of the free parameters (k) were either allowed to vary or were constrained to determine whether dose had no effect on sensitivity to reinforcement (the Null model, H0), affected sensitivity to both reinforcer dimensions (Full model), or selectively affected sensitivity to a single reinforcer dimension. In the Null model, sensitivity to reinforcer magnitude and price (si) in Eq. (1) were constrained (e.g., k = 2; s(BL) = s(Dose 1) = s(Dose)). The Full model placed no restrictions on estimated sensitivity to reinforcement parameters (k = 10). The Magnitude model allowed sensitivity to reinforcer magnitude (sm(BL) / sm(Dose 1) / sm(Dose 2)) to vary across doses while sensitivity to reinforcer price (sp(BL) sp(Dose 1) sp(Dose 2)) was constrained (k = 6). The Price model constrained sensitivity to reinforcer magnitude (sm(BL) = sm(Dose 1) = sm(Dose 2)) while sensitivity to reinforcer price (sp(BL) / sp(Dose 1) / sp(Dose 2)) was allowed to vary across doses (k = 6). + + MESH:C000604007 + Chemical + + sp + + + MESH:C000604007 + Chemical + + sp + + + MESH:C000604007 + Chemical + + sp + + + MESH:C000604007 + Chemical + + sp + + + + METHODS + paragraph + 12548 + The minimum corrected Akaike information criterion (AICc) score indicates the best model after correcting for the number of free parameters (see Supplement for further explanation). Multi-model comparison was used to characterize dose-effects of pharmacological treatments on sensitivity to reinforcement. The corrected Akaike information criterion (AICc) was used to compare different models and DeltaAICc [i.e., DeltaAICc (H0) - DeltaAICc (H1) > 4.0] was used to select the best model for each monkey. The model-comparison approach is analogous to a significance test; therefore, the parameters of the selected model were treated as a significant effect of a specific experimental manipulation for an individual monkey. Because an alternative model was the selected model in 18/19 cases, we focused on the magnitude of systematic parameter changes below. Cocaine vs. food choice and sensitivity to reinforcement dose-effect curves were constructed by averaging the individual, best-fitting parameter estimates. + + MESH:D003042 + Chemical + + Cocaine + + + + RESULTS + title_1 + 13561 + 3. Results + + + RESULTS + paragraph + 13572 + A summary of the multi-model comparison for all treatment drugs and extinction is provided in Table 1 (see Supplemental Table S1 for raw AICc scores1). Overall, the null model was never the selected model for the group data and only for one experimental manipulation was this model selected for an individual monkey. These results indicate that in all but one instance, the treatment drug or extinction condition altered drug choice sufficiently to infer changes in reinforcer sensitivity. In addition, the selected model always fit the individual monkey and group data well, accounting for at least 97% of the variance. + + + RESULTS + paragraph + 14193 + Fig. 2 shows continuous 14-day D-amphetamine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of D-amphetamine dose. The GML model accurately described the cocaine choice dose-effect functions across each D-amphetamine dose and each treatment week (Table 1). The best-fit model for D-amphetamine effects differed somewhat between monkeys with the Full and Price models being selected. In 5/6 monkeys that were sensitive to pharmacological treatment, however, 0.1 mg/kg/h D-amphetamine produced an approximate 10-fold decrease in sensitivity to reinforcer price. Changes were less systematic across monkeys producing a 1.5-fold increase in sensitivity to reinforcer magnitude. + + - + Chemical + + GML + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + + RESULTS + paragraph + 14959 + Fig. 3 shows continuous 14-day (+)-phendimetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of phendimetrazine dose. The GML model provided an accurate description of the cocaine choice dose-effect functions across each (+)-phendimetrazine dose and each treatment week (Table 1). The best-fit model was consistent for four of the five monkeys and indicated that (+)-phendimetrazine produced a selective decrease in sensitivity to reinforcer price. The best-fit model for the other monkey suggested (+)-phendimetrazine increased sensitivity to reinforcer magnitude during 0.32/mg/kg/h treatment and decreased sensitivity to reinforcer magnitude during 1.0/mg/kg/h treatment. The multi-model comparison suggested that sensitivity to reinforcer price decreased dose-dependently and these decreases were more pronounced after 14 days of treatment. + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:C100294 + Chemical + + phendimetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + + RESULTS + paragraph + 15901 + Fig. 4 shows continuous 14-day (+)-phenmetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of phenmetrazine dose The GML model provided an accurate description of the cocaine choice dose-effect functions across each (+)-phenmetrazine dose and each treatment week (Table 1). The best-fit model for three monkeys was the Full model and for the other three monkeys was the price sensitivity model. Decreases in sensitivity to reinforcer magnitude and price were apparent at 0.32 mg/kg/h (+)-phenmetrazine following 14 treatment days. + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + - + Chemical + + GML + + + MESH:D010633 + Chemical + + phenmetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + + RESULTS + paragraph + 16529 + Fig. 5 shows the effects of extinguishing either cocaine- or food-maintained responding for 7 days on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1). Substituting 0.001 for food or cocaine in the GML model during removal of each reinforcer provided an accurate description of the cocaine choice dose-effect functions (Table 1). The best-fit model was consistent between monkeys (note that the AICc values for full and price models are not significantly different for monkey M1414). Cocaine removal produced a greater than 8-fold decrease in sensitivity to reinforcer price, whereas food removal produced an approximately 2-fold increase in sensitivity to reinforcer price. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + Cocaine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + title_1 + 17251 + 4. Discussion + + + DISCUSS + paragraph + 17265 + The aim of the present investigation was to determine monoamine releaser treatment effects on sensitivity to reinforcement parameters of the GML. A multi-model comparison approach was used to evaluate competing hypotheses regarding dose- and time-course effects on sensitivity to relative reinforcer magnitude and price. There were two main findings. First, the GML models provided excellent fits to the cocaine choice dose-effect functions. In addition, in 18/19 fits to individual monkey data models assuming an effect of a manipulation on sensitivity to reinforcement were superior to the null hypothesis model justifying the use of additional model parameters. Second, pharmacological and environmental manipulations altered cocaine vs. food choice in 18/19 monkeys and did so by decreasing sensitivity to reinforcer price in 16/18 monkeys. + + MESH:D012640 + Disease + + fits + + + - + Chemical + + GML + + + - + Chemical + + monoamine + + + - + Chemical + + GML + + + MESH:D012640 + Disease + + fits + + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + title_2 + 18110 + 4.1. The generalized matching law model - comparison approach + + + DISCUSS + paragraph + 18174 + Previous studies have employed the model comparison approach to determine the specificity of pharmacological and neurobiological manipulations on reinforcement-related processes. The present study applied a well-known model of choice that incorporates various reinforcer dimensions to construct a relative value of the available reinforcers. provided rhesus monkeys a choice between alternatives that differed in both reinforcer magnitude and price, and GML model comparison allowed us to determine which reinforcer dimension(s) were affected by pharmacological and environmental manipulations. Taken together, these studies suggest the model comparison provides a unique approach for testing competing hypotheses regarding the behavioral mechanisms of pharmacological and environmental manipulations. + + 9544 + Species + + rhesus monkeys + + + + DISCUSS + paragraph + 18976 + The results of the present study are consistent with prior reports demonstrating GML framework applicability to drug choice procedures with rhesus monkeys (e.g.,). The estimates of sensitivity to reinforcer magnitude obtained in the present study, however, differ markedly from modal values of sensitivity to reinforcer rate established in earlier reviews (e.g.,). Sensitivity to reinforcer magnitude estimates displayed substantial overmatching, often 5-10-fold greater than in previous reports (cf.,). There are at least two possible reasons for this difference. First, this study employed qualitatively different reinforcers, which may have enhanced discrimination of the choice alternatives. A previous study by, however, reported undermatching (s < 1.0) to reinforcer rate in a cocaine vs. food choice procedure employing concurrent variable interval schedules (see also,). Second, the present procedure employed ratio schedules of reinforcement and an additional reset contingency following changeover responses. Concurrent ratio schedules generally produce exclusive preference for the choice alternative associated with the lower ratio requirement (; see also). In studies where choices are made between qualitatively different reinforcers, preference is determined by the relative magnitude and price of each reinforcer. The present findings are consistent with concurrent ratio schedules producing behavior allocation that maximizes relative reinforcer value. + + 9544 + Species + + rhesus monkeys + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + paragraph + 20448 + Two potential limitations from the present GML analysis should be considered. One limitation is the scaling factor for the food reinforcer was held constant across all monkeys and, therefore, did not permit an assessment of individual differences. Although we chose the scaling factor based on previous studies, exploratory modeling with scaling factors of 0.032 +- 0.5 log units produced similar relative changes in parameter estimates, but provided a poorer fit to the data. A second limitation concerns the approach to modeling the effects of extinction. To model data from extinction and facilitate comparison to the pharmacological manipulations, we used non-zero values to predict the observed nonexclusive preference. Moreover, a single reinforcer magnitude (e.g., 0.001 mg/kg/injection for saline) could not predict the cocaine dose dependency still evident in the cocaine choice dose-effect function after 7 days of cocaine and food removal. + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D012965 + Chemical + + saline + + + + DISCUSS + paragraph + 21399 + Therefore, while this ad hoc approach does not provide insight into the mechanisms by which the component-correlated stimuli maintained dose-dependent cocaine choice, model comparison suggests that the effects of extinction on price sensitivity were similar to the effects of pharmacological manipulations. + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + title_2 + 21706 + 4.2. Effects of pharmacological treatments on sensitivity to reinforcement + + + DISCUSS + paragraph + 21781 + Pharmacological manipulations that decreased cocaine vs. food preference did so mostly by decreasing sensitivity to reinforcer price. Changes in sensitivity to reinforcer magnitude were less consistent across monkeys compared to price changes. When changes in magnitude sensitivity did occur in individual monkeys, magnitude sensitivity decreased suggesting that total reinforcer consumption decreased (see below).) proposed that the sensitivity parameter in the GML might serve as an index of economic substitutability between qualitatively different reinforcers (see also). The cocaine vs. food choice procedure employed in the present study systematically varied unit cocaine price (UP, fixed ratio/dose) by increasing the available cocaine dose and provided a measure of the own-price elasticity of cocaine demand and the cross-price elasticity of food demand. Therefore, changes in sensitivity to reinforcer price observed in the present study may be related to parameter values in behavioral economic models of choice. + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + paragraph + 22806 + To examine the relationship between GML and economic approaches, choice data from each 7-day period of the continuous 14-day D-amphetamine (0.032-0.1 mg/kg/h) treatment are presented in Fig. 6 as demand curves; consumption is plotted as a function of the unit price of cocaine. The continuous curves are the predictions of the GML model and the interpolated unit cocaine price (UP50) at which cocaine and food consumption are predicted to be equal is given in each panel. Consistent with previous demand analyses of cocaine vs. food choice procedures, cocaine consumption was a decreasing function of UP of cocaine. At the end of the 14-day treatment, 0.1-mg/kg/h D-amphetamine produced a ten-fold decrease in UP50 and the GML model predicted an increase in cocaine elasticity and a corresponding decrease in food elasticity. Therefore, according to the substitutability interpretation of GML sensitivity to reinforcement, cocaine and food were imperfect economic substitutes under baseline conditions, and the effects of pharmacological and environmental manipulations increased the elasticity of cocaine preference. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + - + Chemical + + GML + + + - + Chemical + + GML + + + + DISCUSS + paragraph + 23926 + There was concordance among the effects of pharmacological treatment and extinction of cocaine-maintained behavior on sensitivity to reinforcer price. Consistent with previous studies, elasticity of demand for saline was greater than demand for drug. Model comparison suggested that sensitivity to reinforcer magnitude was not altered over the 7 days of each extinction manipulation. This result is consistent with the dose-dependent increase in cocaine choice maintained by the component-correlated stimuli throughout extinction. Taken together, these findings suggest that pharmacological treatments and extinction may affect behavior allocation in the presence of cocaine-associated stimuli through a common behavioral mechanism. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D012965 + Chemical + + saline + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + DISCUSS + paragraph + 24659 + Finally, the extension of the GML presented here is closely related to Shizgal and colleagues' 'reinforcement mountain' model of brain stimulation reinforcement in intracranial self-stimulation (ICSS) procedures. Their approach employs a single-operant matching model along with generalized matching reinforcement sensitivity parameters (see also). An intriguing similarity between the present and Shizgal approaches is that matching law analyses suggest that pharmacological manipulations affecting dopaminergic neurotransmission selectively alter sensitivity to reinforcement. For example, reported that the cocaine-induced increases in low baseline rates of responding maintained by low frequencies of brain stimulation were consistent with a decreased sensitivity to the price of brain stimulation. Cocaine-induced facilitation of ICSS also correlates with self-administration procedures and has been interpreted as an abuse-related effect. In turn, the blunted expression of abuse-related cocaine effects on ICSS produced by chronic D-amphetamine treatment may also be attributable to altered reinforcer price sensitivity, consistent with the effects reported here on cocaine self-administration. + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + Cocaine + + + + DISCUSS + title_2 + 25867 + 4.3. Implications for preclinical behavioral studies assessing candidate anti-drug addiction medication efficacy + + + DISCUSS + paragraph + 25980 + Cocaine addiction continues to present a significant public health problem, and the development of effective medications remains a priority of preclinical drug addiction research. The goal of 'agonist-based' medication research is to develop pharmacotherapies that decrease drug use by identifying compounds that share mechanisms of action with cocaine. In a recent review, Haney and Spealman concluded that preclinical self-administration procedures are the most reliable predictor of clinical medication outcomes (; see also). This conclusion suggests the success of preclinical self-administration procedures to accurately predict medication efficacy in humans resides in the neuropharmacological and behavioral mechanisms of reinforcement. Model comparison results of the present study suggest that chronic D-amphetamine and phendimetrazine treatment may function as economic substitutes that selectively increase cocaine demand elasticity by increasing the effective price. Moreover, when a substitutable non-drug reinforcer is concurrently available during chronic D-amphetamine and phendimetrazine treatment, preference for an alternative therapeutic reinforcer will reciprocally increase. + + 9606 + Species + + humans + + + MESH:C100294 + Chemical + + phendimetrazine + + + MESH:D003042 + Chemical + + Cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:C100294 + Chemical + + phendimetrazine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + SUPPL + title_1 + 27181 + Supplementary Material + + + SUPPL + footnote + 27204 + Supplementary material can be found by accessing the online version of this paper at: http://dx.doi.org/10.1016/j.drugalcdep.2014.11.003. + + + SUPPL + footnote + 27342 + + + + SUPPL + footnote + 27368 + Supplementary material can be found by accessing the online version of this paper at http://dx.doi.org/10.1016/j.drugalcdep.2014.11.003. + + + SUPPL + footnote + 27505 + + + + SUPPL + footnote + 27531 + Contributors + + + SUPPL + footnote + 27564 + Banks, Blough, and Negus designed the original study. Hutsell developed the modeling approach. Hutsell, Banks, and Negus wrote or contributed to the manuscript. All authors critically reviewed the content and approved the final version for publication. + + + SUPPL + footnote + 27817 + + + + SUPPL + footnote + 27843 + Conflicts of interest + + + SUPPL + footnote + 27885 + None of the authors have any conflict of interest to declare. + + + SUPPL + footnote + 27947 + + + + 163 + surname:Woolverton;given-names:WL + surname:Velkey;given-names:AJ + REF + surname:Anderson;given-names:KG + 319 + 12373433 + 2002 + Psychopharmacology (Berl) + ref + 326 + 27973 + The generalized matching law as a predictor of choice between cocaine and food in rhesus monkeys + + MESH:D003042 + Chemical + + cocaine + + + + 11 + surname:Woolverton;given-names:WL + REF + surname:Anderson;given-names:KG + 413 + 11103893 + 2000 + Behav Pharmacol + ref + 420 + 28070 + Concurrent variable-interval drug self-administration and the generalized matching law: a drug-class comparison + + + 122 + surname:Shizgal;given-names:P + REF + surname:Arvanitogiannis;given-names:A + 1126 + 18823168 + 2008 + Behav Neurosci + ref + 28182 + The reinforcement mountain: allocation of behavior as a function of the rate and intensity of rewarding brain stimulation + + + surname:Posadas-Sanchez;given-names:D + surname:Sanabria;given-names:F + surname:Reilly;given-names:MP + surname:Avila;given-names:I + 298 + 19073222 + 2009 + Behav Brain Res + ref + 198 + REF + surname:Castaneda;given-names:E + 305 + surname:Killeen;given-names:P + surname:Banerjee;given-names:N + surname:Chavez;given-names:CL + 28304 + Modeling operant behavior in the Parkinsonian rat + + MESH:D010302 + Disease + + Parkinsonian + + + + 231 + surname:Negus;given-names:SS + surname:Banks;given-names:ML + REF + surname:Bauer;given-names:CT + 2461 + 24408209 + 2014 + Psychopharmacology (Berl) + ref + 2470 + 28354 + The effect of chronic amphetamine treatment on cocaine-induced facilitation of intracranial self-stimulation in rats + + MESH:D000661 + Chemical + + amphetamine + + + MESH:D003042 + Chemical + + cocaine 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between behavioral and pharmacological treatment strategies to decrease cocaine choice in rhesus monkeys + + MESH:D003042 + Chemical + + cocaine + + + + 131 + surname:Negus;given-names:SS + surname:Blough;given-names:BE + REF + surname:Banks;given-names:ML + 204 + 23726979 + 2013b + Drug Alcohol Depend + ref + 213 + 28847 + Effects of 14-day treatment with the schedule III anorectic phendimetrazine on choice between cocaine and food in rhesus monkeys + + MESH:C100294 + Chemical + + phendimetrazine + + + MESH:D003042 + Chemical + + cocaine + + + + 22 + REF + surname:Baum;given-names:WM + 231 + 16811782 + 1974 + J Exp Anal Behav + ref + 242 + 28976 + On two types of deviation from the matching law: bias and undermatching + + + 32 + REF + surname:Baum;given-names:WM + 269 + 501274 + 1979 + J Exp Anal Behav + ref + 281 + 29048 + Matching, undermatching, and overmatching in studies of choice + + + 12 + surname:Rachlin;given-names:HC + REF + surname:Baum;given-names:WM + 861 + 16811415 + 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cocaine and opioid abuse using drug self-administration procedures + + MESH:D009293 + Disease + + opioid abuse + + + MESH:D003042 + Chemical + + cocaine + + + + 45 + REF + surname:McDowell;given-names:JJ + 63 + 3950535 + 1986 + J Exp Anal Behav + ref + 74 + 30469 + On the falsifiability of matching theory + + + 139 + REF + surname:McDowell;given-names:JJ + 1000 + 22946881 + 2013 + Psychol Bull + ref + 30510 + On the theoretical and empirical status of the matching law and matching theory + + + 26 + REF + surname:Miller;given-names:HL + 335 + 16811952 + 1976 + J Exp Anal Behav + ref + 347 + 30590 + Matching-based hedonic scaling in the pigeon + + + 27 + surname:Myers;given-names:LE + REF + surname:Myers;given-names:DL + 203 + 16811977 + 1977 + J Exp Anal Behav + ref + 214 + 30635 + Undermatching: a reappraisal of performance on concurrent variable-interval schedules of reinforcement + + + 33 + surname:Woolverton;given-names:WL + surname:Hedeker;given-names:D + REF + 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surname:Cheung;given-names:TH + 31354 + A new criterion for acquisition of nicotine self-administration in rats + + MESH:D009538 + Chemical + + nicotine + + + + 15 + REF + surname:Rachlin;given-names:H + 249 + 16811511 + 1971 + J Exp Anal Behav + ref + 251 + 31426 + On the tautology of the matching law + + + surname:Green;given-names:L + 4 + surname:Kagel;given-names:J + surname:Battalio;given-names:R + REF + surname:Rachlin;given-names:H + 371 + 1981 + Behav Brain Sci + ref + 388 + 31463 + Maximization theory in behavioral psychology + + + 87 + surname:Battalio;given-names:RC + surname:Kagel;given-names:JH + REF + surname:Rachlin;given-names:H + 355 + 1980 + Psychol Rev + ref + 31508 + Substitutability in time allocation + + + 231 + surname:Shizgal;given-names:P + surname:Conover;given-names:K + REF + surname:Trujillo-Pisanty;given-names:I + 1351 + 2014 + Psychopharmacology (Berl) + ref + 1364 + 31544 + A new view of the effect of dopamine receptor antagonism on operant performance for rewarding brain stimulation in the rat + + + 127 + REF + surname:Woolverton;given-names:WL + 195 + 8912397 + 1996 + Psychopharmacology (Berl) + ref + 203 + 31667 + Intravenous self-administration of cocaine under concurrent VI schedules of reinforcement + + MESH:D003042 + Chemical + + cocaine + + + + 186 + surname:Anderson;given-names:KG + REF + surname:Woolverton;given-names:WL + 99 + 16568283 + 2006 + Psychopharmacology (Berl) + ref + 106 + 31757 + Effects of delay to reinforcement on the choice between cocaine and food in rhesus monkeys + + MESH:D003042 + Chemical + + cocaine + + + + SUPPL + title + 31848 + Appendix A. Supplementary data + + + SUPPL + paragraph + 31879 + Supplementary data associated with this article can be found, in the online version, at http://dx.doi.org/10.1016/j.drugalcdep.2014.11.003. + + + FIG + nihms643973f1.jpg + F1 + fig_caption + 32019 + Simulated cocaine vs. food choice dose-effect functions. (A) The effects of changing sensitivity to magnitude with sensitivity to price held constant at 1.0. (B) The effects of changing sensitivity to price with sensitivity to magnitude held constant at 5.0. Ordinates: proportion of cocaine choices. Abscissae: unit cocaine dose (mg/kg/injection). + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + FIG + nihms643973f2.jpg + F2 + fig_caption + 32368 + Continuous 14-day D-amphetamine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of D-amphetamine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: D-amphetamine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + + FIG + nihms643973f3.jpg + F3 + fig_caption + 33047 + Continuous 14-day (+)-phendimetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of (+)-phendimetrazine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phendimetrazine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:C100294 + Chemical + + (+)-phendimetrazine + + + MESH:D003042 + Chemical + + cocaine + + + + FIG + nihms643973f4.jpg + F4 + fig_caption + 33744 + Continuous 14-day (+)-phenmetrazine treatment effects on (A) and (B) cocaine vs. food choice and (C) and (D) best-fitting parameter estimates of Eq. (1) plotted as a function of (+)-phenmetrazine dose. (A) and (B) Ordinates: obtained and predicted (solid curves) proportion cocaine choice. Abscissae: unit dose cocaine in mg/kg/injection (log scale). (C) and (D) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phenmetrazine dose in mg/kg/h. Model predictions (A) and (B) and parameter estimates (C) and (D) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (C) and (D). + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + FIG + nihms643973f5.jpg + F5 + fig_caption + 34435 + Effect of 7-day cocaine or food removal on (A) cocaine vs. food choice and (B) and (C) best-fitting parameter estimates of Eq. (1) plotted as a function of extinction manipulation. (A) Ordinate: obtained and predicted (solid curves) proportion cocaine choice. Abscissa: unit dose cocaine in mg/kg/injection (log scale). (B) and (C) Ordinates: sensitivity to reinforcer magnitude (sm) or price (sp). Abscissae: (+)-phenmetrazine dose in mg/kg/h. Model predictions (A) and parameter estimates (B) and (C) were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Note different ordinate scales in (B) and (C). + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D010633 + Chemical + + (+)-phenmetrazine + + + MESH:D003042 + Chemical + + cocaine + + + + FIG + nihms643973f6.jpg + F6 + fig_caption + 35087 + Continuous 14-day D-amphetamine treatment effects on demand for cocaine and food. Model predictions were derived from averaging the parameter estimates of the best-fitting model for individual monkeys. Ordinates: normalized consumption (reinforcers earned/total reinforcers). Abscissae: unit price of cocaine (UP = FR/dose). The unit price at which cocaine and food consumption are predicted to be equal is indicated in the insets as UP50. + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003913 + Chemical + + D-amphetamine + + + MESH:D003042 + Chemical + + cocaine + + + MESH:D003042 + Chemical + + cocaine + + + + TABLE + table_footnote + 35527 + Multi-model comparison and selection for individual monkeys. The first column specifies the pharmacological or environmental manipulation. Each sub-column gives the AICc of the Null model and the DeltaAICc of competing models. A DeltaAICc > 4 is significantly better than the Null model. The best model (lowest AICc) is indicated in bold, n is the number of observations and R2 is the proportion of variance accounted for by the best model. + + + TABLE + table_footnote + 35968 + + + + TABLE + table + 35989 + + + + TABLE + T1.xml + T1 + table_caption + 36006 + Multi-model comparison and selection for individual monkeys. The first column specifies the pharmacological or environmental manipulation. Each sub-column gives the AICc of the Null model and the DeltaAICc of competing models. A DeltaAICc > 4 is significantly better than the Null model. The best model (lowest AICc) is indicated in bold, n is the number of observations and R2 is the proportion of variance accounted for by the best model. + + + TABLE + T1 + table + T1.xml + 36447 + Mean M1452 M1473 M1504 M1488 M1478 M1480 M1412 M1414 D-Amphetamine AICc (Null, H0) -428.2 -62.4 -99.4 -239.3 -104.5 -57.0 -63.0 Delta (Full, sm, sp) 602.0 22.4 107.1 -12.9 122.8 262.2 129.0 Delta (Magnitude, sm) -4.4 18.0 -24.0 -4.4 13.3 3.1 -3.2 Delta (Price sp) 473.5 15.1 78.2 -1.7 47.1 291.7 167.0 n 15 25 19 25 25 24 R2 0.999 0.983 0.996 0.999 0.991 0.999 0.999 (+)-Phendimetrazine AICc (Null, H0) -276.7 -67.6 -50.8 -136.8 -70.3 Delta (Full, sm, sp) 271.1 57.8 118.1 172.7 46.9 Delta (Magnitude, sm) 63.5 0.2 131.0 -1.1 -3.3 Delta (Price sp) 298.2 93.2 89.3 184.5 58.8 n 20 23 25 25 R2 0.991 0.999 0.999 0.999 0.977 (+)-Phenmetrazine AICc (Null, H0) -425.8 -106.8 -74.4 -73.8 -104.6 -83.5 Delta (Full, sm, sp) 163.7 30.7 170.5 3.0 128.1 145.5 Delta (Magnitude, sm) 58.2 28.2 4.2 4.7 -4.5 144.4 Delta (Price sp) 203.8 30.4 136.0 14.8 139.9 93.1 n 25 25 25 25 25 R2 0.997 0.982 0.999 0.999 0.999 0.997 Extinction AICc (Null, H0) -234.5 -30.4 -29.1 -45.0 -33.9 Delta (Full, sm, sp) 235.6 150.1 83.2 226.4 21.3 Delta (Magnitude, sm) 13.2 -1.6 -8.4 34.2 3.1 Delta (Price sp) 239.1 157.3 90.5 238.0 18.4 n 15 15 15 15 R2 0.996 0.999 0.999 0.999 0.900 + + MESH:D010633 + Chemical + + (+)-Phenmetrazine + + + MESH:C000604007 + Chemical + + sp + + + + + 4272825 + + surname:Esther;given-names:Charles R.;suffix:Jr + surname:Fulcher;given-names:M. Leslie + surname:Quinney;given-names:Nancy L. + 25101886 + surname:Cholon;given-names:Deborah M. + 246ra96 + 246 + 2014 + 4272825 + 10.1126/scitranslmed.3008680 + front + 6 + This file is available for text mining. It may also be used consistent with the principles of fair use under the copyright law. + TITLE + Sci Transl Med; 2014 Jul 23; 6(246) 246. doi:10.1126/scitranslmed.3008680 + surname:Gentzsch;given-names:Martina + surname:Boucher;given-names:Richard C. + 246ra96 + surname:Randell;given-names:Scott H. + surname:Dokholyan;given-names:Nikolay V. + NIHMS623565 + surname:Das;given-names:Jhuma + Cholon DM, Quinney NL, Fulcher ML, Esther CR, Das J, Dokholyan NV, Randell SH, Boucher RC, Gentzsch M + 0 + Potentiator Ivacaftor Abrogates Pharmacological Correction of DeltaF508 CFTR in Cystic Fibrosis + + 1080 + Gene + 55465 + + CFTR + + + MESH:D003550 + Disease + + Cystic Fibrosis + + + MESH:C545203 + Chemical + + Ivacaftor + + + + ABSTRACT + abstract + 96 + Cystic Fibrosis (CF) is caused by mutations in the CF transmembrane conductance regulator (CFTR). Newly developed "correctors" such as lumacaftor (VX-809) that improve CFTR maturation and trafficking and "potentiators" such as ivacaftor (VX-770) that enhance channel activity may provide important advances in CF therapy. Although VX-770 has demonstrated substantial clinical efficacy in the small subset of patients with a mutation (G551D) that affects only channel activity, a single compound is not sufficient to treat patients with the more common CFTR mutation, DeltaF508. Thus, patients with DeltaF508 will likely require treatment with both correctors and potentiators to achieve clinical benefit. However, whereas the effectiveness of acute treatment with this drug combination has been demonstrated in vitro, the impact of chronic therapy has not been established. In studies of human primary airway epithelial cells, we found that both acute and chronic treatment with VX-770 improved CFTR function in cells with the G551D mutation, consistent with clinical studies. In contrast, chronic VX-770 administration caused a dose-dependent reversal of VX-809-mediated CFTR correction in DeltaF508 homozygous cultures. This result reflected the destabilization of corrected DeltaF508 CFTR by VX-770, dramatically increasing its turnover rate. Chronic VX-770 treatment also reduced mature wild-type CFTR levels and function. These findings demonstrate that chronic treatment with CFTR potentiators and correctors may have unexpected effects that cannot be predicted from short-term studies. Combining of these drugs to maximize rescue of DeltaF508 CFTR may require changes in dosing and/or development of new potentiator compounds that do not interfere with CFTR stability. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + human + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:D003550 + Disease + + Cystic Fibrosis + + + MESH:C545203 + Chemical + + ivacaftor + + + MESH:C569105 + Chemical + + VX-809 + + + MESH:C569105 + Chemical + + lumacaftor + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + INTRO + title_1 + 1872 + Introduction + + + INTRO + paragraph + 1885 + The most common autosomal recessive genetic disease of the Caucasian population in the United States and Europe, cystic fibrosis (CF), is characterized by abnormal epithelial ion transport. Mutations in the CF transmembrane conductance regulator (CFTR) result in loss of CFTR-mediated Cl- and HCO3- transport by secretory and absorptive epithelial cells in multiple organs, including lungs, pancreas, liver, and intestine. In the lung, disturbances of airway surface liquid homeostasis produce thick and viscous mucus that leads to mucus stasis, airway obstruction, persistent infection, inflammation, and a progressive decline in lung function. These features are the hallmarks of CF lung disease and result in limited life expectancy. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + MESH:D001639 + Chemical + + HCO3 + + + MESH:D007239 + Disease + + infection + + + MESH:D003550 + Disease + + CF lung disease + + + MESH:D003550 + Disease + + cystic fibrosis + + + MESH:D000402 + Disease + + airway obstruction + + + MESH:D010190 + Disease + + pancreas + + + MESH:D007249 + Disease + + inflammation + + + MESH:D030342 + Disease + + autosomal recessive genetic disease + + + + INTRO + paragraph + 2626 + In 1989, the identification of the CFTR gene on chromosome 7 and its most common mutation, DeltaF508 CFTR, raised hope for a cure that would address the underlying cause of CF. Intense high-throughput screening approaches over the last decade have yielded compounds that modulate mutant CFTR function. Small-molecule compounds that rescue mutant CFTR can be assigned to 2 groups: 1) "corrector" compounds that promote maturation and delivery of CFTR proteins to the apical surface and 2) "potentiator" compounds that activate apical CFTR by increasing the open time of the channel. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + c.508DELTA>CFTR + DNAMutation + + DeltaF508 CFTR + + + + INTRO + paragraph + 3213 + The FDA recently approved the CFTR potentiator compound VX-770 (ivacaftor; trade name Kalydeco) as the first drug that directly restores CFTR activity in CF patients who carry a G551D mutation. G551D CFTR reaches the plasma membrane of epithelial cells, but the protein exhibits a gating defect that abolishes ATP-dependent channel opening and causes severe CF. In patients carrying a G551D mutation, VX-770 has proven to be effective in clinical trials, in which treated patients exhibited marked improvements in sweat chloride values and pulmonary function. The development of a CFTR-targeted drug that benefits CF patients marked a breakthrough in the treatment of CF. Unfortunately, because less than 5% of the CF population have the G551D mutation, this specific therapy helps only a limited number of patients. 90% of CF patients carry the DeltaF508 mutation, which produces a protein that does not mature normally and does not traffic to the plasma membrane. VX-770 treatment did not benefit CF subjects with the DeltaF508 mutation, likely because this compound only acts on protein that has trafficked to the plasma membrane. Based on these findings, an attractive therapeutic strategy for the DeltaF508 CF patient population is to promote transfer of the ER-retained DeltaF508 CFTR protein to the plasma membrane using small-molecule corrector compounds. Studies have estimated that the extent of correction in DeltaF508 airway epithelial cells must approximate 10-25% of wild-type (WT) CFTR function to provide therapeutic benefit. In vitro treatment of CF airway epithelial cultures homozygous for the DeltaF508 mutation with the most promising corrector compound, VX-809 (lumacaftor), resulted in CFTR function of ~14% relative to non-CF ("wild-type") human airway epithelial cells. However, administration of VX-809 did not provide a significant therapeutic benefit for DeltaF508 CF patients in recent clinical trials, most likely because DeltaF508 CFTR correction in vivo was less than 10% of wild-type levels, the lower limit of detection, and thus no mature DeltaF508 CFTR protein was observed. Therefore, a logical next step was to combine corrector and potentiator therapies to rescue DeltaF508 and increase protein function. One of the most promising current clinical trials designed to optimize DeltaF508 CFTR function involved the administration of the corrector VX-809 with the potentiator VX-770. Increases in VX-809-rescued DeltaF508 CFTR function have been demonstrated after acute administration of VX-770 in primary human airway epithelial cells from CF patients and human organoids derived from CF (DeltaF508/DeltaF508) intestinal tissue. Surprisingly, chronic co-administration of VX-809 and VX-770 in Phase 2 and 3 studies produced only small improvements in lung function in CF patients homozygous for the DeltaF508 CFTR mutation. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + MESH:C569105 + Chemical + + VX-809 + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:D002712 + Chemical + + chloride + + + MESH:C545203 + Chemical + + VX-770 + + + c.508DELTA>CFTR + DNAMutation + + DeltaF508 CFTR + + + MESH:C545203 + Chemical + + ivacaftor + + + MESH:D003550 + Disease + + benefits CF + + + MESH:C569105 + Chemical + + VX-809 + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C569105 + Chemical + + lumacaftor + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + INTRO + paragraph + 6075 + The aim of this study was to elucidate the molecular mechanism(s) underlying the limited improvement in DeltaF508 CFTR function when a corrector, VX-809 and a potentiator, VX-770 were co-administered to CF patients. We therefore investigated whether there were unexpected effects of chronically exposing CF cultures in vitro to VX-809 and VX-770, as would be achieved by oral dosing in clinical trials. A combination of CFTR bioelectric and biochemical approaches were utilized to investigate this interaction. Human bronchial epithelial (HBE) cells were used for these studies and exposed for 48 hrs to clinically relevant concentrations of both compounds. In addition, because of the success of VX-770 in CF patients with the G551D mutation, it has recently been suggested that treatment with VX-770 may be a pharmacological approach to enhance CFTR function in patients with chronic obstructive pulmonary disease (COPD). Accordingly, similar experimental approaches were utilized to explore the effects of VX-770 on WT CFTR, which matures normally and traffics to the plasma membrane. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + Human + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 1080 + Gene + 55465 + + CFTR + + + MESH:D029424 + Disease + + COPD + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + MESH:C545203 + Chemical + + VX-770 + + + + RESULTS + title_1 + 7163 + Results + + + RESULTS + title_2 + 7171 + Acute and chronic VX-770 treatments rescue G551D CFTR function + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + RESULTS + paragraph + 7234 + It has been recently demonstrated that acute VX-770 administration increased CFTR function in cell lines expressing G551D CFTR and augmented Cl- secretion in primary human bronchial epithelial (HBE) cells derived from CF patients with the G551D mutation on one allele and the DeltaF508 mutation on the other allele. To compare the effects of chronic versus acute VX-770 drug administration in CF airway epithelia with a G551D mutation, we used well-differentiated primary CF HBE cultures (G551D/DeltaF508) as a model. Cultures were treated chronically for 48 hrs with VX-770 or vehicle in the basolateral medium, and then transepithelial short-circuit currents (ISC) were measured in Ussing chambers (Fig. 1). Cultures were exposed to amiloride to inhibit the epithelial Na+ channel (ENaC) and subsequently forskolin to stimulate Cl- secretion by CFTR. As previously reported, acute administration of VX-770 (aVX770) raised Cl- secretion after forskolin administration (Fig. 1A). Chronic VX-770 (cVX770) administration raised forskolin responsiveness but eliminated subsequent responses to acute VX-770 administration (Fig. 1A,B,C, Table S1). Cultures chronically treated with VX-770 exhibited total CFTR-mediated responses (Fig. 1D) and inhibition with CFTRinh-172 (Fig. 1E) equal to cultures treated with forskolin and acute VX-770. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D005576 + Chemical + + forskolin + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + MESH:D000584 + Chemical + + amiloride + + + MESH:D005576 + Chemical + + forskolin + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:D003550 + Disease + + CF airway epithelia + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + RESULTS + paragraph + 8569 + Chronically VX-770-treated G551D/DeltaF508 HBE cultures also exhibited a decrease in amiloride-sensitive currents (Fig. S1A, Table S2), suggesting decreased ENaC function. This finding is consistent with restoration of CFTR-mediated ENaC inhibitory activity because cleavage of ENaC was diminished in chronically VX-770-treated CF cultures (Fig. S1B). The average UTP responsiveness, an index of Ca2+ activated Cl- channel (CaCC) activity, was reduced with chronic as compared to acute VX-770 administration (Fig. S1C, Table S2). + + 1080 + Gene + 55465 + + CFTR + + + 55107 + Gene + 75079 + + Ca2+ activated Cl- channel + + + 55107 + Gene + 75079 + + CaCC + + + MESH:D014544 + Chemical + + UTP + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + MESH:D000584 + Chemical + + amiloride + + + + RESULTS + paragraph + 9099 + In sum, these results demonstrate that both acute and chronic treatment with VX-770 improved CFTR function in HBE cells with the G551D mutation, consistent with clinical studies. + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + RESULTS + title_2 + 9278 + Chronic VX-770 treatment inhibits functional rescue of DeltaF508 CFTR + + + RESULTS + paragraph + 9348 + CF patients harboring the DeltaF508 CFTR mutation, which produces protein maturation and trafficking defects, have little/no CFTR at the cell surface. Consequently, treatment with a corrector compound, such as VX-809, is required for VX-770 to potentiate surface-localized DeltaF508 CFTR. To mimic clinical administration of VX-809 and VX-770 as a combination therapy for the DeltaF508 CF patients, primary CF HBE cultures (DeltaF508/DeltaF508) were treated with both pharmacological agents for 48 hrs and then Ussing chamber experiments were performed to measure DeltaF508 CFTR function (Fig. 2, Table S3). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + c.508DELTA>CFTR + DNAMutation + + DeltaF508 CFTR + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:C569105 + Chemical + + VX-809 + + + + RESULTS + paragraph + 9956 + Anion efflux across the apical epithelial membrane of airway epithelia in response to cAMP is mediated by CFTR and was not substantial in vehicle-treated DeltaF508/DeltaF508 CF HBE cultures Fig. 2A,B; Vehicle). However, VX-809 administration produced DeltaF508 CFTR correction as evidenced by stimulation of Cl- secretion (Fig. 2). Specifically, correction by VX-809 produced responses to forskolin (Fig. 2A, B; VX809) that were enhanced by acute administration of VX-770 (Fig. 2A,C). These data indicate that acutely applied VX-770 further activated ("potentiated") VX-809-rescued DeltaF508 CFTR. However, the CFTR-mediated ISC increase after addition of VX-770 to corrector-rescued DeltaF508 CFTR was transient. The ISC decrease over time may be indicative of a rapidly decreasing quantity of functional protein at the apical membrane. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C569105 + Chemical + + VX-809 + + + - + Chemical + + cAMP + + + MESH:C569105 + Chemical + + VX-809 + + + + RESULTS + paragraph + 10794 + In contrast, rescue of DeltaF508 CFTR function was dramatically decreased in cultures that had been chronically treated with VX-809 and VX-770 compared to VX-809 alone (Fig. 2A: VX809 vs. VX809+VX770). This loss of "corrected" function was reflected in reduced Cl- secretion responses to forskolin (Fig. 2B) and reduced inhibition of stimulated CFTR Cl- secretion with CFTRinh-172 (Fig. 2D). Thus, these data contrast with the significant (P = 0.0177) acute VX-770 responses in VX-809-treated cultures (Fig. 2C). Again, we noticed that the response to UTP-stimulated ISC decreased upon chronic VX-770 treatment (Figs. 2A and S1D). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + MESH:D014544 + Chemical + + UTP + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + MESH:D005576 + Chemical + + forskolin + + + + RESULTS + paragraph + 11430 + We also tested the impact of chronic VX-770 treatment on DeltaF508 correction in CF HBE cultures (DeltaF508/DeltaF508) by corrector compound VX-661. Similar to VX-809-treated CF cells, VX-661-corrected CF HBE cells showed a drastic reduction in forskolin-mediated CFTR function when VX-770 was chronically added (Fig. S2, Table S4). + + 1080 + Gene + 55465 + + CFTR + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C569105 + Chemical + + VX-809 + + + CVCL:0287 + CellLine + + CF HBE + + + + RESULTS + title_2 + 11763 + Chronic VX-770 administration hinders correction by decreasing the stability of corrected DeltaF508 CFTR + + + RESULTS + paragraph + 11868 + To explore the mechanism(s) mediating the VX-770-induced reduction of VX-809-corrected DeltaF508 CFTR function, we used Western blotting techniques to analyze protein maturation and turnover. In normal HBE cells, we detected a mature, complex glycosylated form, band C (Fig. 3A, NL: *), with a substantially greater molecular weight than the immature band B (Fig. 3A, NL: ). In contrast, only band B could be detected in vehicle-treated DeltaF508/DeltaF508 CF HBE cells (Fig. 3A, CF). As previously reported, treatment with VX-809 alone resulted in formation of a modest amount of mature band C in CF HBE cultures, which was not present in vehicle- or VX-770-treated CF cells (Fig. 3A, CF). However, when CF cells were treated chronically with both VX-809 and VX-770, the amount of mature DeltaF508 CFTR was diminished, and instead the DeltaF508 CFTR protein appeared almost exclusively as immature band B. These data suggest that chronic VX-770 treatment impeded correction of DeltaF508 CFTR by VX-809. We investigated the impact of chronic VX-770 treatment on protein stability by measuring the turnover rate of corrected DeltaF508 CFTR. DeltaF508 CFTR was stably expressed in baby hamster kidney (BHK-21) cells and corrected with VX-809 in the presence and absence of VX-770. Rescue with VX-809 was performed at 27 C for 24 hrs because VX-770 prevented VX-809 mediated correction of DeltaF508 CFTR at 37 C (Fig. 3A). After rescue, cells were shifted to 37 C and protein biosynthesis was inhibited by addition of cycloheximide. The amount of remaining mature DeltaF508 CFTR was then measured after 3 and 6 hrs. The turnover rate of rescued DeltaF508 CFTR band C increased and the half-life accordingly decreased by ~2.5 fold in the presence of VX-770 (Fig. 3B,C, Table S5), whereas the decrease in band B levels was not affected by the presence of VX-770 (Fig. 3B). These data clearly show that VX-770 decreased the stability, and thus increased the turnover rate, of VX-809-rescued DeltaF508 CFTR. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 10034 + Species + + hamster + + + CVCL:1914 + CellLine + + BHK-21 + + + MESH:D003513 + Chemical + + cycloheximide + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:C569105 + Chemical + + VX-809 + + + + RESULTS + title_2 + 13870 + VX-770 affects correction of DeltaF508 CFTR in a dose-dependent relationship + + 1080 + Gene + 55465 + + CFTR + + + + RESULTS + paragraph + 13947 + Lower concentrations of VX-770 were chronically administered to DeltaF508/DeltaF508 CF HBE cells to study the dose effect on VX-809-rescued DeltaF508 CFTR (Fig. 4). To obtain an average measure of CFTR-mediated ISC for the period spanning forskolin stimulation to CFTR inhibition, the area under the curve (AUC) was calculated for this interval. Dividing AUC by time yielded average CFTR DeltaISC between activation and inhibition. When 1 microM VX-770 was administered chronically with VX-809, the forskolin responses of CF HBE cells were intermediate between cells treated with VX-809 alone and cells treated with 5 microM VX-770 and VX-809 (Fig. 4A,B, Table S6). There was a significant reduction in AUC in corrector-treated cells with 1 microM versus 5 microM VX-770 (P =0.0049). Although there was not a significant difference in AUC of VX-809-treated cells with 50 nM VX-770 versus VX-809 alone (Fig. 4B), this low dose of VX-770 caused a rapid decline of the slope after forskolin treatment (Fig 4C, Table S7). Chronic treatment with either 50 nM or 1 microM VX-770 eliminated responses to acute VX-770 (Fig. 4A,C). Western blots to detect mature band C protein in CF HBE cells confirmed that VX-809 rescue was inhibited by VX-770 in a dose-dependent manner. As the concentration of VX-770 increased, the amount of VX-809-corrected DeltaF508 CFTR decreased (Fig. 4D, E, Table S8). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D005576 + Chemical + + forskolin + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + + RESULTS + title_2 + 15335 + Chronic VX-770 treatment decreases function of normal (wild-type) CFTR + + + RESULTS + paragraph + 15406 + To investigate the effects of chronic VX-770 treatment on normal (NL) CFTR, we measured anion secretion of NL primary HBE cultures treated with VX-770 for 48 hrs (Fig. 5). Strikingly, administration of 5 microM VX-770 reduced CFTR-mediated Cl- secretion, as reflected by decreased forskolin responses (Fig. 5A,B,C, Table S9) and decreased inhibition of CFTR-mediated current by CFTRinh-172 (Fig. 5A,D). These functional responses were paralleled by a decrease in CFTR band C (Fig. 5E). In cells chronically treated with VX-770, we also observed a consistent reduction in amiloride-sensitive current (Fig. S3A, Table S2) and a substantial inhibition of UTP-sensitive current (Fig. S3B, Table S2) similar to that observed in CF HBE cells (Fig. S1A,C,D, Table S2). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D000584 + Chemical + + amiloride + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D014544 + Chemical + + UTP + + + MESH:C545203 + Chemical + + VX-770 + + + + RESULTS + title_2 + 16168 + Chronic treatment with VX-770 does not alter HBE cell integrity or barrier functions + + + RESULTS + paragraph + 16253 + As a test for the specificity of chronic VX-770 effects, we analyzed whether fundamental epithelial parameters were altered in HBE cultures chronically treated with VX-770 (Fig. 6). The morphology of highly differentiated ciliated HBE cultures was identical in cells treated with vehicle (DMSO) or VX-770 (Fig. 6A). Transepithelial resistance (Rt) of primary HBE cultures was also not affected by chronic VX-770 exposure (Fig. 6B, Table S10). The inhibition by VX-770 of both Na+ absorption (amiloride-sensitive ISC) and Cl- secretion and currents (CFTR- and CaCC-mediated ISC) raised the possibility that driving forces for ion transport, in part generated by Na+/K+ ATPase activity, were perturbed by VX-770. Nystatin, a polyene antibiotic that enables monovalent cations to permeate biological membranes and raise Na+/K+ ATPase activity, did not produce significantly different ISC responses when applied to vehicle- or VX-770-treated HBE cultures (Fig. 6C, Table S10), suggesting intact Na+/K+ ATPase activity. Measurements of intracellular concentrations of VX-809 and VX-770 by mass spectrometry confirmed the presence of these compounds in treated HBE cultures and indicated that cellular VX-809 concentrations were not affected by the presence of VX-770 (Fig. S4, Table S11). + + 1080 + Gene + 55465 + + CFTR + + + 55107 + Gene + 75079 + + CaCC + + + 6267 + Gene + + S11 + + + 6204 + Gene + 788 + + S10 + + + 6204 + Gene + 788 + + S10 + + + MESH:D004121 + Chemical + + DMSO + + + MESH:D011090 + Chemical + + polyene + + + MESH:D000584 + Chemical + + amiloride + + + MESH:D009761 + Chemical + + Nystatin + + + + RESULTS + title_2 + 17539 + Destabilization by VX-770 is beneficial for G551D, but not for wild-type, or DeltaF508 CFTR function + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + RESULTS + paragraph + 17640 + A high C:B band ratio indicates normal CFTR protein maturation. Biochemical analysis by Western blotting showed that chronic VX-770 treatment dramatically reduced the amount of mature CFTR in both NL cultures expressing wild-type CFTR (Figs. 5E and 7A) and VX-809-rescued DeltaF508 CFTR in CF HBE cells (Figs. 3A and 4E). Indeed, the C:B band ratio decreased with chronic VX-770 exposure by more than 50% in NL cultures (Fig. 7B, NL, Table S12). In contrast, the levels of mature (band C) G551D CFTR detected in G551D/DeltaF508 CF HBE cultures were not significantly diminished by chronic exposure to 5 microM VX-770, suggesting that G551D was resistant to the destabilizing effects of VX-770 (Fig 7A, G551D/DeltaF508). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 6268 + Gene + + S12 + + + CVCL:0287 + CellLine + + CF HBE + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + CVCL:0287 + CellLine + + CF HBE + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + RESULTS + paragraph + 18360 + To explore the relationship between VX-770, VX-809, and CFTR protein stability, we performed calculations of thermodynamic stability of CFTR protein utilizing a structural homology model. This model revealed that CFTR amino acid F508 is located at the nucleotide binding domain 1- cytoplasmic loop 4 (NBD1-CL4) interface (Fig. 7C), and therefore participates in important interdomain interactions. Thus, in DeltaF508 CFTR, the deletion of amino acid F508 not only reduces the stability of the NBD1 domain, but importantly, may destabilize multidomain assembly of CFTR. In contrast, in this structural model of CFTR, amino acid G551 is positioned between the 2 NBDs (Figs. 7C, S5), and the G551D mutation is thought to contort NBD dimer formation and abolish ATP-dependent channel opening by disrupting the signature sequence in NBD1. To evaluate whether the G551D mutation also affects the overall stability of CFTR by inducing conformational restructuring of the protein, we computationally estimated the DeltaDeltaG for G551D CFTR. We found that G551D had a stabilizing effect on the CFTR protein (DeltaDeltaG = -8.1 kcal/mol). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 100862696 + Gene + + CL4 + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + - + Chemical + + amino acid F508 + + + MESH:D000255 + Chemical + + ATP + + + - + Chemical + + amino acid F508 + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + DISCUSS + title_1 + 19490 + Discussion + + + DISCUSS + paragraph + 19501 + Potentiator compounds act on mutant CFTR channels that are on the surface of epithelial cells. VX-770 has been approved as a pharmacological agent to treat CF patients with at least one copy of the G551D mutation. However, the most common mutant protein in CF patients, DeltaF508 CFTR, is not found at the cell surface. DeltaF508 CFTR has a folding defect and is retained in the ER but can be partially rescued by corrector compounds that promote delivery of a small proportion of mutant DeltaF508 proteins to the cell surface. Corrector-rescued DeltaF508 CFTR is reported to have a shorter half-life at the cell surface and exhibits increased thermal inactivation as well as a gating defect when compared to WT CFTR. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + DISCUSS + paragraph + 20219 + The efficacy of orally administered VX-770 was established in clinical trials in G551D CF patients by multiple outcome measurements. The clinical benefit of potentiation of G551D function was predicted from the effectiveness of acute administration of VX-770 in Ussing chambers, which measured rates of Cl- secretion across primary G551D/DeltaF508 CFTR airway epithelial cultures. Our studies confirmed that acute VX-770 administration restored CFTR Cl- secretion activity in HBE cells from patients carrying the G551D mutation. Further, our data demonstrated that G551D/DeltaF508 cultures chronically treated with VX-770 also exhibited increased Cl- secretion via G551D CFTR, but stimulation with forskolin, which raises intracellular cAMP, was required to activate Cl- secretion. This result could suggest a benefit from administering cAMP-raising beta2-adrenergic receptor agonists as a routine part of the treatment for G551D CF patients receiving VX-770. Overall, improvement in G551D CFTR activity with acute VX-770 in vitro was also observed with chronic in vitro VX-770 administration. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D005576 + Chemical + + forskolin + + + - + Chemical + + cAMP + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + - + Chemical + + cAMP + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + DISCUSS + paragraph + 21313 + VX-809 appears to restore approximately 15% of normal function in DeltaF508/DeltaF508 CF HBE cells. However, 10-25% of CFTR function is estimated to be required to overcome CF symptoms. Therefore, a combination of VX-809 with a potentiator compound to further enhance DeltaF508 function may be necessary. Although acute treatment with VX-770 has been reported to enhance VX-809-rescued DeltaF508 activity, our data revealed that chronic application of VX-770 in combination with VX-809 or VX-661 did not. Chronic co-administration of VX-770 with either corrector to DeltaF508/DeltaF508 CF HBE cultures produced Cl- secretory responses that were smaller than responses to corrector alone. Our data suggest that the reduced capacity for Cl- secretion after chronic VX-770/VX-809 exposure reflected an increased turnover rate of corrected DeltaF508 CFTR. The VX-770-induced reduction of DeltaF508 correction observed in primary CF HBE cells was dose-dependent as measured by functional and biochemical approaches. We did not detect alterations in physiological properties of HBE cells that would suggest that toxic effects contributed to CFTR dysfunction after chronic VX-770/VX-809 treatment. Thus, our studies suggest that data describing the effectiveness of acute addition of VX-770 to VX-809-treated DeltaF508 CF HBE cells do not predict the outcome for chronic VX-770/VX-809 administration. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:C569105 + Chemical + + VX-809 + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:C569105 + Chemical + + VX-809 + + + MESH:D006331 + Disease + + dysfunction + + + + DISCUSS + paragraph + 22707 + DeltaF508 CFTR has been shown to exhibit an increased thermodynamic instability of NBD1 and improper assembly of NBD1 into a complex with intracellular loop 4 (ICL4) of the second membrane-spanning domain (MSD2). The recently published data on CFTR domain fragments strongly suggest that VX-809 targets MSD1 of CFTR to suppress folding defects of DeltaF508 CFTR by enhancing interactions among NBD1, MSD1 and MSD2. Thus, VX-809 is predicted to enhance function of DeltaF508 by increasing its stability (Fig. 7D). Importantly, chronic exposure to VX-770 appeared to reverse the stabilization effect of VX-809 in DeltaF508 CFTR. Chronic VX-770 treatment resulted in a severe reduction in rescued DeltaF508 CFTR protein due to destabilization of rescued protein as reflected by a 2.5x increase in turnover rate. In contrast, G551D CFTR was more resistant to destabilization and loss of mature CFTR protein with chronic VX-770 exposure than WT or DeltaF508 CFTR. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + DISCUSS + paragraph + 23666 + Our CFTR computational structural model allows us to speculate how VX-770 may interact with WT or mutant CFTR proteins to alter protein stability. CFTR requires conformational flexibility to function properly. The flexibility and stability of the CFTR protein is finely tuned and precisely balanced, which is a requirement for its ability to function properly as a regulated ion channel. The inherent increase in stability of the G551D protein may render it too rigid and inflexible (stable) for proper channel opening under basal cAMP-stimulated conditions. However, a decrease in stability mediated by VX-770 may render the G551D molecule more flexible and allow it to function as a cAMP-regulated Cl- channel (Fig. 7D). In contrast, destabilization of WT (ideal stability) or VX-809-rescued DeltaF508 CFTR (low stability) by VX-770 bound to the CFTR molecule resulted in decreased WT CFTR function and absent DeltaF508 CFTR function. These considerations reveal that chemical correction of low stability CFTR mutants is complicated and necessitates precision. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + - + Chemical + + cAMP + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + - + Chemical + + cAMP + + + + DISCUSS + paragraph + 24729 + Some airway diseases such as chronic obstructive pulmonary disease (COPD) have been associated with lessened CFTR function and consequently VX-770 has been suggested as a potential therapy. In a study that modeled the effects of VX-770 on COPD patients, reduction of CFTR function was achieved by exposing primary HBE cultures to cigarette smoke extract, and these cultures were subsequently exposed to acute administration of VX-770, which led to augmented CFTR-mediated currents. The finding that acute VX-770 treatment enhanced WT CFTR function contrasts with our finding that chronic treatment with VX-770 reduced WT CFTR function. In addition, forskolin-stimulated ISC in NL HBE cultures chronically treated with VX-770 was not stable over time, as indicated by the downward sloping trace. These functional data, coupled with our observations that the amount of mature WT CFTR was reduced by the continuous presence of VX-770, do not favor VX-770 as a therapy to enhance CFTR function in COPD. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + MESH:D029424 + Disease + + COPD + + + MESH:D029424 + Disease + + COPD + + + CVCL:0287 + CellLine + + NL HBE + + + MESH:C545203 + Chemical + + VX-770 + + + MESH:D029424 + Disease + + COPD + + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + + DISCUSS + paragraph + 25728 + Although potentiation of more rare CFTR mutants with partial defects in CFTR processing was recently detected upon acute VX-770 treatment in Fisher rat thyroid (FRT) cells overexpressing these variants, our data raise the concern that potentiation with chronic VX-770 treatment may not be observed in airway epithelia expressing these rare mutations. To optimize combination therapy for both DeltaF508 and rare CFTR processing mutations, minimizing interference of potentiator with corrector activity is required. One approach may be to finely tune the dosing regimens for potentiator compounds. Studies by Van Goor et al. indicate that the EC50 for acute application of VX-770 differs remarkably in G551D/DeltaF508 (EC50: 236 +- 200 nM) and DeltaF508/DeltaF508 (EC50: 22 +- 10 nM) HBE cultures. These data together with our findings, which demonstrate that inhibition of correction by VX-770 is dose dependent, suggest that drug concentrations are very critical and attempts to optimize potentiator activity on channel function while minimally affecting turnover rate of mutant CFTR should be considered. As a second approach, improved potentiator compounds that do not interfere with DeltaF508 CFTR correction and turnover are needed. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 10116 + Species + + rat + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + + DISCUSS + paragraph + 26965 + After chronic VX-770 treatment, we also observed diminished Cl- secretory responses to additions of the P2Y2 receptor agonist, UTP. UTP-stimulated Cl- secretion is elevated in CF airway epithelia and may compensate for the lack of CFTR function in vivo. Reduction of UTP-stimulated CaCC activity may constitute a disadvantage in CF airways, particularly if insufficient amounts of CFTR have been rescued. Thus, monitoring CaCC activity may be useful in future clinical corrector/potentiator studies. + + 1080 + Gene + 55465 + + CFTR + + + 55107 + Gene + 75079 + + CaCC + + + 1080 + Gene + 55465 + + CFTR + + + 55107 + Gene + 75079 + + CaCC + + + MESH:D014544 + Chemical + + UTP + + + MESH:D003550 + Disease + + CF airway epithelia + + + MESH:D014544 + Chemical + + UTP + + + MESH:D014544 + Chemical + + UTP + + + + DISCUSS + paragraph + 27469 + We observed a decrease in amiloride responses in the presence of chronic VX-770. While the effects in CF cells can be explained by the restoration of CFTR inhibiting activity, the inhibition of ENaC-mediated Na+ absorption in NL HBE cells raises the possibility that VX-770 may have off-target effects. Thus, it may be useful to measure ENaC function in future VX-770 clinical trials. Although a decline in ENaC function may be beneficial for CF patients, diminution of Cl- channels other than CFTR may be a disadvantage for maintaining adequate hydration of CF airways. The recent observation that VX-770 has antimicrobial properties in vitro suggests that it may also display off-target effects in vivo. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + CVCL:0287 + CellLine + + NL HBE + + + MESH:D000584 + Chemical + + amiloride + + + + DISCUSS + paragraph + 28177 + A limitation of our studies is that our experiments were performed in primary HBE cultures and not in vivo. However, primary HBE cultures are a well-established, near-physiologic system that is the most relevant model for studying CFTR function in airway epithelia, the tissue most affected by CF. Drug concentrations, turnover, and formation of metabolites may also differ in vitro and in vivo, which are crucial parameters to consider when extrapolating our data to the clinic. We therefore selected in vitro doses that mimicked clinically measured drug concentrations. For example, 5-day VX-770 treatments with 150 mg or 450 mg (administered as one dose/day) in patients resulted in VX-770 concentrations in blood plasma of 1.4 microg/ml and 5.5 microg/ml, respectively, which are equivalent to ~3.5 microM and ~14 microM. Current clinical trials test VX-770 doses in this range (250 mg taken twice per day). Thus, the concentrations tested in our studies in vitro appear relevant to the clinical experience. + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 29189 + We measured intracellular concentrations of VX-809 and VX-770, which revealed that these compounds (particularly VX-770) accumulated in cells and reached much higher concentrations than in the surrounding media. To determine whether the presence of VX-770 might have a negative impact on the intracellular concentration of VX-809, we obtained measurements of VX-809 concentrations in cell lysates with increasing concentrations of VX-770 and observed that the intracellular concentration of VX-809 was not affected by the presence of VX-770. Thus, comparisons of drug concentrations from in vitro and in vivo tissues may be useful in the future. + + MESH:C545203 + Chemical + + VX-770 + + + MESH:C569105 + Chemical + + VX-809 + + + + DISCUSS + paragraph + 29835 + Because there are no corrector compounds available that provide sufficient rescue of DeltaF508 in CF airways in vivo to alleviate symptoms of CF, potentiation of the small amount of corrected DeltaF508 CFTR is required. However, combination approaches to restore DeltaF508 CFTR function in CF to date have not considered drug-drug interactions of clinically relevant co-administered modulator compounds. Based on our study and the confirmatory data of Veit et al., knowledge of the interactions and interference between corrector and potentiator compounds is essential for successful therapy of the most prevalent mutation in CF patients, most of whom carry at least one allele of the DeltaF508 mutation. Furthermore, understanding the impact that potentiator compounds, such as VX-770, have on the stability of apical WT CFTR may also be important for other airway diseases that would benefit from augmentation of CFTR function. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + + METHODS + title_1 + 30765 + Materials and Methods + + + METHODS + title_2 + 30787 + Study design + + + METHODS + paragraph + 30800 + Simultaneous treatment with small-molecular compounds, VX-809 or VX-661, together with VX-770 (Selleck Chemicals), is currently being examined as therapy for CF patients with the mutation, DeltaF508. Although in vitro studies examining acute treatment with this drug combination have been conducted, we sought to determine the impact of chronic treatment (48 hrs) with these compounds in primary human bronchial epithelial (HBE) cells. Primary HBE cells from normal (NL) or CF patients were obtained from bronchi of human lung tissue, as previously described. To evaluate CFTR function HBE cultures chronically treated with compounds were mounted in Ussing chambers to measure short-circuit currents (ISC). We examined differentiated primary CF HBE cells from at least 3 individuals for each genotype. To visualize the amount of mature and immature CFTR protein from HBE cultures, CFTR from whole-cell lysates was immunoprecipitated as previously described and Western blots were performed. Previously created baby hamster kidney (BHK-21) stably expressing DeltaF508 CFTR, were used in cycloheximide chase studies to examine the rate of protein turnover. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + 9606 + Species + + human + + + 10034 + Species + + hamster + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:1914 + CellLine + + BHK-21 + + + MESH:D003513 + Chemical + + cycloheximide + + + + METHODS + title_2 + 31955 + Cell culture + + + METHODS + paragraph + 31968 + Primary HBE cells were obtained from bronchi of human lung tissue under a protocol approved by the University of North Carolina Medical School Institutional Review Board. Primary NL and CF HBE cells were seeded at passage 2 on collagen-coated Millicell CM inserts (Millipore) and maintained at an air-liquid interface (ALI) at 37 C in 5% CO2 for 3-4 weeks, which allowed the cells to become fully differentiated. + + 9606 + Species + + human + + + CVCL:0287 + CellLine + + CF HBE + + + - + Chemical + + CO2 + + + + METHODS + paragraph + 32384 + BHK-21 cells were obtained from the American Type Culture Collection and grown at 37 C in 5% CO2. BHK-21 cells stably expressing Extope-DeltaF508 CFTR were created previously and maintained as described. + + 101831516 + Gene + + CFTR + + + - + Chemical + + CO2 + + + CVCL:1914 + CellLine + + BHK-21 + + + CVCL:1914 + CellLine + + BHK-21 + + + + METHODS + title_2 + 32588 + Immunoprecipitation and Western blotting + + + METHODS + paragraph + 32629 + Whole-cell lysates were prepared as described previously. CFTR was immunoprecipitated as described previously and isolated using Protein A/G PLUS agarose (Santa Cruz Biotechnology). Samples were separated on 4-20% gradient SDS-PAGE gels (Bio-Rad) and then transferred to nitrocellulose. Blots were probed with mouse monoclonal anti-CFTR antibodies and then with IR Dye 680-goat anti-mouse IgG (Molecular Probes). Anti-actin (Cell Signaling) or anti-tubulin (LI-COR) was used as a loading control. Protein bands were visualized using an Odyssey Infrared Imaging System (LI-COR). + + 1080 + Gene + 55465 + + CFTR + + + 6236 + Gene + 3071 + + Rad + + + 1080 + Gene + 55465 + + CFTR + + + 10090 + Species + + mouse + + + 9925 + Species + + goat + + + 10090 + Species + + mouse + + + MESH:D012967 + Chemical + + SDS + + + MESH:D012685 + Chemical + + agarose + + + + METHODS + title_2 + 33209 + Cycloheximide chase to study turnover of rescued DeltaF508 CFTR + + MESH:D003513 + Chemical + + Cycloheximide + + + + METHODS + paragraph + 33273 + BHK-21 cells expressing Extope-DeltaF508 CFTR were pretreated with compounds (VX-809, 5 microM; VX-770, 5 microM) for 24 hrs at 27 C before treatment with cycloheximide (200 microg/ml; Sigma) in the presence of compounds during chase times at 37 C. Whole-cell lysates were prepared and subjected to Western blotting. + + 1080 + Gene + 55465 + + CFTR + + + CVCL:1914 + CellLine + + BHK-21 + + + MESH:D003513 + Chemical + + cycloheximide + + + MESH:C569105 + Chemical + + VX-809 + + + + METHODS + title_2 + 33590 + Histology and microscopy + + + METHODS + paragraph + 33615 + Primary HBE cultures grown at ALI on Millicell inserts were washed in PBS and fixed in 10% neutral buffered formalin prior to being embedded in paraffin and hematoxylin-eosin stained at the UNC CF Histology Core. Slides were viewed on a Leica DMIRB Inverted Microscope with a 40x 1.0 numerical aperture oil objective. + + MESH:D011535 + Disease + + PBS + + + MESH:D004801 + Chemical + + eosin + + + MESH:D010232 + Chemical + + paraffin + + + MESH:D005557 + Chemical + + formalin + + + MESH:D006416 + Chemical + + hematoxylin + + + + METHODS + title_2 + 33934 + ISC Measurements in Ussing chambers + + + METHODS + paragraph + 33970 + In Ussing chambers (Physiological Instruments) HBE cultures were equilibrated to 37 C in a bilateral bath of Krebs-bicarbonate-Ringer buffer (KBR; in mM: 140 Na+, 120 Cl-, 5.2 K+, 1.2 Ca2+, 1.2 Mg2+, 2.4 HPO42-, 0.4 H2PO4-, 25 HCO3-, and 5 glucose, pH 7.4) and circulated with 95% O2-5% CO2. Short-circuit currents (ISC) were measured as previously described. All NL HBE cultures were measured in bilateral KBR. Unless noted otherwise, CF HBE cultures were measured with high potassium, low chloride (HKLC) buffer applied apically and KBR applied basolaterally, creating a Cl- gradient (5 mM/120 mM). Amiloride (100 microM) was added to block ENaC, followed by forskolin (10 microM), VX-770 (5 microM) and, if applicable, genistein (10 microM) to stimulate CFTR. CFTR was then inhibited with CFTRinh-172 (10 microM) and response to UTP (100 microM) was examined. In some experiments, nystatin (40 microM, apical) was added at the end of the measurements. Data were analyzed using Acquire and Analysis (version 2.3) software (Physiologic Instruments). ISC traces were imported to and processed in Origin 9.0.0. (OriginLab Corporation). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + - + Chemical + + CO2 + + + MESH:D000584 + Chemical + + Amiloride + + + - + Chemical + + Mg2+ + + + CVCL:0287 + CellLine + + CF HBE + + + - + Chemical + + Krebs + + + MESH:D011188 + Chemical + + potassium + + + MESH:D001639 + Chemical + + HCO3 + + + MESH:D014544 + Chemical + + UTP + + + MESH:D019833 + Chemical + + genistein + + + - + Chemical + + HPO42- + + + MESH:D005947 + Chemical + + glucose + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C545203 + Chemical + + VX-770 + + + MESH:D001639 + Chemical + + bicarbonate + + + - + Chemical + + H2PO4 + + + - + Chemical + + HKLC + + + MESH:D000069285 + Chemical + + Ca2+ + + + - + Chemical + + O2 + + + CVCL:0287 + CellLine + + NL HBE + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + MESH:C039004 + Chemical + + KBR + + + MESH:D009761 + Chemical + + nystatin + + + MESH:D002712 + Chemical + + chloride + + + + METHODS + title_2 + 35105 + Detection of VX-770 and VX-809 + + + METHODS + paragraph + 35136 + Mass spectrometric (MS) methods were developed to detect VX-770 and VX-809 using strategies similar to those previously described. VX-770 was detected by monitoring transition of parent to daughter ion of m/z 393.3 171.1 in positive mode MS, with VX-809 detected by monitoring the transition m/z 453.3 197.1. Each compound generated a single peak using previously described liquid-chromatography-tandem MS (LC-MS/MS) methods, with run times of 11.1 and 10.8 minutes, respectively. To quantify drug concentrations in epithelial cells, cell lysates were extracted x 2 with equal volume of MTBE (Sigma), which was then lyophilized to dryness under vacuum centrifugation. Lyophilized samples were resuspended in a volume of 20% methanol in water equal to the original lysate volume, extracted, and 5 microl analyzed by LC-MS/MS as above. To control for matrix effects and variable recovery during extraction, untreated lysates were spiked with known concentrations of VX-770 and VX-809 and extracted in parallel. Concentrations in cell lysates were assessed by examining signal relative to the spiked samples. + + MESH:D014867 + Chemical + + water + + + MESH:D000432 + Chemical + + methanol + + + + METHODS + title_2 + 36244 + Computational stability calculation + + + METHODS + paragraph + 36280 + We computationally estimated the DeltaDeltaG of mutation for G551D mutant CFTR using the Eris suite as described previously. Eris algorithms re-pack the side chains and evaluate the new free energy according to a physical force field upon the substitution of the relevant residue. + + 1080 + Gene + 55465 + + CFTR + + + 340061 + Gene + 18868 + + Eris + + + 340061 + Gene + 18868 + + Eris + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + METHODS + title_2 + 36561 + Statistical analysis + + + METHODS + paragraph + 36582 + Results are presented as means of average response per primary HBE cell donor and error bars are the standard error of the mean (SEM). Statistical analysis was performed by an unpaired two-tailed Student's t test in GraphPad Prism version 6.02 (GraphPad Software). P values of < 0.05 were considered to indicate statistical significance. + + 9606 + Species + + donor + + + + SUPPL + title_1 + 36922 + Supplementary Material + + + SUPPL + footnote + 36945 + Supplementary Materials + + + SUPPL + footnote + 36989 + Supplementary Figures: + + + SUPPL + footnote + 37012 + Fig. S1. Chronic VX-770 treatment alters responses to amiloride and UTP in CF HBE cells. + + MESH:D014544 + Chemical + + UTP + + + MESH:D000584 + Chemical + + amiloride + + + CVCL:0287 + CellLine + + CF HBE + + + + SUPPL + footnote + 37101 + Fig. S2. Chronic VX-770 treatment inhibits functional rescue of DeltaF508 by VX-661. + + + SUPPL + footnote + 37186 + Fig. S3. Chronic VX-770 treatment alters responses to amiloride and UTP in NL HBE cells. + + MESH:D014544 + Chemical + + UTP + + + MESH:D000584 + Chemical + + amiloride + + + CVCL:0287 + CellLine + + NL HBE + + + + SUPPL + footnote + 37275 + Fig. S4. VX-770 and VX-809 concentrations were measured in treated HBE cells. + + + SUPPL + footnote + 37353 + Fig. S5. G551D mutation in the NBD1 stabilizes CFTR protein. + + 1080 + Gene + 55465 + + CFTR + + + + SUPPL + footnote + 37414 + Supplementary Tables: + + + SUPPL + footnote + 37436 + Table S1. VX-770 treatment restores G551D function. + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + SUPPL + footnote + 37488 + Table S2. Chronic VX-770 treatment alters responses to amiloride and UTP. + + MESH:D014544 + Chemical + + UTP + + + MESH:D000584 + Chemical + + amiloride + + + + SUPPL + footnote + 37562 + Table S3. Chronic VX-770 treatment inhibits functional rescue of DeltaF508 by VX-809. + + + SUPPL + footnote + 37648 + Table S4. Chronic VX-770 treatment inhibits functional rescue of DeltaF508 by VX-661. + + + SUPPL + footnote + 37734 + Table S5. VX-770 diminishes biochemical correction by increasing turnover of corrected DeltaF508 CFTR. + + 1080 + Gene + 55465 + + CFTR + + + + SUPPL + footnote + 37837 + Table S6. VX-770-induced hindrance of DeltaF508 correction is dose dependent. + + + SUPPL + footnote + 37915 + Table S7. VX-770 at low dose (50 nM) affects ISC of VX-809-corrected CF HBE cells (DeltaF508/DeltaF508). + + CVCL:0287 + CellLine + + CF HBE + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + + SUPPL + footnote + 38020 + Table S8. VX-770 affects C:B band ratio in CF HBE cultures (DeltaF508/DeltaF508). + + CVCL:0287 + CellLine + + CF HBE + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + + SUPPL + footnote + 38102 + Table S9. Chronic VX-770 treatment decreases the function of wild-type CFTR. + + 1080 + Gene + 55465 + + CFTR + + + + SUPPL + footnote + 38179 + Table S10. Transepithelial resistance and nystatin responses were not altered in HBE cultures chronically treated with VX-770. + + 6204 + Gene + 788 + + S10 + + + MESH:D009761 + Chemical + + nystatin + + + + SUPPL + footnote + 38306 + Table S11. VX-770 and VX-809 concentrations were measured in HBE cell lysates. + + 6267 + Gene + + S11 + + + + SUPPL + footnote + 38385 + Table S12. VX-770 reduces stability of CFTR. + + 1080 + Gene + 55465 + + CFTR + + + 6268 + Gene + + S12 + + + + SUPPL + footnote + 38430 + + + + SUPPL + footnote + 38618 + Author contributions: DMC, NLQ, and MG performed the experiments and analyzed the data. MLF selected and cultured the primary CF and NL HBE cells. MGG, SHR and RCB discussed the overall design of experiments. MGG and DMC wrote the manuscript. CRE measured drug concentrations. JD and NVD determined DeltaDeltaG of the G551D protein. SHR and RCB provided reagents, technical advice and critical evaluation of the manuscript. All authors read and commented on the manuscript. + + MESH:C023025 + Chemical + + DMC + + + CVCL:0287 + CellLine + + NL HBE + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C023025 + Chemical + + DMC + + + + SUPPL + footnote + 39092 + + + + SUPPL + footnote + 39109 + Competing interests: RCB serves as the chairman of the board of Parion Sciences, a company which works on therapies designed to hydrate airway surfaces and also studies CFTR correctors; Parion Sciences does not have a potentiator program. 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transmembrane conductance regulator + + + 1080 + Gene + 55465 + + CFTR + + + + surname:Cui;given-names:L + surname:Aleksandrov;given-names:AA + surname:Kota;given-names:P + surname:He;given-names:L + 536 + 23104983 + 2013 + FASEB J + ref + 27 + REF + 545 + surname:Riordan;given-names:JR + surname:Dokholyan;given-names:NV + surname:Jensen;given-names:T + 43437 + Correctors of DeltaF508 CFTR restore global conformational maturation without thermally stabilizing the mutant protein + + 1080 + Gene + 55465 + + CFTR + + + + surname:Okiyoneda;given-names:T + surname:Xu;given-names:H + surname:Bossard;given-names:F + surname:Rabeh;given-names:WM + 150 + 22265408 + 2012 + Cell + ref + 148 + surname:Roldan;given-names:A + REF + surname:Lukacs;given-names:GL + surname:Konermann;given-names:L + surname:Liu;given-names:Y + surname:di Bernardo;given-names:S + 163 + surname:Du;given-names:K + surname:Mulvihill;given-names:CM + surname:Bagdany;given-names:M + 43556 + Correction of both NBD1 energetics and domain interface is required to restore DeltaF508 CFTR folding and function + + 1080 + Gene + 55465 + + CFTR + + + + surname:Hwang;given-names:TC + 129 + surname:Li;given-names:M + surname:Sohma;given-names:Y + REF + surname:Bompadre;given-names:SG + 285 + 17353351 + 2007 + The Journal of general physiology + ref + 298 + 43671 + G551D and G1349D, two CF-associated mutations in the signature sequences of CFTR, exhibit distinct gating defects + + 1080 + Gene + 55465 + + CFTR + + + p.G1349D;RS#:193922525 + ProteinMutation + + G1349D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + 15 + surname:Dokholyan;given-names:NV + surname:Ding;given-names:F + REF + surname:Yin;given-names:S + 1567 + 18073107 + 2007 + Structure + ref + 1576 + 43785 + Modeling backbone flexibility improves protein stability estimation + + + 4 + surname:Dokholyan;given-names:NV + surname:Ding;given-names:F + REF + surname:Yin;given-names:S + 466 + 17538626 + 2007 + Nature methods + ref + 467 + 43853 + 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Determination of functional half-lives on transfected cells + + 1080 + Gene + 55465 + + cystic fibrosis transmembrane conductance regulator + + + p.508delF + DNAMutation + + delta F508 + + + + surname:Rabeh;given-names:WM + surname:Bagdany;given-names:M + surname:Barriere;given-names:H + surname:Okiyoneda;given-names:T + 805 + 20595578 + 2010 + Science + ref + 329 + REF + surname:Lukacs;given-names:GL + 810 + surname:Young;given-names:JC + surname:Hohfeld;given-names:J + surname:Du;given-names:K + 44090 + Peripheral protein quality control removes unfolded CFTR from the plasma membrane + + 1080 + Gene + 55465 + + CFTR + + + + surname:Lukacs;given-names:GL + 276 + surname:Hu;given-names:W + surname:Benharouga;given-names:M + REF + surname:Sharma;given-names:M + 8942 + 11124952 + 2001 + J Biol Chem + ref + 8950 + 44172 + Conformational and temperature-sensitive stability defects of the delta F508 cystic fibrosis transmembrane conductance regulator in post-endoplasmic reticulum compartments + + 1080 + Gene + 55465 + + cystic fibrosis transmembrane conductance regulator + + + p.508delF + DNAMutation + + delta F508 + + + + surname:Wu;given-names:Y + surname:Cui;given-names:L + surname:Chang;given-names:XB + surname:Gentzsch;given-names:M + 2684 + 15075371 + 2004 + Mol Biol Cell + ref + 15 + REF + surname:Riordan;given-names:JR + 2696 + surname:Pagano;given-names:RE + surname:Choudhury;given-names:A + surname:Ozols;given-names:VV + 44344 + Endocytic trafficking routes of wild type and DeltaF508 cystic fibrosis transmembrane conductance regulator + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:Chen;given-names:L + surname:Jurkuvenaite;given-names:A + surname:Goldstein;given-names:RF + surname:Varga;given-names:K + 555 + 18052931 + 2008 + Biochem J + ref + 410 + REF + surname:Collawn;given-names:JF + 564 + surname:Bebok;given-names:Z + surname:Sorscher;given-names:EJ + surname:Matalon;given-names:S + 44452 + Enhanced cell-surface stability of rescued DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) by pharmacological chaperones + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + cystic fibrosis transmembrane conductance regulator + + + + surname:Jensen;given-names:T + surname:Cui;given-names:L + surname:Kota;given-names:P + surname:Aleksandrov;given-names:AA + 41 + 22406676 + 2012 + J Mol Biol + ref + 419 + surname:Riordan;given-names:JR + REF + surname:Dokholyan;given-names:NV + surname:Aleksandrov;given-names:LA + surname:Gentzsch;given-names:M + 60 + surname:He;given-names:L + surname:Reyes;given-names:S + surname:Alekseev;given-names:AE + 44594 + Allosteric modulation balances thermodynamic stability and restores function of DeltaF508 CFTR + + + surname:Seehausen;given-names:D + surname:Yang;given-names:Z + surname:Protasevich;given-names:I + surname:Wang;given-names:C + 1932 + 20687163 + 2010 + Protein science : a publication of the Protein Society + ref + 19 + surname:Hunt;given-names:JF + REF + surname:Brouillette;given-names:CG + surname:Wetmore;given-names:DR + surname:Spencer Emtage;given-names:J + 1947 + surname:Atwell;given-names:S + surname:Zhao;given-names:X + surname:Skalak;given-names:T + 44689 + Integrated biophysical studies implicate partial unfolding of NBD1 of CFTR in the molecular pathogenesis of F508del cystic fibrosis + + 1080 + Gene + 55465 + + CFTR + + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:Hong;given-names:JS + surname:Tao;given-names:B + surname:Okeyo;given-names:GO + surname:Wang;given-names:W + 41937 + 21965669 + 2011 + J Biol Chem + ref + 286 + REF + 41948 + surname:Kirk;given-names:KL + 44821 + Thermally unstable gating of the most common cystic fibrosis mutant channel (DeltaF508): "rescue" by suppressor mutations in nucleotide binding domain 1 and by constitutive mutations in the cytosolic loops + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:He;given-names:L + surname:Aleksandrov;given-names:LA + surname:Kota;given-names:P + surname:Aleksandrov;given-names:AA + 194 + 20561529 + 2010 + J Mol Biol + ref + 401 + REF + surname:Riordan;given-names:JR + surname:Dokholyan;given-names:NV + 210 + surname:Gentzsch;given-names:M + surname:Cui;given-names:L + surname:Jensen;given-names:T + 45027 + Regulatory insertion removal restores maturation, stability and function of DeltaF508 CFTR + + + 170 + surname:Bridges;given-names:RJ + surname:Frizzell;given-names:RA + REF + surname:Schultz;given-names:BD + 51 + 10398760 + 1999 + The Journal of membrane biology + ref + 66 + 45118 + Rescue of dysfunctional deltaF508-CFTR chloride channel activity by IBMX + + 1080 + Gene + 55465 + + CFTR + + + p.508delF + DNAMutation + + deltaF508 + + + + surname:Jallat;given-names:S + surname:Champigny;given-names:G + surname:Barbry;given-names:P + surname:Dalemans;given-names:W + 526 + 1722027 + 1991 + Nature + ref + 354 + REF + surname:Lazdunski;given-names:M + surname:Lecocq;given-names:JP + surname:Pavirani;given-names:A + 528 + surname:Crystal;given-names:RG + surname:Dreyer;given-names:D + surname:Dott;given-names:K + 45191 + Altered chloride ion channel kinetics associated with the delta F508 cystic fibrosis mutation + + p.508delF + DNAMutation + + delta F508 + + + MESH:D003550 + Disease + + cystic fibrosis + + + + 369 + surname:Plant;given-names:BJ + surname:Murphy;given-names:DM + REF + surname:Harrison;given-names:MJ + 1280 + 24066763 + 2013 + N Engl J Med + ref + 1282 + 45285 + Ivacaftor in a G551D homozygote with cystic fibrosis + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C545203 + Chemical + + Ivacaftor + + + MESH:D003550 + Disease + + cystic fibrosis + + + + 7 + surname:Clancy;given-names:JP + REF + surname:Kotha;given-names:K + 288 + 24004658 + 2013 + Therapeutic advances in respiratory disease + ref + 296 + 45338 + Ivacaftor treatment of cystic fibrosis patients with the G551D mutation: a review of the evidence + + 9606 + Species + + patients + + + MESH:D003550 + Disease + + cystic fibrosis + + + MESH:C545203 + Chemical + + Ivacaftor + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + surname:Hathorne;given-names:H + surname:Hill;given-names:A + surname:Liu;given-names:B + surname:Rowe;given-names:SM + e66955 + 23922647 + 2013 + PLoS One + ref + 8 + surname:Olson;given-names:ER + REF + surname:Clancy;given-names:JP + surname:Stone;given-names:AJ + surname:Ordonez;given-names:CL + surname:Dong;given-names:Q + surname:Accurso;given-names:FJ + surname:Beamer;given-names:JR + surname:Cohen;given-names:M + 45436 + Optimizing Nasal Potential Difference Analysis for CFTR Modulator Development: Assessment of Ivacaftor in CF Subjects with the G551D-CFTR Mutation + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C545203 + Chemical + + Ivacaftor + + + + surname:Skach;given-names:WR + surname:Landstrom;given-names:A + surname:O'Donnell;given-names:N + surname:Liu;given-names:X + 5113 + 22680785 + 2012 + Biochemistry + ref + 51 + REF + 5124 + surname:Dawson;given-names:DC + 45583 + Thermal Instability of DeltaF508 Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Channel Function: Protection by Single Suppressor Mutations and Inhibiting Channel Activity + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + Cystic Fibrosis Transmembrane Conductance Regulator + + + + 86 + surname:Clarke;given-names:DM + surname:Bartlett;given-names:MC + REF + surname:Loo;given-names:TW + 612 + 23835419 + 2013 + Biochem Pharmacol + ref + 619 + 45767 + Corrector VX-809 stabilizes the first transmembrane domain of CFTR + + + surname:Houck;given-names:SA + surname:De La Rosa;given-names:O + surname:Grove;given-names:DE + surname:Ren;given-names:HY + 3016 + 23924900 + 2013 + Mol Biol Cell + ref + 24 + REF + surname:Cyr;given-names:DM + 3024 + surname:Hoffman;given-names:BJ + surname:Van Goor;given-names:F + surname:Sopha;given-names:P + 45834 + VX-809 corrects folding defects in cystic fibrosis transmembrane conductance regulator protein through action on membrane-spanning domain 1 + + 1080 + Gene + 55465 + + cystic fibrosis transmembrane conductance regulator + + + + surname:Hoffman;given-names:BJ + 13 + surname:Burton;given-names:B + surname:Yu;given-names:H + REF + surname:Van Goor;given-names:F + 29 + 23891399 + 2014 + J Cyst Fibros + ref + 36 + 45974 + Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or function + + 1080 + Gene + 55465 + + CFTR + + + MESH:C545203 + Chemical + + ivacaftor + + + + surname:Schreiber;given-names:R + surname:Thomas;given-names:J + surname:Gonska;given-names:T + surname:Mall;given-names:M + 608 + 12612194 + 2003 + Pediatr Res + ref + 53 + REF + surname:Kunzelmann;given-names:K + 618 + surname:Brandis;given-names:M + surname:Kuehr;given-names:J + surname:Seydewitz;given-names:HH + 46090 + Modulation of Ca2+-activated Cl- secretion by basolateral K+ channels in human normal and cystic fibrosis airway epithelia + + 9606 + Species + + human + + + MESH:D003550 + Disease + + cystic fibrosis airway epithelia + + + MESH:D000069285 + Chemical + + Ca2+ + + + + surname:Olsen;given-names:JC + surname:Paradiso;given-names:AM + surname:Loewen;given-names:ME + surname:Tarran;given-names:R + 407 + 12198094 + 2002 + The Journal of general physiology + ref + 120 + REF + surname:Gabriel;given-names:SE + 418 + surname:Boucher;given-names:RC + surname:Argent;given-names:BE + surname:Gray;given-names:MA + 46213 + Regulation of murine airway surface liquid volume by CFTR and Ca2+-activated Cl- conductances + + 12638 + Gene + 55465 + + CFTR + + + 10090 + Species + + murine + + + MESH:D000069285 + Chemical + + Ca2+ + + + + surname:Hardy;given-names:SP + surname:Makhlina;given-names:M + surname:Gabriel;given-names:SE + surname:Thomas;given-names:EJ + C1578 + 11029305 + 2000 + Am J Physiol Cell Physiol + ref + 279 + REF + C1586 + surname:Lethem;given-names:MI + 46309 + Expression of nucleotide-regulated Cl(-) currents in CF and normal mouse tracheal epithelial cell lines + + 10090 + Species + + mouse + + + + surname:Gansemer;given-names:ND + surname:Dohrn;given-names:CL + surname:Abou Alaiwa;given-names:MH + REF + surname:Reznikov;given-names:LR + 2014 + J Cyst Fibros + ref + surname:Welsh;given-names:MJ + surname:Stoltz;given-names:DA + surname:Diekema;given-names:DJ + 46415 + Antibacterial properties of the CFTR potentiator ivacaftor + + 1080 + Gene + 55465 + + CFTR + + + MESH:C545203 + Chemical + + ivacaftor + + + + REF + 203188Orig1s000, http://www.accessdata.fda.gov/drugsatfda_docs/nda/2012/203188Orig1s000OtherRedt.pdf. + Access data FDA + ref + 46474 + + + surname:Phuan;given-names:PW + surname:Perdomo;given-names:D + surname:Avramescu;given-names:RG + surname:Veit;given-names:G + 0 + 2014 + Science Translational Medicine + ref + 0 + surname:Hegedus;given-names:T + REF + surname:Verkman;given-names:AS + surname:Lukacs;given-names:G + surname:Finkbeiner;given-names:WE + surname:Wu;given-names:YS + surname:Szollosi;given-names:D + 0 + surname:Borot;given-names:F + surname:Apaja;given-names:PM + surname:Bagdany;given-names:M + 46475 + Some gating potentiators, including VX-770, diminish DeltaF508-CFTR functional expression + + 1080 + Gene + 55465 + + CFTR + + + + surname:Yankaskas;given-names:JR + surname:Burns;given-names:KA + surname:Gabriel;given-names:S + surname:Fulcher;given-names:ML + 183 + 15492373 + 2005 + Methods in molecular medicine + ref + 107 + REF + 206 + surname:Randell;given-names:SH + 46565 + Well-differentiated human airway epithelial cell cultures + + 9606 + Species + + human + + + + 945 + surname:Randell;given-names:SH + REF + surname:Fulcher;given-names:ML + 109 + 23097104 + 2013 + Methods Mol Biol + ref + 121 + 46623 + Human nasal and tracheo-bronchial respiratory epithelial cell culture + + 9606 + Species + + Human + + + + surname:Gentzsch;given-names:M + surname:Mengos;given-names:A + surname:Gabriel;given-names:SE + surname:Grubb;given-names:BR + 355 + 16284361 + 2006 + Am J Respir Cell Mol Biol + ref + 34 + REF + surname:Boucher;given-names:RC + surname:Riordan;given-names:JR + surname:Drumm;given-names:ML + 363 + surname:Knowles;given-names:MR + surname:Van Heeckeren;given-names:AM + surname:Randell;given-names:SH + 46693 + SERCA pump inhibitors do not correct biosynthetic arrest of deltaF508 CFTR in cystic fibrosis + + 1080 + Gene + 55465 + + CFTR + + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:Tatreau;given-names:JR + surname:Olsen;given-names:JC + surname:Gabriel;given-names:SE + surname:Fulcher;given-names:ML + L82 + 18978040 + 2009 + Am J Physiol Lung Cell Mol Physiol + ref + 296 + REF + surname:Randell;given-names:SH + surname:Salmon;given-names:P + L91 + surname:Saavedra;given-names:MT + surname:Livanos;given-names:E + surname:Gentzsch;given-names:M + 46787 + Novel human bronchial epithelial cell lines for cystic fibrosis research + + 9606 + Species + + human + + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:Cui;given-names:L + surname:Hou;given-names:YX + surname:Mengos;given-names:A + surname:Chang;given-names:XB + 2814 + 18682497 + 2008 + J Cell Sci + ref + 121 + REF + surname:Gentzsch;given-names:M + 2823 + surname:Riordan;given-names:JR + surname:Aleksandrov;given-names:A + surname:Jensen;given-names:TJ + 46860 + Role of N-linked oligosaccharides in the biosynthetic processing of the cystic fibrosis membrane conductance regulator + + - + Chemical + + N-linked oligosaccharides + + + MESH:D003550 + Disease + + cystic fibrosis + + + + surname:Riordan;given-names:JR + surname:Randell;given-names:SH + surname:O'Neal;given-names:WK + surname:Cholon;given-names:DM + L304 + 20008117 + 2010 + Am J Physiol Lung Cell Mol Physiol + ref + 298 + REF + L314 + surname:Gentzsch;given-names:M + 46979 + Modulation of endocytic trafficking and apical stability of CFTR in primary human airway epithelial cultures + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + human + + + + surname:Ansede;given-names:JH + surname:Johnson;given-names:MR + surname:Boucher;given-names:RC + surname:Esther;given-names:CR;suffix:Jr + 76 + 23932897 + 2014 + Pulm Pharmacol Ther + ref + 27 + REF + surname:Hirsh;given-names:AJ + 82 + surname:Ghio;given-names:AJ + surname:O'Riordan;given-names:TG + surname:Donn;given-names:KH + 47088 + Airway drug pharmacokinetics via analysis of exhaled breath condensate + + + surname:Swenberg;given-names:JA + surname:Collins;given-names:LB + surname:Jasin;given-names:HM + surname:Esther;given-names:CR;suffix:Jr + 701 + 18257110 + 2008 + Rapid communications in mass spectrometry : RCM + ref + 22 + REF + 705 + surname:Boysen;given-names:G + 47159 + A mass spectrometric method to simultaneously measure a biomarker and dilution marker in exhaled breath condensate + + + FIG + nihms623565f1.jpg + F1 + fig_caption_title + 47274 + VX-770 treatment restores G551D function + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + FIG + nihms623565f1.jpg + F1 + fig_caption + 47315 + Electrophysiological properties of G551D/DeltaF508 cultures analyzed in Ussing chambers treated chronically with VX-770 (cVX770, 5 microM for 48 hrs) or with vehicle (0.1% DMSO). (A) Representative recording of ISC measured in Ussing chambers. Quantification of response to treatment with (B) forskolin (significant difference between vehicle and cVX770, *P = 0.0009), (C) acute VX-770 (aVX770) (significant difference between vehicle and cVX770, *P = 0.0054), (D) forskolin + aVX770, (E) CFTRinh-172. Primary CF HBE cultures (G551D/DeltaF508) were derived from 3 different patients, 3-4 replicates were performed per patient for a total of 10 measurements per treatment. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D004121 + Chemical + + DMSO + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + + FIG + nihms623565f2.jpg + F2 + fig_caption_title + 47987 + Chronic VX-770 treatment inhibits functional rescue of DeltaF508 + + + FIG + nihms623565f2.jpg + F2 + fig_caption + 48052 + (A) Representative ISC traces of CF HBE cells recorded in Ussing chambers. Primary CF HBE cells (DeltaF508/DeltaF508) were treated with vehicle (DMSO) or VX-809 +/-VX-770 for 48 hrs at 5 microM each. (B) DeltaISC response to forskolin observed in VX-809-treated CF HBE cells (*P = 0.0033, VX809 vs. vehicle) was prevented by chronic VX-770 treatment and significantly different from VX-809-treated cells (#P = 0.0147, VX809 vs. VX809+VX770). (C) CF HBE cells treated with VX-809 responded to acute VX-770 exposure (*P = 0.0177, VX809 vs. vehicle). This response was significantly abrogated in VX-809 + VX-770-treated cells (#P = 0.0031, VX809 vs. VX809+VX770). (D) The response to CFTRinh-172 observed in VX-809-treated cells (*P = 0.0209, VX809 vs. vehicle) was significantly decreased in VX809+VX770-treated cells (#P = 0.0006, VX809 vs. VX809+VX770). Primary HBE cultures (DeltaF508/DeltaF508) were derived from 6 different patients, 2-4 replicates were performed per patient for a total of 15 measurements per condition. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D005576 + Chemical + + forskolin + + + CVCL:0287 + CellLine + + CF HBE + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D004121 + Chemical + + DMSO + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + + FIG + nihms623565f3.jpg + F3 + fig_caption_title + 49077 + VX-770 diminishes biochemical correction by increasing turnover of corrected DeltaF508 CFTR + + + FIG + nihms623565f3.jpg + F3 + fig_caption + 49169 + (A) CFTR Western blot of normal (NL) and CF HBE cultures treated with VX-809 (5 microM) +/-VX-770 (5 microM) for 48 hrs. * indicates the mature, complex glycosylated form of CFTR, band C; indicates the immature band B. (B) Turnover of rescued DeltaF508 in BHK-21 cells. DeltaF508 was rescued at 27 C in the presence of VX-809 +/- VX-770 for 24 hrs. After adding cycloheximide (200 microg/ml, 37 C) cells were lysed at the indicated times and analyzed by Western blotting. (C) Quantification of remaining band C over time, normalized to actin (n = 3). + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + CVCL:1914 + CellLine + + BHK-21 + + + MESH:D003513 + Chemical + + cycloheximide + + + + FIG + nihms623565f4.jpg + F4 + fig_caption_title + 49722 + VX-770-induced hindrance of DeltaF508 correction is dose-dependent + + + FIG + nihms623565f4.jpg + F4 + fig_caption + 49789 + (A) ISC traces of CF HBE cells (DeltaF508/DeltaF508) recorded in Ussing chambers. CF HBE cells were treated as indicated (VX-809: 5 microM, VX-770: 1 or 5 microM) for 48 hrs. (B) CFTR function in VX-809-treated cells decreased as chronic VX-770 concentrations increased. Significant reduction of the area under the curve (AUC)/min calculated from the time period between CFTR stimulation by forskolin and CFTR inhibition by CFTRinh-172 (yields average DeltaIsc (microA/cm2)) was observed in CF cells chronically treated for 48 hrs with VX-809 when compared to VX-809 and 1 microM VX-770, (*P = 0.0352). A further reduction was detected when the chronic VX-770 concentration was increased to 5 microM (#P = 0.0049, VX-809 + 1 microM VX-770 vs. VX-809 + 5 microM VX-770). Primary CF HBE cultures were derived from at least 4 different CF patients; 2-5 replicates were performed per patient for a total of at least 14 measurements per condition. (C) In VX-809-corrected CF HBE cultures (DeltaF508/DeltaF508), the presence of chronic VX-770 at 50 nM caused a significant decline of the slope after forskolin treatment (*P = 0.0353, VX-809 vs. VX-809 + 50 nM VX-770). Primary CF HBE cultures were derived from 4 different patients; 3-5 replicates were performed per patient for a total of at least 15 measurements per condition. (D) Quantification of C:B band ratio in CF HBE cultures (DeltaF508/DeltaF508). CFTR C:B band ratio decreased in CF HBE cells as chronic VX-770 concentrations were increased. The C:B band ratio was significantly reduced in CF cells chronically treated for 48 hrs with VX-809 and 1 microM VX-770 compared to VX-809 alone (*P = 0.0181), and a further reduction was detected when the chronic VX-770 concentration was increased from 1 microM to 5 microM (#P = 0.0151, VX-809 + 1 microM VX-770 vs. VX-809 + 5 microM VX-770). Primary CF HBE cultures (DeltaF508/DeltaF508) from 4 different patients were analyzed. (E) Representative Western blot of CF HBE cells (DeltaF508/DeltaF508) showing decrease of VX-809-corrected DeltaF508 as chronic VX-770 concentrations were increased. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + MESH:D005576 + Chemical + + forskolin + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + CVCL:0287 + CellLine + + CF HBE + + + DeltaF508/DeltaF508 + ProteinMutation + + DeltaF508/DeltaF508 + + + + FIG + nihms623565f5.jpg + F5 + fig_caption_title + 51885 + Chronic VX-770 treatment decreases function of wild-type CFTR + + + FIG + nihms623565f5.jpg + F5 + fig_caption + 51947 + (A) Representative ISC traces of NL HBE cells recorded in Ussing chambers. Cultures were treated with vehicle (DMSO) or 5 microM VX-770 for 48 hrs. HBE cells that were chronically treated with VX-770 showed significantly reduced response to (B,C) forskolin (*P = 0.0198 for forskolin peak and *P = 0.0008 for forskolin plateau) and (D) CFTRinh-172 (*P = 0.0014). Primary HBE cultures were derived from 6 different individuals, 2-4 replicates were performed per individual for a total of 17 measurements per condition. (E) Western blot of HBE cultures treated with VX-809 (5 microM) +/-VX-770 (5 microM) for 48 hrs. Mature CFTR was diminished in HBE cells that were chronically treated with VX770. + + 1080 + Gene + 55465 + + CFTR + + + MESH:D004121 + Chemical + + DMSO + + + MESH:D005576 + Chemical + + forskolin + + + MESH:D005576 + Chemical + + forskolin + + + MESH:C545203 + Chemical + + VX-770 + + + MESH:C482900 + Chemical + + CFTRinh-172 + + + MESH:D005576 + Chemical + + forskolin + + + + FIG + nihms623565f6.jpg + F6 + fig_caption_title + 52644 + Key physiological properties were not altered in chronically VX-770-treated HBE cultures + + + FIG + nihms623565f6.jpg + F6 + fig_caption + 52733 + (A) Microscopy after hematoxylin and eosin (H&E) staining of HBE cultures did not reveal a detectable difference between VX-770- or vehicle-treated cells (bar = 10 m). (B) Transepithelial resistance (Rt) of primary HBE cultures was not altered after chronic treatment with VX-770. (C) Nystatin responses were not significantly different in primary HBE cultures that were treated with vehicle or VX-770 (48 hrs, 5 microM). Nystatin was added to the apical side in Ussing chambers. Primary HBE cultures were derived from 5 different individuals, and 2-4 replicates per individual were performed. + + MESH:D006416 + Chemical + + hematoxylin + + + MESH:D004801 + Chemical + + eosin + + + MESH:D009761 + Chemical + + Nystatin + + + MESH:D009761 + Chemical + + Nystatin + + + + FIG + nihms623565f7.jpg + F7 + fig_caption_title + 53327 + VX-770 reduces stability of CFTR + + + FIG + nihms623565f7.jpg + F7 + fig_caption + 53360 + (A) The amount of mature CFTR was reduced when NL HBE cells were chronically treated with VX-770 (48 hrs, 5 microM). G551D is more stable than NL CFTR and the amount of mature G551D protein in CF cultures (G551D/DeltaF508) was not significantly reduced by 48 hrs treatment with 5 microM VX-770. (B) Quantification of C:B band ratio with chronic treatment of VX-770 at 5 microM. C:B band ratio was significantly decreased in NL cells chronically treated for 48 hrs with 5 microM VX-770, (*P = 0.008) (n = 3, cultures from 3 different NL individuals). The reduction of C:B band ratio in G551D/DeltaF508 cells chronically treated for 48 hrs with 5 microM VX-770 was not statistically significant (n = 3, cultures from 3 different CF patients (G551D/DeltaF508)). (C) Structural model showing positions of G551D and F508 in the CFTR molecule. (D) Illustration representing the proposed relationship between function and stability of CFTR variants. Wild-type CFTR has an intermediate stability that allows for optimal function. G551D CFTR is a more rigid protein that exhibits increased stability compared to wild-type CFTR but lacks sufficient function, presumably due to decreased flexibility. VX-770 decreases G551D CFTR stability and renders it a more flexible protein, resembling the stability and function of wild-type CFTR. However, VX-770 causes destabilization of wild-type CFTR and VX-809-corrected DeltaF508 CFTR, diminishing their function. VX-809 increases the stability of DeltaF508, bringing it closer to resembling wild-type CFTR, but this increased stability is diminished when VX-770 is present. + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 1080 + Gene + 55465 + + CFTR + + + 9606 + Species + + patients + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + MESH:C545203 + Chemical + + VX-770 + + + CVCL:0287 + CellLine + + NL HBE + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + c.551G>D,D;RS#:75527207 + ProteinMutation + + G551D/D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + p.G551D;RS#:75527207 + ProteinMutation + + G551D + + + + + 4272826 + + surname:Reddy;given-names:Poluru L. + surname:Bies;given-names:Juraj + surname:Anastasi;given-names:John + 23910958 + surname:Baron;given-names:Beverly W. + 68 + 1 + 2014 + 4272826 + 10.1016/j.bcmd.2013.07.003 + front + 52 + This file is available for text mining. It may also be used consistent with the principles of fair use under the copyright law. + TITLE + Blood Cells Mol. Dis.; 2014 Jan; 52(1) 68-75. doi:10.1016/j.bcmd.2013.07.003 + BCL6 transgenic mice Cooperating genes Lymphoma development GFI1B EVI5 MYB + surname:Baron;given-names:Joseph M. + surname:Wolff;given-names:Linda + 75 + surname:Wroblewski;given-names:Kristen + surname:Thirman;given-names:Michael J. + NIHMS648290 + surname:Joseph;given-names:Loren + Baron BW, Anastasi J, Bies J, Reddy PL, Joseph L, Thirman MJ, Wroblewski K, Wolff L, Baron JM + 0 + GFI1B, EVI5, MYB:Additional genes that cooperate with the human BCL6 gene to promote the development of lymphomas + + 8328 + Gene + 31223 + + GFI1B + + + 7813 + Gene + 121902 + + EVI5 + + + 4602 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 9606 + Species + + human + + + MESH:D008223 + Disease + + lymphomas + + + + ABSTRACT + abstract + 116 + The BCL6 gene, which is expressed in certain B- and T-cell human lymphomas, is involved with chromosomal rearrangements and mutations in a number of these neoplasms. Lymphomagenesis is believed to evolve through a multi-step accumulation of genetic alterations in these tumors. We used retroviral insertional mutagenesis in transgenic mice expressing the human BCL6 transgene in order to identify genes that cooperate with BCL6 during lymphomatous transformation. We previously reported PIM1 as the most frequently recurring cooperating gene in this model. We now report three newly identified cooperating genes:GFI1B, EVI5, and MYB:that we identified in the lymphomas of retroviral-injected BCL6 transgenic mice (but not in retroviral-injected non-transgenic controls); mRNA and protein expression of GFI1B and EVI5 were decreased in the murine tumors, whereas MYB mRNA and protein expression were increased or decreased. These findings correlated with protein expression in human lymphomas, both B- and T-cell. Improved therapy of lymphomas may necessitate the development of combinations of drugs that target the alterations specific to each neoplasm. + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 18712 + Gene + 11214 + + PIM1 + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + MESH:D009369 + Disease + + neoplasm + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphomas + + + + INTRO + title_1 + 1275 + Introduction + + + INTRO + paragraph + 1288 + It has long been known that chromosomal rearrangements and/or mutations of the BCL6 gene are associated with diffuse large B-cell lymphomas (DLBLs) in humans. BCL6 expression also is known to occur in certain T-cell lymphomas. Because multiple oncogenic hits are believed to accumulate during lymphoma development, we used retroviral insertional mutagenesis in mice containing the human BCL6 transgene in an effort to identify mutated genes that cooperate with BCL6 during lymphoma development. + + 604 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 9606 + Species + + humans + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D016399 + Disease + + T-cell lymphomas + + + + INTRO + paragraph + 1783 + We previously reported that retroviral insertional mutagenesis in mice transgenic for human BCL6 leads to enhanced lymphoma development, and we described the gene proviral integration site for Moloney murine leukemia virus 1 (PIM1) as the most frequently recurring gene cooperating with BCL6 to promote lymphoma development in this model. We now report additional recurring retroviral integration sites that we identified in the lymphomas which developed in retroviral-injected BCL6 transgenic mice but not in lymphomas from retroviral-injected non-transgenic controls. The mRNA and proteins encoded by two of the genes involved, growth factor independent 1B (GFI1B) and ecotropic viral integration site 5 (EVI5), each of which we identified in the lymphomas from three transgenic animals (three B-cell and three T-cell, respectively), showed decreased expression in the lymphomas from transgenic (BCL6-positive) mice as compared with the lymphomas from non-transgenic retroviral injected controls (BCL6-negative) as well as in BCL6-positive (as compared with BCL6-negative) human lymphomas. The mRNA and protein encoded by a third involved gene, myeloblastosis oncogene (MYB), which we also identified in the lymphomas from three transgenic animals (two B-cell, one T-cell) but not in controls, was variably increased or decreased. Although these three genes have been associated with lymphoma development, they have not been reported previously to cooperate with BCL6 in lymphomagenesis. + + 604 + Gene + 7640 + + BCL6 + + + 18712 + Gene + 11214 + + PIM1 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + growth factor independent 1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + ecotropic viral integration site 5 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + myeloblastosis oncogene + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + 11801 + Species + + Moloney murine leukemia virus + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + METHODS + title_1 + 3273 + Materials and methods + + + METHODS + title_2 + 3295 + BCL6 transgenic mice and retroviral insertional mutagenesis + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + + METHODS + paragraph + 3355 + Our transgenic mice (described previously), which express BCL6 constitutively in B- and T-lymphocytes, contain two transgenes: tet-o-BCL6 (human BCL6 cDNA under control of the tetracycline-responsive minimal promoter), and EmuSR-tTA (the tetracycline- transactivating protein under control of the Ig heavy-chain enhancer and the SRalpha promoter). One of our transgenic lines contains four copies of the human BCL6 transgene, the other 20 copies. Under approved institutional protocols, neonatal animals from both transgenic lines and controls (non-transgenic mice of the same background, wild-type or positive for either the EmuSR-tTA or tet-o-BCL6 transgenes, but not both) were injected with 105 infectious units of the retrovirus MOL4070LTR intraperitoneally and followed until the need for euthanasia or natural death. + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D013752 + Chemical + + tetracycline + + + MESH:C010349 + Chemical + + tet + + + MESH:D013752 + Chemical + + tetracycline + + + Disease + + euthanasia + + + + METHODS + title_2 + 4179 + Histology, flow cytometry, inverse PCR, database searches, and real-time quantitative PCR + + + METHODS + paragraph + 4269 + Processing of tissues, FACS analysis, inverse PCR, cloning, sequencing, database searches, total RNA extraction, and real-time RT-PCR methodologies have been described previously. The classification of murine lymphoid neoplasms was according to the Bethesda proposals. Primers included beta-actin as described or glyceraldehyde-3-phosphate dehydrogenase (Real Time Primers, Elkins Park, PA) and primers common to all transcript variants of the murine GFI1B, EVI5, or MYB genes (GFI1B, forward: 5'-CTCTCCAGGCATGGACACTT-3'; reverse: 5'-GACGTGAGTATGCTGCTCCA-3'; EVI5, forward: 5'-CCCATCAAAGTTG AGTCCAG-3'; reverse: 5'-TTCTTCCCCAGAGAATCCAA-3'; MYB, Real Time Primers, forward: 5'-CTGGACAGAAGAGGAGGACA-3'; reverse: 5'-TTGTT CTTCTGGAAGCTCGT-3'). + + 14081 + Gene + 37561 + + FACS + + + 14433 + Gene + 107053 + + glyceraldehyde-3-phosphate dehydrogenase + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 11461 + Gene + 110648 + + beta-actin + + + 10090 + Species + + murine + + + 10090 + Species + + murine + + + MESH:D008223 + Disease + + lymphoid neoplasms + + + + METHODS + title_2 + 5031 + Immunohistochemistry + + + METHODS + paragraph + 5052 + Human lymphoma blocks were retrieved from the surgical pathology archives under an Institutional Review-Board approved protocol. Human and murine tissues were stained with anti-BCL6 as described previously. For additional immunohistochemistry, antigen retrieval was performed in pH 6 solution (Epitomics, Burlingame, CA) in a near boiling waterbath for 40 min. Anti-GFI1B (#HPA007012, for human tissues) and anti-EVI5 (#HPA027339, for human and murine tissues) were affinity-isolated Prestige antibodies produced in rabbit (Sigma-Aldrich, Saint Louis, MO). Anti-GFI1B was diluted 1:20 and anti-EVI5 1:35 for incubation overnight at 4 C. Murine tissues were stained with GFI1B affinity isolated antibody produced in rabbit (Sigma Aldrich, #AV30093), 20 mug/ml, overnight at 4 C. For immunohistochemistry with MYB antibody, human tissues were incubated overnight at 4 C with a 1:20 dilution of a rabbit monoclonal antibody (Epitomics, Burlingame, CA, cat. # 3195-1). Murine tissues were incubated with the same MYB antibody diluted 1:1000 for 1 h at room temperature. For all antibodies, antigen-antibody binding was detected with DAB chromogen, and tissues were counterstained with hematoxylin. Images were taken with a BX41 microscope (Olympus), a DP72 digital camera, and cellSens Standard imaging software (Olympus). + + 12053 + Gene + 7640 + + BCL6 + + + 8328 + Gene + 31223 + + GFI1B + + + 7813 + Gene + 121902 + + EVI5 + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 14582 + Gene + 31223 + + GFI1B + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 9606 + Species + + Human + + + 9606 + Species + + Human + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + 9986 + Species + + rabbit + + + 10090 + Species + + Murine + + + 9986 + Species + + rabbit + + + 9606 + Species + + human + + + 9986 + Species + + rabbit + + + 10090 + Species + + Murine + + + MESH:D008223 + Disease + + lymphoma + + + MESH:C000469 + Chemical + + DAB + + + MESH:D006416 + Chemical + + hematoxylin + + + + METHODS + title_2 + 6377 + Statistical analysis + + + METHODS + paragraph + 6398 + The data from real-time PCR were analyzed by the comparative CT method. Data from the study and control groups were compared by the Wilcoxon rank-sum test. Statistical analyses were performed with Stata software (version 12: StataCorp, College Station, TX). + + + RESULTS + title_1 + 6656 + Results + + + RESULTS + title_2 + 6664 + Lymphoma development in MOL4070LTR-infected mice + + MESH:D008223 + Disease + + Lymphoma + + + + RESULTS + paragraph + 6713 + As previously reported, 92 of 99 transgenic and 32 of 33 control mice infected with retrovirus were analyzable. Lymphomas developed in 24 of 53 animals (45.3%) in the four copy transgenic line (18 T-cell, six B-cell) and in 10 of 39 (25.6%) mice from the 20 copy line (three T-cell, seven B-cell). Additionally, as MOL4070LTR is known to cause lymphomas and leukemias in mice, lymphoid neoplasms also were expected in the control group; 11 of 32 retroviral-injected controls (34.4%) developed lymphomas (5 T-cell, 6 B-cell). The nature of the lymphomas (both B- and T-cell), mouse survival, and murine lymphoma-related mortality rates have been discussed previously. Briefly, the lymphomas usually were aggressive tumors; most of the B-cell lymphomas were mature B-cell, and most T-cell tumors were precursor T-cell lymphoblastic lymphoma/leukemias. The lymphomas from the transgenic mice contained three to 29 insertion sites (average 11), and the lymphomas from the controls contained eight to 20 insertion sites (average 12.7). Only one of the animals studied here (with an insertion site 5' to the GFI1B gene) also had an insertion site 3' to PIM1. + + 14582 + Gene + 31223 + + GFI1B + + + 18712 + Gene + 11214 + + PIM1 + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemias + + + MESH:D007938 + Disease + + leukemias + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphoid neoplasms + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + Lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D001523 + Disease + + aggressive tumors + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + title_2 + 7870 + RNA expression in lymphomas containing retroviral insertions in or near the GFI1B, EVI5, and MYB genes + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + paragraph + 7973 + As previously described, PIM1 was the gene most commonly affected by viral insertion sites in this study. Here we have chosen to study the genes that were the next most commonly affected by viral insertions (after PIM1). The types of lymphomas, insert locations, and relative expression levels following real-time quantitative RT-PCR performed on RNA derived from the lymphomas of transgenic mice as compared with RNA from lymphomas of the same type (B or T) in the same organ from multiple randomly selected non-transgenic controls are shown in Table 1. + + 18712 + Gene + 11214 + + PIM1 + + + 18712 + Gene + 11214 + + PIM1 + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + paragraph + 8528 + The insertion sites near GFI1B were all within 7.9-8.7 kb 5' of the GFI1B gene, and these murine tumors were all large B-cell lymphomas. The transgenic animals were females, ranging in age from 4.2 mos to 7.3 mos (average, 6.2 mos), and included both BCL6 transgenic lines. In each case, relative GFI1B RNA expression level in the lymphomas from transgenic mice was decreased as compared with that in randomly selected B-cell lymphomas from retroviral-injected non-transgenic controls (none of which had inserts in or near GFI1B), see Table 1 (overall decrease, 2.71-fold; mean +- SEM = 0.041 +- 0.018 in the controls as compared with 0.015 +- 0.005 in the transgenics; P < 0.05, Fig. 1a). + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 10090 + Species + + murine + + + 1214577 + Species + + to 7 + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + + RESULTS + paragraph + 9222 + In contrast, the insertion sites involving EVI5 all occurred in T-cell tumors (precursor T-cell lymphoblastic lymphoma/leukemias) from the four copy BCL6 transgenic line. One of these insertions was in the terminal intron, and two were in the 3' untranslated region of the EVI5 gene. One mouse was female, two were males, and these animals ranged in age from 3.8 mos to 5.9 mos (average, 4.6 mos). Relative EVI5 RNA expression levels were decreased in the T-cell lymphomas from the transgenic mice as compared with T-cell tumors from randomly selected controls, none of which had inserts in or near EVI5 (overall decrease, 1.86-fold; mean +- SEM = 0.007 +- 0.001 in the controls as compared with 0.004 +- 0.0007 in the transgenics; P < 0.05, Fig. 2a). + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 10090 + Species + + mouse + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemias + + + + RESULTS + paragraph + 9976 + The retroviral inserts near MYB were all 3' (34.8 to 73.7 kb) and occurred in three females ranging in age from 4.7 mos to 7.8 mos (average, 6.6 mos). Two were large B-cell lymphomas from the 20 copy transgenic line and one was a precursor T-cell lymphoblastic lymphoma/leukemia from the four copy transgenic line. Relative RNA expression in one of the B-cell tumors and in the T-cell lymphoma was increased >=2-fold as compared with that in the three randomly selected control B-cell lymphomas and two randomly selected control precursor T-cell lymphoblastic lymphoma/leukemias, respectively (which did not contain inserts in or near the MYB gene), but relative RNA expression in the other B-cell lymphoma was 3.2-fold decreased as compared with that in the lymphomas from the three randomly selected B-cell controls. These data are depicted in Fig. 3a. + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 1214577 + Species + + to 7 + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemias + + + MESH:D016399 + Disease + + T-cell lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemia + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + title_2 + 10834 + GFI1B, EVI5, and MYB protein levels in lymphomas from retroviral-injected BCL6 transgenic mice and controls, and in human BCL6-positive and negative lymphomas + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + paragraph + 10993 + Protein levels in murine lymphomas detected by immunohisto-chemistry correlated with RNA levels from murine tumors quantified by real-time PCR as described above, and human lymphomas had a similar staining pattern as the murine counterparts (Figs. 1b, 2b, 3b). Whereas GFI1B expression was detected in the cytoplasm, EVI5 and MYB had a nuclear localization. + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + murine + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + + RESULTS + paragraph + 11351 + Tissue sections from the lymphomas of the three transgenic mice containing retroviral inserts near GFI1B (all large B-cell) and from B-cell lymphomas of three randomly selected retroviral-injected nontransgenic controls (no control tumors contained inserts near GFI1B) were studied with anti-GFI1B. Representative sections, shown in Fig. 1b, panel I, indicate that GFI1B staining is decreased in transgenic tumors (nuclear staining is positive for BCL6) as compared with the tumors in non-transgenic retroviral-injected controls (nuclear staining is negative for BCL6). Additionally, sixteen human lymphomas were studied by immunohistochemistry for GFI1B. Nine were B-cell tumors, seven were T-cell neoplasms; eight were BCL6-positive (six B-cell, two T-cell), and eight were BCL6-negative (three B cell, five T-cell). GFI1B expression was decreased in the BCL6-positive lymphomas as compared with the BCL6-negative tumors, which showed strong cytoplasmic expression of GFI1B (Fig. 1b, panel II). Thus, the findings in human lymphomas paralleled those in mice. Additionally, whereas in mice inserts near GFI1B were noted only in BCL6-positive B-cell tumors (which demonstrated decreased expression of GFI1B RNA and protein levels as compared with BCL-negative murine lymphomas), in humans, BCL6-positive T-cell as well as B-cell neoplasms demonstrated decreased expression of GFI1B protein as compared with BCL-negative lymphomas of each cell type (Fig. 1b, panel II). + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 8328 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 604 + Gene + 7640 + + BCL6 + + + 8328 + Gene + 31223 + + GFI1B + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + murine + + + 9606 + Species + + humans + + + 10090 + Species + + transgenic + + + MESH:D016399 + Disease + + T-cell neoplasms + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D009369 + Disease + + transgenic tumors + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + + RESULTS + paragraph + 12820 + Representative immunohistochemical staining of the murine lymphomas containing retroviral inserts in the EVI5 gene (all precursor T-cell lymphoblastic lymphoma/leukemias) is shown in Fig. 2b, panel I and reveals that EVI5 staining (nuclear) is decreased in the BCL6-positive T-cell neoplasms as compared with the non-transgenic retroviral-injected BCL-negative T-cell controls, none of which contained inserts in or near EVI5. Twenty-seven human lymphomas were studied with anti-BCL6. Eight were negative for BCL6 (3 B-cell, 5 T-cell), 17 were BCL6-positive (13 B, 4 T), and two (T-cell) stained weakly with anti-BCL6. Representative sections (Fig. 2b, panel II) indicate that BCL6-positive lymphomas contain less EVI5 than do BCL-negative neoplasms. As in the case of GFI1B, findings for EVI5 in human lymphomas paralleled the findings in murine lymphomas, and, additionally, whereas in the mouse tumors inserts in the EVI5 gene were noted only in BCL6-positive T-cell tumors (which had decreased expression of EVI5 RNA and protein as compared with BCL-negative T-cell lymphomas), in human lymphomas, decreased levels of EVI5 protein were present in BCL6-positive B- as well as T-cell tumors as compared with BCL-negative controls (Fig. 2b, panel II). Non-neoplastic BCL6-positive germinal center cells also contained less EVI5 than the surrounding BCL-negative tumor cells (Fig. 2b, panel II, 4C). + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 604 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 8328 + Gene + 31223 + + GFI1B + + + 7813 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 7813 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + 12053 + Gene + 7640 + + BCL6 + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumor + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemias + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + paragraph + 14220 + Representative immunohistochemistry of MYB staining (Fig. 3b, I) of two murine lymphomas from transgenic mice (a B-cell tumor, panel 1 and a T-cell tumor, panel 2), reveals that these BCL6-positive neoplasms also show nuclear staining for MYB, whereas two murine lymphomas from non-transgenic animals (B-cell, panel 3, and T-cell, panel 4) are BCL-negative (column B) as well as MYB-negative (column C). A third transgenic mouse with a large B-cell lymphoma (not shown) showed little to no staining for MYB. Murine lymphomas stained readily with anti-MYB, which was used in a dilution of 1:1000 for 1 h at room temperature. Human lymphomas were more difficult to stain, requiring a 1:20 dilution and overnight incubation, and even then, positive staining was often focal and noted in only six (24%) of the 25 lymphomas studied. Of the 25 human lymphomas (12 B-cell, 13 T-cell) studied, 17 were BCL6-positive (10 B-cell, seven T-cell), five were BCL-negative (one B-cell, four T-cell), and three stained weakly for BCL6 (one B-cell, two T-cell). In four BCL6-positive lymphomas (two B-cell, two T-cell), the MYB-positive cells were also BCL6-positive (Fig. 3b, II, panels 1 and 2); however, in two T-cell lymphomas, one BCL6-positive, but with variably weak BCL6 staining, and one BCL-negative (Fig. 3b, panels 3 and 4, respectively), the MYB-positive cells were BCL-negative. Our observations in human lymphomas paralleled those in murine tumors in the respect that MYB levels could be increased or decreased in BCL6-positive neoplasms, T- as well as B-cell, and in human tumors, MYB positivity could be demonstrated in BCL6-positive as well as BCL-negative neoplastic cells (Fig. 3b, panel II). + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 4602 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + murine + + + 10090 + Species + + mouse + + + 10090 + Species + + Murine + + + 9606 + Species + + Human + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphomas + + + CVCL:4V27 + CellLine + + 13 T-cell + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D009369 + Disease + + tumor + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + a T-cell tumor + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + RESULTS + title_2 + 15916 + Recurring sites in BCL6-positive transgenic mice not found in nontransgenic controls + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + + RESULTS + paragraph + 16001 + Table 2 lists the genes nearest to recurring sites found in at least two transgenic animals but not in non-transgenic controls. Additional genes that may cooperate with BCL6 occurred in multiple retroviral-injected transgenic mice and also in at least one retroviral-injected non-transgenic (control) animal (e.g., MYC), and thus may have transforming properties on their own. These are not reported here. + + 12053 + Gene + 7640 + + BCL6 + + + 17869 + Gene + 31092 + + MYC + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + + RESULTS + paragraph + 16407 + We previously reported that the most frequent retroviral insertions in our BCL6 transgenic mice occurred in or near the PIM1 gene, and we showed overall higher levels of PIM1 RNA and protein (by immunohistochemistry) in lymphomas (B- or T-cell) containing these insertion sites. Here we have studied the genes involved by the next most common insertion sites (in or near GFI1B, EVI5, and MYB, which were observed in lymphomas from three transgenic mice in each case). + + 12053 + Gene + 7640 + + BCL6 + + + 18712 + Gene + 11214 + + PIM1 + + + 18712 + Gene + 11214 + + PIM1 + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic mice + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + DISCUSS + title_1 + 16875 + Discussion + + + DISCUSS + paragraph + 16886 + Although the BCL6 gene is best known for its association with DLBLs in humans (~40% of these neoplasms are associated with chromosomal rearrangements involving BCL6 and ~16% contain mutations disrupting autoregulation of the BCL6 gene), the role of BCL6 in T-cell development and function has gained increased attention. Through the use of retroviral insertional mutagenesis in BCL6 transgenic mice, we have identified three genes that have not been reported previously as cooperating with BCL6 to promote lymphomagenesis. All of these have been recognized as proto-oncogenes. Two (GFI1B and MYB) are transcription factors and the third (EVI5) is considered an essential regulator of cell membrane trafficking. Additionally, immunohistochemistry performed on B- and T-cell human lymphomas (16 in the case of GFI1B, 27 for EVI5, 25 for MYB), both BCL6-positive and negative, confirmed our observations in murine lymphomas at the RNA and protein level: GFI1B and EVI5 are downregulated in BCL6-positive lymphomas as compared with BCL-negative tumors, and MYB can be either up- or down-regulated in BCL6-positive lymphomas as compared with BCL-negative controls. + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 17863 + Gene + 31311 + + MYB + + + 14020 + Gene + 121902 + + EVI5 + + + 8328 + Gene + 31223 + + GFI1B + + + 7813 + Gene + 121902 + + EVI5 + + + 4602 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 9606 + Species + + humans + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + + DISCUSS + paragraph + 18046 + The GFI1B gene, located on human chromosome 9q34.13, is not a common target in retroviral insertional mutagenesis. It encodes a transcriptional repressor with an N-terminal SNAG (Snail/Gfi1) domain and six C-terminal zinc fingers that are believed to have an important role in hematopoiesis. Of two transcripts in humans, the most frequent is a 330 amino acid protein which binds DNA and suppresses gene expression through recruitment of histone modifying enzymes at target promoters. It is expressed in myeloid progenitor cells as well as in B- and T-cell subsets, is known to control expression levels of genes critical for B-cell development, and is essential for erythroid and megakaryocytic development. It is not expressed in mature thymocytes. + + 8328 + Gene + 31223 + + GFI1B + + + 9606 + Species + + human + + + 9606 + Species + + humans + + + 14581 + Gene + 3854 + + Gfi1 + + + MESH:C536227 + Disease + + hematopoiesis + + + + DISCUSS + paragraph + 18797 + The EVI5 gene, located on human chromosome 1p22, is a common viral integration site in T-cell lymphomas derived from AKXD mice. It encodes a coiled-coil protein that is considered essential for regulation of membrane trafficking and has been believed to be involved in T-cell disease. As we noted in our murine lymphomas, a common viral integration site is in a 3' intron, and integrations are opposite to the transcriptional orientation of the gene. Although real-time RT-PCR revealed decreased RNA expression in all of the lymphomas from the transgenic mice as compared with the nontransgenic controls, the differences were not large; however, it is known that even small perturbations in the expression level of a protein may lead to significant biological effects. Alternatively, as it is known that gene expression can be affected over hundreds of kilobases by retroviral integrations, it is possible that viral insertions in the EVI5 gene could affect another gene, e.g., growth factor independent 1 (GFI1), which is located 18 kb downstream from EVI5 and is known to be a transcription factor that is crucial for normal hematopoietic development. Thus, Schmidt et al., who could not detect a signal with an EVI5 probe on Northern blots prepared from total RNA of lymphomas bearing proviral EVI integrations in MYC/PIM bitransgenic mice, found that the tumors had enhanced GFI1 RNA expression. However, these authors indicate that other targets which are located several hundred kilobases apart also could be activated by EVI5 integrations. + + 7813 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 14581 + Gene + 3854 + + growth factor independent 1 + + + 14581 + Gene + 3854 + + GFI1 + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 68151 + Gene + 11779 + + EVI + + + 14581 + Gene + 3854 + + GFI1 + + + 14020 + Gene + 121902 + + EVI5 + + + 17869 + Gene + 31092 + + MYC + + + 9606 + Species + + human + + + 10090 + Species + + mice + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell disease + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + DISCUSS + paragraph + 20346 + MYB, a site of recurrent retroviral insertional mutagenesis in a number of murine hematopoietic malignancies, is a leucine zipper DNA-binding transcription factor with a short half-life that undergoes post-translational modifications, including ubiquitylation, phosphorylation, acetylation, and sumoylation. Human MYB, located on chromosome 6q23.3, contains 15 exons and encodes a family of related proteins. Alternative splicing leads to splice variants that are predicted to encode proteins with differing transcriptional activities and specificity domains. The MYB protein is expressed in all proliferating hematopoietic cells, is involved in the regulation of proliferation and differentiation of bone marrow progenitor cells (also of colon and adult brain), and is required for normal hematopoiesis, T-cell development, pro-B to pre-B transition, and survival of spleen B cells. In humans, expression of MYB is known to be relevant for the lymph node germinal center phenotype, and that expression is sustained by BCL6 repression of microRNA (miR)-155. Although, traditionally, MYB has been touted as a transcriptional activator, several corepressor molecules that interact through various MYB domains imply that MYB could act also as a repressor. In one study, about half of the genes found to be regulated by MYB were repressed. Its target genes have functions in cell cycle progression, cell differentiation, and survival. The gene is known to be translocated in T-cell acute lymphocytic leukemia, and its protein is overexpressed in that disease, in part through gene duplication. + + 17863 + Gene + 31311 + + MYB + + + 4602 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 4602 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 406947 + Gene + + microRNA (miR)-155 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + murine + + + 9606 + Species + + Human + + + 9606 + Species + + humans + + + 17863 + Gene + 31311 + + MYB + + + MESH:D015179 + Disease + + colon + + + MESH:C536227 + Disease + + hematopoiesis + + + MESH:D054218 + Disease + + T-cell acute lymphocytic leukemia + + + MESH:D019337 + Disease + + hematopoietic malignancies + + + + DISCUSS + paragraph + 21936 + In addition to the three genes described above, additional genes of potential interest are listed in Table 2. Almost all of these are known to be involved in hematopoiesis or hematologic malignancies, lending validity to the notion that they may well cooperate with BCL6 in lymphomagenesis. For example, LMO2 is up-regulated in non-Hodgkin lymphomas and has been noted to be translocated in certain human T-cell acute lymphoblastic leukemias; MN1 is deregulated in human acute myeloid leukemia; AHI1 is an oncoprotein that interacts with BCR-ABL and Janus kinase 2 (JAK2) in chronic myelogenous leukemia cells and is expressed in cutaneous T-cell lymphomas with intermediate to poor prognosis; BCOR is part of a BCL6 repression complex that facilitates survival and proliferation of lymphomas; IKZF1 encodes the IKAROS transcription factor, which drives lymphoid development, and IKZF1 deletions have been noted in childhood B-cell precursor acute lymphoblastic leukemia; SYK was identified as a target in acute myeloid leukemia and, when involved in translocations with the interleukin 2-inducible T cell kinase (ITK) gene, it induces a T-cell lymphoproliferative disease in mice mimicking human disease; DNTT encodes a lymphoid regulator; and, finally, GATA1 encodes an erythroid transcription factor. + + 12053 + Gene + 7640 + + BCL6 + + + 4005 + Gene + 4072 + + LMO2 + + + 4330 + Gene + 37620 + + MN1 + + + 54806 + Gene + 9762 + + AHI1 + + + 25 + Gene + 3783 + + BCR-ABL + + + 3717 + Gene + 21033 + + Janus kinase 2 + + + 3717 + Gene + 21033 + + JAK2 + + + 54880 + Gene + 9809 + + BCOR + + + 604 + Gene + 7640 + + BCL6 + + + 10320 + Gene + 55948 + + IKZF1 + + + 10320 + Gene + 55948 + + IKZF1 + + + 6850 + Gene + 2390 + + SYK + + + 3702 + Gene + 4051 + + interleukin 2-inducible T cell kinase + + + 3702 + Gene + 4051 + + ITK + + + 1791 + Gene + 3014 + + DNTT + + + 2623 + Gene + 1549 + + GATA1 + + + 9606 + Species + + human + + + 9606 + Species + + human + + + 10090 + Species + + mice + + + 9606 + Species + + human + + + 22778 + Gene + 55948 + + IKAROS + + + MESH:D016410 + Disease + + cutaneous T-cell lymphomas + + + MESH:D015470 + Disease + + acute myeloid leukemia + + + MESH:D054198 + Disease + + lymphoblastic leukemias + + + MESH:D007951 + Disease + + myelogenous leukemia + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D015470 + Disease + + acute myeloid leukemia + + + MESH:D054198 + Disease + + lymphoblastic leukemia + + + MESH:D008232 + Disease + + a T-cell lymphoproliferative disease + + + MESH:D019337 + Disease + + hematopoiesis or hematologic malignancies + + + MESH:D008228 + Disease + + non-Hodgkin lymphomas + + + + DISCUSS + paragraph + 23240 + BCL6 is believed to exert its function by repressing hundreds of proteins. B- as well as T-cell lymphomas are aggressive hemato-logic neoplasms resulting from the malignant transformation of B- or T-cell progenitors, respectively. These transformation processes are believed to be multi-step events in which a series of heterogeneous genetic alterations cooperate to induce perturbations in normal lymphocyte growth and differentiation. DLBLs and T-cell acute lymphoblastic leukemias, for example, have been reported to harbor multiple genetic abnormalities (in the case of DLBLs, more than 30 clon-ally represented gene alterations per neoplasm) which differ from tumor to tumor, making approaches to therapeutic intervention diffi-cult. Nonetheless, therapies directed against MYB and the proteins encoded by a number of other genes known to be involved in lymphomatous transformation are currently under investigation. Our studies have added GFI1B, EVI5, and MYB to the list of genes cooperating with BCL6 during lymphomagenesis, thus expanding the arsenal of potential therapeutic targets for DLBL. The finding in this study of several down-regulated cooperating genes suggests that efforts to upregulate them, e.g., by target mRNA manipulation, may have some therapeutic benefit. Identification of the alterations in individual lymphomas, with selection of the appropriate combinations of agents targeting the specific lesions, may hold promise for therapy of these neoplasms. + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D030342 + Disease + + genetic abnormalities + + + MESH:D009369 + Disease + + neoplasm + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D054218 + Disease + + T-cell acute lymphoblastic leukemias + + + MESH:D009369 + Disease + + tumor + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + T-cell lymphomas + + + + DISCUSS + footnote + 24723 + Authorship + + + DISCUSS + footnote + 24734 + Contribution: B.W.B, L.J., and M.J.T. designed research; B.W.B., P.L.R., and J.M.B. performed research; J.B. and L.W. contributed new reagents/ analytic tools; B.W.B., J.A., M.J.T., K.W., and J.M.B. analyzed data; and B.W.B. and J.M.B. wrote the paper. + + + DISCUSS + footnote + 24987 + All authors have approved the final article. + + + DISCUSS + footnote + 25032 + + + + DISCUSS + footnote + 25067 + Conflict-of-interest disclosure + + + DISCUSS + footnote + 25099 + All authors declare no conflict of interests. + + + DISCUSS + footnote + 25145 + + + + DISCUSS + footnote + 25171 + Declaration + + + DISCUSS + footnote + 25183 + The work has not been published previously and is not under consideration for publication elsewhere. The work is approved by all authors. 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Genet + ref + 43 + REF + 837 + surname:Chiarenza;given-names:A + surname:Rossi;given-names:D + 30234 + Analysis of the coding genome of diffuse large B-cell lymphoma + + MESH:D008223 + Disease + + lymphoma + + + + 3 + REF + surname:Vasudevan;given-names:S + 311 + 22072587 + 2012 + Wiley Interdiscip. Rev. RNA + ref + 330 + 30297 + Posttranscriptional upregulation by microRNAs + + + FIG + nihms-648290-f0001.jpg + F1 + fig_caption + 30343 + GFI1B: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative GFI1B RNA expression from the lymphomas (all large B-cell) of three retroviral-injected BCL6 transgenic mice (black bar) containing inserts 7.9 to 8.7 kb 5' to the GFI1B gene as compared with three randomly selected B-cell lymphomas from retroviral-injected non-transgenic controls that did not contain inserts in or near GFI1B (gray bar). Expression in each transgenic animal was decreased as compared with the controls [range, 1.8 to 5.7-fold (Table 1), overall decrease 2.71-fold; mean +- SEM = 0.041 +- 0.018 in the controls as compared with 0.015 +- 0.005 in the transgenics; P < 0.05]. (b) I, Representative murine lymphomas from (1) a transgenic mouse (nuclei positive for BCL6, column B) and (2) a non-transgenic animal (nuclei negative for BCL6). The cytoplasmic staining, evident in the anti-GFI1B (C) column, is decreased in the transgenic mouse as compared with the non-transgenic control. II, Representative human lymphomas: (1) BCL6-positive (B-cell), (2) BCL6-positive (T-cell), (3) BCL6-negative (B-cell), (4) BCL6-negative (T-cell). As in the murine lymphomas, the GFI1B cytoplasmic staining (column C) is decreased in the BCL6-positive lymphomas as compared with the stronger expression in the BCL6-negative neoplasms. The bar (lowest right panel) indicates 50 mum. + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 14582 + Gene + 31223 + + GFI1B + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + mice + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + murine + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D009369 + Disease + + neoplasms + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + FIG + nihms-648290-f0002.jpg + F2 + fig_caption + 31830 + EVI5: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative EVI5 RNA expression from the lymphomas (all T-cell) of three retroviral-injected BCL6 transgenic mice (black bar) containing inserts within the terminal intron or 3'-untranslated region of the EVI5 gene as compared with three randomly selected T-cell lymphomas from retroviral-injected non-transgenic controls that did not contain inserts in or near EVI5 (gray bar). Expression in each transgenic animal was decreased as compared with the controls [range, 1.3 to 2-fold (Table 1), overall decrease, 1.86-fold; mean +- SEM = 0.007 +- 0.001 in the controls as compared with 0.004 +- 0.0007 in the transgenics; P < 0.05]. (b) I, Representative murine lymphomas from (1) a transgenic mouse (nuclei positive for BCL6, column B) and (2) a non-transgenic mouse (nuclei negative for BCL6). The nuclear staining evident in the anti-BCL6 (C) column is decreased in the transgenic mouse as compared with the non-transgenic control. II, Representative human lymphomas: (1) BCL6-positive B-cell, (2) BCL6-positive (T-cell), (3) BCL-negative (B-cell), (4) BCL-negative (T-cell); the cytoplasm of the tumor cells stains strongly positive with the T-cell marker CD3 (II, 4A). A germinal center in 4A (upper left corner) does not stain with CD3 and is BCL6-positive (column B), whereas the tumor cells around it are BCL-negative. As in the murine lymphomas, the EVI5 nuclear staining (column C) is decreased in BCL6-positive cells (tumors 1 and 2, or germinal center cells, tumor 4) as compared with the stronger expression in the BCL-negative tumor cells [tumors 3 and 4 (outside the germinal center)]. The length of the bar in the lowest right panel depicts 50 mum. + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 604 + Gene + 7640 + + BCL6 + + + 12501 + Gene + 586 + + CD3 + + + 12501 + Gene + 586 + + CD3 + + + 12053 + Gene + 7640 + + BCL6 + + + 14020 + Gene + 121902 + + EVI5 + + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + mice + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + murine + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 9606 + Species + + human + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + tumor + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumor + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + FIG + nihms-648290-f0003.jpg + F3 + fig_caption + 33683 + MYB: relative RNA levels in lymphomas from BCL6 transgenic and control mice and immunohistochemistry of BCL6-positive and -negative murine and human lymphomas. (a) The graph depicts average relative MYB RNA expression in the lymphomas of three retroviral-injected BCL6 transgenic mice (black bars, study mice S1, S2, S3) containing inserts ~35 to ~74 kb 3' to the MYB gene as compared with randomly selected B- or T-cell lymphomas (gray bars) from retroviral-injected control mice that did not contain inserts in or near MYB. S1 is a large B-cell lymphoma from a transgenic mouse whose relative expression is 2-fold higher than the mean of three randomly selected B-cell control tumors (S1 expression = 0.54 vs. control B-cell tumors, mean +- SEM = 0.27 +- 0.07), whereas the relative RNA expression of S2 (0.08), also a large B-cell lymphoma from a transgenic animal, is 3.2-fold decreased as compared with the mean of the three randomly selected control B-cell lymphomas (see above). S3 is a precursor T-cell lymphoblastic lymphoma/ leukemia from a transgenic mouse whose relative RNA expression is 2.2-fold higher than the mean of two randomly selected precursor T-cell lymphoblastic lymphoma/leukemia control tumors; S3 expression = 0.42 vs. control T-cell tumors, mean +- SEM = 0.19 +- 0.07. (b) I, Representative murine lymphomas: tumors (1) and (2) depict the B- and T-cell lymphomas, respectively, from transgenic mice (BCL6-positive nuclear staining, column B) with positive MYB nuclear staining (column C); tumors (3) and (4) show B- and T-cell lymphomas, respectively, from non-transgenic mice (nuclei do not stain with anti-BCL6, column B); staining with anti-MYB is also negative in these animals (column C). (b) II, Representative human lymphomas: tumors (1) and (2) show BCL6-positive (column B) B- and T-cell lymphomas, respectively, which show positive nuclear staining for MYB (column C). Tumor (3) is a T-cell lymphoma that is weakly BCL-6 positive (column B), and tumor (4) is a BCL-negative T-cell neoplasm; in tumors 3 and 4, the cells that are MYB positive (column C) are BCL-negative. The bar (lowest right panel) depicts 50 mum. + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL6 + + + 12053 + Gene + 7640 + + BCL6 + + + 17863 + Gene + 31311 + + MYB + + + 604 + Gene + 7640 + + BCL6 + + + 4602 + Gene + 31311 + + MYB + + + 12053 + Gene + 7640 + + BCL-6 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + mice + + + 10090 + Species + + murine + + + 9606 + Species + + human + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + 10090 + Species + + murine + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic mice + + + 9606 + Species + + human + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + 10090 + Species + + transgenic + + + MESH:D009369 + Disease + + tumor + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma/leukemia + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D016399 + Disease + + T-cell lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D007938 + Disease + + leukemia + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D009369 + Disease + + tumors + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell tumors + + + MESH:D009369 + Disease + + neoplasm + + + MESH:D009369 + Disease + + tumors + + + MESH:D016399 + Disease + + T-cell lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + T-cell lymphoblastic lymphoma + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D016399 + Disease + + T-cell lymphoma + + + MESH:D016399 + Disease + + T-cell lymphomas + + + MESH:D016399 + Disease + + T-cell lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + TABLE + table_footnote + 35839 + Nucleotide position 108,201,513. + + + TABLE + table_footnote + 35872 + + + + TABLE + table_footnote + 35904 + Nucleotide position 108,183,744. + + + TABLE + table_footnote + 35937 + + + + TABLE + table_footnote + 35969 + Nucleotide position 108,182,569. + + + TABLE + table_footnote + 36002 + + + + TABLE + table + 36034 + + + + TABLE + T1.xml + T1 + table_caption + 36069 + Location of retroviral inserts in transgenic mice and relative RNA expression levels in lymphomas from BCL6 transgenic mice as compared with lymphomas from retroviral-injected non-transgenic (control) mice. + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + mice + + + 10090 + Species + + transgenic + + + MESH:D008223 + Disease + + lymphomas + + + MESH:D008223 + Disease + + lymphomas + + + + TABLE + T1 + table + T1.xml + 36276 + Mouse gene (chromosome) Lymphoma type Location of insert Expression level vs. control GFI1B (Chr 2) B 8.692 kb 5' to GFI1B 5.7-fold decreased GFI1B B 8.686 kb 5' to GFI1B 2.6-fold decreased GFI1B B 7.877 kb 5' to GFI1B 1.8-fold decreased EVI5 (Chr 5) T Terminal intron* 2.0-fold decreased EVI5 T 3' untranslated region** 1.5-fold decreased EVI5 T 3' untranslated region*** 1.3-fold decreased MYB (Chr 10) T 34.794 kb 3' to MYB 2.2-fold increased MYB B 73.684 kb 3' to MYB 2.0-fold increased MYB B 70.687 kb 3' to MYB 3.2-fold decreased + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + GFI1B + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + EVI5 + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 17863 + Gene + 31311 + + MYB + + + 10090 + Species + + Mouse + + + MESH:D008223 + Disease + + Lymphoma type Location + + + + TABLE + table_footnote + 37634 + Sequences have >=98% identity to the murine databases described in the text. + + 10090 + Species + + murine + + + + TABLE + table_footnote + 37712 + + + + TABLE + table_footnote + 37744 + In one lymphoma, the insert is within SYKb; in the other, SYKb is 61.3 kb 3' to the insert, but there is a gene closer to the insert at the 5' side [40.3 kb away, D/RAS2 (GTP-binding protein DI-RAS2)]. + + 20963 + Gene + 2390 + + SYKb + + + 20963 + Gene + 2390 + + SYKb + + + 68203 + Gene + 56777 + + GTP-binding protein DI-RAS2 + + + MESH:D008223 + Disease + + lymphoma + + + + TABLE + table_footnote + 37950 + + + + TABLE + table_footnote + 37982 + MPPE (5') is closer to the insert in one lymphoma (17.8 kb vs. 50.2 kb), but IMPA2 (3') is closer to the insert in the other lymphoma (1 kb vs. 33.8 kb). + + 114663 + Gene + 22799 + + IMPA2 + + + MESH:D008223 + Disease + + lymphoma + + + MESH:D008223 + Disease + + lymphoma + + + + TABLE + table_footnote + 38140 + + + + TABLE + table + 38172 + + + + TABLE + T2.xml + T2 + table_caption + 38207 + Recurring retroviral integration sites in BCL6 transgenic mice not found in non-transgenic controls.* + + 12053 + Gene + 7640 + + BCL6 + + + 10090 + Species + + transgenic mice + + + 10090 + Species + + transgenic + + + + TABLE + T2 + table + T2.xml + 38309 + Murine genes containing inserts or, if insert not within a gene, the nearest characterized gene (unless indicated) Mouse chromosome Number of mice Lymphoma type PIM1 (proviral integration site for Moloney murine leukemia virus 1) 17 7 6T, 1B GFI1B (growth factor independent 1B) 2 3 B EVI5 (ecotropic viral integration site 5) 5 3 T MYB (transcriptional activator MYB isoform 1) 10 3 2B, 1T LMO2 (LIM domain only 2) 2 2 T MN1 (probable tumor suppressor protein MN1) 5 2 T AHI1 (jouberin isoform 1; Abelson helper integration site-1) 10 2 T BCOR (BCL6 corepressor isoform d) 10 2 B IKZF1 (DNA-binding protein Ikaros isoform a and b) 11 2 T SYKb (tyrosine protein kinase SYK) 13 2** T MPPE (metallophosphoesterase 1); IMPA2 (inositol monophosphatase 2) 18 2*** B DNTT (DNA nucleotidylexotransferase) 19 2 T GATA1 (erythroid transcription factor) X 2 B + + 14582 + Gene + 31223 + + GFI1B + + + 14582 + Gene + 31223 + + growth factor independent 1B + + + 14020 + Gene + 121902 + + EVI5 + + + 14020 + Gene + 121902 + + ecotropic viral integration site 5 + + + 17863 + Gene + 31311 + + MYB + + + 4005 + Gene + 4072 + + LMO2 + + + 433938 + Gene + 37620 + + MN1 + + + 433938 + Gene + 37620 + + MN1 + + + 52906 + Gene + 9762 + + AHI1 + + + 54880 + Gene + 9809 + + BCOR + + + 10320 + Gene + 55948 + + IKZF1 + + + 20963 + Gene + 2390 + + SYKb + + + 20963 + Gene + 2390 + + SYK + + + 114663 + Gene + 22799 + + IMPA2 + + + 1791 + Gene + 3014 + + DNTT + + + 2623 + Gene + 1549 + + GATA1 + + + 10090 + Species + + Murine + + + 10090 + Species + + Mouse + + + 10090 + Species + + mice + + + 11801 + Species + + Moloney murine leukemia virus + + + 17863 + Gene + 31311 + + MYB + + + 22778 + Gene + 55948 + + Ikaros + + + 12053 + Gene + 7640 + + BCL6 + + + MESH:D008223 + Disease + + Lymphoma type PIM1 + + + MESH:D009369 + Disease + + tumor + + + + + 4272827 + + surname:Sevim;given-names:Sezgin + surname:Celik;given-names:Merve + surname:Cevizci;given-names:Sibel + 25568633 + surname:Tanisman;given-names:Beyhan + 329 + 5 + 2014 + 4272827 + MSM-26-329 + 10.5455/msm.2014.26.329-334 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 329-34. doi:10.5455/msm.2014.26.329-334 + Health management trainees mental health stress GHQ-12 + 334 + Tanışman B, Cevizci S, Çelik M, Sevim S + 0 + Work Stress and Risk Factors For Health Management Trainees in Canakkale, Turkey + + MESH:D004194 + Disease + + Stress + + + + ABSTRACT + abstract_title_1 + 81 + Aim: + + + ABSTRACT + abstract + 86 + This study aims to investigate the general mental health situation, work-related stress and risk factors of health management trainees. + + + ABSTRACT + abstract_title_1 + 222 + Methods: + + + ABSTRACT + abstract + 231 + This cross-sectional study was conducted on Health Management Musters students (N=96) in Canakkale Onsekiz Mart University Health Sciences Institute, May-June 2014. A total of 58 students who voluntarily participated in the study were reached (60.42%). Participants completed a 22-question sociodemographic survey form and a 12-item General Health Questionnaire in a face-to-face interview. Data were analyzed using the SPSS software version 20.0. + + 5414 + Gene + 6107 + + Mart + + + 9606 + Species + + Participants + + + + ABSTRACT + abstract_title_1 + 679 + Results: + + + ABSTRACT + abstract + 688 + The average age of participants was 36.4+-6.2 (Min:24-Max:62) years. Thirty five of the participants were female (60.3%), 23 were male (39.7%). The number of people using cigarettes and alcohol were 23 (39.7%) and 9 (15.8%) respectively. In our study group according to GHQ scale 32 people (55.2%) were in the group at risk of depression. Eighty-six percent of participants reported experiencing work stress. The most frequently reported sources of stress were superiors (56.8%), work itself (41.3%), and work colleagues (25.8%). There was no significant difference between those at risk of depression and those not at risk in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family time and vacations (p>0.05). + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + MESH:D000275 + Disease + + depression + + + MESH:D000438 + Chemical + + alcohol + + + - + Chemical + + GHQ + + + MESH:D000275 + Disease + + depression + + + MESH:D002908 + Disease + + chronic disease + + + MESH:D000438 + Chemical + + alcohol + + + + ABSTRACT + abstract_title_1 + 1635 + Conclusions: + + + ABSTRACT + abstract + 1648 + Our study results indicated that majority of participants reported experiencing work stress with more than half at high risk of developing depression. The most reported risk factors were superiors, the work itself and colleagues in the present study. Psychosocial risk factors at work environment should be investigated in terms of psychological, sociological and ergonomics in more detail to reduce the risk of health management trainees experiencing work stress and mental health problems. + + 9606 + Species + + participants + + + MESH:D000275 + Disease + + depression + + + + INTRO + title_1 + 2140 + 1. INTRODUCTION + + + INTRO + paragraph + 2156 + Stress, a psychosocial factor affecting work health and work productivity, is an important public health problem today. Work stress is an employee health problem that negatively affects the health of workers, disturbs public peace, and may cause harmful results for employee and employer. Work stress affects 22% of employees and threatens work health and safety. Physical and mental health problems damage the health of employees, and if necessary precautions are not taken against it, loss of work productivity may cause an increase in health spending. Work stress comprises 21.5% of health spending, 40% of hiring entry-exit costs, 50% of falls in work productivity, 50% of unplanned absences (absences caused by illness, etc.) and 33% of loss of manpower and employee compensation costs. + + MESH:D004194 + Disease + + Stress + + + MESH:C535569 + Disease + + psychosocial + + + + INTRO + paragraph + 2948 + Currently there is a significant increase in the psychological, psychiatric and physical diseases caused by work stress. Previous studies have shown that work stress is the primary cause of work-related health problems and may cause chronic diseases like cardiovascular diseases, musculoskeletal diseases, migraine and depression. Depression developing linked to work is one of the most important causes of long-term chronic diseases. Investigations of the relationship between depression and work environment have found an important increase. The cause of this increase is the continuous presence of depressive disorders in the work environment and it has an excessive negative effect on work productivity, performance, work absences and disability costs. + + MESH:D000275 + Disease + + depressive disorders + + + MESH:D008881 + Disease + + migraine and depression + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + Depression + + + MESH:D009140 + Disease + + musculoskeletal diseases + + + MESH:D001523 + Disease + + psychiatric + + + MESH:D002318 + Disease + + chronic diseases like cardiovascular diseases + + + + INTRO + paragraph + 3705 + Work stress studies of occupational groups at risk have shown that physical, mental and social health problems that may develop after stress can be prevented. A study in Europe found that the occupational group with greatest risk of work stress was health personnel. To reduce the stress experienced linked to work by health personnel, especially nurses, or to prevent negative effects, it is very important to monitor risks in the work environment, increase work fulfilment and provide social support. Reducing the administration and clinical stress load of employees responsible for health management is advantageous to monitor work environment stressors to protect mental health. Studies of health workers and nurses in with work environment frequently use the 12-item General Health Questionnaire (GHQ-12) form to monitor risk of developing depression and anxiety, mental health situation and stress levels. Stress and depression resulting from working conditions are important mental health problems among health professionals. One of the groups at risk of developing this health problem in their working life is health management trainees. + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + MESH:D004194 + Disease + + Stress + + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ-12 + + + + INTRO + paragraph + 4851 + The aim of this study is to investigate the relationship between stress, risk of developing depression and risk factors of health management trainees. + + MESH:D000275 + Disease + + depression + + + + METHODS + title_1 + 5002 + 2. METHODS + + + METHODS + paragraph + 5013 + This was a cross-sectional study which was performed in the Health Science Institute of Canakkale Onsekiz Mart University, Canakkale, covering the period May-Jun 2014. + + 5414 + Gene + 6107 + + Mart + + + + METHODS + paragraph + 5181 + Study population and sampling + + + METHODS + paragraph + 5211 + The population of this cross-sectional study comprised 96 students enrolled in Canakkale Onsekiz Mart University Health Sciences Institute Health Management Department Non-Thesis Masters program. Sampling was not completed as the aim was to reach the whole population. Our study included 58 students (response rate for our study was 60.42%). + + 5414 + Gene + 6107 + + Mart + + + + METHODS + paragraph + 5553 + Data collection + + + METHODS + paragraph + 5569 + Participants completed a 22-question sociodemographic survey form and a 12-item General Health Questionnaire in a face-to-face interview. The sociodemographic survey form included questions about age, gender, educational level, marital status, cigarette and alcohol use, presence of chronic disease, working hours, work stress and causes, regular exercise and nutrition. + + 9606 + Species + + Participants + + + MESH:D002908 + Disease + + chronic disease + + + MESH:D000438 + Chemical + + alcohol + + + + METHODS + paragraph + 5940 + The General Health Questionnaire developed by Goldberg and Hillier (1979) is a short, easy-to-apply, self-evaluating survey for epidemiological research defining mental health problems used for societal scanning and first-stage health services. The Turkish validity and reliability studies were completed by Kilic. The GHQ-12 is widely used to research mental discomfort like depression or anxiety that may be triggered by stress factors. This study used the GHQ point method (0-0-1-1); according to this method a score of 2 points or above is accepted as the "high risk group" for mental health problems. + + - + Chemical + + GHQ + + + - + Chemical + + GHQ-12 + + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + + METHODS + paragraph + 6553 + Ethics + + + METHODS + paragraph + 6560 + The study received written permission from Canakkale Onsekiz Mart University Clinical Research Ethics Committee dated 28.05.2014 and numbered 2014/10-06. + + 5414 + Gene + 6107 + + Mart + + + + METHODS + paragraph + 6714 + Statistical analysis + + + METHODS + paragraph + 6735 + Data were analyzed using the SPSS software version 20.0. Descriptive statistics (frequencies, percentages, means, standard deviations) were used to describe groups of numerical data and the basic features of the data. The variables were investigated using visual (histograms, probability plots) and analytical methods (Kolmogorov-Simirnov/Shapiro-Wilk's test) to determine whether or not they are normally distributed. Chi-square test was used to examine the association between categorical variables. A p-value of less than 0.05 was considered statistically significant. In the present study, the dependent variable was "developing risk of mental health problem according to GHQ-12 scale. Independent variables were sociodemographic characteristics (gender, marital status, education, age), work-related factors (Daily work, computer use, duration of sitting at desk), having a chronic disease, healthy lifestyle behaviors (sleep duration, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations). + + MESH:D002908 + Disease + + chronic disease + + + MESH:D000438 + Chemical + + alcohol + + + + RESULTS + title_1 + 7811 + 3. RESULTS + + + RESULTS + paragraph + 7822 + The average age of participants was 36.4+-6.2 (Min:24-Max:62) years. Thirty five of the participants were female (60.3%), 23 were male (39.7%) with 48 married (82.8%), 10 single (17.2%) and 55 graduates of university (94.8%). When the substance use of participants was examined the number of people using cigarettes and alcohol were 23 (39.7%) and 9 (15.8%) respectively (Table 1). The rate of those considering giving up alcohol was 37.5% with 28.6% feeling guilty for using alcohol. There were 10 people with chronic diseases (17.2%). The most frequently reported chronic diseases were 3 people with hypertension (30%), 3 with asthma (30%), 2 with thyroid function disorders (20%) and 1 with diabetes (10%). When the occupations of participants were examined 22 (37.9%) were nurses, 6 (10.3%) were midwives, 4 (6.9%) were health officers and 26 (44.9%) were health workers from different occupational groups. When lifestyles were examined 16 participated in regular exercise (28.1%), 43 ate regular meals (75.4%) and 27 reported eating fast food (46.6%). Of participants 47 reported spending sufficient time with their families (81.0%). The workplace environmental factors were examined in the working group and the average working day, time at desk, computer use and sleeping durations are presented in Figure 1. + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + MESH:D003920 + Disease + + diabetes + + + MESH:D013959 + Disease + + thyroid function disorders + + + MESH:D006973 + Disease + + hypertension + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D001249 + Disease + + asthma + + + + TABLE + T1.xml + T1 + table_caption + 9138 + Demographic characteristics of the study population, Canakkale, 2014 (*: The percentage of column) + + + FIG + MSM-26-329-g002.jpg + F1 + fig_caption + 9238 + Mean values +- standard deviations of work-related factors among study population, Canakkale, 2014 + + + RESULTS + paragraph + 9337 + There were 39 people in the study group who had weekend holidays (68.4%). The average holiday duration was 1.7+-0.4 days. When the last date of blood pressure reading was questioned 6 people (10.3%) couldn't answer the question, 11 people (18.9%) couldn't remember, 2 people (3.4%) had never measured it, 1 person (1.7%) said 4 years previous, 2 people (3.4%) said 3 years previous, 3 people (5.1%) said 1 year previous, 2 people (3.4%) said 6 months previous, 1 person (1.7%) said 5 months previous, 3 people (5.1%) said 3 months previous, 3 people (5.1%) said 2 months previous, 7 people (12.0%) said 1 month previous, 5 people (8.6%) said 2-3 weeks previous, 5 people (8.6%) said 1 week previous and 7 people (12.0%) said within the last week. The number of the people who had consulted a physician regularly was 24 (41.4%). + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + person + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + person + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + people + + + + RESULTS + paragraph + 10169 + In our study group according to GHQ points from the GHQ scale 32 people (55.2%) were in the group at risk of depression. Statistically, there was no significant difference between those at risk of depression and those not at risk according to the GSA scale in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations (p>0.05). When experience work stress in work environment was questioned, 86.0% of participants reported experiencing work stress. Of participants in the study 33 reported superiors (56.8%), 24 said work itself (41.3%), and 15 said work colleagues (25.8%) had caused them to experience work stress. Insufficient definition of duties, human resources, profession and distance from occupation were other reported stressors result in work stress among participants (Figure 2). + + 2778 + Gene + 55534 + + GSA + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + human + + + 9606 + Species + + participants + + + MESH:D000275 + Disease + + depression + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D002908 + Disease + + chronic disease + + + - + Chemical + + GHQ + + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ + + + + FIG + MSM-26-329-g003.jpg + F2 + fig_caption + 11214 + Stressors reported by study population, Canakkale, 2014. (One more than options were marked). + + + DISCUSS + title_1 + 11308 + 4. DISCUSSION + + + DISCUSS + paragraph + 11322 + Our study aimed to investigate the general mental health and sociodemographic characteristics, work-related factors and factors like work stress in individuals pursuing a masters degree in health management. With this aim students in the Health Management Masters program of Canakkale Onsekiz Mart University, Health Sciences Institute, Health Management Department were included in the study. After the 2-year masters program is completed the health personnel will take the title Health management. The majority of participants were interviewed face-to-face and evaluated for general mental health with the 12-item General Health Questionnaire. Additionally sociodemographic factors and work-related factors that may affect mental health were investigated. + + 5414 + Gene + 6107 + + Mart + + + 9606 + Species + + participants + + + + DISCUSS + paragraph + 12080 + In our working group there were more women than men and more married than single. When the occupations of participants are examined the majority were nurses, midwives, health officers and other health workers. The cigarette use among participants was high (39.7%) while the use of alcohol was lower (15.8%). When other behavior related to healthy lifestyle was examined 16 took regular exercises, 43 ate regular meals and 27 ate fast-food. It has been reported that work stress, high workload and depression can negatively affect the eating habits of workers and may cause obesity, especially in female workers. Sagara et al. showed a significant relationship between increase in weight and development of mental health complaints (high GHQ-12 point) in male employees. In the literature it is reported that increasing exercise and limiting calorie intake can prevent weight gain and development of psychological heatlh problems. In our study, different to the literature, there was no statistically significant difference between those at risk of developing depression according to GHQ-12 scores and those not at risk in terms of regular physical activity. We believe that as health personnel are societal role models their healthy lifestyle and behavior attracts more attention and it may be beneficial, especially for personnel who will play a role in management of health services such as our working group, to participate in internal education on behavior to protect and enhance health. While chronic disease was present in at least 10 people in our study group, the most frequently reported chronic diseases were hypertension, asthma, thyroid function disorders and diabetes. In the literature it is reported that work stress may lead to chronic diseases such as cardiovascular diseases and musculoskeletal diseases. In our study, we also asked to our participants the last date of blood pressure measurement and the last date of general checkup. The minority of the participants reported that they had measured their blood pressure and also consulted a physician regularly. This finding indicated that health management trainees overlooked their health such as measuring blood pressure. Therefore all health professions in this study must be educated to recognize their own health checks and health-promoting opportunities. + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + MESH:D002318 + Disease + + cardiovascular diseases + + + MESH:D009140 + Disease + + musculoskeletal diseases + + + MESH:D001249 + Disease + + asthma + + + MESH:D009765 + Disease + + obesity + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D006973 + Disease + + hypertension + + + MESH:D003920 + Disease + + diabetes + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ-12 + + + MESH:D015430 + Disease + + weight gain + + + MESH:D002908 + Disease + + chronic disease + + + MESH:D013959 + Disease + + thyroid function disorders + + + + DISCUSS + paragraph + 14411 + In a study evaluating the psychological health of health workers in New Zealand, the GHQ-12 was used and while the highest points were for pharmacists, general practitioners and surgeons had similar average points. In our study according to the GHQ socres 32 people (55.2%) were found to have high risk of developing depression and anxiety. Statistically, there was no significant difference between those at risk of depression and those not at risk according to the GSA scale in terms of gender, marital status, educational level, age, work-related factors (daily work, computer use, duration of sitting at desk), sleep duration, presence of chronic disease, substance use (cigarettes, alcohol), regular exercise, regular meals, fast-food consumption, sufficient family timer and vacations (p>0.05). Tabolli et al. in as study of biomedical researchers at a research institute in Rome identified that according to GHQ-12 scores 15.1% of participants were at risk of depression and anxiety, 77.4% were at risk of developing stress and 66% were dissatisfied with work. The most important determinants of work dissatisfaction are reported to be work opportunities, communication between departments, material rewards and participation in departmental organization. Work dissatisfaction related to economic rewards was a risk factor for developing depression independent of gender, age and duration of work. Another Italian study used the GHQ-12 to investigate the psychological status of health workers and found one third of participants were at risk of developing depression and anxiety. The most basic solution to prevent and reduce mental health problems such as development of depression and anxiety linked to psychological stress in the work environment, is to reduce workload while increasing work fulfillment and social supports. A study of personnel managing a hospital in Rome used the GHQ-12 to investigate psychological health and found 37.8% of participants were at high risk of developing depression and anxiety. The same study found that determinants of work dissatisfaction were low control of work and lack of communication between departments. Job dissatisfaction among management personnel was independent of age, gender and duration of work and was reported to be associated with high levels of risk of developing depression/anxiety, low work control and weak communication. + + 2778 + Gene + 55534 + + GSA + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ-12 + + + MESH:D000275 + Disease + + depression + + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + depression + + + - + Chemical + + GHQ-12 + + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + MESH:D004194 + Disease + + anxiety linked to psychological stress + + + MESH:D001008 + Disease + + anxiety + + + - + Chemical + + GHQ-12 + + + - + Chemical + + GHQ-12 + + + MESH:D002908 + Disease + + chronic disease + + + MESH:D001008 + Disease + + anxiety + + + - + Chemical + + GHQ + + + + DISCUSS + paragraph + 16804 + In the health sector, work stress presents as an important risk factor for health workers and at the same time is an important problem negatively affecting the quality of health services. In our study group, the majority of the participants reported that they had experienced work-related stress. Previous studies have emphasized the necessity of researching stress factors linked to work and psychosocial working environment in health workers, a high risk group for developing burnout, role conflict and work dissatisfaction. As a result of the stress factors linked to work in health workers (long working hours, stressful work conditions, etc.) they remain at risk of developing many mental health problems such as work stress, burnout, depression, anxiety and post-traumatic stress disorder. According to a study in an Italian hospital, working environment and ergonomy, shift-work and disruption of workflow are among the factors causing work-related stress. A study of 36 hospitals in America examined the stress factors in nurses and found that the top three stressors were workload, organizational pressure and role conflict. The most frequently reported sources of stress were superiors, the work itself and colleagues. Unclear description of duties, human resources, and distance from occupation and workplace were other reported causes of work stress. A study in Hungary emphasized that there is an important relationship between burnout, role conflict and mental health in the psychosocial work environment of health workers. A study of nurses in Belgium showed that workload, burnout, work satisfaction and expectations were factors affecting the relationship between workers in the work environment and between workers and management. + + 9606 + Species + + participants + + + 9606 + Species + + human + + + MESH:C535569 + Disease + + psychosocial + + + MESH:D001008 + Disease + + anxiety + + + MESH:C535569 + Disease + + psychosocial + + + MESH:D004194 + Disease + + traumatic stress disorder + + + MESH:D000275 + Disease + + depression + + + + DISCUSS + paragraph + 18553 + In our study, workplace factors were examined and the average working day was 9.3+-3.3 hours, with 4.8+-1.9 hours sitting at a desk and 4.9+-3.1 hours computer use. In the literature it is reported that work environment factors like the duration of the working day, time spent sitting at work and time working on a computer may cause psychosocial factors (work stress, depression, anxiety, etc) and musculoskeletal diseases. In our study while the time for sitting at a desk and using computers were at acceptable levels, the average work day of participants was above the legal limit, more than 8 hours. But long working hours negatively affect the health of workers and has been shown to cause harmful results in sensitive individuals. Additionally increased working hours may trigger risk factors for developing depression such as heavy workload, smoking and sleep disorders. In our study the duration of sleep was 6.9+-0.1 hours. When the active working hours of health professionals are considered, heavy workloads, such as being on call, can reduce the duration of sleep and sleep quality. It is reported that middle managers in hospitals could be rewarded for intense workloads, heavy responsibilities and long working hours. However, heavy workloads may cause role conflict, tiredness, burnout and mistakes, interrupting patient care and negatively affecting family life. In our working group 39 of the participants had a weekend holiday and 47 declared they created sufficient time for family. Needs such as weekend holidays and sufficient time with family, which are relaxing and important from the point of view of social support, can reduce work stress and the risk of developing depression, aiding in protecting mental health. + + 9606 + Species + + participants + + + 9606 + Species + + patient + + + 9606 + Species + + participants + + + MESH:D001008 + Disease + + anxiety + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + depression + + + MESH:D000275 + Disease + + depression + + + MESH:C535569 + Disease + + psychosocial + + + MESH:D005221 + Disease + + tiredness + + + MESH:D009140 + Disease + + musculoskeletal diseases + + + MESH:D012893 + Disease + + sleep disorders + + + + DISCUSS + paragraph + 20293 + In a study by Lao, factors affecting work satisfaction among health workers included solving conflict in the workplace, relationships with colleagues and organizational structure. In our working group among the leading causes of work stress were colleagues and superiors. Interpersonal communication and increased social opportunities may solve these problems. Recently new methods of managing hospitals have begun to be developed, and studies have been completed on groups of individuals with high work satisfaction, programs with experienced managers and improving communication levels to increase social support in the workplace and reduce stress factors. A study of managers in Switzerland found that though improving communication between workers and with patients with positive support for widespread communication at all levels, some health organizations may encounter violent consequences linked to not taking sufficient care with communication. According to a study in India, the most important motivation factor not requiring material resources that must be considered by managers and political structures was reported as the opportunity for workers to develop skills in the work environment. + + 259307 + Gene + 22567 + + Lao + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 21496 + The result of this study was that those at risk of developing mental health problems like depression or those with other physical or social health problems were directed to psychiatry and other related branches. To protect work health, an important part of societal health, it is important to monitor risk groups. Additionally employers can periodically monitor and measure work environment factors in the workplace to prevent physical and mental chronic health problems that may accompany the development of work stress risk. Organizations and managers can organise social activities to strengthen inter team communication and reduce the stress levels among workers in the workplace. + + MESH:D000275 + Disease + + depression + + + + CONCL + title_1 + 22181 + 5. CONCLUSION + + + CONCL + paragraph + 22195 + In this study, the majority of participants reported experiencing work stress with more than half at high risk of developing depression according to the GHQ-12 scale. The top three risk factors affecting stress among individuals in the study were superiors, work and colleagues. In our study group while there was no difference found between those at risk of depression and those not at risk in terms of sociodemographic characteristics, work-related factors and health behavior, to reduce the risk of health management trainees experiencing work stress and mental health problems psychosocial risk factors in the work environment should be investigated in more detail. + + 9606 + Species + + participants + + + MESH:D000275 + Disease + + depression + + + MESH:C535569 + Disease + + psychosocial + + + MESH:D000275 + Disease + + depression + + + + CONCL + footnote + 22865 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 22902 + REFERENCES + + + 7 + REF + surname:Parlar;given-names:S + 547 + 2008 + TAF Prev Med Bull + ref + 554 + 22913 + Saglik calisanlarinda goz ardi edilen bir durum: Saglikli calisma ortami + + + surname:Soysal;given-names:A + surname:Kilic;given-names:B + surname:Unal;given-names:B + surname:Demiral;given-names:Y + 11 + 2007 + Toplum Hekimligi Bulteni + ref + 26 + REF + 18 + surname:ve;given-names:ark + surname:Ucku;given-names:R + surname:Bilgin;given-names:AC + 23001 + Isstresi olceginin Izmir Konak Belediyesi' nde calisan erkek iscilerde gecerlik ve guvenilirliginin incelenmesi + + + REF + 2008 + ref + 23129 + Work-related stress: new challenges in a changing workplace. the European Risk Observatory (ERO), part of EU-OSHA, unveiled the emerging risks related to workplaces + + 54719 + Species + + OSHA + + + + surname:Wang;given-names:S + 45 + surname:Ozminkowski;given-names:RJ + surname:Hawkins;given-names:K + REF + surname:Goetzel;given-names:RZ + 5 + 12553174 + 2003 + J Occup Environ Med + ref + 14 + 23294 + The health and productivity cost burden of the "Top 10" physical and mental health conditions affecting six large U.S. employers in 1999 + + + REF + Recruiters World Special Reports + ref + 23435 + Stress in the Workplace. 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+ 21375150 + 2010 + Cah Sociol Demogr Med + ref + 515 + 25974 + Burnout status of interns and associated factors + + + surname:Violante;given-names:B + surname:Manconi;given-names:MP + surname:Gerbaudo;given-names:L + surname:Cortese;given-names:CG + 141 + 2 + 23789520 + 2013 + Med Lav + ref + 104 + REF + 157 + 26023 + Identification of risk factors for work-related stress in a hospital: a qualitative and quantitative approach + + + surname:Sievers;given-names:A + surname:Ehrhart;given-names:MG + surname:Stichler;given-names:JF + surname:Kath;given-names:LM + 1474 + 11 + 23522937 + 2013 + Int J Nurs Stud + ref + 50 + REF + 1480 + 26133 + Predictors of nurse manager stress: a dominance analysis of potential work environment stressors + + + surname:Mondelaers;given-names:M + surname:Willems;given-names:R + surname:Clarke;given-names:S + surname:Van Bogaert;given-names:P + 1515 + 7 + 22989323 + 2013 + J Adv Nurs + ref + 69 + REF + 1524 + 26230 + Nurse practice environment, workload, burnout, job outcomes, and quality of care in psychiatric hospitals: a structural equation model approach + + MESH:D001523 + Disease + + psychiatric + + + + surname:Schaefer;given-names:H + surname:Herrmann-Lingen;given-names:C + surname:Laptinskaya;given-names:D + surname:Blozik;given-names:E + 13 + 19171034 + 2009 + BMC Musculoskelet Disord + ref + 10 + REF + surname:Himmel;given-names:W + surname:Kochen;given-names:MM + 26374 + Depression and anxiety as major determinants of neck pain: a cross-sectional study in general practice + + MESH:D000275 + Disease + + Depression + + + MESH:D001008 + Disease + + anxiety + + + MESH:D019547 + Disease + + neck pain + + + + surname:Rieger;given-names:MA + 9 + surname:Liebers;given-names:F + surname:Gebhardt;given-names:H + REF + surname:Klussmann;given-names:A + 96 + 18588677 + 2008 + BMC Musculoskelet Disord + ref + 26477 + Musculoskeletal symptoms of the upper extremities and the neck: A cross-sectional study on prevalence and symptom-predicting factors at visual display terminal (VDT) workstations + + MESH:D009140 + Disease + + Musculoskeletal symptoms of the upper extremities + + + + surname:Toomingas;given-names:A + surname:Hagman;given-names:M + surname:Floderus;given-names:B + surname:Norman;given-names:K + 201 + 2 + 18413936 + 2008 + Work + ref + 30 + REF + 214 + surname:Tornqvist;given-names:EW + 26656 + Musculoskeletal symptoms in relation to work exposures at call centre companies in Sweden + + MESH:D009140 + Disease + + Musculoskeletal symptoms + + + + surname:Heitmann;given-names:A + surname:Eggerth;given-names:D + surname:Bushnell;given-names:T + surname:Caruso;given-names:CC + 930 + 11 + 16948157 + 2006 + Am J Ind Med + ref + 49 + REF + 942 + surname:Newman;given-names:K + surname:Kojola;given-names:B + 26746 + Long working hours, safety, and health: toward a National Research Agenda + + + surname:Tsutsui;given-names:T + surname:Hoshuyama;given-names:T + surname:Horie;given-names:S + surname:Fujino;given-names:Y + 87 + 4 + 16908956 + 2006 + Sangyo Eiseigaku Zasshi + ref + 48 + REF + 97 + surname:Tanaka;given-names:Y + 26820 + A systematic review of working hours and mental health burden + + + 59 + surname:Theriault;given-names:G + surname:Miyake;given-names:H + REF + surname:Sato;given-names:Y + 14 + 1 + 19001069 + 2009 + Occup Med (Lond) + ref + 19 + 26882 + Overtime work and stress response in a group of Japanese workers + + + surname:Hojou;given-names:M + surname:Takahashi;given-names:M + surname:Nakata;given-names:A + surname:Ikeda;given-names:T + 26 + 1 + 19057116 + 2009 + J Occup Health + ref + 51 + REF + 37 + surname:Nishikido;given-names:N + surname:Haratani;given-names:T + 26947 + Correlates of depressive symptoms among workers in small- and medium-scale manufacturing enterprises in Japan + + MESH:D000275 + Disease + + depressive symptoms + + + + surname:Moore;given-names:C + surname:Gascoigne;given-names:C + surname:Parry;given-names:E + surname:Buchanan;given-names:DA + 646 + 5 + 24341181 + 2013 + J Health Organ Manag + ref + 27 + REF + 664 + 27057 + Are healthcare middle management jobs extreme jobs? + + + surname:Hirosawa;given-names:T + surname:Sarker;given-names:MA + surname:Harun-Or-Rashid;given-names:M + surname:Khamlub;given-names:S + 233 + 3-4 + 24640179 + 2013 + Nagoya J Med Sci + ref + 75 + REF + 241 + surname:Sakamoto;given-names:J + surname:Outavong;given-names:P + 27109 + Job satisfaction of health-care workers at health centers in Vientiane Capital and Bolikhamsai Province, Lao PDR + + 259307 + Gene + 22567 + + Lao + + + + 27 + surname:West;given-names:MA + REF + surname:Buttigieg;given-names:SC + 171 + 2 + 23802397 + 2013 + J Health Organ Manag + ref + 192 + 27222 + Senior management leadership, social support, job design and stressor-to-strain relationships in hospital practice + + + 115 + surname:De Morgan;given-names:PE + surname:Heeb;given-names:JL + REF + surname:Haberey-Knuessi;given-names:V + 8 + 24490450 + 2013 + Rech Soins Infirm + ref + 18 + 27337 + Communication among hospital leaders + + + surname:Steinhardt;given-names:L + 8 + surname:Mahapatra;given-names:P + surname:Chakraborty;given-names:S + REF + surname:Peters;given-names:DH + 27 + 21108833 + 2010 + Hum Resour Health + ref + 27374 + Job satisfaction and motivation of health workers in public and private sectors: cross-sectional analysis from two Indian states + + + + 4272828 + + 25568626 + surname:Cukic;given-names:Vesna + 303 + 5 + 2014 + 4272828 + MSM-26-303 + 10.5455/msm.2014.26.303-305 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 303-5. doi:10.5455/msm.2014.26.303-305 + postoperative pulmonary thromboembolism surgery + 305 + Cukic V + 0 + The Pulmonary Thromboembolism as a Risk of Surgical Treatments and the Role of Anticoagulant Prophylaxiss + + MESH:D011655 + Disease + + Pulmonary Thromboembolism + + + + ABSTRACT + abstract_title_1 + 106 + Objective: + + + ABSTRACT + abstract + 117 + Our objective is to show the number of patients with postoperative pulmonary thromboembolism (PPTE) treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from June 1, 2011 - June 1, 2014 and to indicate the importance of various surgical operations in the development of pulmonary thromboembolism (PTE). + + 9606 + Species + + patients + + + MESH:D008171 + Disease + + pulmonary diseases + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + + ABSTRACT + abstract_title_1 + 478 + Material and methods: + + + ABSTRACT + abstract + 500 + This is the retrospective study which shows the number of patients with PPTE treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from 01.06.2011.-01.06.2014. It represents the number of these patients, per cent of patients with PPTE of total patients with PTE, age and sex of patients, type of surgery, period expressed in days from surgery to diagnosis of PTE, presence of deep venous thrombosis (DVT) of lower extremities, massiveness of PPTE e.g. level of pulmonary artery with embolus. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D004617 + Disease + + embolus + + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D008171 + Disease + + pulmonary diseases + + + OMIM:612862 + Disease + + DVT + + + + ABSTRACT + abstract_title_1 + 1049 + Results: + + + ABSTRACT + abstract + 1058 + In three-year period 232 patients with PTE were treated in Intensive care unit of Clinic "Podhrastovi". 60 of them or 25.86% were patients with 24 males or 40% middle-aged 58.5 years, and 36 females or 60% middle-aged 56.3 years. PPTE developed in 15 patients with abdominal, 11 with urologic, 8 with gynecologic, 15 with orthopedic, 4 with cardiologic, 2 with vascular, 3 with neurosurgical, 1 with glandular and 1 with orl operations. The average period from operation to diagnosis of PPTE was 10.5 days for women, and 13.8 days for men. Only two patients had acute DVT after operation (one man and one woman), and five had amnesias of previous DVT or PTE. The level or the site of pulmonary embolus was different from segmental to main branches of pulmonary artery. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + patients + + + 9606 + Species + + man + + + 9606 + Species + + woman + + + MESH:D000647 + Disease + + amnesias + + + MESH:D004617 + Disease + + pulmonary embolus + + + OMIM:612862 + Disease + + DVT + + + - + Chemical + + PPTE + + + - + Chemical + + PPTE + + + OMIM:612862 + Disease + + DVT + + + + ABSTRACT + abstract_title_1 + 1831 + Conclusion: + + + ABSTRACT + abstract + 1843 + Different surgical operations are the big risk factor for the development of PTE. There is great significance of anticoagulant prophylaxis before surgery even in patients with no anamnesis of previous DVT or PTE. + + 9606 + Species + + patients + + + OMIM:612862 + Disease + + DVT + + + + INTRO + title_1 + 2056 + 1. INTRODUCTION + + + INTRO + paragraph + 2072 + Embolism of the pulmonary artery or one of its branches is the most striking and characteristic appearance of thromboembolic disease. According to textbooks and most of literature in most cases the starting point of the embolus is phlebothrombosis or thrombophlebitis of veins of lower extremities. + + MESH:D011655 + Disease + + Embolism of the pulmonary artery + + + MESH:D004617 + Disease + + embolus + + + MESH:D013924 + Disease + + thrombophlebitis + + + MESH:D013923 + Disease + + thromboembolic disease + + + MESH:D020246 + Disease + + phlebothrombosis + + + + INTRO + paragraph + 2371 + It is often difficult to determine the starting point of pulmonary emboli. Pulmonary embolism is very common nowadays, and favoring factors for its occurrence are the chronic diseases and therefore a long lying (bed rest), which leads to the development of phlebothrombosis and a large number of different surgical operations + + 100653365 + Gene + + bed + + + MESH:D020246 + Disease + + phlebothrombosis + + + MESH:D011655 + Disease + + Pulmonary embolism + + + + INTRO + title_2 + 2698 + 1.1. The etiology of pulmonary thromboembolism: + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + + INTRO + paragraph + 2746 + Phlebothrombosis and thrombophlebitis of lower extremities veins + + MESH:D020246 + Disease + + Phlebothrombosis + + + MESH:D013924 + Disease + + thrombophlebitis of lower extremities veins + + + + INTRO + paragraph + 2815 + Phlebothrombosis of pelvic and abdominal veins usually after surgery + + MESH:D020246 + Disease + + Phlebothrombosis of pelvic + + + + INTRO + paragraph + 2888 + Thrombophlebitis and phlebothrombosis of veins of lower extremities after trauma and various surgical operations + + MESH:D013924 + Disease + + Thrombophlebitis + + + MESH:D014947 + Disease + + trauma + + + MESH:D020246 + Disease + + phlebothrombosis of veins of lower extremities + + + + INTRO + paragraph + 3005 + Intracardiac thrombosis of the right cardiac ventricle + + MESH:D013927 + Disease + + thrombosis + + + + INTRO + paragraph + 3064 + Thrombosis of the pulmonary artery or some of its branches in situ + + MESH:D013927 + Disease + + Thrombosis of the pulmonary artery + + + + INTRO + paragraph + 3135 + There are other, less common types of emboli: e.g. tissue cells, fat, oil, gas embolism and the like + + MESH:D004618 + Disease + + gas embolism + + + MESH:D009821 + Chemical + + oil + + + + INTRO + title_2 + 3241 + 1.2. Risk factors for deep vein thrombosis and pulmonary thromboembolism + + MESH:D020246 + Disease + + deep vein thrombosis + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + + INTRO + paragraph + 3314 + Factors dependent on the patient: + + 9606 + Species + + patient + + + + INTRO + paragraph + 3348 + Age over 40 years + + + INTRO + paragraph + 3370 + Obesity + + + INTRO + paragraph + 3382 + Immobilization / bed rest/- lying in bed for longer than 4 days + + 100653365 + Gene + + bed + + + 100653365 + Gene + + bed + + + + INTRO + paragraph + 3450 + Deep vein thrombosis or pulmonary embolism in history + + MESH:D011655 + Disease + + pulmonary embolism + + + MESH:D020246 + Disease + + vein thrombosis + + + + INTRO + paragraph + 3508 + Thrombophilia, deficiency of antithrombin(AT) III factor, protein C, protein S; lupus anticoagulant, resistance to activated protein C, hemocistinemia + + Disease + + AT + + + Disease + + hemocistinemia + + + MESH:D019851 + Disease + + Thrombophilia, deficiency + + + + INTRO + paragraph + 3664 + Factors dependent on the type of illness or surgery + + + INTRO + paragraph + 3716 + Trauma or surgery: especially the pelvis, hip, leg + + MESH:D014947 + Disease + + Trauma + + + + INTRO + paragraph + 3771 + Malignant processes-particularly in the pelvis, abdomen, primary or metastatic + + + INTRO + paragraph + 3854 + Heart failure + + MESH:D006333 + Disease + + Heart failure + + + + INTRO + paragraph + 3872 + Recent myocardial infarction + + MESH:D009203 + Disease + + myocardial infarction + + + + INTRO + paragraph + 3905 + Hormonal therapy (estrogen, progesterone) + + + INTRO + paragraph + 3951 + Using the contraceptive pills + + + INTRO + paragraph + 3985 + Paraplegia + + + INTRO + paragraph + 4000 + Severe infection + + MESH:D007239 + Disease + + infection + + + + INTRO + paragraph + 4021 + Intestinal inflammations + + + INTRO + paragraph + 4050 + Polycythemia + + MESH:D011086 + Disease + + Polycythemia + + + + INTRO + paragraph + 4067 + Paraproteinemia + + MESH:D010265 + Disease + + Paraproteinemia + + + + INTRO + paragraph + 4087 + Behcet's disease + + + INTRO + paragraph + 4108 + Paroxysmal nocturnal hemoglobinuria + + MESH:D006457 + Disease + + Paroxysmal nocturnal hemoglobinuria + + + + INTRO + paragraph + 4149 + Although etiology and common risk factors for pulmonary thromboembolism are well known there is little data about frequency of postoperative pulmonary thromboembolism after different surgical operations. + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + + INTRO + title_1 + 4353 + 2. OBJECTIVE + + + INTRO + paragraph + 4366 + Our objective is to show the number of patients with postoperative pulmonary thromboembolism (PPTE) treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period : from June 1, 2011 - June 1, 2014 and to indicate the importance of various surgical operations in the development of pulmonary thromboembolism (PTE). + + 9606 + Species + + patients + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + MESH:D008171 + Disease + + pulmonary diseases + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + + METHODS + title_1 + 4727 + 3. MATERIAL AND METHODS + + + METHODS + paragraph + 4751 + This is the retrospective study which shows the number of patients with postoperative pulmonary thromboembolism treated in Intensive care unit of Clinic for pulmonary diseases an TB "Podhrastovi" in three-year period: 01.06.2011.-01.06.2014. It represents the number of these patients, per cent of patients with postoperative pulmonary thromboembolism of total patients with pulmonary thromboembolism, age and sex of patients, type of surgery, period expressed in days from surgery to clinical presence of pulmonary thromboembolism, presence of deep venous thrombosis (DVT) of lower extremities, massiveness of PPTE e.g. level of pulmonary artery with embolus: segmental, lobar, main branches of pulmonary artery; unilateral, bilateral. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D020246 + Disease + + deep venous thrombosis + + + - + Chemical + + PPTE + + + MESH:D008171 + Disease + + pulmonary diseases + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + MESH:D004617 + Disease + + embolus + + + OMIM:612862 + Disease + + DVT + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + + RESULTS + title_1 + 5492 + 4. RESULTS + + + RESULTS + paragraph + 5503 + In three-year period 232 patients with PTE were treated in Intensive care unit of Clinic "Podhrastovi". 60 of them or 25.86% were patients with postoperative PTE. Among them there were 24 males or 40% middle-aged 58.5 years, and 36 females or 60% middle-aged 56.3 years. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + RESULTS + paragraph + 5778 + Only one man had DVT (deep venous phlebothrombosis) (after orthopedic surgery), and five of them had anamnesis about previous DVT (one with abdominal, one with vascular, one with orthopedic and two with cardiology surgery). Only one woman had DVT (after orthopedic surgery), and no one had anamnesis about previous DVT or PTE. + + 9606 + Species + + man + + + 9606 + Species + + woman + + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + MESH:D020246 + Disease + + deep venous phlebothrombosis + + + + RESULTS + paragraph + 6105 + Patients were subjected to different types of surgical operations which are presented on Figure 1. PTE developed in different interval after operation which is presented on the Figure 2. + + 9606 + Species + + Patients + + + + FIG + MSM-26-303-g001.jpg + F1 + fig_caption + 6292 + Type and number of surgical operations before development of PTE. PTE developed in 15 patients with abdominal, 11with urologic, 8 with gynecologic, 15 with orthopedic, 4 with cardiologic. 2 with vascular, 3 with neurosurgical, 1 with glandular (breast) and 1 with orl operations + + 9606 + Species + + patients + + + + FIG + MSM-26-303-g002.jpg + F2 + fig_caption + 6571 + The average period in days from surgery to the development of PTE. PTE developed in one to thirty days after operation, on average for abdominal operations it was 19 days, for urologic 5, for gynecologic 10, for orthopedic 15.5, for cardiologic 13, for vascular 4.5, for neurosurgical 18.5, for glandular 10 and for orl operations 10 days after operation. PPTE developed on average 10 days in women, and 14 days after operation in men. + + 9606 + Species + + women + + + 9606 + Species + + men + + + + RESULTS + paragraph + 7007 + Massiveness of PPTE e.g., the site or level of embolus was different-from segmental to main branches of pulmonary artery which is shown on Figure 3 and Figure 4. + + MESH:D004617 + Disease + + embolus + + + - + Chemical + + PPTE + + + + FIG + MSM-26-303-g003.jpg + F3 + fig_caption + 7169 + The level or site of pulmonary embolus according to the type of surgery in women. The level of pulmonary embolism is different, from segmental to main branches of pulmonary artery with different number of patients according to operation. + + 9606 + Species + + women + + + 9606 + Species + + patients + + + MESH:D004617 + Disease + + pulmonary embolus + + + MESH:D011655 + Disease + + pulmonary embolism + + + + FIG + MSM-26-303-g004.jpg + F4 + fig_caption + 7407 + The level or site of pulmonary embolus according to the type of surgery in men. The level of pulmonary embolism is different, from segmental to main branches of pulmonary artery with different number of patients according to operation + + 9606 + Species + + men + + + 9606 + Species + + patients + + + MESH:D011655 + Disease + + pulmonary embolism + + + MESH:D004617 + Disease + + pulmonary embolus + + + + DISCUSS + title_1 + 7642 + 5. DISCUSSION + + + DISCUSS + paragraph + 7656 + There is notably increasing number of patients with pulmonary thromboembolism in recent years treated in Clinic for pulmonary diseases and TB "Podhrastovi" We are not yet sure whether it is real increasing or it is matter of better diagnostics. In three-year period 232 patients with PTE were treated in Intensive care unit of this clinic. 60 of them or 25.86% were patients where PTE developed after different types of surgical operations. We intended with this study to point out the significance and frequency of postoperative pulmonary thromboembolism, and to indicate that surgical operations of different type are significant etiologic and risk factor for the development of PTE. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + MESH:D008171 + Disease + + pulmonary diseases + + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + + DISCUSS + paragraph + 8346 + This study has several limitations. First of all we did not have real data about regularity of preoperative and postoperative prophylaxis with anticoagulants, early rising from bed after surgery (mechanical prophylaxis), we did not have data about intraoperative or early postoperative deaths caused possibly by PTE. Furthermore, not small number of patients with postoperative PTE was treated in corresponding surgical clinic under the control of pulmonologist from our clinic, and released home to continue anticoagulant therapy under the control of pulmonologist in outpatient department. + + 100653365 + Gene + + bed + + + 9606 + Species + + patients + + + 9606 + Species + + outpatient + + + MESH:D010149 + Disease + + postoperative deaths + + + + DISCUSS + paragraph + 8938 + Although common risk factors for the development of PTE are well known, studied and confirmed in clinical practice there are no enough data in literature about postoperative DVT or PTE out of controlled studies. Many authors indicate the significance of preoperative anticoagulant prophylaxis. + + OMIM:612862 + Disease + + DVT + + + + DISCUSS + paragraph + 9232 + One big prospective study was done by group of authors on 75 771 patients with vascular and orthopedic operations from 1996 to 2001 in Veteran Health Administration Hospital. The average age was 65 years and 96% of patients were males. Major comorbidities included diabetes mellitus, chronic obstructive pulmonary disease, and congestive heart failure. Symptomatic PTE was diagnosed in 805 patients (0.68%) and varied significantly with procedures: 0.14% for carotid endarterectomy to 1.34% for total hip arthroplasty. In our study we dealt with patients of both sex (60% of them women), average age was 58.5 years for men, and aged 56.3 years for women, and patients were subjected to different types of surgery. We did not examine comorbidities before operation. We dealt with patients not prospectively, but only with surgical treated patients with different operations with diagnosed PPTE to indicate the importance of surgery to the development of PTE. These authors indicate the postoperative importance of pneumonia and other infections, including urinary tract infection, myocardial infarction. We did not find such factors of such importance before the development of PPTE. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D006618 + Disease + + hip arthroplasty + + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + MESH:D014552 + Disease + + urinary tract infection + + + MESH:D009203 + Disease + + myocardial infarction + + + MESH:D006333 + Disease + + congestive heart failure + + + MESH:D007239 + Disease + + infections + + + MESH:D011014 + Disease + + pneumonia + + + MESH:D003920 + Disease + + diabetes mellitus + + + + DISCUSS + paragraph + 10415 + Only two patients in our study had DVT after operation (one man and one woman), but in all others were not be able to find the starting point of embolus. So we think that the preoperative and postoperative prophylaxis (either anticoagulant, or mechanical-early rise from bed) is of the most significance, especially in patients with risk factors for DVT or PTE, even it is only the expected lying in bed (bed-rest) longer than 4 days. There is the need for one long prospective study in different surgical clinic for the assessment of more factors (age, sex, preoperative diseases, type and site of operation, postoperative complications, duration of bed-rest, anticoagulant and mechanical prophylaxis etc.) to give the more realistic picture of postoperative pulmonary thromboembolism. + + 100653365 + Gene + + bed + + + 100653365 + Gene + + bed + + + 100653365 + Gene + + bed + + + 100653365 + Gene + + bed + + + 9606 + Species + + patients + + + 9606 + Species + + man + + + 9606 + Species + + woman + + + 9606 + Species + + patients + + + MESH:D011655 + Disease + + postoperative pulmonary thromboembolism + + + OMIM:612862 + Disease + + DVT + + + OMIM:612862 + Disease + + DVT + + + MESH:D004617 + Disease + + embolus + + + + CONCL + title_1 + 11202 + 6. CONCLUSION + + + CONCL + paragraph + 11216 + Different surgical operations are the big risk factor for the development of PTE. There is great significance of anticoagulant prophylaxis before surgery even in patients with no anamnesis of previous DVT or PTE. + + 9606 + Species + + patients + + + OMIM:612862 + Disease + + DVT + + + + CONCL + footnote + 11429 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 11466 + REFERENCES + + + REF + surname:Dordevic;given-names:BS + 563 + 1982 + Medicinska knjiga Beograd-Zagreb + ref + 574 + 11477 + Akutno plucno srce i plucna tromboemblija. U: Plucne bolesti + + Disease + + plucna tromboemblija + + + + 353 + surname:De;given-names:Rosa + surname:Visani;given-names:L + REF + surname:Goldhaber;given-names:SZ + 1386 + 10227218 + 1999 + Lancet + ref + 11541 + Acute pulmonary thromboembolism. Clinical outcomes in the International Cooperative Pulmonary Registry (ICOPER) + + MESH:D011655 + Disease + + pulmonary thromboembolism + + + + 94 + surname:Le Moine;given-names:JR + REF + surname:Moser;given-names:KM + 949 + 1991 + Ann Intern Med + ref + 952 + 11653 + Is embolism risk condition by location of deep venous thrombosis? + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D004617 + Disease + + embolism + + + + 100 + surname:Hales;given-names:CA + surname:Terrin;given-names:ML + REF + surname:Stein;given-names:PD + 598 + 1909617 + 1991 + Chest + ref + 603 + 11719 + Clinical, roendgenographic and electrocardiographic findings in patients with acute pulmonary embolism and no pre-existing cardiac or pulmonary disease + + 9606 + Species + + patients + + + MESH:D011655 + Disease + + acute pulmonary embolism + + + MESH:D008171 + Disease + + pulmonary disease + + + + 330 + surname:Righini;given-names:M + REF + surname:Le Gal;given-names:Teztuz A + 452 + 15684025 + 2005 + BMJ + ref + 455 + 11871 + Reproduction of chest pain by palpation: diagnostic accuracy in suspected pulmonary embolism + + MESH:D002637 + Disease + + chest pain + + + MESH:D011655 + Disease + + pulmonary embolism + + + + 53 + surname:Konjic;given-names:M + surname:Maglajlic;given-names:J + REF + surname:Cukic;given-names:V + 57 + suppl. 3 + 10870628 + 1999 + Med Arh + ref + 58 + 11964 + Postoperative pulmonary thromoboembolism + + MESH:D001261 + Disease + + Postoperative pulmonary thromoboembolism + + + + surname:Wakenfield;given-names:TW + surname:Upcruch;given-names:GR + surname:Rectenvald;given-names:JR + surname:Gangreddy;given-names:C + 335 + 17264013 + 2007 + Journal of Vascular Surgery + ref + 45 + REF + 342 + surname:Kuri;given-names:SH + 12005 + Risk factors and clinical impact of postoperative symptomatic venous thromboembolism + + MESH:D054556 + Disease + + venous thromboembolism + + + + surname:Bergvist;given-names:MR + 126 + surname:Hail;given-names:D + surname:Pineo;given-names:JA + REF + surname:Geerts;given-names:WH + 338s + 15383478 + 2004 + Chest + ref + 400s + 12090 + Prevention of venous thromboembolism: The Seventh ACCP Conference on Antithrombotic and Thrombolytic Therapy + + MESH:D054556 + Disease + + venous thromboembolism + + + + 24 + surname:Pham;given-names:R + surname:Middleton;given-names:G + REF + surname:Tooher;given-names:R + 397 + 2005 + Am Surg + ref + 415 + 12199 + A systematic review of strategies to improve prophylaxis for venous thromboembolism in hospitals + + MESH:D054556 + Disease + + venous thromboembolism + + + + surname:Desjantis;given-names:A + 341 + surname:Darmon;given-names:L + surname:Cohen;given-names:JY + REF + surname:Samama;given-names:MM + 793 + 10477777 + 1999 + N Engl J Med + ref + 800 + 12296 + A comparison of enoxaparin with placebo for the prevention of venous thromboembolism in acutely ill medical patients. Prophylaxis in medical patients with enoxaparin study group + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D017984 + Chemical + + enoxaparin + + + MESH:D054556 + Disease + + venous thromboembolism + + + MESH:D017984 + Chemical + + enoxaparin + + + + + 4272829 + + surname:Thaci;given-names:Shpetim + surname:Qorraj-Bytyqi;given-names:Hasime + surname:Islami;given-names:Hilmi + 25568627 + surname:Hoxha;given-names:Rexhep + 306 + 5 + 2014 + 4272829 + MSM-26-306 + 10.5455/msm.2014.26.306-308 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 306-8. doi:10.5455/msm.2014.26.306-308 + Body mass index weight bone mineral density men + 308 + surname:Bahtiri;given-names:Elton + Hoxha R, Islami H, Qorraj-Bytyqi H, Thaçi S, Bahtiri E + 0 + Relationship of Weight and Body Mass Index with Bone Mineral Density in Adult Men from Kosovo + + 9606 + Species + + Men + + + + ABSTRACT + abstract_title_1 + 94 + Background and objective: + + + ABSTRACT + abstract + 120 + Body weight and body mass index (BMI) are considered strong predictors of osteoporotic fractures, though optimal BMI levels remain unsettled. There are several studies conducted on women about the relationship between BMI and bone mineral density (BMD), and just a few so far on men. Therefore, the objective of current study was to analyze the relationship between weight and BMI and BMD measured in lumbar spine (L1-L4), femur neck and total hip in 64 men from Kosovo. + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + men + + + MESH:D020388 + Disease + + BMD + + + MESH:D058866 + Disease + + osteoporotic fractures + + + MESH:D020388 + Disease + + BMD + + + + ABSTRACT + abstract_title_1 + 591 + Methods: + + + ABSTRACT + abstract + 600 + This cross-sectional study included a population of 64 men divided into three BMI groups. Dual-energy X-ray absorptiometry (DEXA) measurements were done in all the study participants. + + 9606 + Species + + men + + + 9606 + Species + + participants + + + + ABSTRACT + abstract_title_1 + 784 + Results: + + + ABSTRACT + abstract + 794 + Pearson's correlation analysis showed a significant positive correlation between weight and BMI and BMD in femur neck and in total hip, and a significant negative correlation between age and femur neck BMD. Age-adjusted linear regression analysis showed that weight and BMI had a significant positive association with BMD levels. + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + + ABSTRACT + abstract_title_1 + 1124 + Conclusion: + + + ABSTRACT + abstract + 1136 + Although the results show significant relationship between BMI and BMD, the negative relationship between age and femur neck BMD may serve as guidance to initiate early assessment of the BMD in this region as well as preventive measures of osteoporosis and fractures among ageing men population + + 9606 + Species + + men + + + MESH:D050723 + Disease + + fractures + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D010024 + Disease + + osteoporosis + + + + INTRO + title_1 + 1431 + 1. INTRODUCTION + + + INTRO + paragraph + 1447 + Fractures accompanying osteoporosis have become serious health problem. There are many modifiable factors influencing bone mineral density (BMD), which is recognized as a surrogate marker of bone strength. Among them, body weight and body mass index (BMI) are considered strong predictors of osteoporotic fractures. Optimal BMI levels remain still unsettled, though a BMI of 26-28 kg/m2 has been suggested. Overweight and obesity have been considered protective to bone health, while higher BMI levels to be associated with higher BMD score. Beyond the simple mechanism of mechanical load related to body weight on bone density, there are some other assumed mechanisms not yet fully elucidated of interaction between adipose tissue and skeleton through several adipokines. However, recent studies have questioned this traditional view concluding non-linear relationship between BMI and BMD. Some studies reported no association or even lower BMD or increased risk of fractures, sometimes site dependent, in obese patients. + + 9606 + Species + + patients + + + MESH:D009765 + Disease + + obese + + + MESH:D058866 + Disease + + osteoporotic fractures + + + MESH:D010024 + Disease + + osteoporosis + + + MESH:D020388 + Disease + + BMD + + + MESH:D009765 + Disease + + obesity + + + MESH:D020388 + Disease + + BMD + + + MESH:D050723 + Disease + + Fractures + + + MESH:D050723 + Disease + + fractures + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + + INTRO + paragraph + 2470 + The objective of this study was to analyze the relationship between body mass index (BMI) and weight and bone mineral density (BMD) measured in lumbar spine (L1-L4), femur neck and total hip in a population of 64 men from Kosovo. The interest of several studies has been focused on women, but there are only few recent studies worldwide focused on this issue on men and, to our best knowledge, this is the first one undertaken among Kosovo men. + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + men + + + MESH:D020388 + Disease + + BMD + + + + TABLE + T1.xml + T1 + table_caption + 2915 + Descriptive characteristics of the study subjects. BMD bone mineral density, BMI body mass index. *p <0.05, **p<0.01 compared to normal weight, p <0.05, p<0.01 compared to overweight + + MESH:D020388 + Disease + + BMD + + + + METHODS + title_1 + 3106 + 2. MATERIAL AND METHODS + + + METHODS + title_2 + 3130 + 2.1. Study population + + + METHODS + paragraph + 3152 + The study population consisted of 64 men who were divided into three groups according to standard World Health Organization (WHO) Body Mass Index (BMI) classification: Normal Weight group (n=23), Overweight group (n=26) and Obese group (n=15). Subjects were not included in the study if they had history of diseases that might affect bone metabolism or had taken drugs that might influence BMD (glucocortiocids, bisphosphonates). Written informed consent was obtained from all participants and the study protocol was approved by local Ethical Committees. The research was conducted in accordance to guidelines in Declaration of Helsinki. + + 9606 + Species + + men + + + 9606 + Species + + participants + + + MESH:D020388 + Disease + + BMD + + + - + Chemical + + glucocortiocids + + + MESH:D004164 + Chemical + + bisphosphonates + + + + METHODS + title_2 + 3790 + 2.2. Anthropometric and bone mineral density measurements + + + METHODS + paragraph + 3848 + Height (m) and weight (kg) were measured in light clothing and no shoes before performing DEXA, and body mass index (BMI) was calculated as the ratio of subjects' weight to height squared (kg/m2). According to WHO criteria, participants were categorized into three BMI groups: normal weight (BMI <24.9 kg/m2), overweight (BMI=25.0-29.9 kg/m2) and obese (BMI>30 kg/m2). Dual-energy X-ray absorptiometry (DEXA; Hologic QDR-4500, USA) was used to measure bone mineral density (BMD) in lumbar vertebrae (L1-L4), femoral neck and total hip in all study participants. DEXA was calibrated daily using a phantom provided by the manufacturer. The diagnosis of osteoporosis and osteopenia was done according to WHO T-score criteria (osteoporosis if T-score <= -2.5 SD, osteopenia if T-score between -2.5 SD and -1 SD, and normal BMD if T-score > -1SD). + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + MESH:D010024 + Disease + + osteoporosis + + + MESH:D010024 + Disease + + osteoporosis + + + MESH:D020388 + Disease + + BMD + + + MESH:D009765 + Disease + + obese + + + MESH:D001851 + Disease + + osteopenia + + + MESH:D020388 + Disease + + BMD + + + MESH:D001851 + Disease + + osteopenia + + + + METHODS + title_2 + 4694 + 2.3. Statistical analysis + + + METHODS + paragraph + 4720 + SPSS version 16.0 (SPSS Inc., Chicago, IL, USA) was used for statistical analysis. All p-values in statistical analyses were two-tailed with a significance level of 0.05. Continuous variables are expressed as the means +- standard deviations (SD), while categorical variables as percentages. One-way analysis of variance (ANOVA) with Tukey test for Post Hoc Multiple Comparisons for continuous variables and Chi square test for categorical variables were used to compare characteristics of the three groups created on the basis of BMI. Person's correlation test was performed to assess correlation between age, weight, BMI and BMD. Age-adjusted linear regression analysis was calculated with BMI and weight as independent variables and BMD in L1-L4 vertebra, femoral neck and total hip as dependent variables. + + 9606 + Species + + Person + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + + RESULTS + title_1 + 5530 + 3. RESULTS + + + RESULTS + paragraph + 5541 + Descriptive characteristics of the study population in total and according to BMI groups are summarized in The recruited study subjects consisted of 64 males with a mean age of 42.17+-14.86 (SD). There were 23 subjects (36.3+-15.81 years) in normal weight group,26 subjects (42.92+-13.9 years) in overweight group and 15 subjects (49.87+-11.59 years) in obese group. Most men in this study were overweight or obese (n=41 or 64%). + + 9606 + Species + + men + + + MESH:D009765 + Disease + + obese + + + MESH:D009765 + Disease + + obese + + + + RESULTS + paragraph + 5971 + Comparison of BMI groups by using one way ANOVA found no statistically significant difference for L1-L4 BMD, femur neck BMD and total hip BMD levels (p>0.05). Despite the fact that a statistically significant difference was not observed among BMI groups regarding BMD levels, Pearson's correlation analysis shown in table 2 reveals a significant positive correlation between weight and BMI and BMD in femur neck (r=0.445, p<0.01) and in total hip (r=0.502, p<0.01). No statistically significant correlation was shown between weight and BMI and L1-L4 BMD, while a significant negative correlation was observed between age and femur neck BMD (r=-0.302, p<0.05). + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + + TABLE + T2.xml + T2 + table_caption + 6631 + Pearson's correlation analysis between age, weight, BMI and BMD (r) *p <0.05; **p<0.01 + + MESH:D020388 + Disease + + BMD + + + + RESULTS + paragraph + 6718 + Age-adjusted linear regression analysis showed that weight and BMI as explanatory variables had a significant positive association with L1-L4 BMD, FN BMD and TH BMD levels (Table 3). When weight was compared to BMI, it was observed a tendency of differences in L1-L4 BMD and FN BMD (beta=0.308 vs. 0.305 and beta=0.563 vs. beta= 0.559), whereas BMI showed greater difference in total hip BMD (beta=0.577 vs. 0.540). Weight and BMI had highest squared R values for femur neck BMD, where these variables can explain 38.6% and 35.4% of the variation in FN BMD (R2=0.386 and R2=0.354, respectively). + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + FN BMD + + + MESH:D020388 + Disease + + FN BMD + + + MESH:D020388 + Disease + + FN BMD + + + + TABLE + T3.xml + T3 + table_caption + 7314 + Age-adjusted linear regression analysis for BMD with weight and BMI as predictors.*p<0.05;**p<0.01; ***p<0.001 + + MESH:D020388 + Disease + + BMD + + + + DISCUSS + title_1 + 7425 + 4. DISCUSSION + + + DISCUSS + paragraph + 7439 + Body weight and body mass index as potentially modifiable factors are generally considered essential determinants of bone mineral density. Several studies demonstrated positive association of elevated body weight and/or BMI with bone mineral density or even protective effect against osteoporosis and fractures. Principal explanation of this association was heavier mechanical loading on bones with subsequent bone remodelation to resist this loading. Some of other putative mechanisms are increased estrogen synthesis (in women), increased secretion of insulin and increased plasma levels of leptin. However, there are other published studies focused on the impact of fat mass on BMD that demonstrated negative association of obesity with BMD; the assumed mechanisms are secretion of pro-inflammatory cytokines that stimulate bone resorption, decreased levels of adiponectins and increased parathormone levels. + + 3630 + Gene + 173 + + insulin + + + 9606 + Species + + women + + + MESH:D020388 + Disease + + BMD + + + MESH:D050723 + Disease + + fractures + + + MESH:D020388 + Disease + + BMD + + + MESH:D010024 + Disease + + osteoporosis + + + MESH:D009765 + Disease + + obesity + + + + DISCUSS + paragraph + 8351 + In accordance with results of Dogan A et al., Salamat M et al. and Lloyd J et al. results of this cross-sectional study demonstrate a positive correlation of weight and body mass index with both femur neck BMD and total hip BMD in all male subjects included in the study, but, in contrary to them, compared to normal weight, BMD levels of overweight and obese groups were not significantly different; the lack of significant difference between BMI groups might be as limitation of the small sample size. Contrary to results of our study and above mentioned studies, Greco et al. reported low lumbar BMD in obese patients, while a recent study by Emaus N et al. demonstrated a nonlinear relationship between body mass index and bone mineral density. Every unit increase in BMI was associated with an increase of 0.008 g/cm2 in L1-L4 BMD, 0.017 g/cm2 in femur neck BMD and 0.018 g/cm2 in total hip BMD. Interestingly, age was negatively correlated only with femur neck BMD in all subjects. + + 9606 + Species + + patients + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D009765 + Disease + + obese + + + MESH:D009765 + Disease + + obese + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + MESH:D020388 + Disease + + BMD + + + + CONCL + title_1 + 9339 + 5. CONCLUSION + + + CONCL + paragraph + 9353 + In conclusion, we demonstrated a highly significant relationship between body mass index and bone mineral density in femur neck and total hip in this sample of men. Negative relationship between age and femur neck BMD may serve as guidance to focus the evaluation of the bone mineral density in this region and to initiate early preventive measures of osteoporosis and fractures among aging male population. + + 9606 + Species + + men + + + MESH:D020388 + Disease + + BMD + + + MESH:D050723 + Disease + + fractures + + + MESH:D010024 + Disease + + osteoporosis + + + + CONCL + footnote + 9761 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 9798 + REFERENCES + + + 66 + surname:Bianchi;given-names:G + REF + surname:Giusti;given-names:A + 136 + 2 + 25069495 + 2014 + doi: 10.4081/reumatismo.2014.786 + Reumatismo + ref + 43 + 9809 + Male osteoporosis + + MESH:D010024 + Disease + + osteoporosis + + + + surname:Zhang;given-names:H + surname:Wang;given-names:Y + surname:Liu;given-names:Z + REF + surname:Kang;given-names:D + 2014 + Epub ahead of print + J Endocrinol Invest + ref + surname:Wang;given-names:P + surname:Cao;given-names:W + surname:Feng;given-names:X + 9827 + Relationship of body composition with bone mineral density in northern Chinese men by body mass index levels + + 9606 + Species + + men + + + + surname:Khosla;given-names:S + surname:Amin;given-names:S + surname:Beck;given-names:TJ + surname:Melton;given-names:LJ;suffix:3rd + 460 + 5 + 15688123 + 2005 + Osteoporos Int + ref + 16 + REF + 467 + surname:Riggs;given-names:BL + surname:Oberg;given-names:AL + surname:Achenbach;given-names:SJ + 9936 + Contributions of bone density and structure to fracture risk assessment in men and women + + 9606 + Species + + men + + + MESH:D050723 + Disease + + fracture + + + + surname:Ahmed;given-names:LA + surname:Wilsgaard;given-names:T + REF + surname:Emaus;given-names:N + 2014 + doi: 10.1002/jbmr.2232. 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Evaluation of bone mineral density in individuals with high body mass index + + MESH:D009765 + Disease + + obesity + + + MESH:D010024 + Disease + + osteoporosis + + + + surname:Ryu;given-names:OH + surname:Lee;given-names:J + REF + surname:Chung;given-names:W + 2013 + Epub ahead of print + J Bone Miner Metab + ref + 11349 + Is the negative relationship between obesity and bone mineral content greater for olderwomen? + + MESH:D009765 + Disease + + obesity + + + + surname:Cooper;given-names:C + surname:Chapurlat;given-names:R + surname:Watts;given-names:NB + surname:Compston;given-names:JE + 22017783 + 2011 + Am J Med + ref + surname:Siris;given-names:ES + REF + 1050 + 1043 + 11 + 124 + surname:Saag;given-names:KG + surname:Hooven;given-names:FH + surname:Flahive;given-names:J + surname:Netelenbos;given-names:JC + surname:Gehlbach;given-names:S + surname:Lacroix;given-names:AZ + surname:Roux;given-names:C + doi: 10.1016/j.amjmed.2011.06.013 + surname:Adachi;given-names:JD + surname:Lindsay;given-names:R + surname:Rossini;given-names:M + surname:Diez-Perez;given-names:A + surname:Silverman;given-names:S + surname:Pfeilschifter;given-names:J + surname:Sambrook;given-names:PN + surname:Greenspan;given-names:S + surname:Boonen;given-names:S + 11443 + Obesity is not protective against fracture in postmenopausal women: GLOW + + 9606 + Species + + women + + + MESH:D050723 + Disease + + fracture + + + + 1 + surname:Samos;given-names:LF + surname:Malphurs;given-names:JE + REF + surname:Paniagua;given-names:MA + 283 + 3 + 18046882 + 2006 + Clin Interv Aging + ref + 287 + 11516 + BMI and low bone mass in an elderly male nursing home population + + + surname:LeBlanc;given-names:ES + surname:Adams;given-names:AL + surname:Marshall;given-names:LM + surname:Nielson;given-names:CM + 496 + 3 + 20814955 + 2011 + J Bone Miner Res + ref + 26 + REF + doi: 10.1002/jbmr.235 + surname:Orwoll;given-names:ES + surname:Barrett-Connor;given-names:E + 502 + surname:Stefanick;given-names:ML + surname:Ensrud;given-names:K + surname:Cawthon;given-names:PM + 11581 + BMI and fracture risk in older men: the osteoporotic fractures in men study (MrOS) + + 8011 + Gene + + MrOS + + + 9606 + Species + + men + + + 9606 + Species + + men + + + MESH:D058866 + Disease + + osteoporotic fractures + + + MESH:D050723 + Disease + + fracture + + + + surname:Hermosilla;given-names:E + surname:Fina;given-names:Aviles F + surname:Premaor;given-names:MO + surname:Prieto-Alhambra;given-names:D + 294 + 2 + 22095911 + 2012 + J Bone Miner Res + ref + 27 + REF + doi: 10.1002/jbmr.1466 + surname:Diez-Perez;given-names:A + surname:Compston;given-names:JE + 300 + surname:Nogues;given-names:X + surname:Carbonell-Abella;given-names:C + surname:Martinez-Laguna;given-names:D + 11664 + The association between fracture and obesity is site-dependent: a population-based study in postmenopausal women + + MESH:D050723 + Disease + + fracture + + + MESH:D009765 + Disease + + obesity + + + + surname:Pages-Castella;given-names:A + surname:Fina Aviles;given-names:F + surname:Compston;given-names:JE + surname:Premaor;given-names:MO + 1771 + 8 + 23371035 + 2013 + J Bone Miner Res + ref + 28 + REF + doi: 10.1002/jbmr.1878 + 1777 + surname:Prieto-Alhambra;given-names:D + surname:Diez-Perez;given-names:A + surname:Nogues;given-names:X + 11777 + The association between fracture site and obesity in men: a population-based cohort study + + 9606 + Species + + men + + + MESH:D050723 + Disease + + fracture + + + MESH:D009765 + Disease + + obesity + + + + REF + Available at: + ref + 11867 + BMI classification according to WHO + + + surname:Hochberg;given-names:MC + surname:Hawkes;given-names:WG + surname:Alley;given-names:DE + surname:Lloyd;given-names:JT + 175 + 1 + 24664472 + 2014 + Arch Osteoporos + ref + 9 + REF + doi: 10.1007/s11657-014-0175-2. Epub 2014 Mar 25 + surname:Orwig;given-names:DL + surname:Waldstein;given-names:SR + 11903 + Body mass index is positively associated with bone mineral density in US older adults + + + + 4272830 + + surname:Hoxha-Kamberi;given-names:Teuta + surname:Azemi;given-names:Mehmedali + surname:Shala;given-names:Muje + 25568634 + surname:Ismaili-Jaha;given-names:Vlora + 335 + 5 + 2014 + 4272830 + MSM-26-335 + 10.5455/msm.2014.26.335-338 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 335-8. doi:10.5455/msm.2014.26.335-338 + Acute diarrhea rotavirus Kosova + 338 + surname:Jaha;given-names:Luan + surname:Spahiu;given-names:Shqipe + surname:Avdiu;given-names:Muharrem + Ismaili-Jaha V, Shala M, Azemi M, Hoxha-Kamberi T, Avdiu M, Spahiu S, Jaha L + 0 + Characteristics of Rotavirus Diarrhea in Hospitalized Children in Kosovo + + 9606 + Species + + Children + + + MESH:D012400 + Disease + + Rotavirus Diarrhea + + + + ABSTRACT + abstract_title_1 + 73 + Background: + + + ABSTRACT + abstract + 85 + Diarrhea is a leading cause of child mortality worldwide. Rotavirus is one of the most common causes of severe diarrhea and dehydration in children. Authors reviewed epidemiological and clinical data of the rotavirus diarrhea in Kosovo. + + 9606 + Species + + child + + + 9606 + Species + + children + + + MESH:D003967 + Disease + + Diarrhea + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + ABSTRACT + abstract_title_1 + 322 + Methods: + + + ABSTRACT + abstract + 331 + This is a prospective study carried between January 1st and December 31st 2011. All data, comprising demographics, nutrition, clinical presentation, laboratory findings, management and outcome of the rotavirus diarrhea are collected on the specially designed form. + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + ABSTRACT + abstract_title_1 + 596 + Results: + + + ABSTRACT + abstract + 605 + 116 children with rotavirus diarrhea are included in the study. The majority boys (74.4%) and children aged 0 - 12 months (82.75%). Mean age of children in the study was 16.38 months. Almost every third child in the study was hypotrophic (29.2%). More than half of the infants (55.2%) were on mixed food, somewhat more than every third was breast feeding (36.45%), and every twelfth (8.33%) was on artificial milk (animal or formula). Apart from diarrhea, present in all patients, vomiting (97.41%) and fever (43.96%) were characteristics of the clinical presentation of the diarrhea. Two thirds of the children had mild grade dehydration (70.7%). All patients recovered with no sequels. + + 9606 + Species + + children + + + 9606 + Species + + boys + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + infants + + + 9606 + Species + + patients + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D003681 + Disease + + dehydration + + + MESH:D014839 + Disease + + vomiting + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D005334 + Disease + + fever + + + MESH:D003967 + Disease + + diarrhea + + + + ABSTRACT + abstract_title_1 + 1295 + Conclusion: + + + ABSTRACT + abstract + 1307 + Rotavirus continues to be responsible for a significant portion of acute diarrhea in Kosovo. Clinical features, epidemiological data and the agglutination test are safe enough to establish the diagnosis. Treated correctly rotavirus diarrhea has a favorable outcome. + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + + INTRO + title_1 + 1573 + 1. BACKGROUND + + + INTRO + paragraph + 1587 + Since 1973 when discovered by Bishop, rotavirus was found responsible for one third of the children hospitalization for acute diarrhea worldwide. Rotavirus diarrhea is a cause of 440 thousand deaths, 2 million hospitalizations, 25 million outpatients visit and 111 million episodes of domiciliary diarrhea among children. The aim of this study is to analyze the epidemiology of the acute diarrhea inflicted by rotavirus, demographics of the patients and clinical presentation of the disease in children hospitalized at the Department of Pediatric Gastroenterology of the University Clinical Center of Kosovo in Prishtina, Republic of Kosovo. + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + 9606 + Species + + children + + + MESH:D012400 + Disease + + Rotavirus diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + + INTRO + paragraph + 2229 + It is a busy metropolitan center serving population of around 2 million inhabitants of the Republic of Kosovo. Country itself a decade ago emerged from the devastating war and is the poorest in Europe. As found by the Kosovo Agency of Statistics, 46% of the population in country is younger than 18 years, 28% is younger than 14 years and 8% younger than 5%. + + + METHODS + title_1 + 2588 + 2. METHODS + + + METHODS + paragraph + 2599 + This is a prospective study carried between January 1st and December 31st 2011. All data, comprising demographics, nutrition, clinical presentation, laboratory findings, management and outcome of the rotavirus diarrhea are collected on the specially designed form. + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + METHODS + paragraph + 2864 + Retrovirus is confirmed in the stool of patients using Latex agglutination test LTA s.r.l. - Via Milan with sensitivity and specify of 96%. + + 9606 + Species + + patients + + + + RESULTS + title_1 + 3006 + 3. RESULTS + + + RESULTS + paragraph + 3017 + Of 1011 children admitted at the Hospital for acute diarrhea during the study period, every third is tested for rotavirus. Of 337 tested, 116 were positive on rotavirus (34.42%). + + 9606 + Species + + children + + + MESH:D003967 + Disease + + diarrhea + + + + RESULTS + paragraph + 3196 + The majority of the children in the study were boys (74.4) and infants (82.75%). Average age of children in the study was 16.38 months. Almost every third child in study was hypotrophic (29.2%). The hypotrophy was more frequent in girls than in boys (41% vs. 23.5%). + + 9606 + Species + + children + + + 9606 + Species + + boys + + + 9606 + Species + + infants + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + girls + + + 9606 + Species + + boys + + + + RESULTS + paragraph + 3463 + Only 20 children were not infants (17.2%). More than half of the infants (55.2%) were on mixed food, somewhat more than every third was breast feeding (36.45%), and every twelfth (8.33%) was on artificial milk (animal or formula). + + 9606 + Species + + children + + + 9606 + Species + + infants + + + 9606 + Species + + infants + + + + RESULTS + paragraph + 3694 + Data on demographics, nutritional status and feeding habits are presented in table 1. + + + TABLE + T1.xml + T1 + table_caption + 3780 + Demographics, nutritional status and mode of feeding of the patient with rotavirus diarrhea in the study + + 9606 + Species + + patient + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + RESULTS + paragraph + 3885 + Apart from diarrhea, present in all patients, vomiting (88.8%) and fever (43.96%) were characteristics of the clinical presentation of the diarrhea. Vomiting was more frequent in boys than in girls (97.4% vs. 71.8%) as well as temperature (46.7% vs. 38.5%). Two patients in the study (1.7%), one boy and one girl have had a convulsion at the admission The frequency of the convulsions was equal (each 1.7%) + + 9606 + Species + + patients + + + 9606 + Species + + boys + + + 9606 + Species + + girls + + + 9606 + Species + + patients + + + 9606 + Species + + boy + + + 9606 + Species + + girl + + + MESH:D012640 + Disease + + convulsions + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D012640 + Disease + + convulsion + + + MESH:D014839 + Disease + + Vomiting + + + MESH:D014839 + Disease + + vomiting + + + MESH:D005334 + Disease + + fever + + + MESH:D003967 + Disease + + diarrhea + + + + RESULTS + paragraph + 4292 + All patients in the study had moderate or severe dehydration with the later present in almost every third patient in the study (29.3%). There are no significant differences in the degree of the dehydration between girls and boys. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + girls + + + 9606 + Species + + boys + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + + RESULTS + paragraph + 4522 + Clinical presentation of the rotavirus diarrhea and the degree of dehydration at the admission are presented in table 2. + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D003681 + Disease + + dehydration + + + + TABLE + T2.xml + T2 + table_caption + 4643 + Symptoms and degree of dehydration of the patient with rotavirus diarrhea at the admission + + 9606 + Species + + patient + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D003681 + Disease + + dehydration + + + + RESULTS + paragraph + 4734 + Only about one third of the patients with rotavirus diarrhea in our study had associated diseases (36.2%). The most frequent one was bronchopneumonia that was present in 31.9% of our patients. Less common were upper urinary tract infections (1.7%), upper respiratory tract infections (0.9%) and gastro esophageal reflux (1.7%). Both urinary tract infections in our study were in girls (5.1% of the girls). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + girls + + + 9606 + Species + + girls + + + MESH:D012141 + Disease + + respiratory tract infections + + + MESH:D012141 + Disease + + tract infections + + + MESH:D005764 + Disease + + gastro esophageal reflux + + + MESH:D001996 + Disease + + bronchopneumonia + + + MESH:D014552 + Disease + + upper urinary tract infections + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + RESULTS + paragraph + 5140 + Data on associated conditions and diseases in patients with rotavirus diarrhea are presented in table 3. + + 9606 + Species + + patients + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + TABLE + T3.xml + T3 + table_caption + 5245 + Associated conditions and diseases in patient with rotavirus diarrhea in our study + + 9606 + Species + + patient + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + RESULTS + paragraph + 5328 + Paralytic ileus was the most common complication of the rotavirus diarrhea. In our study it occurred in 9.5% of patients or almost in every tenth patient. The incidence of this complication was for a little more frequent in girls than in boys (15.4% vs 6.5%). There was also a single case of ascites among our patients (0.86%). One patient presented with neurological symptoms (seizures). + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + girls + + + 9606 + Species + + boys + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D007418 + Disease + + Paralytic ileus + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D012640 + Disease + + seizures + + + MESH:D009422 + Disease + + neurological symptoms + + + MESH:D001201 + Disease + + ascites + + + + RESULTS + paragraph + 5717 + Little more than two of three patients in our study were treated without antibiotics (36.2%) and only eight patients (6.9%) were given concentrated erythrocytes or fresh frozen plasma. Ceftriaxone was the most commonly used antibiotic in our practice. The drug was given to 50% of the patients as the only antibiotic and in another 11.2% of patients combined with aminoglycosides - amikacin or gentamicin. Amikacin was given as the only drug in 2.6% of patients and Bactrim (sulfamethoxazole and trimethoprim) in 0.9% of patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000617 + Chemical + + aminoglycosides + + + MESH:D002443 + Chemical + + Ceftriaxone + + + MESH:D000583 + Chemical + + amikacin + + + MESH:D005839 + Chemical + + gentamicin + + + MESH:D013420 + Chemical + + sulfamethoxazole + + + MESH:D015662 + Chemical + + Bactrim + + + MESH:D000583 + Chemical + + Amikacin + + + MESH:D014295 + Chemical + + trimethoprim + + + + RESULTS + paragraph + 6250 + Data on the use of antibiotics and blood derivates are presented in table 4. + + + TABLE + T4.xml + T4 + table_caption + 6327 + The use of antibiotics, blood and blood derivates for the treatment of patients with rotavirus diarrhea in study. *Ceftriaxone in combination with amikacin or gentamicin + + 9606 + Species + + patients + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D000583 + Chemical + + amikacin + + + MESH:D005839 + Chemical + + gentamicin + + + MESH:D002443 + Chemical + + Ceftriaxone + + + + RESULTS + paragraph + 6497 + Assessment of the seasonal distribution of the rotavirus diarrhea has shown that the disease most commonly occurred in autumn, winter and early spring with peaks in September, November and January (Graph 1). All patients recovered without sequels. + + 9606 + Species + + patients + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + FIG + MSM-26-335-g005.jpg + F1 + fig_caption + 6745 + Seasonal distribution of the rotavirus diarrhea + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + DISCUSS + title_1 + 6793 + 4. DISCUSSION + + + DISCUSS + paragraph + 6807 + Rotavirus is the common cause of diarrhea of the infant worldwide. According to Parashar and associates, who reviewed the Global mortality associated with Rotavirus disease among children, rotavirus is responsible for 527,000 deaths (95% confidence interval, 475,000-580,000 deaths) annually or 29% of all deaths due to diarrhea among children <5 years of age. This translates into approximately 1440 deaths due to rotavirus disease per day (1 of 237 children born each year would die of rotavirus disease by 5 years of age). + + 9606 + Species + + infant + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D012400 + Disease + + Rotavirus disease + + + MESH:C564120 + Disease + + 000-580,000 deaths + + + MESH:D012400 + Disease + + rotavirus disease + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D003643 + Disease + + 000 deaths + + + MESH:D012400 + Disease + + rotavirus disease + + + + DISCUSS + paragraph + 7333 + Although rotavirus is ubiquitous and the incidence of disease is similar among children in industrialized and developing countries, there is a great disparity in mortality associated with rotavirus. Whereas very few deaths due to rotavirus disease occurred in affluent countries, countries with the greatest level of child mortality accounted for approximately 86% of all deaths due to rotavirus disease. In fact, 6 countries (India, Nigeria, China, Pakistan, Ethiopia, and Democratic Republic of Congo) accounted for >50% of all deaths due to rotavirus disease, with India alone accounting for one-fourth of the deaths. + + 9606 + Species + + children + + + 9606 + Species + + child + + + MESH:D012400 + Disease + + rotavirus disease + + + MESH:D012400 + Disease + + rotavirus disease + + + MESH:D012400 + Disease + + rotavirus disease + + + MESH:D006479 + Disease + + Democratic Republic of Congo + + + + DISCUSS + paragraph + 7954 + Papers discussing the prevalence of diarrhea caused by rotavirus in the Republic of Kosovo are lacking. At the moment, there are only two papers, one presented by Azemi alone and another presented by Azemi with coworkers discussing this issue. According to the paper in year 1987, 24.27% of children hospitalized at the Department of Pediatric Gastroenterology of the University Clinical Center of Kosovo had acute diarrhea associated with severe dehydration (toxicosis), and, of them, 7% inflicted by rotavirus. In 2002, according to the second paper, rotavirus is found responsible for 16.8% of acute diarrhea. + + 9606 + Species + + children + + + MESH:C565846 + Disease + + toxicosis + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + + DISCUSS + paragraph + 8567 + In our study, the percentage is found to be significantly higher - 34.42%. It is rather reflection of better living standard of the Kosovars and health care provision than of the change in epidemiological behavior of the virus. Better feeding habits, better education of the mothers and better health care, no doubt, have made for other causes of the diarrhea to diminish. + + MESH:D003967 + Disease + + diarrhea + + + + DISCUSS + paragraph + 8942 + With one third of acute diarrhea due to rotavirus (34.42%), prevalence of the disease in Kosovo is similar to the prevalence in other European countries. It is similar to the prevalence in Central and Southeastern Europe, that varies between 22% (Czech Republic) and 55.3% (Russia), with Bosnia and Hercegovina (23.9%), Hungary (27.1%), Albania (32.4%), Bulgaria (32.4%), Romania (42%) and Ukraine (42%) in between, as found by Ogilvie and coauthors in their study of burden of rotavirus gastroenteritis in the pediatric population in this region. But, also similar to the prevalence found in REVEAL study that have found prevalence ranging between 27.8% in Germany and 52% in Sweden, with Spain (31.2%), France (33.5%), Great Britain (35.9%), Italy (43.6%) and Belgium (44.7%) in between. + + MESH:D012400 + Disease + + rotavirus gastroenteritis + + + MESH:D003967 + Disease + + diarrhea + + + Disease + + Bosnia + + + Disease + + Romania + + + + DISCUSS + paragraph + 9732 + The majority of children with acute diarrhea in our study were infants (48%) and boys (66%). The higher infliction of the boys with rotavirus diarrhea is noted in several other studies, both local and worldwide, but, up to day, there are no known reasons established. Furthermore, there are studies as well, stating the opposite. + + 9606 + Species + + children + + + 9606 + Species + + infants + + + 9606 + Species + + boys + + + 9606 + Species + + boys + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D003967 + Disease + + diarrhea + + + + DISCUSS + paragraph + 10062 + Vomiting, diarrhea and temperature usually occurred at the second day after exposition, following an exposition period of less than 48 hours. Consequently, in our study, dehydration was present in all children. In almost every third child, this dehydration was severe. + + 9606 + Species + + children + + + 9606 + Species + + child + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + MESH:D014839 + Disease + + Vomiting + + + MESH:D003967 + Disease + + diarrhea + + + + DISCUSS + paragraph + 10331 + The REVEAL study reported that the proportion of children with dehydration due to acute RVGE varied between 11.1% (Spain) and 71.4% (Sweden), and in most countries, was considerably higher than for those with rotavirus-negative disease. In Belgium and the UK, the prevalence of dehydration was comparable among children with or without RVGE, whereas the dehydration prevalence ratio of rotavirus versus non-rotavirus disease was 1.82, 5.54, 3.27, 3.47, and 2.18, respectively, in France, Germany, Italy, Spain, and Sweden. A second prospective multicenter study from France, Germany, Italy, Spain, and the UK reported that dehydration was evident in 75.7% of patients with RVGE, and was severe in 11.3% of them. In comparison, only 54.5% of children with non-RVGE were dehydrated, while 4.7% had severe dehydration (25). Another study from France confirmed this trend, showing significant differences in dehydration between rotavirus-positive and negative gastroenteritis (26.8% vs.14.7%, P < .0001) as did a study from Greece. Further, one Italian community-based study showed that dehydration at initial presentation in primary care was associated with a higher likelihood of RVGE (OR: 1.8; 95% CI, 1.1-3; P = 0.02). + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + 9606 + Species + + children + + + MESH:D003681 + Disease + + dehydration + + + MESH:D012400 + Disease + + rotavirus versus non-rotavirus disease + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + MESH:D005759 + Disease + + gastroenteritis + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + MESH:D003681 + Disease + + dehydration + + + + DISCUSS + paragraph + 11550 + It is interesting to find that about every third patients in our study has bronchopneumonia as associated disease and every tenth had paralytic ileus. + + 9606 + Species + + patients + + + MESH:D007418 + Disease + + paralytic ileus + + + MESH:D001996 + Disease + + bronchopneumonia + + + + DISCUSS + paragraph + 11701 + Although affection of the central nervous system is not common in children with rotavirus diarrhea, it occurred in two our patients. First to report association of this disease with rotavirus diarrhea were Day and coauthors. They reported neurological symptoms and stool virology positive for rotavirus in four children, presented within 4 weeks in Barnet and Chase Farm Hospitals NHS Trust, London, UK. Similar to our patients, all four made a quick recovery and were discharged home after a few days without medication. At 6 weeks all were well, with no further neurological episodes. + + 9606 + Species + + children + + + 9606 + Species + + patients + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D009422 + Disease + + neurological symptoms + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + DISCUSS + paragraph + 12288 + Later, Fuchigami and coauthors reported a case of encephalopathy in 4 year old girl at the Department of General Pediatrics, Nihon University Nerima Hikarigaoka Hospital, Tokyo. Probably there will be more such reports in the future. + + 9606 + Species + + girl + + + MESH:D001927 + Disease + + encephalopathy + + + + DISCUSS + paragraph + 12522 + Use of antibiotics, blood and blood derivates in rotavirus diarrhea is a matter of a worldwide discussion. In out study almost two thirds of the patients were treated with antibiotics and about every twentieth was given blood or blood derivates. High association with bronchopneumonia may explain why. + + 9606 + Species + + patients + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + MESH:D001996 + Disease + + bronchopneumonia + + + + DISCUSS + paragraph + 12824 + Rotavirus diarrhea most commonly occurred in autumn, winter and early spring with peaks in September, November and January. + + MESH:D012400 + Disease + + Rotavirus diarrhea + + + + CONCL + title_1 + 12948 + 5. CONCLUSION + + + CONCL + paragraph + 12962 + Rotavirus diarrhea continues to be a serious health problem in Kosovo. Epidemiological and clinical features of the disease and Latex agglutination test are essential tools to establish the diagnosis at the time. + + MESH:D012400 + Disease + + Rotavirus diarrhea + + + + CONCL + footnote + 13175 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 13212 + REFERENCES + + + surname:Ruck;given-names:BJ + 1 + surname:Holmes;given-names:IH + surname:Davidson;given-names:GP + REF + surname:Bishop;given-names:RF + 1281 + 4127639 + 1973 + Lancet + ref + 1284 + 13223 + Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis + + 9606 + Species + + children + + + MESH:D005759 + Disease + + gastroenteritis + + + + 4 + REF + surname:Parashar;given-names:U + 4 + 1998 + Emerging Infectious Diseases + ref + 13333 + Rotavirus + + + 63 + surname:Feachem;given-names:RV + REF + surname:de Zoysa;given-names:I + 569 + 3876173 + 1985 + Bull World Health Organ + ref + 683 + 13343 + Interventions for the control of diarrhoeal diseases among young children: rotavirus and cholera immunisation + + 9606 + Species + + children + + + MESH:D004194 + Disease + + diarrhoeal diseases + + + + REF + ref + 13453 + Kosovo Agency of Statistics + + + 200 + REF + surname:Parashar;given-names:U + S9 + Supplement 1 + 19817620 + 2009 + J Infect Dis + ref + S15 + 13481 + Global Mortality Associated with Rotavirus Disease among Children in 2004 + + 9606 + Species + + Children + + + MESH:D012400 + Disease + + Rotavirus Disease + + + + REF + surname:Azemi;given-names:M + 1987 + article in Albanian + PHD tesis, Faculty of Medicine + ref + 13555 + The corelation of the etiological agent and acidobasic status in children with toxicosis + + 9606 + Species + + children + + + MESH:C565846 + Disease + + toxicosis + + + + surname:Begolli;given-names:M + surname:Koci;given-names:R + surname:Zejnullahu;given-names:M + REF + surname:Azemi;given-names:M + 2002 + Book of abstracts (article in Albanian) + surname:dhe;given-names:Gashi N + ref + surname:Gojani;given-names:Xh + surname:Ismaili-Jaha;given-names:V + surname:Avdiu;given-names:M + 13644 + Clinical and biochemical features of the acute diarrhea from rotavirus + + MESH:D003967 + Disease + + diarrhea + + + + surname:Goetghebeur;given-names:M + surname:El Khoury;given-names:A + surname:Khoury;given-names:H + surname:Ogilvie;given-names:I + 523 + 5 + 21422818 + 2011 + Hum Vaccin + ref + 7 + REF + 533 + 13715 + Burden of rotavirus gastroenteritis in the pediatric population in Central and Eastern Europe. Serotype distribution and burden of illness + + MESH:D012400 + Disease + + rotavirus gastroenteritis + + + + 195 + REF + S4 + Supplement 1 + 17387650 + 2007 + J Infect Dis + ref + S16 + 13854 + Multicenter Prospective Study of the Burden of Rotavirus Acute Gastroenteritis in Europe 2004 - 2005: The Reveal Study + + MESH:D012400 + Disease + + Rotavirus Acute Gastroenteritis + + + + surname:Begolli;given-names:M + surname:Koci;given-names:R + surname:Zejnullahu;given-names:M + REF + surname:Azemi;given-names:M + 2002 + Book of abstracts (article in Albanian) + surname:dhe;given-names:Gashi N + ref + surname:Gojani;given-names:Xh + surname:Ismaili-Jaha;given-names:V + surname:Avdiu;given-names:M + 13975 + Clinical and biochemical features of the acute diarrhea from rotavirus + + MESH:D003967 + Disease + + diarrhea + + + + surname:Raic;given-names:F + REF + surname:Votava;given-names:A + 1982 + article in Croatian + PHD thesis + ref + 14046 + Impact of diseases afecting gastrointestinal system in pediatric toxicosis + + MESH:D005767 + Disease + + diseases afecting gastrointestinal system + + + MESH:C565846 + Disease + + toxicosis + + + + surname:Apostolov;given-names:D + surname:Sosev;given-names:B + REF + surname:Stojanovski;given-names:N + 444 + 1979 + article in Bosnian + ref + 445 + 14121 + Acidobasic amd electrolitic disorders in pediatric toxicosis and their role in treatment + + MESH:D063766 + Disease + + pediatric toxicosis + + + + surname:Predokevic;given-names:L + surname:Anic;given-names:O + REF + surname:Hodzic;given-names:L + 147 + 1983 + article in Serbian + Novi Sad, Republic of Serbia + ref + 148 + 14210 + Prognosis of the children with diarrhea toxicosis.12th Congress of the pediatritian of the Yugoslavia + + 9606 + Species + + children + + + MESH:D003967 + Disease + + diarrhea toxicosis.12th Congress + + + + 99 + surname:de Villiers;given-names:GK + surname:Sawyerr;given-names:TN + REF + surname:de Villiers;given-names:FP + 249 + 4 + 19588778 + 2009 + S Afr Med J + ref + 252 + 14312 + The incidence and clinical presentation of infantile rotavirus diarrhoea in Sierra Leone + + MESH:D012400 + Disease + + infantile rotavirus diarrhoea + + + + surname:Sarikaya;given-names:S + surname:Gurol;given-names:Y + surname:Izbirak;given-names:G + surname:Akan;given-names:H + 8 + 1 + 19943964 + Asia Pac Fam Med + ref + 8 + REF + surname:Vitrinel;given-names:A + surname:Hayran;given-names:O + surname:Yilmaz;given-names:G + surname:Gunduz;given-names:TS + 14401 + Rotavirus and adenovirus frequency among patients with acute gastroenteritis and their relationship to clinical parameters: a retrospective study in Turkey + + 9606 + Species + + patients + + + MESH:D005759 + Disease + + acute gastroenteritis + + + + 31 + surname:Unicomb;given-names:LE + surname:Tzipori;given-names:SR + REF + surname:Jarecki-Khan;given-names:K + 484 + 3 + 8458940 + 1993 + J Clin Microbiol + ref + 489 + 14557 + Enteric adenovirus infection among infants with diarrhea in rural Bangladesh + + 9606 + Species + + infants + + + MESH:D003967 + Disease + + diarrhea + + + MESH:D000257 + Disease + + adenovirus infection + + + + surname:Catal;given-names:F + surname:Avci;given-names:Z + surname:Acikgoz;given-names:ZC + surname:Karadag;given-names:A + 269 + 4 + 15804663 + 2005 + Scand J Infect Dis + ref + 37 + REF + 275 + surname:Uras;given-names:N + surname:Gamberzade;given-names:S + surname:Gocer;given-names:S + 14634 + Childhood diarrhoea in Ankara, Turkey: epidemiological and clinical features of rotavirus-positive versus rotavirus-negative cases + + 9103 + Species + + Turkey + + + MESH:D003967 + Disease + + diarrhoea + + + + surname:Lwin;given-names:T + surname:Oo;given-names:WM + surname:Hummelman;given-names:EG + surname:Moe;given-names:K + S111 + Suppl 1 + 16088793 + 2005 + J Infect Dis + ref + 192 + REF + 113 + surname:Htwe;given-names:TT + 14765 + Hospital-based surveillance for rotavirus diarrhea in children in Yangon, Myanmar + + 9606 + Species + + children + + + MESH:D012400 + Disease + + rotavirus diarrhea + + + + surname:Marek;given-names:A + surname:Ratajczak;given-names:B + surname:Bako;given-names:W + surname:Rytlewska;given-names:M + 117 + 1 + 11208297 + 2000 + Med Sci Monit + ref + 6 + REF + article in Polish + surname:Korzon;given-names:M + surname:Tyl;given-names:J + 122 + surname:Swiatkowska;given-names:H + surname:Czarnecka-Rudnik;given-names:D + surname:Gwizdek;given-names:A + 14847 + Epidemiological and clinical characteristics of rotaviral diarrhoea in children from Gdansk, Gdynia and Sopot + + 9606 + Species + + children + + + Disease + + Gdynia + + + MESH:D003967 + Disease + + rotaviral diarrhoea + + + + surname:Mathew;given-names:MM + surname:Mathan;given-names:MM + surname:Zhen;given-names:LG + surname:Huilan;given-names:S + 549 + 5 + 1659953 + 1991 + Bull World Health Organ + ref + 69 + REF + surname:Sami;given-names:Z + surname:Khan;given-names:MA + surname:Ghafoor;given-names:MA + 555 + surname:Khin;given-names:Maung U + surname:Espejo;given-names:R + surname:Olarte;given-names:J + 14958 + Etiology of acute diarrhoea among children in developing countries: a multicentre study in five countries + + 9606 + Species + + children + + + MESH:D003967 + Disease + + diarrhoea + + + + surname:King;given-names:BR + surname:Awad;given-names:SH + REF + surname:Nguyen;given-names:DD + 2005 + Ne: eMedicine Specialties, Emergency Medicine, Pediatrics + ref + 15064 + Rotavirus gastroenteritis + + MESH:D012400 + Disease + + Rotavirus gastroenteritis + + + + 12 + REF + surname:Ogilvie + 62 + 22429601 + 2012 + BMC Infectious Diseases + ref + 15090 + Burden of community-acquired and nosocomial rotavirus gastroenteritis in the pediatric population of Western Europe: a scoping review + + MESH:D012400 + Disease + + nosocomial rotavirus gastroenteritis + + + + surname:Bryant;given-names:PA + surname:Jackson;given-names:C + REF + surname:Day;given-names:TG + 2012 + BMJ Case Rep, 2012 + ref + 15224 + Cluster of neurological manifestations of rotavirus infection in children + + MESH:D012400 + Disease + + rotavirus infection + + + + surname:Ismaili-Jaha;given-names:V + surname:Avdiu;given-names:M + surname:Azemi;given-names:M + surname:Hoxha;given-names:TF + 300 + 5 + 2014 + Med Arh + ref + 68 + REF + doi: 10.5455/medarh.2014.68.300-303 + 303 + surname:Petrela;given-names:E + surname:Gragjevci;given-names:V + 15298 + The Usefulness of Clinical and Laboratory Parameters for Predicting Severity of Dehydratation in Children with Acute Gastroenteritis + + 9606 + Species + + Children + + + MESH:D005759 + Disease + + Gastroenteritis + + + + + 4272831 + + surname:Lepara;given-names:Zahid + surname:Hasanbegovic;given-names:Munira + surname:Junuzovic;given-names:Dzelaludin + 25568625 + surname:Prstojevic;given-names:Jelena Kovacevic + 297 + 5 + 2014 + 4272831 + MSM-26-297 + 10.5455/msm.2014.26.297-302 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 297-302. doi:10.5455/msm.2014.26.297-302 + stone in urinary tract ESWL method of urinary stone treatment + 302 + surname:Selimovic;given-names:Mirsad + Prstojevic JK, Junuzovic D, Hasanbegovic M, Lepara Z, Selimovic M + 0 + Characteristics of Calculi in the Urinary Tract + + + ABSTRACT + abstract_title_1 + 48 + Introduction: + + + ABSTRACT + abstract + 62 + Elimination of stone is determined by size and its localization. Stone from the ureter in 80% of cases can be eliminated spontaneously. If the stone by its characteristics is not spontaneously eliminated, taken are further steps and therapeutic protocols to solve this problem. + + + ABSTRACT + abstract_title_1 + 340 + Material and methods: + + + ABSTRACT + abstract + 362 + The study was prospective, open and comparative. It was conducted at the Urology Clinic Clinical Center of Sarajevo University in the period from 2007 to 2013. The study included 404 patients with urinary tract lithiasis treated by ESWL. ESWL treatment is performed on the machine Siemens Model Lithostar Multiline, which has a combined ultrasonographic and fluoroscopic display, large energy density in order to obtain optimum focus (without damaging surrounding tissue) and minimal pain that on rare occasions requires for mild sedation-sedation. + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + MESH:D052878 + Disease + + urinary tract lithiasis + + + + ABSTRACT + abstract_title_1 + 911 + Results: + + + ABSTRACT + abstract + 920 + From a total of 404 patients included in the study there were 234 (57.92%) male and 170 (42.08%) female patients. The most common type of stone both in female and male patients was calcium type. From a total of 262 calcium stones, 105 of them (40.07%) was present in female patients and 157 (59.92%) in male. Share of infectious type of stone in female patients was 63 (49.60%) and 64 among males (50.39%). Other stones were less abundant in both the gender groups and their total number was only 17. In women their frequency was 2 (13.33%) and 13 among males (86.67%). There was a significant difference in the frequency of different types of stones by gender (chi2 = 11.47, p = 0.009). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + women + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + + ABSTRACT + abstract_title_1 + 1608 + Conclusion: + + + ABSTRACT + abstract + 1620 + There was no statistically significant correlation between the number of treatments and localization of stones in the ureter, as well as a statistically significant correlation between the size of the stone and the localization of calculus in the ureter. + + MESH:D002137 + Disease + + calculus + + + + INTRO + title_1 + 1875 + 1. INTRODUCTION + + + INTRO + paragraph + 1891 + Every year between the 1200 to 1400 persons per million develop urinary calculosis with a male to female ratio of 3:1. Urology is probably the oldest branch of surgery, knowing that one of the first operations was safe and resolve stone jammed on the outer opening of the urethra. + + 9606 + Species + + persons + + + MESH:D020347 + Disease + + calculosis + + + + INTRO + paragraph + 2172 + Creation of stones in the urinary organs (urolithiasis) is a very common disease. Urinary tract stones are sixth health problem. Epidemiological data suggest an increased prevalence of upper urinary tract calculi in developed countries. Various authors suggest that urinary calculi constitute 1-3 per thousand total hospitalized patients which makes 15-24% of patients in the urology departments. This disease have several names: nephrolithiasis-meaning kidney stone, nephrolithiasis and the word is derived from the Greek word nephros (kidney) and lithos (stone). Word of urolithiasis-urinary (urine) stone is derived from the French word urine and Greek ouron meaning urine. It is also used as renal calculus from the Latin word calculus (plural-calculi), which means gravel. Urolithiasis also affects animals: the stones were found in dogs, cats, rabbits, minks, etc., and the components are the same as in humans. Urolithiasis occurs during the most productive human age from 30 to 50 years of age, but has been described in all age groups. It was found in newborns and fetuses. However, urolithiasis in children is rare. Only 2-3% of cases occur in the pediatric age. It is believed that 12% of men and 4% of women in developed countries suffer from urinary stones. In case of a positive family history, this number doubles. One of the reasons for its frequent occurrence in men is explained by the increased endogenous production of oxalate in the liver under the influence of testosterone, while in the urine of women is on average higher concentration of citrate, otherwise, increase the solubility of calcium. Calculosa with women predominantly induced urinary infections and metabolic disorders, while in men predominate idiopathic lithiasis or calcium lithiasis and uric acid. Percentage of recurrent calculosis is 50% within 5 years and 70% over 10 years. Stones less than 2 mm easily pass through the ureter. Most calculi (90%) from 4 to 5 mm are eliminated spontaneously. Percentage decreases to 50% at stones of 4 to 6 mm and 20% at stones larger than 6 mm. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9615 + Species + + dogs + + + 9685 + Species + + cats + + + 9986 + Species + + rabbits + + + 9666 + Species + + minks + + + 9606 + Species + + humans + + + 9606 + Species + + human + + + 9606 + Species + + children + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + women + + + 9606 + Species + + men + + + MESH:D008569 + Disease + + Word + + + MESH:D007669 + Disease + + nephrolithiasis-meaning kidney stone + + + MESH:D052878 + Disease + + urolithiasis-urinary + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D008659 + Disease + + metabolic disorders + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002137 + Disease + + plural-calculi + + + MESH:D053040 + Disease + + nephrolithiasis + + + MESH:D010070 + Chemical + + oxalate + + + MESH:D014545 + Disease + + upper urinary tract calculi + + + MESH:D008569 + Disease + + word calculus + + + MESH:D020347 + Disease + + calculosis + + + MESH:D020347 + Disease + + lithiasis + + + MESH:D001745 + Disease + + urinary infections + + + MESH:D052878 + Disease + + Urolithiasis + + + MESH:D013739 + Chemical + + testosterone + + + MESH:D007669 + Disease + + renal calculus + + + MESH:D020347 + Disease + + idiopathic lithiasis + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D019343 + Chemical + + citrate + + + MESH:D008569 + Disease + + word nephros + + + MESH:D052878 + Disease + + Urolithiasis + + + + INTRO + title_1 + 4251 + 2. PROBLEM FORMULATION + + + INTRO + paragraph + 4274 + One of the most common pathological conditions in human medicine is the presence of a stone in the urinary tract, characterized more often as urolithiasis. In countries with developed industry this disease affects approximately 1500 to 2000 patients per million inhabitants. The disease usually affects people in the fourth and fifth decade of life who are working, and their absence from work due to healing has significant social and economic implications on the society. Stones can be localized in different areas of the urinary system. Localization of the stone is determined by several factors, including even the demographic characteristics. In developed countries, 97% of the stones are localized in the kidney and ureter where in 59% of cases in the ureter. Urethral stone is in 75% of cases located in the iliac and pelvic part of the ureter. Elimination of the calculi depends on size and its localization. Stones from the ureter in 80% of cases can be eliminated spontaneously. If the calculi in their characteristics are not spontaneously eliminated, taken are further steps and therapeutic protocols to solve this problem. Asperitan stone and stone larger than 7 mm in diameter, which practically cannot be spontaneously eliminated, threatening development of kidney urostasis and infection. Indications for an active therapeutic approach to ureter stones is stone diameter over 7 mm with low (below 20%) probability of spontaneous elimination or absence of spontaneous stone elimination of any size for a period longer than 30 days from the first renal colic, urinary infection develop, sepsis, calculous anuria, as well as the request of patients. Treatment of urolithiasis includes conservative, surgical treatment, and treatment with extracorporeal shock wave lithotripsy (ESWL) depending on the evaluation. Recently, endoscopic procedures, such as eretheroscopy (URS) and percutaneous lithotripsy (PCNL), together with the aforementioned ESWL, have been almost entirely replaced open surgery, so in just 30 years, radically changed the approach and functional outcome of this disease. + + 9606 + Species + + human + + + 9606 + Species + + patients + + + 9606 + Species + + people + + + 9606 + Species + + patients + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D001745 + Disease + + urinary infection + + + MESH:D001002 + Disease + + calculous anuria + + + MESH:D014516 + Disease + + ureter stones + + + MESH:D007674 + Disease + + kidney urostasis and infection + + + MESH:D018805 + Disease + + sepsis + + + + INTRO + paragraph + 6377 + ESWL treatment is the first-line treatment of urinary tract stones, but there are still no clearly defined limits and recommendations for its use in the treatment of urinary calculi, depending on its location, size and morphological structure. This raises the question of efficiency of ESWL depending on the characteristics of urinary calculi. + + + INTRO + title_1 + 6721 + 3. GOALS + + + INTRO + paragraph + 6730 + Determine characteristics of the patients with calculi in the urinary system, possibilities of disintegration of stones and its spontaneous elimination dependent on morphological structure of stones, their size and location in the urinary tract. + + 9606 + Species + + patients + + + + METHODS + title_1 + 6976 + 4. MATERIAL AND METHODS + + + METHODS + paragraph + 7000 + The study was prospective, open and comparative. It was conducted at the Urology Clinic Clinical Center of Sarajevo University in the period from 2007 to 2013. The study included 404 patients with urinary tract lithiasis. All patients prior to initiation of therapy-ESWL treatments were subjected to the following diagnostic procedures: anamnesis, clinical examination of patients, laboratory tests, ultrasound examination of the urinary tract and urinary tract X-ray, from which was derived chemical qualitative analysis of morphological stone composition. Diagnostics are used in the detection of complications such as obstruction with dilatation of the renal colorectal system, reduction of the renal parenchyma and in monitoring of renal obstruction during treatment. This type of diagnosis is not suitable for stones in the ureter and can make the difference between calcified and radiolucent concrement. ESWL treatment is performed on the machine Siemens Model Lithostar Multiline, which has a combined ultrasonographic and fluoroscopic display, large energy density in order to obtain optimum focus (without damaging surrounding tissue) and minimal pain that on rare occasions requires for mild sedation-sedation. For nominal and ordinal variables chi-square test was used. In cases when the frequency was lower than expected was used the Fisher's exact test. The degree of correlation was determined by means of the Spearman. P value of <0.05 was considered statistically significant. Statistical analysis was performed using SPSS computer software for statistical analysis (SPSS Statistical Package for the Social Sciences) version 13.0. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + MESH:D052878 + Disease + + urinary tract lithiasis + + + MESH:D007674 + Disease + + renal obstruction + + + MESH:D007674 + Disease + + dilatation of the renal colorectal system + + + MESH:D000402 + Disease + + obstruction + + + MESH:D007674 + Disease + + reduction of the renal parenchyma + + + + RESULTS + title_1 + 8649 + 5. RESULTS + + + RESULTS + paragraph + 8660 + From a total of 404 patients included in the study there were 234 (57.92%) male and 170 (42.08%) female patients. The most common age group in the sample was at age from 35 to 45 years and consisted of 110 respondents (27.09%). The minimum number of respondents had the age over 65 years 19 respondents (4.67%). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + RESULTS + paragraph + 8972 + The most common type of stone both in female and male patients was calcium type. From a total of 262 calcium stones, 105 of them (40.07%) was present in female patients and 157 (59.92%) in male. Share of infectious type of stone in female patients was 63 (49.60%) and 64 among males (50.39%). Other stones were less abundant in both the gender groups and their total number was only 17. In women their frequency was 2 (13.33%) and 13 among males (86.67%). There was a significant difference in the frequency of different types of stones by gender (chi2 = 11.47, p = 0.009). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + women + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + + RESULTS + paragraph + 9546 + Due to the very low prevalence of other types of stones and inability to perform the chi-square test to a group of other stones are grouped stones which, by virtue were cystine, xanthine stones and uric acid stones. The incidence of cystine calculi was 4 (0.9%), frequency of xanthine stones 3 (0.7%) and uric acid 8 (1.9%). In the group of female respondents 74 (40.88%) had calculus size up to 10 mm, while in the group of male patients stone size up to 10 mm had 107 (59.12%). In the group of female respondents 96 of them (43.05%) had a size of stone exceeding 10 mm, while in the group of male patients stone size over 10 mm had 127 (56.95%). There was no statistically significant difference in the incidence of stones with sizes up to 10 and over 10 mm by gender (chi2 = 0.192, p = 0.661) + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D019820 + Chemical + + xanthine + + + MESH:D002137 + Disease + + cystine calculi + + + MESH:D002137 + Disease + + calculus + + + MESH:D003553 + Chemical + + cystine + + + MESH:D019820 + Chemical + + xanthine + + + MESH:D014527 + Chemical + + uric acid + + + + RESULTS + paragraph + 10342 + Number of stones localized in the upper pole of the kidney in women was 43 (48.86%), and among men was slightly lower and amounted to 45 (51.13%). Number localized in the lower pole of the kidney in women was 22 (61.11%), and among men was slightly lower and amounted to 14 (38.88%). Number of stones localized medially among women was 38 (45.78%), and among men was slightly lower and amounted to 45 (54.21%). Number of localized in the renal pyelon in females was 27 (40.39%), and among men was slightly lower and amounted to 40 (59.70%). There was no statistically significant difference in the frequency of localization of stones by gender. + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + men + + + + RESULTS + paragraph + 10987 + The incidence of urolithiasis at the site of physiological narrowing of the ureter in female subjects was 22 (27.5%) and 58 in males (72.5%). The incidence of urolithiasis at the site of the physiological enlargement of the ureter in female subjects was 8 (50%), and the same among men 8 (50%). There was no statistically significant difference in the frequency of localization of calculi to physiological narrowing and widening of the ureter in relation to gender differences (p = 0.086). + + 9606 + Species + + men + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + + RESULTS + paragraph + 11477 + The most common size of the calculi in the sample was 15 mm. This size had 21.04% of respondents. The second most common size of the calculi in the sample was 9 mm, which had 14.6% of respondents, followed by the size of 20 mm, which had 14.38% of the respondents. The least frequent size calculi were 17 mm, which was only 0.25% of the respondents. + + + RESULTS + paragraph + 11827 + Size of the calculi in women was 12 (8-15 mm), while in men it was 12 (8-15 mm). There was no statistically significant difference in the size of calculi between males and females (p = 0.557). + + 9606 + Species + + women + + + 9606 + Species + + men + + + + RESULTS + paragraph + 12020 + There was a significant mild positive correlation between age and size of the stone in the total sample (rho = 0.240, p <0.01), i.e. Increase with age slightly increases the size calculi. + + + RESULTS + paragraph + 12208 + In the group of patients who had a negative family history of calculus size to 10 mm were present in 108 (59.67%), while in the group of subjects who had a family history size of stones up to 10 mm had 73 (40.33%). In the group of patients who had a negative family history of calculus size over 10 mm was registered in 160 (71.75%), while in the group of subjects who had a positive family history of calculus size over 10 mm was registered in 63 (28.25%). There was a statistically significant correlation between the size of stones and positive / negative family history (chi2 = 6.529, p = 0.011), respectively, in patients with a positive family history more often were present small stones (to 10 mm). Size of the calculi in patients who had a positive family history was 12 (9-15 mm), while the size of stones in patients who had a positive family history was 10 (8-15 cm). There was no statistically significant difference in the size of the stones between the groups of patients (p = 0.013). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002137 + Disease + + calculus + + + MESH:D002137 + Disease + + calculus + + + MESH:D002137 + Disease + + calculus + + + + RESULTS + paragraph + 13208 + Size of calculus in patients without recurrent urolithiasis was 11 (9-15 mm), while the size of the calculus in patients who have had recurrent urolithiasis was 12 (8-15 mm). There was no statistically significant difference in the size of calculi between groups of patients (p = NS). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002137 + Disease + + calculus + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D002137 + Disease + + calculus + + + MESH:D052878 + Disease + + urolithiasis + + + + RESULTS + paragraph + 13493 + Frequency of localization in the upper pole of kidney which have the size of 10 mm was 30 (34.09%), while the frequency of localization in the top half of kidney s which had more than 10 mm in size was 58 (65.91%) (Table 1). + + + TABLE + T1.xml + T1 + table_caption + 13718 + The dependence of the calculus size and localization in relation to poles. Fisher exact test; p=0.003 + + MESH:D002137 + Disease + + calculus + + + + RESULTS + paragraph + 13820 + Frequency of localization in the lower pole of the calculi that had the size of 10 mm was in 3 cases (8.33%), while the frequency of localization in the lower pole of the calculi that had the size over 10 mm was present in 33 cases (91.67%). There was a significant dependence between the size of the stone and the same localization in renal pole (Fisher's exact test, p = 0.003). + + + RESULTS + paragraph + 14201 + Looking at the frequency of stones it is a lot bigger on the top half, but looking at their size, larger stones (over 10 mm) were significantly more often present in the lower half. + + + RESULTS + paragraph + 14383 + Size of calculi localized medially was 14 (10-15 mm), while the localization pyelon was 15 (12-18 mm). There was a significant difference in size between the mentioned calculus localization (p = 0.022). Stones that occur in the renal pyelon are higher than the stones that are found in our study, in medial position. + + MESH:D002137 + Disease + + calculus + + + + RESULTS + paragraph + 14700 + Frequency medial localization of calculi which size was less than 10 mm was 33 (39.75%), and frequency of 50 (60.25%) for stones ranging in size over 10 mm. Pyelon localization of calculi which size was less than 10 mm was 5 (7.46%), and frequency of 62 (92.53%) for stones ranging in size over 10 mm. There was a significant correlation between the size of the stone and localization of stones (medial / pyelon) (chi2 = 20.443, p <0.001). + + + RESULTS + paragraph + 15140 + Frequency of calculi localization at sites of physiological stricture site that had the size of 10 mm was 68 (85%), while the frequency of localization at sites of calculi physiological stricture site which had the size over 10 mm was 12 (15%). Frequency of localization at sites of calculi enlargement physiological ureter which had the size of 10 mm was 15 (93.75%), while the frequency of localization at sites of calculi enlargement physiological ureter which had the size over 10 mm, was 1 (6.25%). There was no statistically significant dependence between the size of tartar and calculus localization to physiological constriction and expansion of the ureter (p = 0.688) (Table 2). + + MESH:D002137 + Disease + + calculus + + + MESH:D002137 + Disease + + calculi enlargement + + + MESH:D002137 + Disease + + calculi enlargement + + + + TABLE + T2.xml + T2 + table_caption + 15828 + Dependence between the size of the stone and its localization in the ureter. Fisher exact test; p=0.688. E1- physiological narrowing of the ureter, E2 physiological enlargement of the ureter + + + RESULTS + paragraph + 16019 + In the group of patients under 35 years the incidence of calcium stones was 88 (33.58%), while in the age group of 35-55 years the incidence of calcium type of calculi was 135 (51.52%). The frequency of this type of calculi in subjects older than 55 years was 39 (14.88%). In the group of patients under 35 years the incidence of infectious stones was 26 (20.47%), and at the age of 35 to 55 years the incidence of infectious types of calculi was 63 (49.60%). The frequency of this type of calculi in subjects older than 55 years was 38 (29.92%). In the group of patients under 35 years the incidence of other types of stones was 4 (26.66%), while in the age group of 35-55 years the incidence of other types of calculi was 7 (46.66%). The incidence of other types of calculi in subjects older than 55 years was 4 (26.66%). Statistically significant dependence between age and type of calculi (chi2 = 15.170, p = 0.004) (Table 3). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + + TABLE + T3.xml + T3 + table_caption + 16950 + Dependence of age and types calculus. chi2=15.170 df=4; p=0.004 + + MESH:D002137 + Disease + + calculus + + + + DISCUSS + title_1 + 17014 + 6. DISCUSSION + + + DISCUSS + paragraph + 17028 + Renal lithiasis is a disease in which the stones were formed in the collecting tubules, cups and pyelon. The incidence was similar in both kidneys, and about 40% of patients had bilateral stones. There is a wide range of risk factors that may be associated with the disease, including local and general factors. Of local risk factors that favor the emergence of this disease are: trail of urine, disorders of innervation, anomalies of drainage pathways, anatomical abnormalities (sponge kidney, horseshoe kidney), and recurrent infection. General risk factors are categorized as: metabolic (calciuria, cystinuria), hormonal (primary hyperparathyroidism, hyperthereodosis and hypovitaminosis) and other factors such as climatic conditions of life, feeding, pH of urine excretion of concentrated urine, prolonged immobilization, etc.. + + 9606 + Species + + patients + + + MESH:D049950 + Disease + + primary hyperparathyroidism, hyperthereodosis and hypovitaminosis + + + MESH:D020347 + Disease + + Renal lithiasis + + + MESH:D007239 + Disease + + infection + + + MESH:D003555 + Disease + + cystinuria + + + + DISCUSS + paragraph + 17861 + Formation of kidney stones attempted to explain many theories but none fully explains the mechanism of occurrence. Urine is in the usual conditions supersaturated solution in which the particles are held in solution influence crystallization inhibitor and colloids. If these factors are disrupted leads to precipitation and aggregation of crystals. The organic matrix, which consists of cellular detritus, blood and bacteria may be the main factor that leads to nucleation and crystal growth. One theory in the spotlight puts deficit of crystallization inhibitors. There is a theory which states that the formation of stones occurs when the crystals, which are constantly being created in the urine oversaturated with salts, are not washed away in the urine. This may occur due to damage to the epithelial duct to which it adheres crystal. + + MESH:D007669 + Disease + + kidney stones + + + + DISCUSS + paragraph + 18701 + Changes in the socio-economic conditions have influenced the changes in the frequency and type of urolithiasis in terms of localization and physico-chemical properties of stones. Major variations on the occurrence of urolithiasis in the world are presented to the public in terms of geographical areas. Annual incidence in England is 22 cases per 100,000 inhabitants, in Kuwait 23.9 per 100,000 population. However, in some countries there are significant differences in the incidence, such as Sweden, where the incidence of 140 per 100,000 inhabitants, Italy 168 per 100,000 inhabitants and United States with high prevalence of 277 per 100,000 population. In Europe, urinary stones are found mainly in the upper urinary tract, while the proportion of stones in the bladder does not exceed 10.0%. It is shown that the urinary bladder calculi more common in the elderly. + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D001744 + Disease + + urinary bladder calculi + + + MESH:D052878 + Disease + + type of urolithiasis + + + + DISCUSS + paragraph + 19572 + The disease is more common in men and usually occurs after the third decade of life. It is shown that Caucasians are more likely to develop kidney stones than African Americans, and men more often than women patients. A possible explanation why men are more frequent in patients than in females lies in the fact that men in performing physical activities much sweat and lose a lot of fluid, which is also the main reason for the formation of a kidney stone or a stone in the bladder. + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + patients + + + 9606 + Species + + men + + + 9606 + Species + + patients + + + 9606 + Species + + men + + + MESH:D007669 + Disease + + kidney stones + + + MESH:D007669 + Disease + + kidney stone + + + + DISCUSS + paragraph + 20056 + Some new studies are mostly directed of change in the relationship of occurrence calculi in men and women. In the United States-and the data showed that the overall incidence of urolithiasis 10.6% for men and 7.1% for women. This is explained by balancing risks for both men and women today, compared to the past when they were significantly different. + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + MESH:D052878 + Disease + + urolithiasis + + + + DISCUSS + paragraph + 20411 + Results of this study showed that the total number of subjects included higher percentage occupied by male respondents and 57.92%, while the remaining 42.08% are occupied by the respondents are female. The results are consistent with published data showing a higher incidence of this disease in male patients compared to female respondents. + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 20752 + Greater tendency of men to concentrate urine compared to the opposite sex, may be the reason for higher incidence of this disease in men. It is shown that the higher the osmolality in men also work super saturation which is responsible for the crystallization of poorly soluble compounds. Even if the concentration of poorly soluble compounds reaches the threshold of saturation of the total 24-h urine, then it can exceed during transient episodes, the way to increase the intake of foods rich in protein, at night, or during intense exercise, especially in the summer months, or season in which men showed a significant decrease in urine volume. Although urolithiasis 2-3 times more common in men, there are no studies that clearly define the reasons for variations incidence of this disease. + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + men + + + MESH:D052878 + Disease + + urolithiasis + + + + DISCUSS + paragraph + 21547 + Daudon and colleagues show that men older than 80 years constitute 40.0% of the analyzed patients with urinary bladder calculi. Prostatic hyperplasia is more common in the elderly and causes an obstruction in the urinary tract. This may be a possible explanation for the high incidence of stones in the bladder in the elderly. Women are also at risk calculus appears in the lower parts of the urinary tract. This suggests the existence of other risk factors, such as changes in bladder function associated with relaxation of smooth muscle tone in the elderly, with an efficiency reduction bladder emptying favoring stagnation of urine and the occurrence of tartar. + + 9606 + Species + + men + + + 9606 + Species + + patients + + + 9606 + Species + + Women + + + MESH:D000402 + Disease + + obstruction + + + MESH:D001744 + Disease + + urinary bladder calculi + + + MESH:D011470 + Disease + + Prostatic hyperplasia + + + MESH:D001745 + Disease + + efficiency reduction bladder + + + MESH:D002137 + Disease + + calculus + + + + DISCUSS + paragraph + 22212 + When it comes to the size calculi, the results of our study showed a statistically significant positive correlation between age and size of the stone in the total sample (rho = 0.240, p <0.01). Classifying respondents into three age groups, our results showed that the highest incidence of stones measuring greater than 10 mm was represented at the age between 35 and 55 years. Chi-square test showed a statistically significant correlation between age and size of the stone (chi2 = 40.287, p <0.00005). + + + DISCUSS + paragraph + 22716 + The study by Alaya and associates, who analyzed the sample of 1301 urinary tract stones, it has been proven that the highest percentage share of 58.6% occupied stones of calcium type, that type of calcium oxalate, but they also recorded an increase in the incidence and types of stones uric acid. The authors believe that in the last 50 years there has been a change in eating habits associated with an increase in foods rich in purines (animal proteins projections and seafood), which correlates the increase in uric acid in the urine, and increasing incidence of this type of calculi. Their results show a clear increase in uric acid stones in both sexes. + + MESH:D002118 + Chemical + + calcium + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D002129 + Chemical + + calcium oxalate + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D011687 + Chemical + + purines + + + + DISCUSS + paragraph + 23374 + The results of our study showed that the group of patients under 35 years the incidence of calcium stones was 33.58%, the incidence of infectious stones 20.47%, while the incidence of other types of stones was 26.66%. At the age of 35 to 55 years the incidence of a calcium type of calculi was 51.52%, the frequency of the infectious type of calculi was 49.60%, and the prevalence of other types of stones was 46.66%. The frequency of this type of calcium calculi in subjects older than 55 years was 14.88%, infectious stones were 29.92%, while the incidence of other types of calculi in subjects older than 55 years was 26.66%. There was a statistically significant correlation between age and type of calculi (chi2 = 15.170, p = 0.004), i.e. among younger patients are the most common types of calcium stones, while the incidence of infectious types of stones rarer. Increasing age leads to equalization of the frequency of calcium and infectious stones. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + + DISCUSS + paragraph + 24331 + Differences in the incidence of age and stones were analyzed through a large number of epidemiological studies. There are certain variations according to geographical areas. In 1993, Baker et al reported that in Australia peak incidence of urolithiasis calcium oxalate sampled observed in individuals between 50 and 60 years of age. In Europe, research shows that the stones which are chemically calcium oxalate more common in people between 40 and 50 years of age. In Asia, the highest prevalence of calcium oxalate stone formation occurs at an earlier age range 30 to 50 years. + + 9606 + Species + + people + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D002129 + Chemical + + calcium oxalate + + + MESH:D002129 + Chemical + + calcium oxalate + + + MESH:D002129 + Chemical + + calcium oxalate + + + + DISCUSS + paragraph + 24911 + Certain hereditary disorders that run in the family increases the risk of recurrent kidney stones. A rare hereditary disease, renal tubular acidosis, increases the acidity of urine, which is favorable for the occurrence of kidney stones. Cystinuria is a hereditary disorder of the metabolism of amino acids, which results in high levels of cystine in the urine and blood, leading to frequent formation of cystine kidney stones. Second, a hereditary disorder of metabolism, hyperoxaluria, is resulting in high levels of oxalate salt in the body, which is combined with calcium in the form of kidney stones. Hypercalciuria causes high accumulation of calcium in the body, which increases the incidence of kidney stones. Hiperuricosuria increased level of uric acid in the urine, which leads to the formation of uric acid stones. + + MESH:D002118 + Chemical + + calcium + + + Disease + + Hiperuricosuria + + + MESH:D002118 + Chemical + + calcium + + + MESH:D007669 + Disease + + cystine kidney stones + + + MESH:D007669 + Disease + + kidney stones + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D007669 + Disease + + kidney stones + + + MESH:D030342 + Disease + + hereditary disease + + + MESH:D003553 + Chemical + + cystine + + + MESH:D030342 + Disease + + hereditary disorder + + + MESH:D030342 + Disease + + hereditary disorder + + + MESH:D006959 + Disease + + hyperoxaluria + + + MESH:D030342 + Disease + + hereditary disorders + + + MESH:D007669 + Disease + + kidney stones + + + MESH:D000141 + Disease + + renal tubular acidosis + + + MESH:D003555 + Disease + + Cystinuria + + + MESH:D014527 + Chemical + + uric acid + + + MESH:D007669 + Disease + + kidney stones + + + MESH:D053565 + Disease + + Hypercalciuria + + + MESH:D010070 + Chemical + + oxalate salt + + + + DISCUSS + paragraph + 25738 + Diet, in terms of animal protein (52 g / day), sodium (50 mg / day) and the oxalate (200 mg / day) with a normal intake of calcium (1,200 mg / day), reduced recurrent stones for almost 50% and more, within five years, compared to a diet low in calcium (400 mg / day) and oxalate. + + MESH:D010070 + Chemical + + oxalate + + + MESH:D012964 + Chemical + + sodium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D010070 + Chemical + + oxalate + + + + DISCUSS + paragraph + 26018 + Our results show that the proportion of respondents who did not have positive personal and family history of urolithiasis was 44.94%, and patients who had a positive family history of urolithiasis, but he had a personal history was 50.05%. The share of respondents, who had a family history, and negative personal history of urolithiasis, was 42 (30.88%), while the number of respondents who had a positive to families and personal history was 136 (33.74%). There was a significant frequency dependence of family and personal history (chi2 = 7.41, p = 0.006), i.e. Higher incidence of recurrence in patients who have a positive family history of urolithiasis. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + MESH:D052878 + Disease + + urolithiasis + + + + DISCUSS + paragraph + 26678 + Similar results were presented by the study Koyuncu and associates, which showed a significant correlation recurring calculi and positive family history. The authors also found that the time interval between the onset of recurrence was significantly shorter in patients who had a positive family history of urolithiasis. By analyzing the patients according to sex, the authors noted that the incidence of recurrence was more frequent in males than females respondents. The authors believe that the information positivity family history is very important and can provide valuable information about the possibility of future attacks as well as the severity of the disease. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D052878 + Disease + + urolithiasis + + + + DISCUSS + paragraph + 27349 + Epidemiological and randomized studies have shown greater security ESWL treatment methods in breaking stones when it starts with applying lower energy sequences of the same, with a gradual increase energy sequences, resulting in a vasoconstriction which prevents renal damage and the difference in the fragmentation is not significant despite the fact that whether amplification is carried out or not. + + MESH:D007674 + Disease + + renal damage + + + + DISCUSS + paragraph + 27751 + Previous clinical and epidemiological studies have shown that as an indication of ESWL treatment of urolythiasis depends on several factors, including the size, localization, consistency and other histological characteristics of calculi. Pregnancy and specific internship and urological diseases, with an emphasis on acute urinary infection, contraindications to perform of ESWL. + + MESH:D014570 + Disease + + urological diseases + + + MESH:D001745 + Disease + + urinary infection + + + MESH:D019966 + Disease + + urolythiasis depends + + + + CONCL + title_1 + 28131 + 7. CONCLUSION + + + CONCL + paragraph + 28145 + From a total of 404 patients included in the study there was 57.92% male and 42.8% female respondents, or male : female ration of 1.2 : 1, while the most common age group in a sample of patients with urolithiasis was between 35 and 45 years(27%). The mean size of the stones in men and in women was 12 mm. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + men + + + 9606 + Species + + women + + + MESH:D052878 + Disease + + urolithiasis + + + + CONCL + paragraph + 28451 + Most frequent type of calculi both in female and male subjects was of calcium type. In younger patients the most common are calcium stones, with increase in respondents age calcium and infectious stones frequencies were equalized. + + 9606 + Species + + patients + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + MESH:D002118 + Chemical + + calcium + + + + CONCL + paragraph + 28682 + There was a significant correlation between the size of the stone and its localization in renal pole. The incidence of stones was significantly greater in the top half, but the size of large stones (over 10 mm) were significantly more present in the lower kidney pole. + + + CONCL + paragraph + 28951 + There was no statistically significant relationship between the size of stones and personal history, but statistically significant correlation is determined between the size of the stone and family history of urolithiasis. + + MESH:D052878 + Disease + + urolithiasis + + + + CONCL + paragraph + 29174 + Also there was significant correlation between the size of the stone and its localization (medial / pyelon). Stones up to 10 mm are more frequently localized medially, and those larger than 10 mm are somewhat more common in the pyelon. + + + CONCL + footnote + 29410 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 29447 + REFERENCES + + + REF + surname:Kovacevic-Prstojevic;given-names:J + 2014 + Magistarski rad + ref + 29458 + Broj vantjelesnih razbijanja kamenaca u odnosu na velicinu kamenca, njegovu lokalizaciju u urinarnom traktu, te morfolosku strukturu kamenca + + + 22 + surname:Kovacevic-Prstojevic;given-names:J + REF + surname:Junuzovic;given-names:Dz + 309 + 5 + 2014 + doi: 105455/aim.2014.22.309-314 + Acta Inform Med + ref + 314 + 29601 + Evaluation of Extracorporeal Shock Wave Lithotripsy (ESWL): Efficasy in Treatment of Urinary System Stones + + + surname:Tanagho;given-names:EA + surname:McAninch;given-names:JW + surname:Tanagho;given-names:EA + REF + surname:Marshall;given-names:LS + 2004 + Smith's General Urology + ref + 29708 + Urinary Stone Disease + + + REF + Dostupno na: + ref + 29730 + European Association of Urology. Guidelines 2012 edition + + + surname:Urologija;given-names:I + REF + surname:Markovic;given-names:V + 1997 + ref + 29787 + Hirurgija mokracnih organa + + + 16 + surname:Perkovic;given-names:J + REF + surname:Galic;given-names:J + 180 + 91 + 2010 + MEDIX + ref + 184 + 29815 + Pristup lijecenju urolitijaze + + + surname:Zhang;given-names:SL + surname:Zhang;given-names:Q + surname:Fan;given-names:YZ + surname:Li;given-names:LP + 295 + 4 + 23927804 + 2013 + Zhonghua Er Ke Za Zhi + ref + 51 + REF + 297 + 29846 + Clinical analysis of 41 children's urinary calculus and acute renal failure + + 9606 + Species + + children + + + MESH:D002137 + Disease + + calculus + + + MESH:D058186 + Disease + + acute renal failure + + + + 350 + REF + surname:Teichman;given-names:JMH + 684 + 7 + 14960744 + 2004 + N Engl J Med + ref + 693 + 29922 + Acute Renall Colic from Ureteal Calculus + + MESH:D002137 + Disease + + Ureteal Calculus + + + + surname:Vrhovac;given-names:B + surname:Radonic;given-names:M + REF + surname:Hadzic;given-names:N + 1990 + Prirucnik interne medicine + ref + 29963 + i sur + + + surname:Partin;given-names:AW + surname:Novick;given-names:AC + surname:Kavoussi;given-names:LR + REF + surname:Wein;given-names:AJ + 2011 + ref + 29969 + Campbell-Walsh Urology + + + 47 + surname:Alken;given-names:P + surname:Kohrmann;given-names:KU + REF + surname:Kalble;given-names:T + 537 + 5 + 18379747 + 2008 + doi: 10.1007/s00120-008-1732-1733 + Urologe A + ref + 29992 + Clinically relevant facets of urolithiasis + + MESH:D052878 + Disease + + urolithiasis + + + + surname:Saad;given-names:H + surname:Belgith;given-names:M + surname:Nouri;given-names:A + surname:Alaya;given-names:A + 177 + 3 + 22563551 + 2012 + Ann Lab Med + ref + 32 + REF + 183 + surname:Najjar;given-names:MF + surname:Jouini;given-names:R + 30035 + Changes in urinary stone composition in the Tunisian population: a retrospective study of 1,301 cases + + + surname:Saigal;given-names:CS + 62 + surname:Hanley;given-names:JM + surname:Smith;given-names:AC + REF + surname:Scales;given-names:CD;suffix:Jr + 160 + 2012 + Eur Url + ref + 165 + 30137 + Urologic Diseases in America Project. Prevalence of kidney stones in the United States Source + + MESH:D007669 + Disease + + kidney stones + + + + 239 + surname:Jungers;given-names:P + REF + surname:Daudon;given-names:M + 37 + 2001 + Feuillets de Biologie + ref + 39 + 30231 + Diabetes and calculations + + MESH:D003920 + Disease + + Diabetes + + + + surname:Rofe;given-names:AM + surname:Bais;given-names:R + surname:Coyle;given-names:P + surname:Baker;given-names:PW + 390 + 6 + 8377690 + 1993 + Med J Aust + ref + 159 + REF + 392 + 30257 + Influence of season, age, and sex on renal stone formation in South Australia + + + 363 + surname:Coe;given-names:FL + REF + surname:Worcester;given-names:EM + 954 + 10 + 20818905 + 2010 + N Engl J Med + ref + 963 + 30335 + Clinical practice. Calcium kidney stones + + MESH:D002118 + Chemical + + Calcium + + + MESH:D007669 + Disease + + kidney stones + + + + surname:Sarica;given-names:K + surname:Eryildirim;given-names:B + surname:Yencilek;given-names:F + surname:Koyuncu;given-names:HH + 105 + 2 + 20077110 + 2010 + Urol Res + ref + 38 + REF + 109 + 30376 + Family history in stone disease: how important is it for the onset of the disease and the incidence of recurrence? + + MESH:D007669 + Disease + + stone disease + + + + + 4272832 + + surname:Agincic;given-names:Alija + surname:Cvorak;given-names:Alemko + 25568583 + surname:Rustempasic;given-names:Nedzad + 329 + 5 + 2014 + 4272832 + AIM-22-329 + 10.5455/aim.2014.22.329-332 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 329-32. doi:10.5455/aim.2014.22.329-332 + varicose veins EVLA surgical treatment comparison + 332 + Rustempasic N, Cvorak A, Agincic A + 0 + Outcome of Endovenous Laser Ablation of Varicose Veins + + + ABSTRACT + abstract_title_1 + 55 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 64 + Introduction: + + + ABSTRACT + abstract + 78 + In Bosnia and Herzegovina according to available data, treatment of incompetent superficial lower extremity varicose veins by endovenous laser ablation (EVLA) has been introduced two years ago and so far no paper has been published regarding results of EVLA treatment of patients from our country. We wanted to present our results with EVLA treatment. + + 9606 + Species + + patients + + + - + Chemical + + EVLA + + + Disease + + Bosnia + + + - + Chemical + + EVLA + + + - + Chemical + + EVLA + + + + ABSTRACT + abstract_title_1 + 430 + Aim of study: + + + ABSTRACT + abstract + 444 + to evaluate and compare primary posttreatment outcomes of endovenous laser ablation (EVLA) with classical surgical method of varicose vein treatment. + + + ABSTRACT + abstract_title_1 + 594 + Patients and methods: + + 9606 + Species + + Patients + + + + ABSTRACT + abstract + 616 + The study was clinical and prospective. It was carried out at Clinic for vascular surgery in Sarajevo where fifty-eight (58) patients received surgical treatment for varicose veins and in Aesthetic Surgery Center "Nasa mala klinika" in Sarajevo were sixty-one (61) patients with varicose veins were treated by endovenous laser ablation. Total 119 patients (limbs) with pathologic reflux only in great saphenous vein were evaluated between 1st of January 2013 and 31st of April 2014. Following primary outcome endpoints were evaluated smean day of return to normal everyday activities, patient subjective quantification of pain during first seven days after intervention, incidence of deep venous thrombosis (DVT), incidence of wound bleeding requiring surgical intervention, incidence of peri-saphenous vein hematoma and infection rate. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D006470 + Disease + + bleeding + + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D006406 + Disease + + vein hematoma and infection + + + MESH:D010146 + Disease + + pain + + + + ABSTRACT + abstract_title_1 + 1457 + Results: + + + ABSTRACT + abstract + 1466 + Mean of return to normal activities (expressed in days after intervention); EVLA vs. stripping (surgery) =1.21vs12.24, T test 13,619; p=0, 000, p<0,05. T test was used for comparing Mean value of visual pain analog scale for the first 7 days between groups, for all seven days pain was significantly higher in surgical group of patients as compared to EVLA group; p<0,05. Incidence of hematoma greater than 1% of total body surface area was significantly higher in patients receiving surgical treatment; Pearson Chi Square=23,830, p<0,05; odds ratio:10,453. Incidences of infection, deep venous thrombosis and posttreatment bleeding were not statistically different between analyzed groups; EVLA vs Surgery (Pearson Chi Square =3,237; p>0,05; Pearson Chi Square=2,139, p>0,05, Pearson Chi Square=2,139, p>0,05, respectively.) + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D020246 + Disease + + deep venous thrombosis + + + - + Chemical + + EVLA + + + MESH:D014786 + Disease + + visual pain + + + MESH:D007239 + Disease + + infection + + + MESH:D010146 + Disease + + pain + + + MESH:D006406 + Disease + + hematoma + + + MESH:D006470 + Disease + + bleeding + + + + ABSTRACT + abstract_title_1 + 2292 + Conclusion: + + + ABSTRACT + abstract + 2304 + EVLA offers better patient recovery in terms of significantly lower post treatment pain, faster return to everyday activities and lower incidence of bruising (hematomas). + + 9606 + Species + + patient + + + MESH:D003288 + Disease + + bruising + + + MESH:D006406 + Disease + + hematomas + + + - + Chemical + + EVLA + + + MESH:D010146 + Disease + + pain + + + + INTRO + title_1 + 2475 + 1. INTRODUCTION + + + INTRO + paragraph + 2491 + In the recent past the gold standard in the treatment of lower extremity varicose veins with incompetent great saphenous vein (GSV) or small saphenous vein (SSV) was classical surgicall method of ligation and stripping of incompetent saphenous vein along with varicectomies. However, during the last decade other minimal invasive techniques have been invented and introduced to treat the incompetent saphenous vein and varicosities. Endovenous laser ablation (EVLA) is one of them and it has been used as minimally invasive replacement to classical surgical way of treating varicose veins for many years (Figure 1, Figure 2). Avoiding surgical incisions, mechanical disruption of the sapheno-femoral junction (SFJ) as well as aggressiveness of avulsion of saphenous vein, EVLA may offer reduced postoperative pain, post intervention bleeding and peri-saphenous vein hematoma along with decreased rate of wound infection and potentially shorter periods of absenteeism from work. In addition neo-vascularisation is significantly reduced which in turn might cause decreased incidence of varicose vein recurrence. + + MESH:D006406 + Disease + + vein hematoma + + + MESH:D006470 + Disease + + bleeding + + + MESH:D010149 + Disease + + postoperative pain + + + MESH:D001523 + Disease + + aggressiveness of avulsion + + + MESH:D007239 + Disease + + infection + + + + FIG + AIM-22-329-g001.jpg + F1 + fig_caption + 3601 + Varicose veins before treatment + + + FIG + AIM-22-329-g002.jpg + F2 + fig_caption + 3633 + Same patient after EVLA treatment + + 9606 + Species + + patient + + + - + Chemical + + EVLA + + + + INTRO + paragraph + 3667 + In Bosnia and Herzegovina according to available data, endovascular laser ablation of incompetent superficial lower extremity veins has been introduced two years ago and so far no paper has been published regarding EVLA treatment of patients from our country. Since many papers on this topic have certain contradictory results we wanted to present our experience with these two methods of varicose vein tretament. + + 9606 + Species + + patients + + + - + Chemical + + EVLA + + + Disease + + Bosnia + + + + INTRO + title_1 + 4081 + 2. AIM OF STUDY + + + INTRO + paragraph + 4097 + To evaluate and compare primary posttreatment outcomes of endovenous laser ablation (EVLA) with classical surgical method of varicose vein treatment + + + METHODS + title_1 + 4246 + 3. PATIENTS AND METHODS + + 9606 + Species + + PATIENTS + + + + METHODS + paragraph + 4270 + The study was clinical and prospective. It was carried out at Clinic for vascular surgery in Sarajevo where patients received surgical treatment for varicose veins and in Aesthetic Surgery Center "Nasa mala klinika" were endovenous laser treatment of varicose vein was performed. Total 119 patients with pathologic reflux only in great saphenous vein were evaluated between 1st of January 2013 and 31st of April 2014. Sixty-one (61) patients who received endovenous laser treatment for varicose veins in Aesthetic Surgery Center "Nasa mala klinika" were assigned to group A, while fifty-eight (58) patients that received surgical treatment of varicose veins at the Clinic for Vascular surgery were assigned to Group B. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + METHODS + paragraph + 4998 + Primary outcome endpoints were evaluated: mean day of return to normal everyday activities, patient subjective quantification of pain during first seven days after intervention, incidence of deep venous thrombosis (DVT), incidence of wound bleeding requiring surgical intervention, incidence of peri-saphenous vein hematoma and infection rate. Every day activities were defined as ability for self-care and all other professional, recreational, household or outdoor physical activities that used to be carried out before varicose vein treatment. Return to those activities was advised to all patients to be as soon as possible and the information about the day of return to those activities were recorded when patient resumed them without any difficulty ie pain. Infection was defined as redness of skin around wound (phlebectomy site) along with presence of pus in the wound (site of intervention) with or without patient being febrile. Hematoma along the direction of treated saphenous vein was examined on 7th day and its surface area estimated by comparing its total surface to the surface area of the palm of individual patient (1% of total body surface area)-procedure similar to the method used in assessment of burned skin surface area. Patients were then categorized either as having hematoma less than 1% or having hematoma >= 1% of total body surface area. Pain score was measured using a visual analogue scale ranging from 0 (no pain) to 5 (most severe pain) (Figure 3). + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + Patients + + + MESH:D006470 + Disease + + bleeding + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D006406 + Disease + + Hematoma + + + MESH:D006406 + Disease + + vein hematoma and infection + + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + Pain + + + MESH:D006406 + Disease + + hematoma + + + MESH:D010146 + Disease + + pain + + + MESH:D006406 + Disease + + hematoma + + + + FIG + AIM-22-329-g003.jpg + F3 + fig_caption + 6484 + Visual analog scale pain estimation + + MESH:D010146 + Disease + + pain + + + + METHODS + paragraph + 6520 + In case of presence of clinical signs of lower extremity deep venous thrombosis (DVT) Color Doppler examination was performed. Any bleeding after intervention that required surgical intervention either in the form of surgical revision of bleeding or need for additional surgical suture were noted and defined as postintervention bleeding. Return to normal activities were recorded taking 30 days after intervention as maximum period. This period was sufficient for full mobilization of patient even in the case of unwanted outcome endpoints like DVT. Each patient had a pre-operative duplex ultrasound examination that was performed in order to identify the site of pathologic reflux and evaluate suitability for intervention on superficial venous system. Ultrasonography was performed by vascular surgeon using Mindray 6 apparatus for group of patients treated surgically or GE ultrasound for patients treated by endovascular laser ablation. Ultrasonographic mapping of the venous systems was performed in all patients in standing position. Incompetence of the GSV was defined as reflux > 0.5 s in duration following calf compression. Exclusion criteria were: previous surgical treatment of the GSV, GSV diameter >= 2 cm, pregnancy, allergy to lidocaine, active superficial phlebitis, presence of any dermatologic phlebostatic changes in leg like hyperpigmentation or skin ulcer, positive history of previous deep venous thrombosis, diabetes mellitus and arterial disease. Mechanical stripping of great saphenous vein was performed in general anesthesia. After a groin incision, high ligation of the GSV and ligation of all tributaries were performed. Access to distal part of GSV was achieved through a small incision at the level of knee joint. GSV stripping was performed by metalic stripper. Varicosities were removed by minimal stab avulsion technique (Klapp/Smetana method). In the EVLA group of patients, endovascular access for laser fiber introduction into GSV was ultrasonically guided and performed at the knee level. The laser fiber tip was positioned 1 cm distal to the SFJ. Before laser ablation was started tumescent anesthetic (25 ml Lidocaine 2% in 500 ml of normal saline 0,9%) was injected all around GSV under ultrasound guidance. Thermal ablation of great saphenous vein was performed by Biolitec 1470 nm radial fiber with energy 70J/cm at 10 watts. Varicosities were removed by microphlebectomy technique. Post procedurally elastic venous compression was applied to both group of patients for 30 days in surgically treated group and for only 7 days in group of patients that received laser ablation treatment. Wound dressings were changed every three days and wounds were disinfected with povidone-iodine solution and sutures removed on 10th postoperative day in surgical group of patients. Wound dressing in EVLA group of patients was left intact for total 7 days and then removed along with compressive stocking. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9913 + Species + + calf + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D004342 + Disease + + allergy + + + MESH:D002539 + Disease + + arterial disease + + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D010689 + Disease + + phlebitis + + + MESH:D006470 + Disease + + bleeding + + + MESH:D008012 + Chemical + + Lidocaine + + + MESH:D012883 + Disease + + skin ulcer + + + MESH:D017495 + Disease + + hyperpigmentation + + + MESH:D020246 + Disease + + deep venous thrombosis + + + MESH:D006470 + Disease + + bleeding + + + MESH:D011206 + Chemical + + povidone-iodine + + + MESH:D006470 + Disease + + bleeding + + + MESH:D008012 + Chemical + + lidocaine + + + MESH:D003920 + Disease + + diabetes mellitus + + + + RESULTS + title_1 + 9461 + 4. RESULTS + + + RESULTS + paragraph + 9472 + Statistical data analysis was performed using the IBM SPSS version 22.0. In this study we analyzed continuous and categorical (dichotomous) variables. Statistical data analysis was performed using mainly methods of inferential statistics. For continuous variables, parametric independent-samples t-test was performed. Non-parametric chi-square test was performed for testing hypothesis for categorical variables. Both tests were calculated on the basis of 95% confidence level. Odds ratio was calculated only for the incidence of hematoma. For other variables odds ratio calculation was not possible because of zero frequencies in the cross tabulation cells. Mean of return to normal activities was expressed in days after intervention ei EVLA vs surgery (stripping) =1.21vs12.24, T test 13,619; p=0, 000, p<0,05 (Table 1). T test was used for comparing Mean value of visual pain analog scale (figure 3) for first 7 days between groups, for all seven days pain was significantly higher in surgical group of patients as compared to EVLA group; EVLA vs Surgery: 1st day T test= -17.900, p<0.05; 2nd day T test=-19,817, p<0.05; 3rd day T test=-21.986, p<0.05; 4th day T test=-4.898, p<0.05; 5th day T test=-4.355, p<0.05; 6th day T test -3,643, p<0.05; 7th day T test=2.386, p<0.05. + + 9606 + Species + + patients + + + MESH:D006406 + Disease + + hematoma + + + MESH:D010146 + Disease + + pain + + + MESH:D014786 + Disease + + visual pain + + + + TABLE + T1.xml + T1 + table_caption + 10752 + Mean of return to normal activities in days after intervention + + + TABLE + T2.xml + T2 + table_caption + 10815 + Mean value of visual pain scale for first seven days after intervention + + MESH:D014786 + Disease + + visual pain + + + + RESULTS + paragraph + 10887 + Incidence of hematoma greater than 1% of total body surface area was significantly higher in patients receiving surgical treatment; Pearson Chi Square=23,830, p<0,05; odds ratio:10,453 (Table 3). Incidences of infection, deep venous thrombosis and posttreatment bleeding were not statistically different between analyzed groups; EVLA vs Surgery: (Pearson Chi Square =3,237; p>0,05; Pearson Chi Square=2,139, p>0,05, Pearson Chi Square=2,139, p>0,05, respectively (Table 4, Table 5 and Table 6). + + 9606 + Species + + patients + + + MESH:D006406 + Disease + + hematoma + + + + TABLE + T3.xml + T3 + table_caption + 11382 + Incidence of hematoma greater than 1% of total body surface area + + MESH:D006406 + Disease + + hematoma + + + + TABLE + T4.xml + T4 + table_caption + 11447 + Incidence of infection + + MESH:D007239 + Disease + + infection + + + + TABLE + T5.xml + T5 + table_caption + 11470 + Incidence of wound bleeding after intervention + + MESH:D006470 + Disease + + bleeding + + + + TABLE + T6.xml + T6 + table_caption + 11517 + Incidence of deep venous thrombosis after intervention + + MESH:D020246 + Disease + + deep venous thrombosis + + + + DISCUSS + title_1 + 11572 + 5. DISCUSSION + + + DISCUSS + paragraph + 11586 + Return to normal everyday activities in EVLA group was on first day after intervention (mean) while in surgical group it was on 12th day (Mean: 1.23 vs. 12. 24); (Table 1). The difference was statistically significant p<0.05. All patients immediately after EVLA procedure was finished, left Clinic walking without any difficulties and regular every day walking was advised as only measure for prevention of DVT in this group. + + 9606 + Species + + patients + + + - + Chemical + + EVLA + + + + DISCUSS + paragraph + 12012 + Pain was quantified every day (for all seven days after intervention/operation) by each patient on scale from 0-5 (Figure 1) based on mean value of visual pain scale. In EVLA group for all seven days after intervention mean value of pain intensity was significantly less than in surgical group (Table 2). Less pain is probably related to avoiding trauma to skin (groin incisions) and trauma to subcutaneous tissue caused by avulsion (mechanical stripping of great saphenous vein). These results are opposite to results published by Pronk et al. where EVLA patient had more pain in second week as compared to surgical group of patients. Underlying reason for greater pain in their EVLA group is probably the type of laser apparatus they used (older type) because new Laser systems with higher wavelengths like 1470 nm have a better absorption in water and cause less pain and bruising after the procedure because they cause less vein wall perforation. This is in accordance with recommendation that low-energy EVLA with the use of a 1,470-nm laser fiber with linear endovenous energy density (LEED) of 80 J/cm or less is advisable option for the treatment of incompetent saphenous veins. Regarding post intervention hematoma, in our study there were 5 cases of hematoma in EVLA group and 28 cases being >= 1% of total body surface area in surgical group. This difference was statistically significant, p<0.05 (Table 3). Similar results were obtained by study of Siribumrungwong et al in which they had statistically significant advantage of EVLA method over surgery in hematoma appearance. Postoperative hematomas were also significantly smaller after EVLA than those after stripping in the study of Kalteis et al.. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D006406 + Disease + + hematoma + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D003288 + Disease + + bruising + + + MESH:D010149 + Disease + + Postoperative hematomas + + + MESH:D014947 + Disease + + trauma + + + - + Chemical + + EVLA + + + MESH:D010146 + Disease + + Pain + + + MESH:D014786 + Disease + + visual pain + + + MESH:D014867 + Chemical + + water + + + - + Chemical + + EVLA + + + MESH:D010146 + Disease + + pain + + + - + Chemical + + EVLA + + + MESH:D006406 + Disease + + hematoma + + + MESH:D010146 + Disease + + pain + + + MESH:D014947 + Disease + + trauma + + + MESH:D006406 + Disease + + hematoma + + + MESH:D010146 + Disease + + pain + + + + DISCUSS + paragraph + 13728 + Incidence of infection in this study was smaller in EVLA group (0 patients) as compared to surgical group of patients (3 patients) but this difference was not statistically significant, p>0.05 (Table 4). These findings have similar correlation with results of meta-analysis on 2245 limbs by Pan et al. which reported also fewer complications with EVLA as compared to high ligation and stripping in case of bleeding and hematoma (1.28% versus 4.83%) as well as wound infection (0.33% versus 1.91%). Striping itself is mechanically tissue destructive procedure which after removal of saphenous vein leaves avulsed tributaries that are left to bleed into subcutaneous tissue and amount of bleeding can be controlled only by subsequent compression. In that regard we had to additionally control bleeding after operations in 2 patients while there was no need to control any unwanted bleeding in patients after EVLA, (Table 5). However, this difference was not statistically relevant, p>0.05. We had no cases of DVT in EVLA group while 2 cases of crural DVT were recorded in surgical group of patients that were confirmed by color Doppler. Both of them received low molecular weight heparin therapy with switch to oral anticoagulation according to standard protocol for DVT treatment. Recovery was uneventful. This difference in incidence of DVT between groups was not statistically significant, (Table 6), p>0.05. Incidence of deep vein thrombosis (DVT) after varicose vein surgery vary between 0.4% historically and 5.7% while after EVLA from 0 to 7.7%. We administered LMWH prevention for surgical group of patients during their hospital stay although according to some authors there is no superiority of a short-term regimen of LMWH, early ambulation and compression therapy as compared with early ambulation and compression therapy alone. This decision was guided by idea that mobilization of patient after surgery due to general anesthesia was delayed as compared to EVLA group. EVLA treatment is rarely related to possible endovenous heat-induced thrombosis (EHIT) that leads to clinically significant DVT. EHIT may be caused by heat that is delivered by laser catheter placed in the saphenous vein. This brings about closure of the vein by denaturing the vessel wall with consequent thrombus formation. There are no relevant guidelines regarding advice for routine anticoagulation for EVLA patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 1214577 + Species + + to 7 + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + - + Chemical + + EVLA + + + MESH:D006406 + Disease + + hematoma + + + - + Chemical + + EVLA + + + MESH:D006470 + Disease + + bleeding + + + - + Chemical + + EVLA + + + MESH:D006470 + Disease + + bleeding + + + MESH:D006470 + Disease + + bleeding + + + - + Chemical + + EVLA + + + Disease + + EHIT + + + MESH:D006470 + Disease + + bleeding + + + - + Chemical + + EVLA + + + MESH:D006495 + Chemical + + LMWH + + + MESH:D006493 + Chemical + + heparin + + + MESH:D006495 + Chemical + + LMWH + + + MESH:D010291 + Disease + + crural DVT + + + MESH:D007239 + Disease + + infection + + + Disease + + EHIT + + + MESH:D007239 + Disease + + infection + + + MESH:D013927 + Disease + + thrombosis + + + - + Chemical + + EVLA + + + MESH:D020246 + Disease + + deep vein thrombosis + + + + CONCL + title_1 + 16131 + 6. CONCLUSION + + + CONCL + paragraph + 16145 + According to obtained results this endovenous laser ablation (EVLA) enables patients after treatment of varicose veins with better recovery in terms of significantly lower post treatment pain, faster return to everyday activities and lower incidence of bruising (hematomas). + + 9606 + Species + + patients + + + MESH:D006406 + Disease + + hematomas + + + MESH:D003288 + Disease + + bruising + + + MESH:D010146 + Disease + + pain + + + + CONCL + paragraph + 16420 + It has also shown that EVLA had offered no advantages in terms of significantly lower incidence of post intervention bleeding, infection rate or DVT. + + MESH:D006470 + Disease + + bleeding + + + MESH:D007239 + Disease + + infection + + + - + Chemical + + EVLA + + + + CONCL + paragraph + 16570 + LIMITATIONS: Study was carried out on patients with varicose veins without dermatologic complications so further evaluation is required especially regarding need for inclusion of patients with C3 level of disease or higher according to CEAP classification. In addition patient expectations might have had influence on subjective pain score scale since they have received information about so far published advantages of EVLA before intervention was performed. + + 282991 + Gene + 45479 + + CEAP + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D010146 + Disease + + pain + + + - + Chemical + + EVLA + + + + CONCL + footnote + 17030 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 17067 + REFERENCES + + + 9 + REF + surname:McBride;given-names:KD + 150 + 21550521 + 2011 + Surgeon + ref + 159 + 17078 + Changing to endovenous treatments for varicose veins: how much more evidence is needed? + + + surname:Forrestal;given-names:M + 2 + surname:Isaacs;given-names:M + surname:Zimmet;given-names:SE + REF + surname:Min;given-names:RJ + 167 + 2001 + J Vasc Interv Radiol + ref + 1171 + 17166 + Endovenous laser treatment of the incompetent greater saphenous vein + + + surname:Lissens;given-names:P + 21 + surname:Audenaert;given-names:EA + surname:Vanden;given-names:Bussche D + REF + surname:Vuylsteke;given-names:M + 80 + 2006 + Phlebology + ref + 87 + 17235 + Endovenous laser obliteration for the treatment of primary varicose veins + + + surname:Mavor;given-names:AID + surname:Dellagrammaticas;given-names:D + surname:Theivacumar;given-names:N + surname:Darwood;given-names:RJ + 294 + 18278775 + 2008 + Br J Surg + ref + 95 + REF + 301 + surname:Gough;given-names:MJ + 17309 + Randomized clinical trial comparing endovenous laser ablation with surgery for the treatment of primary great saphenous veins + + + surname:Gaastra;given-names:MTW + surname:Mooij;given-names:MC + surname:Gauw;given-names:SA + surname:Pronk;given-names:P + 649 + 5 + 20888274 + 2010 + European Journal of Vascular and Endovascular Surgery + ref + 40 + REF + 656 + surname:Lawson;given-names:JA + 17435 + Randomised Controlled Trial Comparing Sapheno-Femoral Ligation and Stripping of the Great Saphenous Vein with Endovenous Laser Ablation (980 nm) Using Local Tumescent Anaesthesia: One Year Results + + + 40 + surname:Demirkilic;given-names:U + REF + surname:Doganci;given-names:S + 254 + 2 + 20547079 + 2010 + European Journal of Vascular and Endovascular Surgery + ref + 259 + 17632 + Comparison of 980 nm Laser and Bare-tip Fibre with 1470 nm Laser and Radial Fibre in the Treatment of Great Saphenous Vein Varicosities: A Prospective Randomised Clinical Trial + + + surname:Mauriello;given-names:J + surname:Javier;given-names:J + surname:Mackay;given-names:E + surname:Almeida;given-names:J + 467 + 5 + 19628516 + 2009 + Vasc Endovascular Surg + ref + 43 + REF + 472 + surname:Raines;given-names:J + 17809 + Saphenous laser ablation at 1470 nm targets the vein wall, not blood + + + surname:Hwang;given-names:JJ + surname:Chang;given-names:IS + surname:Park;given-names:SW + surname:Park;given-names:JA + 2014 + J Vasc Interv Radiol + ref + REF + Epub ahead of print + surname:Yun;given-names:IJ + surname:Chee;given-names:HK + surname:Kim;given-names:JS + surname:Lee;given-names:SA + 17878 + The 1470-nm Bare-Fiber Diode Laser Ablation of the Great Saphenous Vein and Small Saphenous Vein at 1-Year Follow-up Using 8-12 W and a Mean Linear Endovenous Energy Density of 72 J/cm + + + surname:Attia;given-names:J + surname:Wilasrusmee;given-names:C + surname:Noorit;given-names:P + surname:Siribumrungwong;given-names:B + 214 + 2 + 22705163 + 2012 + Eur J Vasc Endovasc Surg + ref + 44 + REF + 223 + surname:Thakkinstian;given-names:A + 18063 + A systematic review and meta-analysis of randomised controlled trials comparing endovenous ablation and surgical intervention in patients with varicose vein + + 9606 + Species + + patients + + + + surname:Pistrich;given-names:R + surname:Messie-Werndl;given-names:S + surname:Berger;given-names:I + surname:Kalteis;given-names:M + 822 + 4 + 18295441 + 2008 + J Vasc Surg + ref + 47 + REF + 829 + surname:Hieller;given-names:F + surname:Polz;given-names:W + surname:Schimetta;given-names:W + 18220 + High ligation combined with stripping and endovenous laser ablation of the great saphenous vein: early results of a randomized controlled study + + + surname:Shao;given-names:M + surname:Mei;given-names:J + surname:Zhao;given-names:J + surname:Pan;given-names:Y + 109 + 2 + 23390218 + 2014 + Phlebology + ref + 29 + REF + 119 + surname:Zhang;given-names:J + 18364 + Comparison of endovenous laser ablation and high ligation and stripping for varicose vein treatment: a meta-analysis + + + surname:Harrison;given-names:C + surname:Holdstock;given-names:J + surname:Price;given-names:BA + surname:Marsh;given-names:P + 521 + 4 + 20655773 + 2010 + European Journal of Vascular and Endovascular Surgery + ref + 40 + REF + 527 + surname:Whiteley;given-names:MS + 18481 + Deep Vein Thrombosis (DVT) after Venous Thermoablation Techniques: Rates of Endovenous Heat-induced Thrombosis (EHIT) and Classical DVT after Radiofrequency and Endovenous Laser Ablation in a Single Centre + + MESH:D020246 + Disease + + Deep Vein Thrombosis + + + Disease + + EHIT + + + MESH:D013927 + Disease + + Thrombosis + + + + surname:Vizcaino;given-names:I + surname:Taylor;given-names:JH + surname:Merino;given-names:B + surname:San Norberto;given-names:Garcia EM + 940 + 7 + 23993109 + 2013 + Ann Vasc Surg + ref + 27 + REF + 946 + surname:Vaquero;given-names:C + 18687 + Low-molecular-weight heparin for prevention of venous thromboembolism after varicose veinsurgery in moderate-risk patients: a randomized, controlled trial + + 9606 + Species + + patients + + + MESH:D054556 + Disease + + venous thromboembolism + + + MESH:D006493 + Chemical + + heparin + + + + surname:Christie;given-names:RA + surname:Hill;given-names:GB + surname:Chai;given-names:J + surname:Van Rij;given-names:AM + 1582 + 12 + 15386324 + 2004 + Br J Surg + ref + 91 + REF + 1585 + 18842 + Incidence of deep vein thrombosis after varicose vein surgery + + MESH:D020246 + Disease + + deep vein thrombosis + + + + + 4272833 + + surname:Spahovic;given-names:Hajrudin + surname:Avdagic;given-names:Edin + surname:Schefe;given-names:L. + 25568577 + surname:Jaganjac;given-names:Suad + 302 + 5 + 2014 + 4272833 + AIM-22-302 + 10.5455/aim.2014.22.302-305 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 302-5. doi:10.5455/aim.2014.22.302-305 + kidney preoperative tumor embolization + 305 + surname:Hiros;given-names:Mustafa + Jaganjac S, Schefe L, Avdagić E, Spahović H, Hiros M + 0 + Preoperative Kidney Tumor Embolization as Procedure for Therapy of Advanced Kidney Cancer + + MESH:D007674 + Disease + + Kidney Tumor + + + MESH:D007680 + Disease + + Advanced Kidney Cancer + + + + ABSTRACT + abstract_title_1 + 90 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 99 + Introduction: + + + ABSTRACT + abstract + 113 + Preoperative kidney tumor embolization is standard procedure for therapy in advanced kidney cancer. Preoperative embolization has a goal to reduce intraoperative bleeding and also to shorten the time of surgery. + + MESH:D016063 + Disease + + intraoperative bleeding + + + MESH:D007680 + Disease + + kidney cancer + + + MESH:D007674 + Disease + + kidney tumor + + + + ABSTRACT + abstract_title_1 + 325 + Materials and methods: + + + ABSTRACT + abstract + 348 + We retrospectively observed 50 patients between 2000-2011, in which the preoperative embolization was performed. Mean age of patients was 64 years. All patients with preoperative embolization were compared with the group of 51 patients from Urology Sarajevo, who underwent nephrectomy without preoperative embolization. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + ABSTRACT + abstract_title_1 + 668 + Results: + + + ABSTRACT + abstract + 677 + Symptoms that are dominating among patients were haematuria and pain. Analysis of mean size of tumors based on CT evaluation showed statistically significance in between the biggest size of tumors in group from Hamburg (9.11+-3cm) and the smallest size of tumors in Sarajevo group (4.94+-1.6cm) p=0.0001. Reason for this is difference in selection of patients for treatment in Hamburg from Sarajevo. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumors + + + MESH:D006417 + Disease + + haematuria + + + MESH:D010146 + Disease + + pain + + + + ABSTRACT + abstract_title_1 + 1077 + Conclusion + + + ABSTRACT + abstract + 1088 + Kidney as functional finishing organ is extremely suitable for transcatheter therapeutic procedures. The gold standard in the treatment of advanced and metastatic tumor is the nephrectomy. As preparation for nephrectomy in metastatic cancer total capillary embolization is performed. After embolization, surgery is shorter, procedure can be done 24-48 hours after embolization or delayed nephrectomy done 2-3 weeks after the intervention. + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + cancer + + + + INTRO + title_1 + 1527 + 1. INTRODUCTION + + + INTRO + paragraph + 1543 + Kidney cancer is the most common kidney tumor and the third one by the frequency in urology. Comparing to other malignancy, 2% of all tumors is the kidney cancer. In end stage kidney cancer therapy of choice is radically nephrectomy, which means and adrenalectomy with removing fat kidney capsule and proximal urethra. + + MESH:D007676 + Disease + + end stage kidney cancer + + + MESH:D007680 + Disease + + kidney cancer + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + malignancy + + + MESH:D007674 + Disease + + kidney tumor + + + MESH:D007680 + Disease + + Kidney cancer + + + + INTRO + paragraph + 1862 + Kidney tumor embolization is the standard procedure in therapy of kidney cancer, due to embolization whole kidney or its parts should be exclude from the function. First experimental embolization was performed by lang in 1971. He used radioactive particles to induce tumor necrosis. On humans, first embolization was performed in 1973, by Almagard. From that time the technique has been changed a lot, also diagnostic procedures for detecting the tumors were fulfilled with CT and MRI. In the same time the new operative techniques were introduced laparoscopic nephrectomy, organoprotective resection and extracorporal Work-Bench procedures. + + 9606 + Species + + humans + + + MESH:D007674 + Disease + + Kidney tumor + + + MESH:D009369 + Disease + + tumors + + + MESH:D007680 + Disease + + kidney cancer + + + MESH:D009336 + Disease + + tumor necrosis + + + + INTRO + paragraph + 2504 + Preoperative tumor embolization is performed in patients with: + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + tumor + + + + INTRO + paragraph + 2567 + Advanced stage of tumor + + MESH:D009369 + Disease + + tumor + + + + INTRO + paragraph + 2591 + Tumor rich in retroperitoneal collateralize + + + INTRO + paragraph + 2635 + Infiltration of venous system + + + INTRO + paragraph + 2665 + Tumor in close range of kidney hillus + + + INTRO + paragraph + 2703 + Metastatic tumors and + + MESH:C538445 + Disease + + Metastatic tumors + + + + INTRO + paragraph + 2725 + Patients who are not treatable to become treatable. + + 9606 + Species + + Patients + + + + INTRO + paragraph + 2777 + Preoperative embolization has the goal to reduce intraoperative bleeding and to reduce the time of surgical procedure, by making preparation of tumor easier. This is particularly important for T3 and T4 tumors, with infiltration of renal vein and vena cava inferior. + + MESH:D009369 + Disease + + tumor + + + MESH:D007674 + Disease + + infiltration of renal vein + + + MESH:D016063 + Disease + + intraoperative bleeding + + + MESH:D009369 + Disease + + tumors + + + + METHODS + title_1 + 3044 + 2. MATERIALS AND METHODS + + + METHODS + paragraph + 3069 + We retrospectively observed 50 patients between 2000-2011, in which the preoperative embolization was performed. The majority of patients were treated in Eilbek Clinic in Hamburg and few in Clinical Center Sarajevo. Mean age of patients was 64 years, range between 3 and 86 years. 19 patients were female and 31 patients were male. Patients that were enrolled on study were the patients with the tumor larger than 8cm on the upper pole of left kidney, and patients wit proven bilateral kidney tumor. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + tumor + + + MESH:D007674 + Disease + + kidney tumor + + + + METHODS + paragraph + 3569 + Patients that were excluded from study were the patients with tumor smaller than 8cm, if the tumor was good for surgical procedure. Embolised patient were compared with the group of 51 patients from Urology Clinic Sarajevo, which were performed surgical nephrectomy without embolization. This group has been concised from 19 female and 32 male. Mean age was 61 years, range between 32 and 80 years. The total number of patients was 101. Patients were diagnosed with CT or MRI, as outpatients and small amount of patients we diagnosed in our Institution (CT siemens Somatom duo and MRI Unit 1.5T Symphonie). + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + tumor + + + + METHODS + paragraph + 4176 + Conventional angiography was performed on Siemens Polystar, 2 frames/sec. Angiography was performed by Seldinger technique, with contrast agent Xenetix 300. Angiography was performed to determine the blood supply of tumors in attacked kidney; we performed first aortography followed by selective angiography of renal arteries. Once done selective angiography, we approached embolization. Embolization is done first on the capillary level. As embolization material at the capillary level in the course of the study we used alcohol or Ivalon 150-250 mu particles. Alcohol is the cheapest and most efficient means of embolization. The disadvantage is that alcohol is the only means of embolization during one embolization must be applied in a blocked blood vessel in order to prevent reflux. We used 96% alcohol. In some patients inhibit the supply vessel is impossible, and then we performed a combined embolization with alcohol and Ivalon or just Ivalon particle embolization. The advantage in using Ivalon that supply blood vessel may not be blocked. Upon completion of the capillary embolization, we performed control angiography, and if the tumor / kidney is embolised we performed the central embolization of supply vessel with metal spirals. In the course of the study, Tornado spirals (COOK) were used. After that is done aortography to confirm complete embolization and showed situation surrounding blood vessels that were not aimed by embolization. + + 9606 + Species + + patients + + + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + Alcohol + + + MESH:C026699 + Chemical + + Ivalon + + + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D009369 + Disease + + tumors + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D009369 + Disease + + tumor + + + + METHODS + paragraph + 5632 + Besides angiographies findings, we were processed operating protocols and protocols of anesthesia. To test the existence of the correlation between the groups for each parameter was used Pearson correlation test. In all tests of statistical significance was considered at p <0.05 or at the level of 95%. All data were analyzed using statistical software SPSS version 13.0 for Windows, while the resulting tables and graphs subsequently processed using Microsoft Word 2003. + + MESH:D008569 + Disease + + Word + + + + RESULTS + title_1 + 6105 + 3. RESULTS + + + RESULTS + paragraph + 6116 + Symptomatology that led the patient's physician is shown in Figure 1 and 2. A small number of patients reported the loss of body weight, 11 patients from the Hamburg group (H) and 7 patients in the Sarajevo group (S). Infiltration of the kidney hollow system has been registered with 5 (H) patients and in 7 (S) patients. Infiltration of veins, tumor thrombus was found in a total of 16 patients (10 H and 6 S) and the infiltration of adjacent organs was found in 15 patients (8 H 7 and S). AV-shunt was diagnosed in only one patient at Hamburg group. Analysis of the average size of the tumor based on CT findings showed a statistically significant difference in terms of the maximum average size of tumors in patients from Hamburg (9.11 +- 3 cm) and minimum size in patients in Sarajevo (4.94 +- 1.6 cm), p = 0.0001. The reason for this is in the selection of patients in Hamburg preoperative group compared to Sarajevo operative group. The presence of metastases in regional lymph nodes showed statistically significant differences between groups p = 0.030. It is evident that the invasion of local lymph nodes at least recorded in patients in Sarajevo (7.8%), and patients from Hamburg (22%). Distant metastases were found in 4 (H) and in 2 (S) patients. + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009362 + Disease + + metastases + + + MESH:D009369 + Disease + + tumors + + + MESH:D009369 + Disease + + tumor + + + MESH:D013927 + Disease + + tumor thrombus + + + MESH:D009362 + Disease + + metastases + + + + FIG + AIM-22-302-g001.jpg + F1 + fig_caption + 7375 + Number cases of Haematuria in our sample + + MESH:D006417 + Disease + + Haematuria + + + + FIG + AIM-22-302-g002.jpg + F2 + fig_caption + 7416 + Presence of pain in our patients + + MESH:D010146 + Disease + + pain + + + + RESULTS + paragraph + 7449 + Embolization was repeated in two patients. Patohistological classification is shown in Figure 3. + + 9606 + Species + + patients + + + + FIG + AIM-22-302-g003.jpg + F3 + fig_caption + 7546 + Pathohistological classifi cation + + + RESULTS + paragraph + 7580 + The most common t stage was pT3a with 28% (H) and pT1a (S), and the least represented pT4 and pT2 with 6% in the Hamburg group and pT4 in Sarajevo group with 2%. + + + RESULTS + paragraph + 7742 + Histological determination of tumor size confirms previous statement on CT tumor size that the same was higher in patients from Hamburg (8.2 +- 3.4 cm), compared to patients in Sarajevo (3.89 +- 1.5 cm). Analysis of variance also showed statistically significant differences p = 0.0001 in favor of more specific histopatological tumor size in patients from Hamburg. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + tumor + + + + RESULTS + paragraph + 8108 + The average length of surgery was very similar and it was 105.6 +- 23 minutes in patients from Hamburg, and 101.4 +- 38 minutes in patients in Sarajevo. Minimum duration of the procedure was recorded in Sarajevo on 45 minutes, which was recorded as well as the maximum duration of treatment of 195 minutes. There were no statistically significant differences. The Hamburg group's surgery was done on average 23 days after embolization. Blood transfusion during and after surgery are shown in Table 1. With statistical significance, the difference can be seen that the operations carried out in Sarajevo was used more units of blood during the same. Figure 4 presents the case of preoperative embolization, which was done in so called sandwich-technique alcohol mixed with lipiodol and Ivalon particle. Central occlusion was performed with Tornado spirals. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D004998 + Chemical + + lipiodol + + + MESH:D015356 + Disease + + Central occlusion + + + + TABLE + T1.xml + T1 + table_caption + 8964 + Blood transfusion intra-post op. chi2=5,492, p=0,040 + + + FIG + AIM-22-302-g005.jpg + F4 + fig_caption + 9017 + a), b) present angiographic findings of kidney tumor; c) and d) present angiographic findings after embolisation. + + MESH:D007674 + Disease + + kidney tumor + + + + DISCUSS + title_1 + 9131 + 4. DISCUSSION + + + DISCUSS + paragraph + 9145 + Treatment of choice for kidney cancer is surgical resection. In early stage of TU and with favorable localization is sufficient to perform enucleation or partially nephrectomy, and in advanced disease should be done radical nephrectomy. + + MESH:D007680 + Disease + + kidney cancer + + + + DISCUSS + paragraph + 9382 + By age, group of our patients who were preoperatively embolised, was very heterogeneous. We performed one embolization of Wilms tumor. Wilms tumor occurs at the age of 2-4 years. After 10 years of appearing less frequently. In addition to Wilms tumor in children is possible occurrence of renal cancer. Incidence of 3- 6%. + + 9606 + Species + + patients + + + 9606 + Species + + children + + + MESH:D009396 + Disease + + Wilms tumor + + + MESH:D009396 + Disease + + Wilms tumor + + + MESH:D007680 + Disease + + renal cancer + + + MESH:D009396 + Disease + + Wilms tumor + + + + DISCUSS + paragraph + 9705 + Reasons for blood loss during nephrectomy are: hypervascularisation of tumor, atypical blood vessels and hilar lymph nodes that hamper visibility of blood vessels. + + MESH:D006473 + Disease + + blood loss + + + MESH:D009369 + Disease + + tumor + + + + DISCUSS + paragraph + 9869 + For decisive loss of blood atypical blood vessels are the main reason, which are often fulfilled with the blood. In violation of these vascular structures, Intraoperative bleeding is abundant because these vessels are less contractile. Another disadvantage is that these blood vessels are retroperitoneal positioned which makes preparation of large tumors very difficulty. With preoperative embolization, we have to occlude not only renal artery and capillary circulation, but must be able to occlude all atypical arteries to reduce blood loss to a minimum. + + MESH:D016063 + Disease + + Intraoperative bleeding + + + MESH:D006473 + Disease + + blood loss + + + MESH:D009369 + Disease + + tumors + + + + DISCUSS + paragraph + 10427 + Craven study showed minimal blood loss in delayed nephrectomy after embolization even with advanced stages of cancer with vascularized thrombus in the inferior vena cava. Delayed nephrectomy stimulates tumor regression as well as venous thrombosis, which significantly facilitates op procedure. In our group's average delay was 23 days. Some authors recommend a delay of less than 48 hours between embolization and surgery because postinfarctus syndromes do not develop fully. The same authors as Lanigian et al. suggest that a longer interval between embolization and surgery lead to the development of collateral that hamper preparation. These data from the literature do not agree with our experience. In most of our patients, we performed delayed nephrectomy. In these patients, there was no collateral development in the periphery. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + cancer + + + MESH:D020246 + Disease + + venous thrombosis + + + MESH:D006473 + Disease + + blood loss + + + MESH:D009369 + Disease + + tumor + + + + DISCUSS + paragraph + 11264 + Our first embolization we performed only with the alcohol, in 96%. Alcohol is very potent and very cheap embolisaton material. If it is used in combination with distal balloon occlusion, the rate of complications such as migration is very low. Alcohol is very powerful embolization material, and destruction of capillary system is very powerful, better than any other embolization material. Destruction of capillary system lead to tumor necrosis. Tolle and Cramer performed 20 embolisations with very low rate of the complications. Those authors claim that alcohol is very safe to use, and percent of recanalization is almost zero. We also performed majority of our embolisations with same techniques. The volume of alcohol that we used was determined for every patient individually. After balloon occlusion, angiography was performed. The volume of contrast material that was necessary for demarcation of whole kidney is the volume of alcohol that we used. Occlusion was performed 40 minutes after alcohol injection. In our study the rate of complication was zero. With lack of occlusion catheters we abandoned this technique. Some authors reported severe complications after alcohol embolization. Laarmann at all. reported two cases of testicular damage, as reason of anatomical variation where art. testiculariswas the branch of renal artery. Same group of authors performed alcohol embolization in 44 patients with no distal balloon occlusion. The volume of alcohol that was used was 14-22ml. Timmerman performed capillary embolization in combination of alcohol and Etiblock. + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D054549 + Disease + + balloon occlusion + + + - + Chemical + + Etiblock + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + Alcohol + + + MESH:D054549 + Disease + + balloon occlusion + + + MESH:D000438 + Chemical + + Alcohol + + + MESH:D009336 + Disease + + tumor necrosis + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D054549 + Disease + + balloon occlusion + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D007674 + Disease + + branch of renal artery + + + + DISCUSS + paragraph + 12845 + Several publications confirmed severe complications after alcohol embolization, so the use of alcohol was diminished and Etiblock was introduced as main embolization material. Initial results with Etiblock determined vasospasm after its use, so results were incomplete embolization. After another embolization materials were introduced, such as Ivalon 150-250 microns. Ivalon embolization does not need use of occlusion balloons, and application is very simple over microcatheter. Ivalon also has very good X ray visibility. In our series we had two case of recanalization after Ivalon embolization, so embolization had to be repeated. After this we started with so called sandwich embolization technique alcohol and Ivalon particles 150-250 microns. Central embolization was performed with Tornado colils. We had no complication with these technique. + + MESH:C026699 + Chemical + + Ivalon + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D020301 + Disease + + vasospasm + + + MESH:D054549 + Disease + + occlusion balloons + + + MESH:C026699 + Chemical + + Ivalon + + + + DISCUSS + paragraph + 13697 + With preoperative embolization there is significant reduce of blood transfusion during surgery, comparing to patients with no embolization. Volume of blood in group of embolised patients was around 250ml, and in group of non embolised patients was around 800ml. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 13959 + Zielinski et al. in their work compare rate of surveillance of five and ten years between group of 118 patients that were embolised and group of 116 patients with no embolization prior to surgery. The both groups had same demographics. Five and ten year's surveillance was statistically significant in first group 62%, and in the second group 35%. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 14307 + Kauffmann in his work showed reduction of blood loss during surgery in the group of patients preoperatively embolised. Vogel et al. published the same results as Kauffmann with loss of blood of 600ml in patients that were preoperatively embolised. Embolization was performed with 2-6ml Histoacril mixed with 50 % Lipiodol. Danish group of authors with Christensen published that they had smaller loss of blood during surgery in group of patients who were embolised prior to surgery, and the time of surgery was shorter, comparing with the group of patients with no embolization prior to surgery. Embolization was performed only with coils. The surgery was performed five days after embolization. Surveillance rate of embolised patients was better than in patients with no embolization. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D006473 + Disease + + reduction of blood loss + + + MESH:D004998 + Chemical + + Lipiodol + + + - + Chemical + + Histoacril + + + + DISCUSS + paragraph + 15093 + Stosslein at all. published that blood volume loss during surgery was in average 300ml. They had 92 patients. There were no significant shortening it time of surgery between two groups of patients. Surveillance rate was much higher in group of patient with embolization than in another group. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + + DISCUSS + paragraph + 15387 + Latal at all published that there were no statistically significance in surveillance rate between patients who were embolised prior to surgery comparing with patients who had only surgical nephrectomy. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + CONCL + title_1 + 15589 + 5. CONCLUSION + + + CONCL + paragraph + 15603 + Kidney cancer is the rare tumor considering other malignancies. Kidney as the organ is very suitable for all transcatheter procedures as for percutaneous procedures. The gold standard for treatment of kidney cancer in advanced stage is surgical procedure with nephrectomy, and for small tumor is enucleation of tumor. Full capillary embolization is method of choice to prepare surgical nephrectomy. The most potent embolization material is alcohol, but its is the most dangerous one. Because of that, alcohol should be administered over the occlusion or in small portions with conjunction of Lipiodol to be visible. The goal of this embolization is to make surgical procedure easier, thru less bleeding during the surgical procedure. On other goal is to make surgery less time consuming. The surgery can be made 24-48 h after the embolization, or it can be made as late nephrectomy after 2-3 weeks. Some percent of patients who were inoperable before the embolization, become operative after the procedure. It is a fact that the cost of treatment of those patients are bigger than with patients who are treated only with the surgery. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D007680 + Disease + + Kidney cancer + + + MESH:D009369 + Disease + + tumor + + + MESH:D007680 + Disease + + kidney cancer + + + MESH:D006470 + Disease + + bleeding + + + MESH:D058405 + Disease + + small tumor + + + MESH:D009369 + Disease + + malignancies + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D000438 + Chemical + + alcohol + + + MESH:D004998 + Chemical + + Lipiodol + + + MESH:D009369 + Disease + + tumor + + + + CONCL + footnote + 16737 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 16774 + REFERENCES + + + surname:Becker;given-names:N + REF + surname:Holzheimer;given-names:S + 1998 + ref + 16785 + Krebsatlas + + + 98 + REF + surname:Lang;given-names:EK + 391 + 5541057 + 1971 + Radiology + ref + 399 + 16796 + Superselective arterial catheterization as a vehicle for delivering radioactive infarct particles to tumors + + MESH:D007238 + Disease + + infarct + + + MESH:D009369 + Disease + + tumors + + + + 45 + surname:Haverling;given-names:M + surname:Fernstrom;given-names:I + REF + surname:Almgard;given-names:LE + 474 + 4748393 + 1973 + Br J Urol + ref + 479 + 16904 + Treatment of renal adenocarcinoma by embolic occlusion of the renal circulation + + MESH:C538614 + Disease + + renal adenocarcinoma by embolic occlusion of the renal circulation + + + + 39 + surname:Aretz;given-names:K + surname:Hallscheidt;given-names:P + REF + surname:Hansmann;given-names:HJ + 783 + 10525637 + 1999 + Radiologe + ref + 789 + 16984 + Nierentumoremboisation + + + 162 + surname:Takeda;given-names:M + surname:Nakai;given-names:H + REF + surname:Asanuma;given-names:H + 1402 + 10492224 + 1999 + J Urol + ref + 1405 + 17007 + Renal cell carcinoma in children: experience at a single institution in Japan + + 9606 + Species + + children + + + MESH:C538614 + Disease + + Renal cell carcinoma + + + + 155 + surname:Stewart;given-names:T + surname:Vates;given-names:TS + REF + surname:Freedman;given-names:AL + 1708 + 8627866 + 1996 + J Urol + ref + 1710 + 17085 + Renal cell carcinoma in children: the Detroit experience + + 9606 + Species + + children + + + MESH:C538614 + Disease + + Renal cell carcinoma + + + + surname:Rubben;given-names:H + 5 + surname:Rubben;given-names:H + surname:Goeppel;given-names:M + REF + surname:Hawers;given-names:W + 568 + 2001 + (Hrsg) Uroonkologie + ref + 587 + 17142 + Nephroblastom + + + 42 + surname:Stumpfl;given-names:M + surname:Gravou;given-names:C + REF + surname:Ebert;given-names:A + 263 + 12733509 + 2003 + Urologe A + ref + 268 + 17156 + Das Nierenzellkarzinom im Kindesalter + + + 17 + surname:Belgrano;given-names:E + surname:Carmignani;given-names:G + REF + surname:Giuliani;given-names:L + 431 + 7233655 + 1981 + Urology + ref + 434 + 17194 + Usefulness of preopertive transcatheter embolisaton in kidney tumors + + MESH:D007674 + Disease + + embolisaton in kidney tumors + + + + 146 + surname:Mendez;given-names:R + surname:Choop;given-names:RT + REF + surname:Singsaas;given-names:MW + 704 + 1979 + Urology + ref + 708 + 17263 + Preopertive renal embolization as adjunct to radical Nephrectomy + + MESH:D007674 + Disease + + Preopertive renal embolization + + + + 146 + surname:Kumpe;given-names:DA + surname:Redmond;given-names:Pl + REF + surname:Craven;given-names:WM + 704 + 1875477 + 1991 + J Urol + ref + 708 + 17328 + Planned delayed nephrectomy after ethanol embolization of renal carcinoma + + MESH:C538614 + Disease + + renal carcinoma + + + MESH:D000431 + Chemical + + ethanol + + + + 33 + surname:Varenhorst;given-names:E + REF + surname:Kalman;given-names:D + 162 + 10452291 + 1999 + Scand J Urol Nephrol + ref + 170 + 17402 + The role of arterial embolization in renal cell carcinoma + + MESH:C538614 + Disease + + renal cell carcinoma + + + + 46 + surname:Hammonds;given-names:JC + surname:Jurrians;given-names:E + REF + surname:Lanigan;given-names:D + 176 + 1395422 + 1992 + Clin Radiol + ref + 178 + 17460 + The current status of embolization in renal cell carcinoma - a survey of local and national practice + + MESH:C538614 + Disease + + renal cell carcinoma + + + + 22 + surname:Cramer;given-names:BM + REF + surname:Tolle;given-names:E + 208 + 6684344 + 1983 + Urologe A + ref + 212 + 17561 + Klinische Erfahrungen mit der arteriellen Embolization von Nierentumoren mit Athanol + + MESH:D004617 + Disease + + Embolization von + + + MESH:D000431 + Chemical + + Athanol + + + + 26 + surname:Timmermann;given-names:J + surname:Straube;given-names:W + REF + surname:Laarmann;given-names:S + 94 + 3590441 + 1987 + Urologe A + ref + 95 + 17647 + Alkoholtoxische Gonadenschadigung durch Alkoholembolisation bei Nierenmalignomen + + + 23 + surname:Straube;given-names:W + surname:Schlake;given-names:W + REF + surname:Timmermann;given-names:J + 275 + 6542268 + 1984 + Urologe A + ref + 277 + 17729 + Der absolute Alkohol, ein steuerbares Embolisationsmaterial bei Nierentumoren + + - + Chemical + + steuerbares + + + MESH:D000431 + Chemical + + Alkohol + + + + 24 + surname:Schlake;given-names:W + surname:Straube;given-names:W + REF + surname:Timmermann;given-names:J + 260 + 2414892 + 1985 + Urologe A + ref + 263 + 17807 + Klinische Beobachtungen bei dem Gefassverschluss des Nierentumors durch absoluten Alkohol + + + 21 + surname:Schwartmann;given-names:KP + surname:Greipel;given-names:V + REF + surname:Boeminghaus;given-names:F + 201 + 1982 + Urologie A + ref + 204 + 17898 + Analyse der Langzeitergebnisse nach Nephrektomie wegen Hypernephrom anhand 442 Fallen + + MESH:D014890 + Disease + + Nephrektomie wegen Hypernephrom + + + + 46 + surname:Hammond's;given-names:JC + surname:Jurriaans;given-names:E + REF + surname:Lanigan;given-names:D + 176 + 1395422 + 1992 + Clinical Radiology + ref + 178 + 17985 + The current status of embolization in renal cell carcinoma-a survey of local and national practice + + MESH:C538614 + Disease + + renal cell carcinoma + + + + 4 + surname:Lakritz;given-names:PS + surname:Cynamon;given-names:J + REF + surname:Bakal;given-names:CW + 727 + 8280991 + 1993 + J Vasc Interv Radiol + ref + 731 + 18084 + Value of preoperative renal artery embolization in Reducing blood transfusion requirements during nephrectomy for renal cell carcinoma + + MESH:C538614 + Disease + + renal cell carcinoma + + + MESH:D007674 + Disease + + renal artery embolization + + + + 33 + surname:Pomer;given-names:S + surname:Dhremer;given-names:I + REF + surname:Staehler;given-names:G + 116 + 8178405 + 1994 + Urologe A + ref + 121 + 18219 + Tumorbefall der Vena cava beim Nierenzellkarzinom. Operationstechnik, Ergebnisse und Prognose + + + 23 + surname:Petrovich;given-names:Z + surname:Szmigelski;given-names:S + REF + surname:Zielinski;given-names:H + 6 + 1 + 10683065 + 2000 + Am J Clin Oncol + ref + 12 + 18313 + Comparison of preoperative embolization followed by radical nephrectomy with radical neprectomy alone for renal cell carcinoma + + MESH:C538614 + Disease + + renal cell carcinoma + + + + 29 + surname:Richter;given-names:GM + REF + surname:Kauffmann;given-names:GW + 205 + 2426999 + 1986 + Ann Radiol + ref + 207 + 18440 + Palliative capillary embolization renal carcinoma + + MESH:C538614 + Disease + + Palliative capillary embolization renal carcinoma + + + + 5 + surname:Ahlen;given-names:H + surname:Gorich;given-names:Jv + REF + surname:Vogel;given-names:J + 301 + 1995 + Akt Radiol + ref + 304 + 18490 + Ergebnisse nach Katheterembolization bei Nierenzellkarzinom + + + 12 + surname:Rohrbach;given-names:R + surname:Richter;given-names:GM + REF + surname:Kauffmann;given-names:GW + 22 + 1989 + Cardiovas Interv Radiol + ref + 28 + 18550 + Prolonged survival following palliative renal tumor embolization by capillary occlusion + + MESH:D007674 + Disease + + renal tumor + + + + 133 + surname:Andersen;given-names:JF + surname:Dyreborg;given-names:U + REF + surname:Christensen;given-names:K + 191 + 2 + 3968730 + 1985 + J Urol + ref + 193 + 18638 + The value of transvascular embolization in the treatment of renal carcinoma + + MESH:C538614 + Disease + + renal carcinoma + + + + 80 + surname:Munster;given-names:W + surname:Schwenke;given-names:A + REF + surname:Stosslein;given-names:F + 577 + 3433986 + 1987 + Z Urol Nephrol + ref + 586 + 18714 + Verbesserung der Prognose des Nierenzellkarzinoms durch perkutane transvasale Embolization + + + 83 + surname:Wanek;given-names:R + surname:Kautzky;given-names:W + REF + surname:Latal;given-names:D + 27 + 2330762 + 1990 + Z Urol Nephrol + ref + 31 + 18805 + Einfluss der Nierenocclusion auf die Uberlebensrate beim Nierenzellkarzinom: Eine retrospektive 10-Jahres-Studie + + MESH:C535733 + Disease + + Einfluss der Nierenocclusion auf + + + + 28 + surname:Wojtowicz;given-names:J + REF + surname:Karwowski;given-names:A + 533 + 4 + 1987 + Radiol Diagn + ref + 535 + 18919 + Long-term results of embolization in renal tumors + + MESH:D007674 + Disease + + renal tumors + + + + 99 + surname:Trost;given-names:DW + surname:Smith;given-names:EB + REF + surname:Schwartz;given-names:MJ + 881 + 2006 + BJU + ref + 886 + 18969 + Renal artery embolization: clnical indication and experience from over 100 cases + + MESH:D007674 + Disease + + Renal artery embolization + + + + + 4272834 + + surname:Krstevska;given-names:Svetlana + surname:Genadieva-Stavric;given-names:Sonja + surname:Stojanovic;given-names:Aleksandar + 25568624 + surname:Sotirova;given-names:Tatjana + 292 + 5 + 2014 + 4272834 + MSM-26-292 + 10.5455/msm.2014.26.292-296 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 292-6. doi:10.5455/msm.2014.26.292-296 + myelodysplastic syndromes prognostic factors overall survival + 296 + surname:Balkanov;given-names:Trajan + surname:Spasovski;given-names:Dejan + Sotirova T, Stojanovic A, Genadieva-Stavric S, Krstevska S, Spasovski D, Balkanov T + 0 + Influence of Prognostic Factors on OverallSurvival in Myelodysplastic Syndromes + + MESH:D009190 + Disease + + Myelodysplastic + + + + ABSTRACT + abstract_title_1 + 80 + Background: + + + ABSTRACT + abstract + 92 + Accurate prediction of a patient's prognosis is useful to define the risk posed by the disease. Age, gender, peripheral blood cytopenia, proportion of bone marrow (BM) blasts, performance status, comorbidities, transfusion dependence, specific karyotype abnormalities and molecular biomarkers can refine the prediction of prognosis in MDS. + + 9606 + Species + + patient + + + MESH:D009190 + Disease + + MDS + + + MESH:D006402 + Disease + + peripheral blood cytopenia + + + + ABSTRACT + abstract_title_1 + 432 + Aim: + + + ABSTRACT + abstract + 437 + to assess the influence of the some prognostic factors like age, gender, cytopenia, BM blast percentage, transfusion dependence, ferritin, hemoglobin (Hb), lactate dehydrogenase (LDH), albumin and specific karyotype abnormalities in myelodysplastic syndromes on overall survival (OS). + + MESH:D006402 + Disease + + cytopenia + + + MESH:D009190 + Disease + + myelodysplastic syndromes + + + + ABSTRACT + abstract_title_1 + 722 + Patients and methods: + + 9606 + Species + + Patients + + + + ABSTRACT + abstract + 744 + we retrospectively analyzed the cohort of 108 patients diagnosed between 1.1.2011 and 31.12.2013 at the University Clinic of Hematology, Ss Cyril and Methodius University, Skopje, Macedonia. They were evaluated for clinical and hematologic features at diagnosis and at leukemic transformation. + + 9606 + Species + + patients + + + MESH:D007938 + Disease + + leukemic transformation + + + + ABSTRACT + abstract_title_1 + 1038 + Results: + + + ABSTRACT + abstract + 1047 + in the study group 62 were man and 46 women. Male to female ratio was 1.35 to 1. The differences in OS between men and women were significant (p = .03015). The mean age at diagnosis was 66,6 years. According to the age OS was 16,4 months. FAB subtypes influenced OS significantly (p = .03015). OS inversely correlated with BM blast percentage (p= .02327). Cytopenia had no impact on OS (p=.33755). Hb as a whole and groups with different levels of Hb had no influence on OS (p = .12142) and (p= .07535), respectively. The group with ferritin <500 microg/L had better OS (p=.04720). Transfusion dependence, LDH and albumin had no impact on OS. Leukemic transformation was noticed in 10 (9,3%) patients. Mortality was 36,1%. + + 9606 + Species + + man + + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + patients + + + MESH:D007938 + Disease + + Leukemic + + + - + Chemical + + OS + + + MESH:D006402 + Disease + + Cytopenia + + + + ABSTRACT + abstract_title_1 + 1770 + Conclusion: + + + ABSTRACT + abstract + 1782 + gender, FAB subtypes, BM blast percentage and the serum levels of ferritin had an influence on OS, while age, hemoglobin level, transfusion dependence, LDH and albumin had no impact on OS. + + + INTRO + title_1 + 1971 + 1. INTRODUCTION + + + INTRO + paragraph + 1987 + Myelodysplastic syndromes (MDSs) are a heterogeneous group of disorders characterized by dysplasia in one or more cell lines, followed by a progressive impairment in the ability to differentiate, manifested with peripheral cytopenia and an increasing risk of evolution into acute myeloid leukemia (AML). + + MESH:D015470 + Disease + + acute myeloid leukemia + + + MESH:D009190 + Disease + + Myelodysplastic syndromes + + + MESH:D006402 + Disease + + peripheral cytopenia + + + MESH:D004476 + Disease + + dysplasia + + + MESH:D015470 + Disease + + AML + + + + INTRO + paragraph + 2291 + The clinical course is variable so much effort has been focused on methods for predicting prognosis. Accurately predicting the prognosis once a malignancy has been diagnosed is of great importance to both patients and their physicians alike. From a patient's perspective, the prognosis helps define the severity of disease and sets expectations as to how it is likely to impact them. In contrast, prognostic information from a physician's standpoint is essentially a means of staging the disease in a manner that can be used to help direct therapy. For both patients and physicians, the estimation of prognosis is a continual process that does not happen just at the time of diagnosis. Reevaluating the prognosis may be useful when a patient shows signs of progression or after they have become refractory to standard treatment. The vast majority of cases (80% to 90%) occur 'de novo', whereas 10% to 20% of cases are secondary. French-American British (FAB) classification for MDS is still in use, based on morphologic findings. The standard prognostic tool for prognosis in MDS is the International Prognostic Scoring System (IPSS) which classifies patients into low-, intermediate-1, intermediate-2 and high-risk categories. IPSS takes into account BM blast percentage, number of cytopenia and cytogenetic abnormalities. Beside the known prognosis factors included in the IPSS, other variables that have an impact on prognosis in MDS were studied: age, gender, FAB subtypes, degree of anemia and RBC transfusion dependence, ferritin (, LDH, albumins, mutations and comorbidities. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D006402 + Disease + + cytopenia + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D009369 + Disease + + malignancy + + + + INTRO + paragraph + 3878 + Diagnosis requires BM examination and cytogenetic studies, and lately molecular studies. The consensual minimum criterion for diagnosis is the presence of erythroid, granulocyte or megakaryocyte dysplasia in 10% or more of informative cells. One must exclude the possibility of erythroid dysplasia associated with vitamin B12/folate/copper deficiency, viral infections, chemotherapy, or lead/arsenic poisoning. + + 4709 + Gene + 37630 + + B12 + + + MESH:D001102 + Disease + + viral infections + + + MESH:D004476 + Disease + + dysplasia + + + MESH:C535468 + Disease + + copper deficiency + + + MESH:D005492 + Chemical + + folate + + + MESH:C538442 + Disease + + erythroid dysplasia + + + MESH:D020261 + Disease + + arsenic poisoning + + + + INTRO + paragraph + 4289 + The incidence of MDS increases with age (median about 70 years). MDS is rare in children Age had a significant effect on OS of the MDS population analyzed as a whole and stratified by subgroups-the older the age, the worse the prognosis. During the analysis on the subgroups, the effect of age was statistically relevant within RA and RARS patients, whereas it was not significant within and RAEB subgroup. + + 5917 + Gene + 68281 + + RARS + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + + INTRO + paragraph + 4698 + A significant influence of sex on OS was observed in several studies. The overall incidence of MDS is slightly higher in males than in females (1.5-2 to 1). Male patients had worse prognosis. Considering subgroups, the effect of sex was relevant in refractory cytopenia, but did not affect the outcome in patients with RAEB. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D006402 + Disease + + cytopenia + + + MESH:D009190 + Disease + + MDS + + + + INTRO + paragraph + 5025 + According to the multivariate analysis peripheral cytopenia did not reach a statistical significance on OS in MDS patients. FAB subtypes can worsen the prognosis in MDS in terms of OS, with differences among subtypes. Refractory anemia had the best and RAEB-T the worst prognosis. + + 9606 + Species + + patients + + + MESH:D000754 + Disease + + RAEB-T + + + MESH:D000740 + Disease + + anemia + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D006402 + Disease + + cytopenia + + + + INTRO + paragraph + 5306 + Blast count showed a significant predictive value on OS in MDS patients. More than 5% blasts in BM are considered as a bad prognostic factor for OS and leukemic transformation. + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + MESH:D007938 + Disease + + leukemic + + + + INTRO + paragraph + 5483 + Red blood cell (RBC) transfusions are a commonly used therapy to treat symptomatic anemia that affects most patients with MDS. Transfusion dependence is defined by the MDS International Study Group (2000) as requiring transfusion of at least one RBC at 8 weeks for 4 months. Transfusion-dependent patients had shorter survival rate than those who received less than 18 units of blood over a period of 36 months (37). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000740 + Disease + + anemia + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + + INTRO + paragraph + 5900 + Serum ferritin level >=500 microg/l at diagnosis was a strong independent predictor of survival. Serum ferritin was significantly correlated with OS in Chinese patients. These patients suffer from comorbidities such as heart failure, diabetes, infections, disorders of the thyroid gland and liver, shortening their survival. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D003920 + Disease + + diabetes + + + MESH:D006333 + Disease + + heart failure + + + MESH:D007239 + Disease + + infections + + + + INTRO + paragraph + 6225 + Lactate dehydrogenase (LDH) is a parameter that should be recognized as a prognostic factor in MDS. One German study pointed out the correlation between LDH level and OS. Serum albumin is an independent prognostic factor that influences OS in patients with MDS. Hypoalbuminemia is a marker for shorter OS in MDS. + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D034141 + Disease + + Hypoalbuminemia + + + + INTRO + paragraph + 6538 + Recurrent chromosomal abnormalities have been identified in 40-70% of the 'de novo' MDS and 95% of secondary MDS. These chromosomal aberrations include 5q-, 7q-/-7, +8, 20q-, 12p-, abnormalities in 17p, 11q23 and chromosome 3. Favorable prognostic markers according to IPSS include: a normal karyotype, 5q-as an isolated anomaly, 20q-as an isolated anomaly and -Y chromosome. Karyotype findings associated with poor prognosis include complex karyotype and abnormalities of chromosome 7. Other cytogenetic abnormalities confer an intermediate prognosis. Abnormal karyotype correlates with poor prognosis and shorter OS. According to the multivariate analysis, cytogenetics showed a significant predictive value on OS. + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + MESH:D000014 + Disease + + anomaly + + + MESH:D000014 + Disease + + anomaly + + + MESH:D002869 + Disease + + chromosomal abnormalities + + + + INTRO + paragraph + 7263 + Somatic mutations are identified in more than 70% of patients with MDS, including more of the patients with normal karyotype. These mutations are major predictors of the clinical phenotype, and could also be predictors of prognosis. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + + INTRO + paragraph + 7496 + The incidence of MDS increases with the age, so does the prevalence of comorbidities. About 50% of MDS patients have one or more comorbidities. Congestive heart disease, hypertension, lung diseases, diabetes, liver failure, bleeding and solid tumors are cited as the most often reasons for non-leukemic dead. Comorbidities are significant and independent predictors in MDS. Congestive heart disease and chronic obstructive lung disease are associated with shorter OS, while diabetes and cerebro-vascular diseases do not change prognosis in MDS. + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + solid tumors + + + MESH:D009190 + Disease + + MDS + + + MESH:D029424 + Disease + + chronic obstructive lung disease + + + MESH:D006973 + Disease + + hypertension + + + MESH:D009190 + Disease + + MDS + + + MESH:D003920 + Disease + + diabetes + + + MESH:D006333 + Disease + + Congestive heart disease + + + MESH:D017093 + Disease + + liver failure + + + MESH:D000783 + Disease + + cerebro-vascular diseases + + + MESH:D009190 + Disease + + MDS + + + MESH:D007938 + Disease + + non-leukemic + + + MESH:D006333 + Disease + + Congestive heart disease + + + MESH:D006470 + Disease + + bleeding + + + MESH:D008171 + Disease + + lung diseases + + + MESH:D009190 + Disease + + MDS + + + MESH:D003920 + Disease + + diabetes + + + + INTRO + paragraph + 8041 + The median OS in MDS is 2.5 years. Prognosis is poor for patients with MDS, with 3-year survival rates estimated at less than 50%. The median survival rates according to IPSS are estimated at 8, 5.3, 2.2, and 0.9 years, respectfully. + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + MDS + + + + INTRO + title_1 + 8275 + 2. AIM + + + INTRO + paragraph + 8282 + The aim of this study was to assess the influence of the some prognostic factors in myelodysplastic syndromes (age, gender, cytopenia, BM blast percentage, transfusion dependence, serum levels of hemoglobin, ferritin, lactate dehydrogenase and albumin, and specific karyotype abnormalities) on OS. + + MESH:D006402 + Disease + + cytopenia + + + MESH:D009190 + Disease + + myelodysplastic syndromes + + + + METHODS + title_1 + 8580 + 3. PATIENTS AND METHODS + + 9606 + Species + + PATIENTS + + + + METHODS + paragraph + 8604 + We retrospectively analyzed the cohort of 108 patients (62 male and 46 female) diagnosed between 1.1.2011 and 31.12.2013 at the University Clinic of Hematology, Ss Cyril and Methodius University, Skopje, Macedonia. They were evaluated for clinical and hematologic features at diagnosis and leukemic transformation. Observation time was 36 months. Diagnosis was made upon dysplastic changes in peripheral blood smear and bone marrow aspirate. Confirmation analysis included bone marrow aspirate, cytochemical stain, immunohistochemical tests, cytochemical stain of medullar iron and karyotype analysis in some cases. We evaluated some parameters that could influence OS: age, gender, cytopenia, BM blast percentage, transfusion dependence, serum levels of Hb, ferritin, LDH and albumin as well as specific karyotype abnormalities. Chromosomal analysis was performed only in 5 patients using BM aspirate, according to laboratory procedures. IPSS was not calculated in the vast majority of patients, because we could not perform cytogenetic analysis in all patients. Most patients (pts) received supportive care: transfusion of RBC and platelets, red cell and granulocyte growth factors (21 pts), vitamins, corticosteroids, iron chelatation therapy (7 pts), some received chemotherapy (4pts) and few patients allogeneic transplantation (3 pts). OS was estimated in months including the period from the date of diagnosis to the time of death / time of last visit. Leukemic transformation was noticed in 10 (9,3%) patients. Mortality was 36,1%. Infectious and hemorrhagic complications as well as BM failure were considered causes of MDS-related deaths. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + pts + + + 9606 + Species + + pts + + + 9606 + Species + + pts + + + 9606 + Species + + pts + + + 9606 + Species + + pts + + + MESH:D007501 + Chemical + + iron + + + MESH:D007501 + Chemical + + iron + + + MESH:D007938 + Disease + + leukemic transformation + + + MESH:D009190 + Disease + + MDS + + + MESH:D007938 + Disease + + Leukemic + + + MESH:D006470 + Disease + + hemorrhagic complications + + + MESH:D017093 + Disease + + BM failure + + + MESH:D006402 + Disease + + cytopenia + + + MESH:D003643 + Disease + + death + + + MESH:D004416 + Disease + + dysplastic + + + + RESULTS + title_1 + 10253 + 4. RESULTS + + + RESULTS + paragraph + 10264 + Mean age at diagnosis was 66,6 years (range 18-89) (SD 14.2). Mean OS depending on age was 16,4 months, and it was not statistically significant (p= .46375). In our study, 62 (57.4%) patients were men, 46 (42.6%) women. Male to female ratio was 1.35 to 1. Women had better OS than men (p = .00819). (Figure 1). According to the FAB classification patients were classified as follows: 75 patients as having refractory anemia (RA; 69.4%), 1-RA with ringed sideroblasts (RARS; 0.9%), 21-RA with excess of blasts (RAEB; 19.4%), 3-RAEB in transformation (RAEB-T 2.8%), and 8 as having chronic myelomonocytic leukemia (CMML 8.2%). (Table 1) + + 5917 + Gene + 68281 + + RARS + + + 9606 + Species + + patients + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + Women + + + 9606 + Species + + men + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D054429 + Disease + + myelomonocytic leukemia + + + MESH:D000754 + Disease + + RAEB-T + + + MESH:D000740 + Disease + + anemia + + + + FIG + MSM-26-292-g001.jpg + F1 + fig_caption + 10905 + OS depending on gender + + + TABLE + T1.xml + T1 + table_caption + 10928 + Distribution of patients according to the FAB classification + + 9606 + Species + + patients + + + + RESULTS + paragraph + 10989 + Statistically significant differences in OS were found among patients with RA, RAEB, RAEB-T and CMML (p = .03015). (Figure 2) + + 9606 + Species + + patients + + + MESH:D000754 + Disease + + RAEB-T + + + + FIG + MSM-26-292-g003.jpg + F2 + fig_caption + 11115 + OS depending on FAB subtypes + + + RESULTS + paragraph + 11144 + Cytopenia had no influence on OS (p=.09340). In our group 23 patients (21.3%) presented with one cytopenia, 46 patients (42.6) with bicytopenia and 39 patients (36.1%) with pancytopenia. There was no statistically significant difference in OS among patients with different cytopenia (p = .33755). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D006402 + Disease + + Cytopenia + + + MESH:D010198 + Disease + + pancytopenia + + + MESH:D006402 + Disease + + different cytopenia + + + Disease + + bicytopenia + + + MESH:D006402 + Disease + + cytopenia + + + + RESULTS + paragraph + 11441 + Percentage of BM blasts correlated with OS (p=.02327). Mean OS in the group with < 5% blasts was 17,8 months, in the group with blasts 5-9%-17.7 months, with blasts 10-19%-9.7 months and in the group with 20-30% blasts - 9.5 months. The differences among groups were statistically significant (p<0.05). + + + RESULTS + paragraph + 11750 + Mean Hb level in our cohort was 85,1 g/L (range 41-150). Hb had no impact on OS (16.4 months) (p=.12142). The patients were grouped in three categories according to the severity of anemia: severe anemia (Hb <70 g/L), moderate anemia (Hb 70-100 g/L) and patients with Hb >100 g/L. OS was highest in patients with Hb >100 g/L-21.6 months, in moderate anemia-14,8 months and in severe anemia-15.2 months, being not statistically significant (p=0.07535). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000740 + Disease + + anemia + + + MESH:D000740 + Disease + + anemia-15 + + + MESH:D000740 + Disease + + anemia-14 + + + MESH:D000740 + Disease + + anemia + + + MESH:D000740 + Disease + + anemia + + + + RESULTS + paragraph + 12207 + Mean level of serum ferritin in our group was 3826.1microg/L. Serum ferritin had no impact on OS - 16,5 months (p=.13178). But, groups with ferritin <500 microg/L (17 months) and >500 microg /L (16 months) had significant differences in OS (p= .04720). (Figure 3). + + + FIG + MSM-26-292-g004.jpg + F3 + fig_caption + 12472 + OS in groups with different serum ferritin levels + + - + Chemical + + OS + + + + RESULTS + paragraph + 12522 + Transfusion dependence is defined by the MDS International Group (2000) study as requiring transfusion of at least one packed red blood cells (RBC) at 8 weeks for 4 months OS depending on transfusion as a whole was 16.2 months, being not statistically significant (p= .16298). OS in the group without RBC was 16.5 months, in the group that received <= 18 RBC had OS 15.4 months, and the group that received >18 RBC had OS 19.8 months. Mean transfused doses in this cohort were 13.2 RBC (St.dev.-16.8). Transfusion dependent patients with >18 doses RBC had shorter survival than patients who required <= 18 doses RBC, being not statistically significant (p=.72185). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + + RESULTS + paragraph + 13189 + Mean serum level of LDH was 809 IU/L. There was no statistical significance in OS between the two groups with different LDH (<423 IU/L and >423 IU/L) (p = .48310). + + + RESULTS + paragraph + 13353 + Mean albumin level in the study group was 39g/L. Albumins had no influence on OS - 20 months (p= .25087). There was no statistical significance in OS between the two groups with different levels of albumins (<40 g/L and >40 g/L) (p= .18986). + + + RESULTS + paragraph + 13597 + When testing mutual influence of several parameters on OS, we found that age, gender, FAB subtypes, bone marrow blast percentage, hemoglobin level and transfusion dependence had statistically significant impact on OS (p=.01628). + + + RESULTS + paragraph + 13826 + We were unable to perform cytogenetic analysis in all patients. From those 5 patients who underwent cytogenetic tests, 3 had normal karyotype, one patient had +8 and one inv16. We could not show survival curves due to the small number of patients in whom cytogenetic analysis was performed. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + + METHODS + title_1 + 14117 + 5. STATISTICAL ANALYSIS + + + METHODS + paragraph + 14141 + The statistical analysis was performed using statistical package SSPS 16.0. For the parameters or variables used in this paper, the range, mean and standard deviation were presented. Differences among variables were evaluated by the Chi-square test. The determinations of correlations between different variables was based on the Pearson correlation coefficient in all cases where the variables had a normal distribution, and a certain correlation was considered statistically significant if p <0.05. Overall survival was defined as the time interval between diagnosis date and death date. Patients who were alive were censored at the last follow-up date. The probabilities of OS were estimated using the method of Kaplan and Meier. Cox proportional hazards regression models were used to assess the association between prognostic factors and OS. + + 9606 + Species + + Patients + + + MESH:D003643 + Disease + + death + + + - + Chemical + + OS + + + + DISCUSS + title_1 + 14988 + 6. DISCUSSION + + + DISCUSS + paragraph + 15002 + Our results considering age (66.6 years) correspond with those in the literature. Age in this group did not affect OS neither as a whole nor stratified by subgroups, similar with some studies. Considering gender, men had worse OS than women, corresponding with the data in the literature. FAB subtypes had a significant effect on OS. Considering subtypes, the worst results were observed in RAEB-T, than in CMML, RAEB and RA subtype, similar with those in the literature. There was only one patient with RARS and he was not taken in consideration. Cytopenia had no impact on OS, corresponding with some data in the literature. But, others showed opposite results. OS correlated inversely with BM blast percentage, showed in other studies. Hemoglobin as a whole and stratified into subgroups did not show any impact on OS. Transfusion dependence could be an independent prognostic factor for bad prognosis, although in our study group the influence on OS was not significant. Serum ferritin level >=500 microg/L at presentation is an independent poor predictor for OS. The group with ferritin <500 microg/L had better OS. LDH is a factor sited as having predictive value, although we could not prove that. Hypoalbuminemia is an independent poor prognostic factor in MDS patients, but in our study group it was not significant. + + 5917 + Gene + 68281 + + RARS + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D000754 + Disease + + RAEB-T + + + - + Chemical + + OS + + + MESH:D009190 + Disease + + MDS + + + - + Chemical + + OS + + + MESH:D034141 + Disease + + Hypoalbuminemia + + + MESH:D006402 + Disease + + Cytopenia + + + + DISCUSS + paragraph + 16328 + Cytogenetics in other studies showed a significant predictive value on OS and patients were accordingly stratified in groups with different IPSS score. Only 5 patients were analyzed for cytogenetic abnormalities, three of them with good prognosis, and two of them (+8, inv16) with intermediate prognosis. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 16633 + Limitations in our study include retrospective analysis, insufficient data on cytogenetic analysis and impossibility to stratify patients according to IPSS score, lack of data on comorbidities, insufficient data on ferritin, LDH and albumins, that probably reflected on the results making some of them different than those cited in the literature. + + 9606 + Species + + patients + + + MESH:D000309 + Disease + + insufficient + + + MESH:D000309 + Disease + + insufficient + + + + DISCUSS + footnote + 16981 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 17018 + REFERENCES + + + 96 + REF + surname:Cazzola;given-names:M + 349 + 21357714 + 2011 + Haematologica + ref + 351 + 17029 + Risk assessment in myelodysplastic syndromes and 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States 2001-2004, using data from the NAACCR and SEER programs + + MESH:D009196 + Disease + + myeloproliferative disorders + + + MESH:D009190 + Disease + + myelodysplastic syndromes + + + + 19 + surname:Pfeilstocker;given-names:M + surname:Hildebrandt;given-names:B + REF + surname:Germing;given-names:U + 2223 + 16193087 + 2005 + Leukemia + ref + 2231 + 20052 + Refinement of the international prognostic scoring system (IPSS) by including LDH as an additional prognostic variable to improve risk assessment in patients with primary myelodysplastic syndromes (MDS) + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + MDS + + + MESH:D009190 + Disease + + primary myelodysplastic syndromes + + + + surname:Sperr;given-names:WR + surname:Germing;given-names:U + surname:Tuchler;given-names:H + surname:Nosslinger;given-names:T + 120 + 1 + 19605511 + 2010 + Ann Oncol + ref + 21 + REF + surname:Valent;given-names:P + surname:Giagounidis;given-names:A + surname:Stauder;given-names:R + 125 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Disease + + myelodysplastic syndrome + + + MESH:D009190 + Disease + + Myelodysplastic + + + MESH:D013921 + Disease + + thrombocytopenia + + + + 2 + REF + surname:Cheson;given-names:BD + 28 + 1 + 10388027 + 1997 + The Oncologist + ref + 39 + 20481 + The Myelodysplastic Syndromes + + MESH:D009190 + Disease + + Myelodysplastic + + + + surname:Dreyfus;given-names:F + surname:Quesnel;given-names:B + surname:Thepot;given-names:S + surname:Itzykson;given-names:R + 403 + 2 + 20940414 + 2011 + Blood + ref + 117 + REF + 411 + surname:Turlure;given-names:P + surname:Beyne-Rauzy;given-names:O + 20511 + Prognostic factors for response and overall survival in 282 patients with higher-risk myelodysplastic syndromes treated with azacitidine + + 9606 + Species + + patients + + + MESH:D009190 + Disease + + myelodysplastic syndromes + + + MESH:D001374 + Chemical + + azacitidine + + + + surname:Invernizzi;given-names:R + surname:Pascutto;given-names:C + surname:Della;given-names:Porta MG + surname:Malcovati;given-names:l + 7594 + 30 + 2005 + JCO + ref + 23 + surname:Lazzarino;given-names:M + REF + surname:Cazzola;given-names:M + surname:Bernasconi;given-names:P + surname:Maffioli;given-names:M + surname:Arcaini;given-names:L + 7603 + surname:Passamonti;given-names:F + surname:Travaglino;given-names:E + surname:Boni;given-names:M + 20648 + Prognostic Factors and Life Expectancy in Myelodysplastic Syndromes Classified According to WHO Criteria: A Basis for Clinical Decision Making + + MESH:D009190 + Disease + + Myelodysplastic + + + + surname:Lupu;given-names:AR + surname:Stanca;given-names:O + surname:Ciobanu;given-names:A + surname:Triantafyllidis;given-names:I + 295 + 4 + 23483702 + 2012 + Maedica (Buchar) + ref + 7 + REF + 302 + 20791 + Prognostic factors in Myelodysplastic syndromes + + MESH:D009190 + Disease + + Myelodysplastic syndromes + + + + + 4272835 + + surname:Vrecko;given-names:Helena + 25568575 + surname:Klemenc-Ketis;given-names:Zalika + 292 + 5 + 2014 + 4272835 + AIM-22-292 + 10.5455/aim.2014.22.292-296 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 292-6. doi:10.5455/aim.2014.22.292-296 + medical ethics medical students qualitative study + 296 + Klemenc-Ketis Z, Vrecko H + 0 + The Perceptions of Professionalism by 1st and 5th Grade Medical Students + + + ABSTRACT + abstract_title_1 + 73 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 82 + Introduction: + + + ABSTRACT + abstract + 96 + Professionalism is essential for the development of mature physicians but not much education is devoted to that theme. Aim: We aimed to determine the views of undergraduate medical students on medical professionalism. + + + ABSTRACT + abstract_title_1 + 314 + Methods: + + + ABSTRACT + abstract + 323 + This was a qualitative study, based on focus groups of the first and fifth-year undergraduate medical students. Transcripts of the focus groups were independently evaluated by two researches. Segments of transcripts, identified as important, were marked as verbatims. A grounded theory method with open coding was applied. A list of codes was developed and reviewed by both researchers until the consensus was reached. Then, the codes were reviewed and put into the categories and dimensions. + + + ABSTRACT + abstract_title_1 + 816 + Results: + + + ABSTRACT + abstract + 825 + Students recognized 10 main medical professionalism dimensions (empathy, respect, responsibility, autonomy, trust, communication, difference between professional and private life, team work, partnership) and two dimensions associated with it (physician's characteristics, external factors). Slight change of the attitudes towards a more self-centred future physicians' figure was observed in the fifth-year medical students. + + Disease + + empathy + + + + ABSTRACT + abstract_title_1 + 1252 + Conclusion: + + + ABSTRACT + abstract + 1264 + The students have an appropriate picture of the physicians' figure even at the beginning of their medical studies but still needs an education in professionalism. It seems that the fifth-year students perceive physicians as more self-centred when compared to their first-year colleagues. + + + INTRO + title_1 + 1554 + 1. INTRODUCTION + + + INTRO + paragraph + 1570 + In the era of sophisticated medical technology, family physicians should use technological advances but continue to practice comprehensive medicine in spite of health policy demands for more focus on public than individual outcomes. Consequently, the medical professionalism is slowly finding its way back to medical education. It is widely acknowledged that medical professionalism can be learned and taught. However, usually it is a part of a so called hidden curriculum: it is incorporated into teaching different subjects without setting clear learning objectives. There are voices that medical students besides communication skills, team work, practice organization, and knowledge management also should be taught in humanism, accountability and altruism. However, not much is known about the role of medical education in building a future physician who will possess the appropriate professional values, attitudes and behaviors. Even a unifying theoretical or practical model to use as a format for developing a professionalism teaching curriculum does not exist yet. + + Disease + + altruism + + + + INTRO + paragraph + 2643 + There is neither an unified definition nor theoretical model of medical professionalism at the moment. The American Board of Internal Medicine suggested that professionalism has six components: altruism, accountability, excellence, duty, honour/integrity and respect. A model from USA and Europe declared that professionalism consists of three fundamental principles (patient welfare, patient autonomy and social justice) with ten professional responsibilities. The model from UK suggests that medical professionalism consists of integrity, compassion, altruism, continuous improvement, excellence and working in partnership with team members. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + Disease + + altruism + + + Disease + + altruism + + + + INTRO + paragraph + 3287 + So far, several studies have tried to determine the views of undergraduate medical student regarding the professionalism. They showed that undergraduate students in general have a clear perception of what medical professionalism was. On the other hand, a recent qualitative study from Australia showed that undergraduate students have had some difficulties with perceiving what a professional physician was; they differentiated between the term "good" doctor and professional doctor as they perceived professionalism as an external and imposed construct. It also seems that some differences exist regarding the perceptions of medical professionalism in undergraduate students of different study years. Students in higher years of education reported the decline of professionalism in terms of idealism, empathy, the perception of physicians' status and income and the rise of the negative views on primary care and the importance of lifestyle and family. This change of attitudes seems to be largely dependent on the curriculum and the teaching method applied. The shift from informal to formal teaching of professionalism resulted in higher sensitivity to professionalism and the early patient contact and small groups in professionalism enhanced the understanding of professionalism. + + 9606 + Species + + patient + + + Disease + + empathy + + + + INTRO + paragraph + 4578 + Medical faculty Maribor is a young faculty which was established in 2003. The study of medicine lasts for six years and is consistent with the Bologna principles. No formal subject in the undergraduate study is devoted to professionalism therefore we can say that professionalism is a part of a hidden curriculum. + + + INTRO + title_1 + 4892 + 2. GOAL + + + INTRO + paragraph + 4900 + The aim of this study was to determine the views of undergraduate medical students on medical professionalism and recognise any potential qualitative differences in their views according to the study year. + + + METHODS + title_1 + 5106 + 3. METHODS + + + METHODS + title_2 + 5117 + 3.1. Study design and settings + + + METHODS + paragraph + 5148 + This was a qualitative study, based on focus groups. It took part on the Faculty of Medicine in Maribor, Slovenia and was performed in June 2014. The study was approved by the National Ethics Board (No. 226/09/13). + + Disease + + Slovenia + + + + METHODS + title_2 + 5363 + 3.2. Participants and focus groups + + 9606 + Species + + Participants + + + + METHODS + paragraph + 5398 + The first and fifth-year undergraduate medical students took part in this study. They were recruited by the researchers through the personal contact. In the study year 2013/2014, there were 89 students in the first year and 90 students in the fifth year. + + + METHODS + paragraph + 5653 + The students were included in two focus groups. The first one consisted of the five first-year medical students (three women and two men), each 19 years old. The second one consisted of seven fifth-year medical students (two men and five women), each 23 years old. Both focus groups were moderated by one of the researchers (HV) and observed by the second researcher (ZKK). The focus groups participants discussed upon the several questions (Box 1). + + 9606 + Species + + women + + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + participants + + + + FIG + AIM-22-292-g001.jpg + F1 + fig_caption + 6103 + The questions discussed in focus groups + + + METHODS + paragraph + 6143 + Focus groups were audio-recorded. Oral informed consent was sought from the participants prior to the session. + + 9606 + Species + + participants + + + + METHODS + title_2 + 6254 + 3.3. Analysis + + + METHODS + paragraph + 6268 + The interviews were typed to word documents. Transcripts were independently evaluated by both researches (HV and ZKK). Segments of transcripts, identified as important, were marked as verbatim. A grounded theory method with open coding was applied. A list of codes was developed and reviewed by both researchers until the consensus was reached. Then, the codes were reviewed and put into the categories and dimensions. + + + RESULTS + title_1 + 6687 + 4. RESULTS + + + RESULTS + paragraph + 6698 + Students recognized 10 main medical professionalism dimensions (Table 1) and two dimensions associated with it (Table 2). + + + RESULTS + title_2 + 6820 + 4.1. Objectivity + + + RESULTS + paragraph + 6837 + Students of both study years expressed the importance of setting a clear line between objectivity and subjectivity in order not to let the subjective feelings influence the management of the patients. + + 9606 + Species + + patients + + + + RESULTS + paragraph + 7038 + "The sympathy towards a patient should not blur your professional objectivity." (Fifth-year female student) + + 9606 + Species + + patient + + + + RESULTS + paragraph + 7150 + "...exactly here...in the attitude towards the patient...to set the line between subjective and objective." (First-year female student) + + 9606 + Species + + patient + + + + RESULTS + title_2 + 7290 + 4.2. Empathy + + Disease + + Empathy + + + + RESULTS + paragraph + 7303 + First-year students perceived empathy as something you could learn but not in order to really become an empathic person but in to make an impression that you are an empathic person. + + 9606 + Species + + person + + + 9606 + Species + + person + + + Disease + + empathy + + + + RESULTS + paragraph + 7485 + "I think it is important that the physicians do not feel sorry for the patient as their primary goal is to cure the patients" (First-year female student) + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + + RESULTS + paragraph + 7643 + "I think it is important to ask the patients how they are feeling. Not because you are really interested but to show them that you care. (First-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 7810 + "...(empathy) is learned not innate...you can act that you care for someone but you really don't." (First-year male student) + + Disease + + empathy + + + + RESULTS + paragraph + 7941 + Fifth-year students were more concerned with how to draw a line between being too empathic and being not empathic at all. + + + RESULTS + paragraph + 8063 + "It is necessary to find a healthy line between taking the emotional stuff home and not being able to sleep and between being a misanthrope and a cold person." (Fifth-year female student) + + 9606 + Species + + person + + + + RESULTS + paragraph + 8255 + "Not only that we write in the patient's record that we will keep him for another week in the hospital because he has nowhere to go but also to understand his situation and to adjust the management and communication." (Fifth-year male student) + + 9606 + Species + + patient + + + + RESULTS + title_2 + 8503 + 4.3. Respect + + + RESULTS + paragraph + 8516 + Students of both study years recognized the importance of a respectful relationship towards coworkers and patients. + + 9606 + Species + + patients + + + + RESULTS + paragraph + 8632 + "I cannot imagine the relationship my colleague will have towards patients if he is not able to say hi to me when we meet even though he has been my classmate for the last five years. " (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 8850 + "To show respect to the patients by addressing them with sir or madam." (First-year female student) + + 9606 + Species + + patients + + + + RESULTS + title_2 + 8954 + 4.4. Responsibility + + + RESULTS + paragraph + 8974 + Students expressed the need for responsibility in form of continuous professional education and being able to recognize your limits. + + + RESULTS + paragraph + 9107 + "...to engage in continuous education, to try to follow guidelines, not to rest on laureates." (Fifth-year female student) + + + RESULTS + paragraph + 9234 + "...being responsible to your patients in a way to be able to help them as much as you can." (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 9359 + "If the physician does not know what is wrong with you he refers you to other physicians." (First-year female student) + + + RESULTS + title_2 + 9482 + 4.5. Autonomy + + + RESULTS + paragraph + 9496 + The first-year students perceived autonomy as the right of patients to make their own decisions and also as their obligation for taking more responsibility. The fifth-year students referred also to the autonomy of the physicians. + + 9606 + Species + + patients + + + + RESULTS + paragraph + 9726 + "I think that nowadays patients have way to much autonomy and very little responsibility and this is not right." (First-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 9871 + "...that you do not let the patient to control you i.e. to tell you which drug to prescribe, to set this line." (Fifth-year female student) + + 9606 + Species + + patient + + + + RESULTS + paragraph + 10015 + "That the explanatory duty/informed consent does not become some kind of a blackmail and a form of persuasion that your truth is the only truth for this patient but that you only try to explain and present different treatment options." (Fifth-year female student) + + 9606 + Species + + patient + + + + RESULTS + title_2 + 10283 + 4.6. Trust + + + RESULTS + paragraph + 10294 + The students agreed that trust and confidentiality were essential parts of medical professionalism. + + + RESULTS + paragraph + 10394 + "I perceive professionalism as the relationship where patient's problems stay between you and him. That you do not talk about them outside your office." (First-year male student) + + 9606 + Species + + patient + + + + RESULTS + paragraph + 10577 + "You have to gain confidence, that he trust you, that your knowledge is sufficient and he can put his health into your hands." (First-year female student) + + + RESULTS + paragraph + 10736 + Communication + + + RESULTS + paragraph + 10750 + Students perceived communication as an essential part of professionalism practice. They considered that patient should understand physician and also physician should understand patient. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + + RESULTS + paragraph + 10936 + "It is important to now that nowadays patients read a lot and then they can mislead their physician that they understand something but they really don't. They just talk about something they've read somewhere and do not understand it." (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 11207 + "I think that medicine has become a service activity ad people seek service when they go to the doctor. They sometimes forget how good it felt when physicians took the time and listened to them." (Fifth-year male student) + + 9606 + Species + + people + + + + RESULTS + paragraph + 11433 + "Professionalism is being able to communicate in such a way that the patient understands you. If he comes from low social class you communicate in a more lay language and if he comes from a high class you can communicate in a more sophisticate way." (First-year male student) + + 9606 + Species + + patient + + + + RESULTS + paragraph + 11713 + Difference between professional and private life + + + RESULTS + paragraph + 11762 + The students recognized the importance of drawing the line between professional and private life. First-year students were very strict about it, whereas fifth-year students allowed the possibility to do so but being aware of the consequences. + + + RESULTS + paragraph + 12005 + "...if you take this home as a luggage then this can limit your ability to perform your work. Which also affects the patients." (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 12165 + "When you come home you must leave your work at work. There is no other way." (First-year female student) + + + RESULTS + title_2 + 12275 + 4.7. Team work + + + RESULTS + paragraph + 12290 + Team work as a part of professionalism was only recognized by fifth-year students. + + + RESULTS + paragraph + 12373 + "...meaning that there is not any gossiping and malicious imputation in the team so you can focus on work. This is what patients deserve." (Fifth-year male student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 12542 + "For example, the patients can see if there are some disputes within the team and this is a sign for them that they will not receive a quality management." (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + title_2 + 12730 + 4.8. Partnership + + + RESULTS + paragraph + 12747 + Partner relationship with patients was only recognized by fifth-year students. + + 9606 + Species + + patients + + + + RESULTS + paragraph + 12826 + "That you do not give the patient the feeling that he has no clue and that you do not place yourself above him." (Fifth-year female student) + + 9606 + Species + + patient + + + + RESULTS + paragraph + 12971 + "I think that the patient has to feel as he has the possibility to participate in the decisions but that is our duty to present out professional opinion in such a way that he will agree with it and still think that it was solely his decision." (Fifth-year female student) + + 9606 + Species + + patient + + + + RESULTS + title_2 + 13247 + 4.9. Physician's characteristics + + + RESULTS + paragraph + 13280 + Physician's personality + + + RESULTS + paragraph + 13304 + Students perceived the personality of the physician to be an important part of professionalism. However, they thought it was greatly a matter of family patterns. + + + RESULTS + paragraph + 13466 + "I think that this is a thing that people bring it from their childhood, from their families." (Fifth-year female student) + + 9606 + Species + + people + + + + RESULTS + paragraph + 13593 + "Some people always ignore those who for example fall on the street while others always stop and help. This is not something you could learn." (Fifth-year female student) + + 9606 + Species + + people + + + + RESULTS + paragraph + 13768 + "Professionalism largely depends on the personality. Some are more collegial and help their colleagues even now and lend them their notes while others keep the things to themselves. You know that they will be like that with their patients : they will not be willing to listen to anybody or to recognize their mistakes." (First-year male student) + + 651272 + Species + + the things + + + 9606 + Species + + patients + + + + RESULTS + paragraph + 14120 + Role modelling + + + RESULTS + paragraph + 14135 + This was recognized only by fifth-year students. + + + RESULTS + paragraph + 14184 + "To be a good mentor to students and trainees and others you ought to educate. Nowadays, many are involved in the education process as teachers but they fail to remember that." (Fifth-year male student) + + + RESULTS + paragraph + 14391 + Expertise + + + RESULTS + paragraph + 14401 + Students perceived expertise to be a part of medical professionalism but not necessary a positive part. + + + RESULTS + paragraph + 14505 + "Someone can be a great expert but has a very low professional relationship towards patients." (Fifth-year female student) + + 9606 + Species + + patients + + + + RESULTS + paragraph + 14632 + "I think that there are also doctors who are not very professional but are great experts. They can diagnose a patient very good but are not considered to be good doctors by patients." (First-year female student) + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + + RESULTS + title_2 + 14848 + 4.10. External factors + + + RESULTS + paragraph + 14871 + Education + + + RESULTS + paragraph + 14881 + According to fifth-year students, professionalism can be learned and the education system can affect it, especially in a negative way. + + + RESULTS + paragraph + 15016 + "I think that professionalism or at least one part of it, can be learned." (Fifth-year male student) + + + RESULTS + paragraph + 15121 + "I think that the medical profession can soon become inhuman. So, the humanistic part of it should be constantly a part of education. Otherwise, we can quickly start regarding patients as objects and not humans." (Fifth-year male student) + + 9606 + Species + + patients + + + 9606 + Species + + humans + + + + RESULTS + paragraph + 15364 + "I think that you change a lot during your study. You start to hate people." (Fifth-year female student) + + 9606 + Species + + people + + + + RESULTS + paragraph + 15473 + Health care system + + + RESULTS + paragraph + 15492 + Students regarded health care system as an obstacle to professionalism. + + + RESULTS + paragraph + 15564 + "The problem is the health care system itself. You do not have time to engage in a humanistic and professional relationship." (Fifth-year female student) + + + RESULTS + paragraph + 15722 + "I still think that the health care system made the people to perceive the health care workers as their slaves or as salesmen in the store - they expect to get some kind of drug each time they go to the doctor." (First-year female student) + + 9606 + Species + + people + + + + DISCUSS + title_1 + 15968 + 5. DISCUSSION + + + DISCUSS + paragraph + 15982 + Students in this study recognized the majority of professionalism components as recognized in several theoretical models. The also recognized some external and internal factors that were associated with or could in their opinion affect the medical professionalism. Although there were no significant differences in the attitudes towards medical professionalism between first-and fifth-year students, a change in the perceptions of the fifth-year medical students was observed in terms of recognizing more professionalism categories and more self-centred perceptions about the physician figure. + + + DISCUSS + paragraph + 16576 + As previously stated, there is no unified theoretical model of medical professionalism and it is therefore difficult to relate our results to the theoretical concepts. If we compare the medical professionalism categories recognized by the undergraduate students to the aforementioned theoretical models (Table 1), we can see that the main professionalism components have been recognized: altruism, autonomy, integrity, relationship, empathy and confidentiality. Of course, the wording and the terms of some components are sometimes different but the meaning is similar, i.e. objectivity and responsibility relate to altruism and autonomy relates to integrity. However, the students failed to recognize some professionalism components, i.e. continuous improvement, quality of care, access to care, distribution of finite resources and scientific knowledge. If we take a look at some studies that dealt with professionalism attitudes in undergraduate students, we can see that students recognized a very wide range of professionalism components and some of them are comparable to the categories in our study. For example, five out of seven components reported by Blackall et al were recognized in our study: accountability, altruism, duty, honour and integrity and respect. In another study, similar to our study, respect, relationship and partnership were recognized. Yet another study put forward confidentiality - similar to our study. However, the students in our study also recognized empathy, communication, team work and difference between professional and private life which other studies failed to recognize. + + Disease + + empathy + + + Disease + + altruism + + + Disease + + altruism + + + Disease + + altruism + + + Disease + + empathy + + + MESH:D008569 + Disease + + wording + + + + DISCUSS + paragraph + 18194 + Some factors which are not professionalism components but rather relate to professionalism or even affect it were recognized by the students in our study. These were physician's characteristics and external factors (Table 2). These factors act either as facilitators or inhibitors of professionalism. They are also not recognized in the aforementioned theoretical models. However, some of them were recognized in some other studies on students' attitudes to professionalism, i.e. physician's self-development. Also, some of them were recognized in some studies as the possible influences on professionalism (i.e. role modelling). In our study, the students recognized the expertise of the physicians to be among possible influential factors whilst in some other studies and/or theoretical models this entity was one of professionalism components. In our study, the students also expressed the role of external factors. One of them is the role of education which has already been studies and agreed that professionalism can be learned and taught. Also, the students recognized a very important role of the health care system which can have a (mainly) negative influence on professionalism. + + + DISCUSS + paragraph + 19385 + In this study, some differences were detected between the first-and fifth-year students' attitudes towards professionalism. First of all, some categories were recognized only by the fifth-year students: teamwork, partnership, role modelling and education. It seems that students do learn about professionalism through their medical studies. Also, they seem to grasp the importance of education in developing a figure of a professional physician. Additionally, there were some qualitative differences in perception of several professionalism components. Although the students of both study years recognized empathy as one of the components, first-year students expressed a very pragmatic attitude towards it whereas fifth-year students were concerned about how to cope with too much empathy. Similar findings were reported regarding drawing the line between personal and professional life. Several studies reported on the decline of empathy through the medical studies. Although our study was qualitative and hence the differences are hard to detect, our results suggest that the students become more sensible to empathy as they move towards the end of their studies. Based on the results of our study, we cannot claim that they become more or less empathic; we can only say that they become aware of potential harm empathy could have on their personal lives. This finding relates also to the finding about the autonomy. Namely, the fifth-year students also recognized the autonomy of the physician not only of the patient. + + 9606 + Species + + patient + + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + + + Disease + + empathy + + + + DISCUSS + paragraph + 20910 + The finding that the first-year students recognized the majority of the professionalism components points to the fact that the students have an appropriate picture of the physicians' figure even at the beginning of their medical studies. The finding that the fifth-year students recognized more components then their first-year colleagues confirms the need for professionalism education. A slight change of the attitudes detected in our study might point to the development of a more self-centred future physicians' figure which is also consistent with other studies. There is a need for quantitative prospective studies which will evaluate these qualitative findings and critically assess the role of education. + + + DISCUSS + paragraph + 21627 + This study had a qualitative methodology which was appropriate for determination of the attitudes but less appropriate for the evaluation of differences between first-and fifth-year medical students. With qualitative methodology, we cannot detect quantitative differences in attitudes but only the substantive differences. However, the study gave us the insight to the attitudes of students towards professionalism and will serve for the development of a quantitative tool for measuring these attitudes. + + + CONCL + title_1 + 22131 + 6. CONCLUSION + + + CONCL + paragraph + 22145 + The students seem to have an appropriate picture of the physicians' figure even at the beginning of their medical studies. It seems that the fifth-year students perceive physicians as more self-centred when compared to their first-year colleagues. Therefore, an education on professionalism is essential also at the undergraduate level. 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2014 Oct; 26(5) 309-12. doi:10.5455/msm.2014.26.309-312 + assessment socioeconomic status status of oral health school children + surname:Panjeta;given-names:Mirsad + surname:Ridic;given-names:Goran + 312 + surname:Coric;given-names:Jozo + surname:Dzananovic;given-names:Nejra + surname:Delic-Sarac;given-names:Marina + Saban A, Ridic O, Karamehic J, Saban O, Delic-Sarac M, Dzananovic N, Coric J, Ridic G, Panjeta M + 0 + Assessments of the Socioeconomic Status and Diet on the Prevalence of Dental Caries at School Children in Central Bosnian Canton + + 9606 + Species + + Children + + + + ABSTRACT + abstract_title_1 + 129 + Aim: + + + ABSTRACT + abstract + 134 + The main aim of this research was to determine the influence of socioeconomic status and residence/living conditions on the status of oral health (e.g. health of mouth and teeth) in primary school students residing in Canton Central Bosnia. + + MESH:D012607 + Disease + + Canton Central Bosnia + + + + ABSTRACT + abstract_title_1 + 375 + Methods: + + + ABSTRACT + abstract + 384 + The study was designed as a cross-sectional study. Our research included two-phased stratified random sample of 804 participants. The quantitative research method and newly designed survey instrument were utilized in order to provide data on the oral health of the examined children. The alternate hypothesis foresaw that "there were significant statistical differences between the levels of incidence of dental caries in comparison to the incidence in children of different socioeconomic status. + + 9606 + Species + + participants + + + 9606 + Species + + children + + + 9606 + Species + + children + + + + ABSTRACT + abstract_title_1 + 883 + Results: + + + ABSTRACT + abstract + 892 + The Chi square of 22.814, degree of freedom (Df) = 8, coefficient of contingency of 0.163 and T-test (Stat) of-0.18334 showed that there were no significant statistical differences at p < 0.05 level between the primary school children from urban and rural areas. The obtained results showed that the caries indexes in elementary schools in Central Bosnia Canton were fairly uniform. Research showed that there were a difference in the attitudes towards a regular dental visits, which correlated with social-educational structure of the children's' families. + + 9606 + Species + + children + + + 9606 + Species + + children + + + MESH:D012607 + Disease + + Central Bosnia Canton + + + + ABSTRACT + abstract_title_1 + 1457 + Conclusion: + + + ABSTRACT + abstract + 1469 + According to the results, we can see that the socioeconomic status of patients had an effect on the occurrence of dental caries and oral hygiene in patients in relation to the rural and urban areas, because we can see that by the number of respondents, the greater unemployment of parents in both, rural and urban areas, caused a host of other factors, which were, either, directly or indirectly connected with the development of caries. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + INTRO + title_1 + 1907 + 1. INTRODUCTION + + + INTRO + paragraph + 1923 + Oral health (i.e. health of mouth and teeth) of school children represents a significant social and economic value of the modern world. Retrospective research data show that there is a difference in attitudes of examined children regarding the regular control of oral health, which is in correlation with social and educational structure of their families. The opinion that by practicing a regular preventive controls diseases of mouth and teeth can be prevented, obviously and unfortunately is not prevalent as life practice in many of primary school children's families Even though the children are exposed to the oral health at the fairly young age and although the parents are increasingly aware of the importance of the oral health for the entire well-being of their children, in many instances the role of parents and children in maintaining health of mouth and teeth is not sufficient, so the necessity arises to seek the professional help. It is best to seek help of dental professionals preventively, even then when everything seems to be well with the health of mouth and teeth. + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + MESH:D009059 + Disease + + diseases of mouth + + + + INTRO + paragraph + 3012 + In 2005, as an answer to the rather poor state of primary school children's oral health in Federation entity of Bosnia and Herzegovina (F.B&H), within all of its ten Cantons, a promotional-preventive activities have been carried out with the main objective to familiarize the matriculated first grade children with the importance of proper oral hygiene and its correct application. This ambitious program was carried out in cooperation with Federal Ministry of Health, Federal Institute for Public Health, Federal Institute for Insurance and Re-insurance and primary health care institutions in F.B&H. In the academic school year 2004/2005 this program included 99.5 percent of all primary schools in F.B&H (i.e. 398 of central primary schools and majority of regional schools), while, at the same time education 89.2 percent of first grade children. In the academic school year 2006/2007 this program was additionally carried out to the first and second grade primary school pupils in Federation of Bosnia and Herzegovina. + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + Disease + + Federation of Bosnia + + + Disease + + Bosnia + + + + INTRO + paragraph + 4036 + The progress of the infection process is exemplified by the penetration into the tissue around the tooth's root and development of periodontitis. Each hole in the tooth represents an additional place for keeping food, plaque formation and propagation of a large number of bacteria. + + MESH:D007239 + Disease + + infection + + + MESH:D010518 + Disease + + periodontitis + + + + INTRO + paragraph + 4318 + The treatment actually consists in detecting caries damage and removing softened dentin, followed by the appropriate treatment and disinfection of the oral cavity and eventually sealing. + + MESH:D003731 + Disease + + caries damage + + + + INTRO + paragraph + 4505 + While there are many theories about the origin of caries none of them give a complete answer to all the questions that come to mind when it comes to the etiopathogenesis of dental caries. According to contemporary views, some factors have greater importance in its origination, namely: + + + INTRO + paragraph + 4791 + a) The structure and roughness of enamel; + + + INTRO + paragraph + 4833 + b) Dental plaque; + + + INTRO + paragraph + 4851 + c) Properties of saliva; and + + + INTRO + paragraph + 4880 + d) Carbohydrates. + + MESH:D002241 + Chemical + + Carbohydrates + + + + INTRO + paragraph + 4898 + Dental plaque is of great importance, both in the development of caries, as well as in the pathogenesis of parondotopathy. It consists of desquamated epithelium of the oral mucosa, microorganisms and mucin (constituents of saliva). + + 100508689 + Gene + + mucin + + + Disease + + parondotopathy + + + + INTRO + paragraph + 5130 + The accumulation of plaque on the tooth surfaces on which the physiological self-cleaning and cleaning with a brush is rather inefficient, is preceded and followed by caries. A special attention should be paid to microorganisms that are found in plaque, especially cariogenic strains of streptococci (Streptococcus mutans and sangus). Their biochemical activities carry out decomposition of carbohydrates to simple acids and synthesize different polysaccharides (i.e. dextran, levan and amylopectin). + + 1309 + Species + + Streptococcus mutans + + + MESH:D003911 + Chemical + + dextran + + + MESH:D011134 + Chemical + + polysaccharides + + + MESH:D002241 + Chemical + + carbohydrates + + + + METHODS + title_1 + 5631 + 2. MATERIALS AND METHODS + + + METHODS + paragraph + 5656 + The study was designed as a cross-sectional study. The survey instrument was used for the survey interview and clinical examination of the mouth and teeth to provide data on oral health of the examined children. The survey was conducted in all units of observation, in selected schools in the stage of choice within these departments-selected groups of students. This also meant that two phases stratified sample of compatible groups, where 804 students were surveyed in elementary schools and where 355 students were from urban and 449 from rural schools. + + 9606 + Species + + children + + + + METHODS + paragraph + 6213 + Creating questionnaires and publicity materials; + + + METHODS + paragraph + 6266 + Working team members' education; + + + METHODS + paragraph + 6305 + Members were familiarized with the available data on oral hygiene habits and their impact on oral health, various techniques and tools for proper oral hygiene, and tools and techniques for the determination of OHI (e.g. index hygiene oral). + + + METHODS + paragraph + 6550 + Data were collected by conducting surveys in several classrooms (i.e. one class from each age group selected among primary schools) and filling out questionnaires with the assistance of members of the survey team, in order to help to determine the status and oral hygiene index. + + + METHODS + paragraph + 6833 + At the end, the data analysis and evaluation of the project was carried out. + + + RESULTS + title_1 + 6914 + 3. RESULTS + + + RESULTS + paragraph + 6925 + Below is presented the analysis of the data that we received after interviewing 804 students in elementary schools, where 355 students were from urban and 449 students from rural schools, the following results were obtained: + + + RESULTS + paragraph + 7150 + Assessment of the socioeconomic status in children from urban and rural areas was carried out on the basis of the group questions in the application form, which were related to employment and qualifications of father / mother, the place of residence, and on the basis of their financial status. The Chi squared had the value of 22.814, the Level of Significance of P = 0.0001 and Contingency Coefficient of 0.163. + + 9606 + Species + + children + + + + RESULTS + paragraph + 7564 + The obtained responses from this group of questions are presented in the table 1 below: + + + TABLE + T1.xml + T1 + table_caption + 7652 + t-Test according to the Socioeconomic status in examinees residing in rural and urban areas. + + + RESULTS + paragraph + 7745 + No statistical significance was observed at p < 0.05, and therefore there is no statistically significant difference between urban and rural primary school children according to the socioeconomic status. + + 9606 + Species + + children + + + + RESULTS + paragraph + 7949 + From total amount of 72.37 percent of surveyed children listed that their father was employed, (i.e. 44.00 percent in urban and 28.37 percent in rural areas), 25.75 listed that their father was unemployed (i.e. 14.00 percent from urban and 11.37% from rural areas). Situation with mothers' employment status was quite different; whereas 52.5 percent of surveyed children listed (i.e. 22.25 percent in urban and 30.25 percent from rural areas) listed that their mother was employed, while 66.62 percent (i.e. 18.5 percent from urban and 48.125 percent from rural areas) listed that their mother was unemployed. 86.5 percent of examinees listed to reside in their own house or apartment (e.g. 34.37 percent in urban and 52.12 percent in rural areas). 40.50 percent of surveyed answered to have at disposal its own room, while 53.75 percent listed to share the room with brother or sister. By taking into consideration the answers regarding employment and socioeconomic status of examined children, where are clearly visible changed in rural and urban areas, we can ascertain that socioeconomic status (as an independent variable) was in causal relationship with the causation and development of dental caries and adverse changes in oral hygiene of examinees. Poorer socioeconomic status seems to have caused the incidence of the increasing number of examinees from rural areas who have visited dentist(s) more than four times; (i.e. 19.75 percent) in comparison to the examinees from the urban areas measured at 16.5 percent. At the same time there were not a single examinee residing in the urban areas who had reported to have visited a dentist more than ten times, while there were 1.5 percent of surveyed from the rural areas who visited a dentist more than ten times. + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + + FIG + MSM-26-309-g002.jpg + F1 + fig_caption + 9722 + Graphical representation of prevalence of CAP Index (includes the prevalence of: caries, filling and extractions) + + + DISCUSS + title_1 + 9836 + 4. DISCUSSION + + + DISCUSS + paragraph + 9850 + When defining the socioeconomic status we can clearly see that the majority of respondents from rural and urban areas stated that their father was employed. In a small number of answers the unemployment was viewed as a direct impact on the socioeconomic status of respondents, as well as the position of the mother's employment where it was clear that there was a difference between respondents in urban and rural, where a much larger number of respondents answered that the mother was not employed, which was not the case in the urban environment. Accordingly, the city center had a much larger number of respondents who reported that their parents were employed compared to rural population. + + + DISCUSS + paragraph + 10544 + Most of the respondents in urban and rural areas stated to have their own house / apartment, while in the middle of both had the same number of responses for respondents who were living as tenants. The largest number of respondents shared his/her room with some brother / sister, while the rural areas had a higher number of respondents who had their own room. In the city, although the higher the number was higher than in the rural areas, statistically speaking there was some difference in favor of the respondents' rural areas. + + + DISCUSS + paragraph + 11078 + The second group of questions related to hygiene and diet and nutrition in relation to the hypotheses of this study gave some interesting answers. When asked how often they were having breakfast on weekends most of the respondents answered that they had breakfast on both days of the weekend, in both the city and rural areas. + + + DISCUSS + paragraph + 11405 + Most of the respondents stated that they were pleased with his/her weight and that there was no need or feeling the need to dedicate to a diet, and that a large number of respondents regularly consumed meals. A large number of respondents stated that they most commonly consumed fast food, and that having in mind its composition it was felt as a predictor for the occurrence of various diseases, including dental caries. + + + DISCUSS + paragraph + 11827 + It is important to note that this factor had an impact on the status of oral hygiene in our patients, and that our sample consisted of a very large number of respondents who consumed sweets and snacks, each day and several times a day. Encouraging responses were in the forms of answers where the fruits were consumed more than three times in the last three days, although it was directly linked with socioeconomic status of respondents. + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 12265 + The high prevalence of dental caries was caused by certain safe factors, so in this case we consider diet and place of residence (e.g. rural and urban environment), where the respondents from urban areas were more susceptible to the influence of factors that lead to tooth decay, such as fast food and sugary treats, with 64.48% of the urban environment usually taking fast food during the big break at school, but also, a large number of children from rural areas at 80.95%, which meant that the predominant factors were no longer tied to regional-geographical determination (e.g. in rural and urban communities). + + 9606 + Species + + children + + + + DISCUSS + paragraph + 12882 + In our sample, 45.25% of respondents from urban areas have been consuming candy daily, while 68.5% of them from rural areas also consumed sweets every day, which indicated that the predominant factors were not specific geographical locality. + + + CONCL + title_1 + 13124 + 5. CONCLUSION + + + CONCL + paragraph + 13138 + On the basis of this research the following conclusions can be drawn: + + + CONCL + paragraph + 13208 + Conditions of oral hygiene habits' among school children was on a satisfactory level and no large differences were observed between respondents in urban and rural areas; + + 9606 + Species + + children + + + + CONCL + paragraph + 13384 + The age group that comes to improving oral hygiene in the second half of elementary education, with a number of information, etc. + + + CONCL + paragraph + 13518 + Factors that were predominant in relation to residence and caries were: eating poor quality food during the big break at school, and in urban areas more than in rural areas, one of the essential factors of consuming sweets and snacks was equally represented in all areas. Regular brushing was the factor that had an impact on the formation of cavities, but it could not be correlated with the geographic distribution. Taking diet that was supposed to improve tooth nutrition, such as drinking glasses of milk, gave no statistically significant results in both groups, but, rather favored the rural areas. + + + CONCL + paragraph + 14127 + According to the results, we can see that the socioeconomic status of patients was affected by the occurrence of dental caries and oral hygiene, in patients, in relation to their rural and urban residence areas, because we could observe that by the number of respondents and greater unemployment in both parents in rural areas than in the city, which caused a host of other factors that were directly and indirectly connected with the development of caries. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + + CONCL + paragraph + 14585 + According to the type of food consumed we could see that in urban and rural areas the responses were fairly balanced, with the city where the larger number of respondents ate foods that posed a risk factors for tooth 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+ + + ABSTRACT + abstract_title_1 + 59 + Aim: + + + ABSTRACT + abstract + 64 + The main objective of this descriptive and development research was to introduce a design protocol to develop radiology dashboards. + + + ABSTRACT + abstract_title_1 + 196 + Material and methods: + + + ABSTRACT + abstract + 218 + The first step was to determine key performance indicators for radiology department. The second step was to determine required infrastructure for implementation of radiology dashboards. Infrastructure was extracted from both data and technology perspectives. The third step was to determine main features of the radiology dashboards. The fourth step was to determine the key criteria for evaluating the dashboards. In all these steps, non-probability sampling methods including convenience and purposive were employed and sample size determined based on a persuasion model. + + + ABSTRACT + abstract_title_1 + 792 + Results: + + + ABSTRACT + abstract + 801 + Results showed that there are 92 KPIs, 10 main features for designing dashboards and 53 key criteria for dashboards evaluation. As well as, a Prototype of radiology management dashboards in four aspects including services, clients, personnel and cost-income were implemented and evaluated. Applying such dashboards could help managers to enhance performance, productivity and quality of services in radiology department. + + + INTRO + title_1 + 1222 + 1. INTRODUCTION + + + INTRO + paragraph + 1238 + The field of radiology is continuously growing. Since imaging services are widely used and affect on patient care in the hospital, much attention has been paid to quality assurance in radiology departments. Since, measuring the quality of imaging services is inherently difficult, standard sets of KPIs should be used as key tools for evaluating and improving departmental processes. In traditional method, the monitoring of indicators involved the review of patient records to document the effect of diagnostic imaging on the care outcome is costly in staff time and labor. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D048949 + Disease + + labor + + + + INTRO + paragraph + 1813 + With the modernization of radiology departments, along with using electronic information systems such as PACS, RIS, speech recognition systems, preliminary report systems, hospital information systems, and electronic health records, workflow and informatics infrastructure supporting them became very complex. This is because it requires combining many computerized systems with different user environments to provide quality efficient services in less time. + + + INTRO + paragraph + 2272 + Since within this complexity, close monitoring the operational and financial data is an essential element for the management; Traditional methods of data involving review of Key Performance Indicators become quickly outdated and changed to use new technologies. In recent year, a variety of management tools have been introduced to digitally monitor essential data under the general name of dashboards. + + + INTRO + paragraph + 2675 + Dashboards is an intellectual and visual tool for monitoring of KPI which can capture key data from different systems and represent them summarized and useful in real-time with easier reading and interpretation for users. This business intelligence tool can applied in radiology department as used in other sections such as operation rooms, emergency rooms, diabetes, and hospital infection control. + + MESH:D003920 + Disease + + diabetes + + + MESH:D056987 + Disease + + operation rooms + + + MESH:D003428 + Disease + + hospital infection + + + + INTRO + paragraph + 3075 + In radiology department, workflow are optimized when radiologists have real-time information to make informed decisions, and the capacity to efficiently act on that information. Dashboards technology can facilitate informed, optimized decisions about workflow by link directly to the electronic information systems and provide the right information in a right format at the right time to monitor productivity in order to improve in performance and effectiveness of activity. In this research, a design protocol to develop radiology dashboards in order to enhance performance, productivity and quality of services in this department, has been proposed. As well, a prototype of radiology management dashboards were implemented and evaluated using data obtained from a selected hospital affiliated to Tehran University of Medical Sciences. + + + METHODS + title_1 + 3912 + 2. RESEARCH METHOD + + + METHODS + paragraph + 3931 + This descriptive and development research has been done in 4 steps during 2012 up to 2014. + + + METHODS + paragraph + 4022 + Determine Key Performance Indicators for radiology department: This descriptive study was conducted using the Delphi technique. In this step, study population was including 30 radiologists who were members of Iranian Radiology Association and had professional experience in managing of radiology department. Non-probability sampling method including convenience and purposive were employed. Electronic questionnaire was used to collect information. At the end of the two-stage Delphi technique 92 key indicators to assess the performance of the radiology department into 7 main categories including "safety, service, internal and external customers, teaching and research, resource utilization, financial performance and excellence in the workplace" was determined. + + + METHODS + paragraph + 4792 + Determine a comprehensive model for designing of dashboards: This step of the research is descriptive. The study population was composed of 15 specialists in the field of informatics who had experience in creating dashboards. Non-probability sampling method including convenience and purposive were employed. Electronic questionnaire was used to collect information. At the end of this stage, 10 main features including" determining the goal of dashboards design, aligning with organizational goals, determining KPIs, setting a time frame, extracting the accurate data, flexibility, drilling-down capabilities, security, how to representation and display alert" were determined as a model for designing dashboards. + + + METHODS + paragraph + 5511 + Determine required infrastructure for implementation of Radiology Management Dashboards: Infrastructure was extracted from both data and technology perspectives and the following steps are performed: + + + METHODS + paragraph + 5711 + Determining the goals of the dashboards design in order to determine the KPIs + + + METHODS + paragraph + 5789 + Defining the target group who are authorized to use dashboards + + + METHODS + paragraph + 5852 + Determining whether the dashboards show the retrospective data or real-time data? + + + METHODS + paragraph + 5934 + Determining KPIs to achieve defined goals and how to calculate them + + + METHODS + paragraph + 6002 + Identifying the required data to achieve these indicators and sources of data + + + METHODS + paragraph + 6080 + Setting intervals to update the information + + + METHODS + paragraph + 6124 + Identifying the reporting format of the data resource + + + METHODS + paragraph + 6178 + Specifying how to extract data + + + METHODS + paragraph + 6209 + Determine the key criteria for assessing the dashboards: In this step, a total of 46 specialists from medical informatics, health information management, software engineering, and radiology field who had experience in the field of business intelligence were chosen as study population. Non-probability sampling method including convenience and purposive were employed. Electronic questionnaire was used to collect information. At the end of this stage, 53 key criteria in seven categories including"user-customization, knowledge discovery, security, information delivery, visual design, alerts, connectivity and integration" were determined for dashboards evaluation. + + + RESULTS + title_1 + 6881 + 3. RESULTS + + + RESULTS + paragraph + 6892 + The findings based on the research steps are the following: In first step, as shown in Table, 92 key indicators to assess the performance of the radiology department at the seven categories were determined. In the second step, a comprehensive model for the design of the dashboards as shown in Table was determined. In the third step, to prepare prototype radiology management of dashboards, QlikView software as open source integration software that is executable in a hospital operating systems including Windows XP, Windows 7 and Windows 8 are from Microsoft, was used. As well as, some KPI as shown in table was selected from table to display in radiology management dashboards, the data was captured form radiology information system, financial information system and personnel information system and the time interval for updating was monthly. In the fourth step, the dashboards evaluation criteria were determined which are presented in Tables 1-4. + + + TABLE + T1.xml + T1 + table_caption + 7848 + Key performance indicators in radiology department + + + TABLE + T2.xml + T2 + table_caption + 7899 + A Comprehensive Model for Designing Radiology Dashboards + + + TABLE + T3.xml + T3 + table_caption + 7956 + Selected KPIs for radiology department management dashboards + + + TABLE + T4.xml + T4 + table_caption + 8017 + key criteria for the dashboards evaluation + + + DISCUSS + title_1 + 8060 + 4. DISCUSSION + + + DISCUSS + paragraph + 8074 + Given the above it can be concluded that the field of radiology is faced with the challenges of increasing service use, declining reimbursements, and staff shortages. In such an economically challenging environment, it becomes crucial for radiology departments to monitor their performance so that they can provide a high quality of services while staying within operational boundaries. Accordingly, in this research, enhancement of performance, productivity and quality of services in radiology department as the aims of dashboards designing was considered. + + + DISCUSS + paragraph + 8633 + In this field, Performance improvement is a process that differs from analyses made in commercial companies, where increased revenue is the major goal. In healthcare, the benefit to the individual patient is the main goal, with less emphasis being placed on economic implications. So to improve the quality required to ensure the proper planning done. A key element to planning is to select required data, and then adopt appropriate methods to collect data actively, consistently and effectively and represent them in performance indicator format. + + 9606 + Species + + patient + + + + DISCUSS + paragraph + 9181 + Therefore it is necessary to instill a culture to determine necessary data to calculate the KPIs, the ways to collect these data and appropriate tools should be used for data analysis and trending. However, it should be noted that quality is not a goal at the radiology department but it is responsibility and accountability of and necessary to create a shared vision by hospital and radiology practice leadership that centers on patient. + + 9606 + Species + + patient + + + + DISCUSS + paragraph + 9620 + About Dashboards design, because of radiology department is complex environment and has a variety of services, clients, personnel, equipment, technologies, data and information which are generated, the following key points need to be considered: + + + DISCUSS + paragraph + 9866 + Organizational culture: Organizational culture is the normal behaviors dominating an organization and can be an important factor in successful implementation of technology. Implementing dashboards requires organizational culture to accept objective data as the basis for decision making. + + + DISCUSS + paragraph + 10154 + Determining the goal of dashboards design: Experts believe that dashboards objectives should be limited in order for the managers to allocate the resources to critical issues and divide the objectives into three groups: financial, operational and quality. Therefore, the goals of dashboards design for the department should be clear; whether it is for workflow consolidation, workload distribution, urgency evaluation, capacity and workflow management, surveillance, or financial reporting. + + + DISCUSS + paragraph + 10645 + Involving users: in designing a comprehensive dashboards, it is necessary to involve users in the design and development processes because expectations will closely match to functionality and users provide suggestions based on their needs which can reduce the time of development and dashboards redesign after implementation as feedbacks and evaluations are provided. + + + DISCUSS + paragraph + 11013 + Aligning with organizational goals: Dashboards should be designed based on overall goals and objectives of organization in such a way that each department by monitoring its KPIs facilitates achieving its objectives besides the strategic goals of organization. + + + DISCUSS + paragraph + 11273 + Determining KPIs and benchmark standards: Selecting the type of indicators which should be included in dashboards is important. Depending on its situation, each department needs to measure the indicators related to its own area of performance, not all the indicators, in other words, indicators which are critical and special to the quality of their performance. Each indicator is selected according to business purpose and the ability to be gathered easily through standard business functions and besides accurate measurement of the indicator, we should identify dashboards metrics from regulatory standards which we need for accreditation or credentials. In designing an effective dashboards, the number of measurements should be limited, at least 15 to 25, in order to be viewed at a glance. + + + DISCUSS + paragraph + 12068 + Data: A well designed dashboards should include accurate data which is related to business process from different sources and based on acceptable and standard definitions in order to be used in benchmarking process. In addition, data storage should be based on standard format and coding to be able to manipulate them easily, to facilitate sharing them, and reduce their processing time while updating and querying from dashboards. + + + DISCUSS + paragraph + 12500 + Knowledge discovery: It is necessary that the structure of data storage be in a logical hierarchy and with required granularity for data mining and online analytical processing (OLAP) in order to extract knowledge from the data in dashboards. This will enable the user to drill down data with clicking on operational indicator to gain deep analysis of that indicator to prospectively predict the effects of decisions and preparing the reports in the form of pdf and Excel. Regarding knowledge extraction, considering the time of search, number of clicks, and the precision and accuracy of data are critical. + + 9568 + Species + + drill + + + + DISCUSS + paragraph + 13108 + Flexibility: According to dashboards should have the ability to be optimized, customized, workflow-integrated, and context-specified. This feature is also important for determining indicators such that the user can have access to the information for specific purposes and can organize them according to his or her preferences and save them to reuse in future. In designing dashboards it is also important that dashboards would have the capability to add required metrics or make change in it. + + + DISCUSS + paragraph + 13601 + Security: It is important to make sure about the security of data at all times because in radiology department, dashboards data is the result of integration of a number of information systems and also based on the internet or intranet. According to it is necessary to consider a unique accession number for connecting data parts together and a secure log-in to have access to pages containing personal health information. In addition, utilizing 'single sign -on' technology for the user to have access to each application is suggested. + + + DISCUSS + paragraph + 14141 + Time frame: The frequency of measurement of indicators depends on the nature of indicator and its effectiveness and importance on operation of department. The focus of measurement should change from period to period. Updating information based on users' view, type of use, and importance of task can be from every second to every month. + + + DISCUSS + paragraph + 14480 + Representation: Since dashboards is a tool for quick, brief, and real-time representation of data, organizing and displaying data should be such that it could be easily read and interpreted and the user does not need to scroll the pages. Depending on the type of indicators and their numbers, we can use tables or graphs to display for users to understand and make decisions. It is better to provide visualization in the form of graphs according to the type of indicator to show benchmarking comparison because tables cannot be easily and quickly read. What is critical is that the direct representation should be able to alert by color coding to show levels of threats. These alerts are based on performance target thresholds which are derived from the yearly goals and objectives. According to user interface dashboards in radiology department should be divided into three groups: User, Division, and System. They can be represented in different colors based on the priority of alerts like the traffic light (a red, yellow, or green circle). + + + DISCUSS + paragraph + 15524 + Dashboards Assessment: assessment should be considered based on two aspects. One is the verification of dashboards and other side is evaluation of dashboards itself to find out its effect on the department performance which can be done through questionnaires given to users. The results can be published in the form of newsletter in order for the users to make a general evaluation of their own and the department's performance over time. + + + DISCUSS + paragraph + 15963 + About dashboards implementation, for achieve to a comprehensive set of dashboards that contain managerial and clinical operations of the radiology department So as to be able to create the accountability and strategic value in all aspects of financial, quality and clinical, the following technical infrastructure is necessary to be considered: + + + DISCUSS + paragraph + 16308 + Designing data warehouse for gathering data from a variety of systems such as clinical information system and financial information system in order to get a comprehensive view of operations + + + DISCUSS + paragraph + 16499 + Using back-end algorithm of indicators or statistics for providing general and detailed reports + + + DISCUSS + paragraph + 16596 + Accessibility to Internet or Intranet in order to facilitate sharing data and maintain data dynamism + + + DISCUSS + paragraph + 16698 + Utilizing SOA architecture to encapsulate data in middle ware layer to be of use in different systems + + + DISCUSS + paragraph + 16801 + Using API to create a hyperlink in dashboards alert to load a study when clicked by the user + + + DISCUSS + paragraph + 16895 + Using relational database with a web service interface for modifying and querying state + + + DISCUSS + paragraph + 16984 + Utilizing interoperability standards such as HL7 messaging standards and DICOM to make connections and data exchange among different systems + + CVCL:2492 + CellLine + + HL7 + + + + DISCUSS + paragraph + 17126 + Using agents in data models for registering and managing alerts if alerts are to be shown in groups or individual + + + DISCUSS + paragraph + 17241 + Designing dashboards in. Net environment because it will result in flexibility, ease and integration with the system + + + DISCUSS + paragraph + 17359 + Utilizing web based interfaces for visualizing information + + + DISCUSS + paragraph + 17419 + Displaying graphics using server based web control which is much faster than the client based control + + + CONCL + title_1 + 17522 + 5. CONCLUSION + + + CONCL + paragraph + 17536 + In this research, the key performance indicators were determined in order to performance assessment of academic radiology departments. On top of this, a comprehensive model for designing of dashboards and a checklist including key criteria for assessing of dashboards were provided. It should be noted that the model and assessment checklist could be used for all types of dashboards. + + + CONCL + paragraph + 17921 + Finally a prototype of radiology management dashboards was implemented in four aspects including services, clients, personnel and cost-income. Applying such dashboards could help managers to evaluate radiology department from desirable or undesirable stance of performance, resource consumption and distribution of manpower at a glance, as well as identifying problems, root cause analysis and solving them using valid information driven from dashboards. 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Dashboards lowers cost per case, improves patient satisfaction and minimizes staff frustration + + 9606 + Species + + patient + + + + surname:Toland;given-names:C + surname:Daly;given-names:M + surname:Warnock;given-names:MJ + surname:Nagy;given-names:PG + 1897 + 7 + 19734469 + 2009 + Radiographics + ref + 29 + REF + 1906 + surname:Mezrich;given-names:RS + surname:Meenan;given-names:CD + 21208 + Informatics in radiology: automated Web-based graphical dashboards for radiology operational business intelligence + + + surname:Linder + surname:Jeffrey;given-names:A + surname:Eunice;given-names:Jung + surname:Maya;given-names:Olsha-Yehiav JSE + 1052 + 2006 + AMIA + ref + REF + Symposium Proceedings + surname:Blackford;given-names:Middleton + surname:Schnipper + surname:Jeffrey;given-names:L + surname:Qi;given-names:Li + surname:Julie;given-names:Greim RN + 21323 + Quality Dashboardss: Technical and Architectural Considerations of an Actionable Reporting Tool for Population Management + + + 9 + surname:Singer;given-names:PA + REF + surname:Masum;given-names:H + e32 + 4 + 17951216 + 2007 + J Med Internet Res + ref + 21445 + A visual dashboards for moving health technologies from "lab to village" + + + REF + surname:Few;given-names:S + 2006 + ref + 21522 + Information Dashboards Design: The Effective Visual Communication of Data + + + 27 + REF + surname:Woodcock;given-names:EW + 70 + 1 + 2006 + Dermatology Times + ref + 77 + 21596 + Practice dashboards + + + + 4272838 + + 25568636 + surname:Radic-Hozo;given-names:Endica + 343 + 5 + 2014 + 4272838 + MSM-26-343 + 10.5455/msm.2014.26.343-347 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 343-7. doi:10.5455/msm.2014.26.343-347 + Preschool education of children communication patterns + 347 + Radic-Hozo E + 0 + Communication Patterns in Preschool Education Institutions - Practical Examples + + + ABSTRACT + abstract_title_1 + 82 + Introduction: + + + ABSTRACT + abstract + 96 + Proper communication in pre-school institutions for education is undeniable importance to the development of the child, as evidenced by numerous studies. After the child's birth follows the most complex phase in its early phases - preschool education. Only high-quality, synergistic relationship triad: parent-child-educator and the modern postulates of preschool child education, warrants successful preschool child education. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + ABSTRACT + abstract_title_1 + 524 + Methods and materials: + + + ABSTRACT + abstract + 547 + Description, with examples from daily practice in a large institution for preschool education, marked were the critical points on the complex way in child education, many pitfalls encountered by both parents and educators. Considered are the errors in communication with the proposed solution to avoid the same in practice. + + 9606 + Species + + child + + + Disease + + pitfalls + + + + ABSTRACT + abstract_title_1 + 871 + Conclusion: + + + ABSTRACT + abstract + 883 + Proper, daily communication in the preschool institution for education, within a relationship between parent-child-educator, mutual consultation, respect, acceptance, facilitation, resulting in successful common goal - the proper education and socialization of children in institutions for preschool education. + + 9606 + Species + + child + + + 9606 + Species + + children + + + + INTRO + title_1 + 1194 + 1. INTRODUCTION + + + INTRO + paragraph + 1210 + Children speak the truth ... + + 9606 + Species + + Children + + + + INTRO + paragraph + 1239 + :Chinese saying + + + INTRO + paragraph + 1257 + Regarding the structure of communication models related to the whole system of preschool education, especially the institutions in which the system is implemented (Kindergartens or "Child House") are the basic communication patterns within and outside the institutions of preschool education. In addressing communication models in kindergarten, it is necessary to consider the needs of all participants are meeting within the communication in kindergarten. Communication in the context of human needs means meeting the needs together with other people, and with child population. Communication can be improved by the satisfaction of their needs, but not at the expense of others to meet their needs. The concept of communicative competence implies the possibility of achieving personal and relational objectives of communication, so that the objectives are achieved alone or together with others. + + 9606 + Species + + Child + + + 9606 + Species + + participants + + + 9606 + Species + + human + + + 9606 + Species + + people + + + 9606 + Species + + child + + + + INTRO + paragraph + 2158 + It is important to mention Waclavek rules on the communication process: + + + INTRO + paragraph + 2230 + It is impossible not to communicate: + + + INTRO + paragraph + 2271 + Every communication has content and relational aspects. + + + INTRO + paragraph + 2331 + Relationship characteristics are determined by the interpretation of the communication relations. + + + INTRO + paragraph + 2433 + Interpersonal intelligence is the ability to understand others, what motivates them, how they work, how to cooperate with them. Interpersonal (internal) intelligence (is the basis of interpersonal): balance in changes of personal thoughts and emotions (personal satisfaction and self-acceptance) so that we can access others without prejudice; empathize with someone else; help another person in a way that it appropriate for that person. + + 9606 + Species + + person + + + 9606 + Species + + person + + + + INTRO + title_1 + 2872 + 2. COMMUNICATION BETWEEN PARENTS-CHILD-EDUCATOR + + 9606 + Species + + CHILD + + + + INTRO + paragraph + 2924 + If we now shift the focus to the area of educating a child, there is no doubt that the most important link in the educational process of the child is parental care, communication between parent and child. Ideally, the child should grow in a family surrounded by parental care, in the company of brothers and sisters. This, idyllic, image was possible previously, before the industrial revolution when there was no institution of kindergartens, and when the kids all the time was at home under the supervision of other family members. In accordance with the new circumstances, today's parents are preoccupied with work to provide needs for the family, there is less time to socialize with the child (children). Therefore, the logical replacement for parental home and parental care, is the institution in which the child (children) stay while their parents are occupied with work in their workplaces. Their new home is a preschool (kindergarten) in which teacher/educator takes parental powers and becomes a "surrogate parent" for a group of children entrusted with their education and care. History of kindergarten just coincides with the development of society and the era of industrialization, so the first kindergartens were organized in Western countries. It also showed a critical need for them because the workers' families during their stay on the job had to find a substitute for the care of the child. On the other hand, communication cooperation and participation of parents in the program, early childhood education for long time has been questionable because of Hawkins paradigm "that parents will offer sooner the subjective assessment than expert opinions or objective observations". Finally, the fact that parents are responsible for their child, according to some eminent experts, parents should also have a voice in program evaluation. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 4787 + Bronfenbrenner argued that for better efficiency of the early childhood programs for upbringing and education and their lasting impact on children, there is the need to involve parents of children and the community, and that they should work together to promote same goals. + + 9606 + Species + + children + + + 9606 + Species + + children + + + + INTRO + paragraph + 5061 + In practice, until the beginning of the 90' school partnerships in relation kindergarten-family was more the exception than the rule. Since then, however, provided there is growing evidence that parental involvement has a positive effect on the quality of education of children and their success in school. Higher levels of parental involvement in their child's upbringing is not due to the legal framework and regulations, but also due to the simple realization that parents are best acquainted with all the characteristics of their child. Reform moves from the mid-eighties, parents communicate daily with preschools staff, which are entitled to inform them about child development while choosing the right kindergarten. Some studies confirm that parental involvement also contributes to better relations between children and between children and educators. Finally, good communication between staff and parents is a prerequisite for quality care and education of young children. Growth of the child, after which family care and communication in the family, transforms into kindergarten or preschool, some new communication relationships arises so now is recognized triad parent-child-educator. + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + title_1 + 6260 + 3. COMMUNICATION RELATIONS IN KINDERGARTEN + + + INTRO + paragraph + 6303 + This relation reflects the essence of communication processes in institutions where child, "little man" is for the first time faced with the collective ("children's house"), in which applies some new rules in relation to life in home. Synergy of proper parental and teachers approach to the child, communication of the parents, educators and child care, between children, is of crucial importance for the further development of the child. A child who comes from a healthy, unencumbered, families in which it grows with love and attention relative to adapt quickly to new rules of community life. In contrast, a child that carries the frustration of the home from many, different, reason will have great difficulty to adjust. Especially for such children is reflected knowledge and experience of educators to identify such children and select them adequate (preferably individualized) approach. Important component is communication with children, hard work on the "opening" of the introverted child, learning to live in a collective, rules of life and finally, open communication with other children. The child is at the center of all communication patterns in pre-school institutions, and it focused all other relations, the primary communication applies in kindergarten are shown in Diagram 1. + + 9606 + Species + + child + + + 9606 + Species + + man + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + + INTRO + paragraph + 7610 + If we are consider coming to kindergarten from child's perspective, first visit to the nursery for them is a violent separation from parents, therefore, from the environment in which to feel safe, in a new, unknown space and handed over to strangers. It is understandable that a child in these circumstances is experiencing the frustrations that can manifest in different forms of behavioral changes and mood (energetic protest, crying and withdrawal, denial of communication with the teacher or, paradoxically, "stick" to teacher, refusing food, frantically holding objects which are associated to the house, waking, not sleeping, express anger and disobedience to the new conditions, manifest aggression towards the environment...). + + 9606 + Species + + child + + + 9606 + Species + + child + + + MESH:D001523 + Disease + + aggression + + + + INTRO + paragraph + 8349 + The first days of the child's stay in the kindergarten are stressful for children and parents, and of course, for educators. Since kindergarten, is surrogate home and family environment, the child educator needs to develop a sense of security and belonging to a new family that is kindergarten. The task of the kindergarten, therefore, is to provide a safe and stable environment, an atmosphere which understand, accept and support the child's feelings + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 8802 + If these communication relationships are observed from the parent's point of view (whether or not the educator take care of my child in the way I do, to understand its personality and its needs, "bugs", will have patience with my child, along with many other children, whether to accept it, finally, whether the child will be fine in the new environment), it is clear what is the responsibility of educators as well as the difficulties they will encounter during the upbringing of children. To respond to the complex requirements of parent educators will use an appropriate form of communication in order to achieve a harmonious relationship, with communication as a fundamental feature of mutual trust and understanding. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + + INTRO + paragraph + 9528 + Finally, a view of the educators on parents (monitors the extent to which parents are under stress due to separation from the child, how to recognize stress and frustration of parents, how to handle a parent who is frustrated because of the child's frustration, the way of feed and removal of the child, views and nonverbal communication, which sometimes speak louder than words, restraint in communication, (not) a rare and aggression by parents...) are all signs that the teacher should be recognized and interpreted in order to be properly set according to each parent by building specific communication form. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + MESH:D001523 + Disease + + aggression + + + + INTRO + paragraph + 10141 + The parent is the first and most important link in the upbringing of the child. For the child's emotional stability and proper development it is most important that parents establish proper communication with the child from its earliest years. A warm family environment in the home, harmonious parental relations, relationships among siblings and other family members (grandparents) contributes to the proper development of the child, without the stress and frustration. The aforementioned departure of a child to kindergarten is the first stress to the child's psyche, which is why parents need to be prepared to that milestone in a child's life. In the first place, it is essential that their positive attitude towards kindergarten and confidence in educators, shortening the stay of a child in kindergarten the first day, parent involvement in all activities related to kindergarten, establishing a precise rhythm of arrival and departure from the kindergarten, and in the interim period adaptations allow the child to the brings his/hers toys, pacifier, bottle, etc.., which remind the child to home. Period of adjustment of the child is different and depends on the age group, of course, to quickly adapt to children older groups. A sudden separation from home child interprets as treason by parents and often reacts with anger, disapproval and aggression, often regressive behavior (crawling, bedwetting, etc.). For all these options parent should be ready to answer them with stronger affection, conversation and rationalization, of course, to the extent that to the child is understandable. Before kindergarten, a child needs to prepare and get used to this act, an occasional visit to a kindergarten, observing children at play, talking with staff, to be able to eat independently and possibly self-dresser. It is useful to occasionally leave the child with her grandparents or people who kept the diet for a few hours or a whole day to get the child used to the occasional separation from parents. Also, it is very important to build a relationship of trust and mutual respect with educators, proper communication and collaborative partnership because they are for parents and educators directed towards a common goal-successful adaptation of the child in the new circumstances. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + people + + + 9606 + Species + + child + + + 9606 + Species + + child + + + MESH:D001523 + Disease + + aggression + + + Disease + + psyche + + + + INTRO + paragraph + 12432 + Educator is the next link in the educational process of the child. To educator belongs a fundamental and very responsible role, "surrogate parent" for parents employed because they due to the workload have less time to socialize with the child, which are increasingly leaving the work in kindergarten (full day, 10-hour programs). + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 12767 + Certainly in the educational process in the kindergarten equally affect the other participants: psychologist, pedagogue, special education teacher, speech therapist, and others who are important to the upbringing of the child (assistant, cooks, other staff), therefore all complementary to achieve complex missions in child-rearing. + + 9606 + Species + + participants + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + title_1 + 13100 + 4. COMMUNICATION PATTERNS - WAYS OF COMMUNICATION + + + INTRO + paragraph + 13151 + As stated above word, verbal communication is the foundation of every communication, including with pre-school children, some of which are still learning to speak. Proper verbal communication seeks calm approach to child, educator's equidistance towards other children in kindergarten (not favoring one child, so the other would not feel neglected) and finally individual approach, if necessary (e.g., children with special needs). Apart from this, regular communication, according to psychological divisions, there are also non-verbal communication ("body language"), and paraverbal communication (mode of speech, facial expressions, tone of voice). Like adults, children are very well understood by all three forms of communication, so that the educator in her work has extremely careful in all three forms (in) direct communication with the child. Child communication seeks educator's attention, equal relationship and the need for him to tutor his full attention to the conversation, i.e. that it is not in any way neglected. + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + Child + + + + INTRO + paragraph + 14185 + So, if the educator is occupied some other business (e.g. preparation of workbooks, or other activities) and the child spoke, educator should stop working and devote its full attention. This means, not with some other work, it is necessary to look into the eyes of a child, to patiently hear its question or comment. + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 14502 + Also, body language-gestures that betray impatience (tapping a pencil, interrupting the conversation, watching over the child, non-listening...) and the desire to talk or request as soon as possible, lead to poor communication between educators and child. + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 14760 + Finally, the raised voices in conversation with the child, threatening expression (paraverbal communication) at the start eliminate child's desire to communicate, to which particular must watch every educator. + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 14970 + It should be noted that the child during the entire stay in the kindergarten assess teacher's person, personality, way of communicating, and vice-versa, in a similar way to educator evaluate children in behavior and adoption of knowledge. + + 9606 + Species + + child + + + 9606 + Species + + person + + + 9606 + Species + + children + + + + INTRO + paragraph + 15209 + The most important part of a successful adaptation of the child to kindergarten makes teachers. During the period of adjustment child educator needs to know, as a group, parents and children, recognize the individual needs of each child. Also, as a token of good communication relations educators need to be accepted by the child, through their activities, providing support to the child and parents. Educators monitor, observe the child during his stay in kindergarten take notes, plan activities and design child room to learn, play and other activities. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 15766 + What is the key moment in relation educator-child is that the everything that child is going through in kindergarten transmits to parents, or independently of its, spontaneous narratives, parents questioned in detail about (every)day stay in kindergarten and events (the ratio of educators to child, other children, food hygiene, etc.). Follow every move, every educator's raised voice, or some other form of punishment of the child (rough handling, pushing, pulling hair, etc..) Which may lead to very poor communication form educator-child. From such, disturbed, relations subsequently followed poor communication parent-educator, because any change in the relationship between child communications signals to parents. Of course, the relationship between educators and parents (the child's testimony parents often exaggerates!) Two vital components in preschool, thereby permanently disrupted. According to the findings of misfortunate events in kindergarten, and after talking with the child, the parents immediately, ultimately the director of the institution seeking punishment or removal of educators for the sake of the child's testimony. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 16914 + The above triad is the subject of numerous studies, psychological, pedagogical, as well as in the field of communication sciences, and they all agree with the fundamental postulates of communication, which in relation to the child has its specificities. So, parents and educators, spending time with your child should adhere to the following postulates: + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 17268 + Talk with child-teach introvert child to talk, encourage it to conversation, avoid conversation with the answers yes and no. + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 17397 + Call child by name-if you call them by the name, the child interprets this intimacy with a sense of responsibility, contrary to anonymous communication which means indifference. + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 17581 + Look child in the eyes: a direct sight means that you are interested in it (not too long, because fixed look achieve the opposite effect, watching manner means indifference or its absence). + + 9606 + Species + + child + + + + INTRO + paragraph + 17775 + Touch of a hand, touch child's hair: intimacy, commitment, "reward" for a completed task. + + 9606 + Species + + child + + + + INTRO + paragraph + 17873 + Diction, voice tonality of voice, facial expression-bland calm voice, without elevated tones, soothing facial expression. + + + INTRO + paragraph + 18001 + Short, simple communication, understandable words. + + + INTRO + paragraph + 18056 + Repetition-repeat your request, if you are ignored. + + + INTRO + paragraph + 18114 + Award him (your choice award) if it met your expectations. + + + INTRO + paragraph + 18177 + Deprive him an award if you it does not fulfill their task, it was his worst punishment Teach him the eighth decency, "please" and "thank you"-mandatory in communication. + + + INTRO + paragraph + 18362 + Point out errors, teach the child conversation. + + 9606 + Species + + child + + + + INTRO + paragraph + 18414 + If you mad tell him that you are sorrowful and sad because he did, do not allow yourself to be provoked or shout at a child. + + 9606 + Species + + child + + + + INTRO + title_1 + 18543 + 5. EXAMPLES FROM DAILY PRACTICE IN THE INSTITUTION FOR PRESCHOOL EDUCATION + + + INTRO + paragraph + 18618 + Although Chinese proverb (quoted at the beginning of the text) says that children are telling the truth, we know from experience that it cannot be true. Reason false accusations may be because of relations between parents, (the so-called. Said syndrome sexual allegations in divorce) but also false allegations that the child expresses (invent). What happened was that children of parents accuse educators for (not) work that the same is not really so. + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + MESH:D016097 + Disease + + Said syndrome sexual allegations + + + + INTRO + paragraph + 19071 + Example. (Statement of an educator). "Peter was lively and happy child often would come in my lap. He was not always willing to fulfill the tasks, but with a little persuasion he gets them done. One morning he came with his father, corpulent worker from shipyard and very rudely attacked me, because if I had yesterday beaten child. I told him it was not true, and that such a thing has never happened in our kindergarten, but the father was insistent, saying that his Peter never lies. Immediately I called Peter and asked him in front of his father: "Did I beat you yesterday?" He embarrassingly said that I did not, and that he invented it so he does not have to do a task that for him was so hard. Parent immediately pounced the child, but now with a story that is a little liar and that his house does not believe anything. " + + 7939 + Gene + + lap + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + + INTRO + paragraph + 19910 + In some cases the child, out of fear, hides a traumatic event in kindergarten fearing retaliation of the educators. Example: (testimony of parents) ... My Igor, a boy with signs of mild MCD, which required a special, individual approach because it is often obstinately persisting in their demands. Educator obviously had no patience with him, because the lady who earlier came by my child said that he is an educator led him to the bathroom and well beaten because he in a loud voice yelled. I have noticed that since that day my son did not want to go to kindergarten, he was afraid, he was crying but he would not say why, I guess out of fear. Of course ... I'm also reported the director of the kindergarten "... + + 9606 + Species + + child + + + 9606 + Species + + boy + + + 9606 + Species + + child + + + MESH:D012514 + Disease + + MCD + + + MESH:D014947 + Disease + + traumatic + + + + INTRO + paragraph + 20630 + These cases are sporadic, rare, but should be assiduously investigated, because the truth always eventually came to light. For example after talks with parents who report changes in child behavior (frustration, fear, anxiety ...) followed by psychological testing (processing, drawing, etc..) That reveal educators / woman who got lost in the water for their upbringing and that systematically punish children in kindergarten. These cases are rare, but there are some, an example of a case where educators are parents set up a hidden camera that has discovered what all the kids did and how they were punished. + + 9606 + Species + + child + + + 9606 + Species + + woman + + + 9606 + Species + + children + + + MESH:D001008 + Disease + + anxiety + + + MESH:D014867 + Chemical + + water + + + + INTRO + paragraph + 21242 + However, in most cases by children honestly convey to parents all events, was pleasant and unpleasant, and the knowledge and application relevant to kindergarten every case non pedagogic work with the child should be thoroughly investigated and, if necessary, sanctioned. + + 9606 + Species + + children + + + 9606 + Species + + child + + + + INTRO + paragraph + 21514 + Without a doubt, these situations are extreme, because the vast majority of educators do their job responsibly, and finally themselves have chosen the honorable invitation educators. Therefore, any case report violations of the child's integrity and personality (login parents or third parties, public or anonymously) should be accepted, checked thoroughly investigated and, if necessary, conduct psychological testing or other actions that would not have occurred cases which, needless, throw filth the honorable job of educators. A child who is developing normally, during his stay in kindergarten, collective, rapidly improves your vocabulary, speech impediments, talking with other children and with the teacher. However, the unconscious child can take on some negative qualities of speech and facial expressions (drawl, mispronounced words, tics) of children with these problems, imitating them. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + + INTRO + paragraph + 22415 + Example: Mirko child from highly educated family, whose parents are alternately brought and taken away from the nursery. (testimony of parents)... "In the beginning was staying in kindergarten flowed really well, Mirko quickly adapted and played with children. The problem arose when we noticed that Mirko weird talking, delaying and modulating voice in a special way, which has never before been done. I'm a little bothered, questioned and found that Mirko most endeared and hung out with the boy John, (a boy with Down syndrome), which, when we checked spoke in the same manner as is now saying our Mirko! " + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + boy + + + 9606 + Species + + boy + + + + INTRO + paragraph + 23031 + All these are the moments that must be observed educator who, as a corrective, talking with parents, seeks to correct the child's behavior, to direct it and to give special attention to such children. Because once mistakenly learned, it is difficult to correct. Kids play, learn unencumbered through the game, absorbing every word of educators, even if they do not know the true meaning of the word, but according to the sense and context are decrypted. + + 9606 + Species + + child + + + 9606 + Species + + children + + + + INTRO + paragraph + 23485 + Every word spoken, remembered the child may return like a boomerang, if a parent mechanically in a situation or someone send a curse, the child will remember and reproduce in approximately similar situation (anxiety, anger, resentment). Child resonates: If dad (or mom) so saying it is wrong and can also say in front of other people. So can it happen in a completely unmanageable situation juicy cursed child, even if you do not know the meaning of swearing, because it is a situation where it occurs associated with parental cursing in a similar situation. + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + Child + + + 9606 + Species + + people + + + 9606 + Species + + child + + + MESH:D001008 + Disease + + anxiety + + + + INTRO + paragraph + 24044 + Example 1: (testimony of parents) "The car journey with us was a little Ida, in her chair tied in last place. While driving in front of us carelessly, high speed from the side street crashed driver who almost hit us. I admit I slipped a juicy curse, but it all ended well. Next time driving, when I slammed on the brakes, Ida repeated cursing in the same way as I did I previously spoke". + + 3425 + Gene + 170 + + Ida + + + 3425 + Gene + 170 + + Ida + + + + INTRO + paragraph + 24437 + Example 2 (testimony of parents). "We looked at (wife, little Emma) TV with an entertainment show in which he participated in one of our show biz stars be rampant. I quietly (not to hear me daughter!) Told the woman to switch the channel because I cannot watch that bitch..... Ok, a woman had done a few days back on TV appeared the same stars from the entertainment industry. Emma immediately cried loudly: There goes that son of a ... immediately shut down the TV." + + 9606 + Species + + woman + + + 9606 + Species + + woman + + + + INTRO + paragraph + 24909 + This will also happen in relation to the teacher: child will house use phrases that he heard from educators, was in communication with him or with other children. + + 9606 + Species + + child + + + 9606 + Species + + children + + + + INTRO + title_1 + 25072 + 6. IMPORTANCE OF PROPER COMMUNICATION WITHIN THE FAMILY + + MESH:D001929 + Disease + + WITHIN + + + + INTRO + paragraph + 25128 + Immeasurable importance of proper communication relation child and parents. Famous aphorism Earl of Rochester's (I had six theories about raising children when I had no child, now I have six of them and no theories ...) confirms this. Parents with a debut child learn communication and education on the forms that they experienced in their youth when they were raised by their parents. Parents who grew up as an only child, unconsciously accepted patterns of his youth, and almost identical to convey the paradigm of education to their child. Parents who grew up in a large family with different forms of education and communication with the child. What is now clear, the economic situation in the society, parents are reluctant to have another child, let alone more: partly out of conformism, but more for economic reasons. In families with one child, a kindergarten is a surrogate for a large family, and as such is necessary in the education of an individual, especially as parents occupied a job cannot be adequately measured participate in the upbringing of the child. Of course it is necessary nursery with large families in which the parents work, and the parents were all agree that education is the first child toughest. This is partly from inexperience and partly because of a family with one child unconsciously, exaggerated care, such children spared all possible situations in which a child could possibly be the endangered (e.g. Child is not sent to the nursery or day care due to the risk of transmission of infections, keeping the cold, rain, presvalacenje several times a day, especially feeding, mixed foods, avoiding the society of other children in the building where they live or work outside of a possible infection, avoidance of vaccination for the possible consequences, avoid rooms which housed more people, etc ...). 14 Because over care and isolation of the child, in such families are left only for parents, lonely, eager to socialize with other children. Company other kids did milieu in which the child is developing normally, realizes communication relationships with children their own age, and through the game with the children a feeling of satisfaction and happiness. Children who are only children in the family, "condemned" the only parents that if he wanted to, I cannot provide what the child wants, subconsciously are lonely and dissatisfied, because it is the day in the house monotonous and uneventful, until now the prevailing mood state. All knowledge about the life of children from their parents and in the best case of the second generation (today's increasingly rare!) Grandparents, who take the role of the nursery. So evident is the discrepancy between the generations, the generation gap, it is questionable as a good solution in education of a child. That is why the kindergarten indeed the solution for their situation, the postulation of kindergarten (kindergarten), which was established in 1840 by Frobel and remains very topical today, of course, in the new, modern conditions. + + 54922 + Gene + 9847 + + rain + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + Child + + + 9606 + Species + + children + + + 9606 + Species + + people + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + Children + + + 9606 + Species + + children + + + 9606 + Species + + child + + + 9606 + Species + + children + + + 9606 + Species + + child + + + MESH:D007239 + Disease + + infections + + + MESH:D007239 + Disease + + infection + + + + INTRO + footnote + 28168 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 28205 + REFERENCES + + + 54 + REF + surname:Sindik;given-names:J + 9 + 2008/2009 + Dijete, Vrtic, Obitelj + ref + 11 + 28216 + Jesu li profesionalci socijalno kompetentni? + + + REF + surname:Goleman;given-names:D + 1995 + ref + 28261 + Emocionalna inteligencija + + + 24 + REF + surname:Hawkins;given-names:Robert P + 205 + 2 + 1890041 + 1991 + Journal of Applied Behavior Analysis + ref + 213 + 28287 + Is social validity what we are interested in? Argument for a functional approach + + + 9 + REF + surname:Guralnick;given-names:MJ + 1 + 3 + 1989 + Topics in Early Childhood Special Education + ref + 17 + 28368 + Recent developments in early intervention efficacy research: Implications for family involvement + + + REF + surname:Bronfenbrenner;given-names:U + 1979 + ref + 28465 + The ecology of human development: Experiments by design and nature + + 9606 + Species + + human + + + + surname:Downey;given-names:G + surname:Caspi;given-names:A + surname:Bolger;given-names:N + REF + surname:Bronfenbrenner;given-names:U + 25 + 1988 + Persons in context: Developmental processes + ref + 49 + surname:Moorehouse;given-names:M + 28532 + Interacting systems in human development. Research paradigms: Present and future + + 9606 + Species + + human + + + + 10 + surname:Walters;given-names:LH + REF + surname:Amirali;given-names:Jinnah H + 1 + 1 + 2008 + ERCP + ref + 7 + 28613 + Including Parents in Evaluation of a Child Development Program: Relevance of Parental Involvement + + 9606 + Species + + Child + + + + 1 + surname:MacNaughton;given-names:G + REF + surname:Hughes;given-names:P + 241 + 3 + 2000 + Contemporary Issues in Early Childhood + ref + 258 + 28711 + Consensus, dissensus or community: The politics of parent involvement in early childhood education + + + surname:Milkie;given-names:MA + surname:Robinson;given-names:JP + REF + surname:Bianchi;given-names:SM + 2006 + ref + 28810 + Changing Rhythms of American Family Life + + + surname:Ney;given-names:T + REF + surname:Walker;given-names:Perry N + 73 + 1995 + True and False Allegations of Child Sexual Abuse: Assessment and Case Management + ref + 98 + 28851 + Childrens's comprehension od truths, lies and false belifs + + + 8 + REF + surname:Faller;given-names:KC + 473 + 6542819 + 1984 + Child Abuse and Neglect + ref + 481 + 28910 + Is the child victim of sexual abuse telling the truth? + + 9606 + Species + + child + + + MESH:D012735 + Disease + + sexual abuse telling + + + + REF + surname:Ney;given-names:T + 3 + 1995 + An owrwiew + ref + 23 + 28965 + True and False Allegations of Child Sexual Abuse: Assessment and Case Management + + 9606 + Species + + Child + + + + surname:Reece;given-names:RM + surname:Drotar;given-names:D + REF + surname:Frank;given-names:DA + 298 + 1994 + Child abuse: Medical diagnosis and management + ref + 324 + 29046 + Failure to thrive + + + 156 + surname:Dodge;given-names:K + REF + surname:Lansford;given-names:JE + 824 + 8 + 12144375 + 2002 + Arch Pediatr Adolesc Med + ref + 830 + 29064 + A 12-Year Prospective Study of the Long-term Effects of Early Child Physical Maltreatment on Psychological, Behavioral, and Academic Problems in Adolescence + + 9606 + Species + + Child + + + + REF + surname:Connell;given-names:HL + 2000 + ref + 29221 + The Childcare Answer Book + + + surname:Giesecke;given-names:J + surname:Hermanssonn;given-names:G + surname:Tegnell;given-names:A + surname:Dannetun;given-names:E + 149 + 3 + 16162466 + 2005 + Scandinavian Journal of Primary Health Care + ref + 23 + REF + 153 + 29247 + Parents' reported reasons for avoiding MMR vaccination. A telephone survey + + + 35 + REF + surname:Taylor;given-names:Allen A + 173 + 2006 + Journal of the History of Education Society + ref + 188 + 29324 + The Kindergarten in Germany and the United States, 1840 - 1914: A Comparative Perspective, History of Education + + + + 4272839 + + surname:Spahic;given-names:Emina + surname:Biscevic;given-names:Mirza + surname:Krupic;given-names:Ferid + 25568584 + surname:Tiric-Campara;given-names:Merita + 333 + 5 + 2014 + 4272839 + AIM-55-333 + 10.5455/aim.2014.22.333-340 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 333-40. doi:10.5455/aim.2014.22.333-340 + technological diseases carpal tunnel syndrome mouse shoulder cervical pain syndrome + surname:Masic;given-names:Izet + 340 + surname:Zunic;given-names:Lejla + surname:Masic;given-names:Zlatan + surname:Maglajlija;given-names:Kerima + Tiric-Campara M, Krupic F, Biscevic M, Spahic E, Maglajlija K, Masic Z, Zunic L, Masic I + 0 + Occupational Overuse Syndrome (Technological Diseases): Carpal Tunnel Syndrome, a Mouse Shoulder, Cervical Pain Syndrome + + 10090 + Species + + Mouse + + + MESH:D010146 + Disease + + Pain + + + MESH:D012090 + Disease + + Occupational Overuse Syndrome + + + + ABSTRACT + abstract_title_1 + 121 + ABSTRACT + + + ABSTRACT + abstract + 130 + Technological diseases are diseases of the modern era. Some are caused by occupational exposures, and are marked with direct professional relation, or the action of harmful effects in the workplace. Due to the increasing incidence of these diseases on specific workplaces which may be caused by one or more causal factors present in the workplace today, these diseases are considered as professional diseases. Severity of technological disease usually responds to the level and duration of exposure, and usually occurs after many years of exposure to harmful factor. Technological diseases occur due to excessive work at the computer, or excessive use of keyboards and computer mice, especially the non-ergonomic ones. This paper deals with the diseases of the neck, shoulder, elbow and wrist (cervical radiculopathy, mouse shoulder and carpal tunnel syndrome), as is currently the most common diseases of technology in our country and abroad. These three diseases can be caused by long-term load and physical effort, and are tied to specific occupations, such as occupations associated with prolonged sitting, working at the computer and work related to the fixed telephone communication, as well as certain types of sports (tennis, golf and others). + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D011843 + Disease + + cervical radiculopathy + + + + INTRO + title_1 + 1382 + 1. INTRODUCTION + + + INTRO + paragraph + 1398 + Computers are considered as an integral part of everyday life in today's work and life. They are used in a wide variety of professions from banking, health, communication to entertainment and leisure time. "Thanks to the" great use of computers, occurs the diseases of the modern era, which one of the authors of this article named "Technological diseases" - Carpal Tunnel Syndrome (CTS), a Mouse Shoulder (MS) and Cervical Pain Syndrome (CPS). Within past a few decades rapidly increased automation of offices and very few authors in the scientific literature have reported about positive association between computer use and musculoskeletal symptoms as consequences. Under term of"Technological diseases" we mean the diseases caused by the harmful influence of the job in the first place, the working position in which it is involved excessive work on a computer, such as the banking sector, the health sector and many others who are directly or indirectly connected with the work on the computer and overuse of keyboard and computer mice, which are in large number of cases of non-ergonomic, and inappropriate for a hand. It is therefore recommended to use ergonomic computer mice. Using them can prevent diseases of the modern era, technological diseases associated with prolonged sitting and working on a computer such as Carpal Tunnel Syndrome, Cervical syndrome, and certainly the most interesting for the many unknown, but not less important disease, mouse shoulder an increasingly common diagnosis in clinics of family medicine. These diseases are described in more detail in this paper. In Bosnia and Herzegovina to overuse injuries has not paid enough attention from the medical nor the social and economic aspects, although it is evident that large amounts of funds are allocated for treatment and rehabilitation of these diseases. In Clinical Center University of Sarajevo in the cabinet for EMNG of the Neurology clinic, monthly are diagnosed 10 cases with carpal tunnel syndrome, while cases of cervical syndrome is in a significant expansion during last 5 years which more and more represent not only a medical but also social economic problem. + + 7276 + Gene + 317 + + CTS + + + 10090 + Species + + Mouse + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mouse + + + MESH:D002575 + Disease + + cervical syndrome + + + MESH:D020165 + Disease + + CPS + + + MESH:D019547 + Disease + + Cervical Pain + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D012090 + Disease + + Bosnia and Herzegovina to overuse injuries + + + MESH:D002575 + Disease + + Cervical syndrome + + + + INTRO + title_1 + 3573 + 2. CARPAL TUNNEL SYNDROME + + + INTRO + paragraph + 3599 + Carpal tunnel syndrome (CTS) is the most common canaliculus syndrome resulting from pressure on the central nerve (median nerve) in the carpal tunnel (lat. canalis carpi) and reflected in reduction of sensation, pain, paresthesia (numbness, tingling), and muscle weakness in the hands and forearms. + + 27040 + Gene + 7811 + + lat + + + 7276 + Gene + 317 + + CTS + + + MESH:D018908 + Disease + + muscle weakness + + + MESH:D006987 + Disease + + numbness + + + MESH:D010146 + Disease + + pain + + + MESH:D002349 + Disease + + Carpal tunnel syndrome + + + MESH:D013577 + Disease + + canaliculus syndrome + + + + INTRO + paragraph + 3898 + The median nerve in its course passes through the carpal canal (Figure 1). The bones of the hand (lat. ossa carpi) make gutter channels, and rectangular connection (lat. retinaculum flexorum) covers the channel. In the channel along with the nerve are located tendon flexor muscle of fingers, m. pronator teres, m. pronator quadrates, m. flexor carpi radialis, m. palmaris longus, m. flexor carpi ulnaris, m. flexor digitorum superficialis et profundus and m. flexor policis longus. All these muscles supplying the median nerve. Carpi ulnaris M.flexor the inner half, m. Flexor digitorum profundus is innervated by n. ulnaris. + + 27040 + Gene + 7811 + + lat + + + 27040 + Gene + 7811 + + lat + + + + FIG + AIM-22-333-g001.jpg + F1 + fig_caption + 4525 + The muscles of the forearm. + + + INTRO + paragraph + 4553 + Carpal tunnel syndrome (CTS) was first described in the mid-1800 by James Paget's (Figure 1). The world's best known as peripheral compressive neuropathy and one of the fastest growing technology diseases in the United States. At least 1 out of 10 people develop this disorder or suffering from the symptoms of this syndrome. This is one of the most common causes of absenteeism in the workplace and disability in the United States + + 7276 + Gene + 317 + + CTS + + + 9606 + Species + + people + + + MESH:D010523 + Disease + + peripheral compressive neuropathy + + + MESH:D002349 + Disease + + Carpal tunnel syndrome + + + Disease + + absenteeism + + + + INTRO + paragraph + 4986 + Signs and symptoms are: a) pain in the palm of the hand, especially near the thumb and first two fingers; b) numbness and tingling often occur in the thumb and first two fingers; and c) weakness, usually thenar eminence (muscles at the base of the thumb), can occur when a severe clinical picture (Figure 2). + + MESH:D006987 + Disease + + numbness + + + MESH:D018908 + Disease + + weakness + + + MESH:D010146 + Disease + + pain + + + + FIG + AIM-22-333-g002.jpg + F2 + fig_caption + 5295 + Sensory innervation of the median nerve in a hand + + + INTRO + paragraph + 5345 + Compression of the nerve in the carpal tunnel (carpal tunnel syndrome) is recognized as the impact of trauma and attributed to tenosynovitis of the rheumatoid arthritis. Even since 1954, Albert and his colleagues concluded that there are numerous reports describing the spontaneous development of carpal tunnel syndrome of unknown pathogenesis. + + MESH:D014947 + Disease + + trauma + + + MESH:D001172 + Disease + + tenosynovitis of the rheumatoid arthritis + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + INTRO + paragraph + 5690 + In case of a longer duration of illness and long-term nerve compression, there is an unnecessary prolongation of the patient discomfort, severe nerve damage, prolonged recovery time after surgery, and generally reduced chances for successful surgical treatment. Thereby reducing the working ability of the patient, leading both to the individual and to the socioeconomic consequences. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D004194 + Disease + + nerve damage + + + MESH:D002908 + Disease + + illness + + + + INTRO + paragraph + 6075 + At the Department of Orthopedic Surgery, Faculty of Medicine, University of Zagreb, and Zagreb University Hospital Center is conducted a retrospective study on 114 patients and 154 hands with CTS that were surgically treated between 1999 and 2004. In this study, it was found that patients in only 52% of cases within one year since reporting to the physician were referred to an orthopedist. In the period prior to surgical treatment, 96.1% of patients were treated by physical therapy, although all eventually underwent a surgical procedure, and 42% of patients in the same time period it was on sick leave, usually from three to six months. The results demonstrate that the timeliness of surgical treatment is essential to the success of the outcome of the procedure, and that the time which elapses from entering the patient's health care system to the surgical treatment of unnecessary cost. + + 7276 + Gene + 317 + + CTS + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + + INTRO + paragraph + 6972 + This situation, besides being detrimental for the patient, and leads to unnecessary economic losses include the expense of physical therapy and absenteeism. The patients involved in the study in question is estimated that these economic losses, in the period prior to surgical treatment, for several hundred thousand Euros higher than in the postoperative period. + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9319 + Species + + Euros + + + Disease + + absenteeism + + + + INTRO + paragraph + 7336 + From this stems the need for shortening the period of time that has elapsed since entering the patient in the health care system to its referral to an orthopedic clinic, so as to accelerate the processing and diagnosis of the patient, i.e. shortened unnecessarily long period of nonsurgical treatment. Accordingly, physical therapy is helpful in the treatment. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + + INTRO + paragraph + 7697 + In the diagnosis of CTS we also use provoking tests of which we will mention four that are suitable for use in general practice. These are the Tinel, Phalen, Bilic and Tourniquet test. Tinel's test is positive if the percussion in the carpal ligament is followed with occurrence of pain and/or paresthesia. Phalen's test is performed so that the wrist is placed in the second forced palmar flexion for 60 seconds, and is positive if within this time span occurs pain and/or parasthesias (Figure 2). Compression of median nerve in this position with carpal tunnel syndrome causes the characteristic symptoms of burning and tingling. + + 7276 + Gene + 317 + + CTS + + + MESH:D010146 + Disease + + pain + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 8329 + Bilic test is performed in palmar flexion of the wrist by 45 degrees by pressing above the median nerve in the distal ridges and mid wrist for 30 seconds. The test is positive if within this time span occurs pain and/or paresthesia. Tourniquet test is performed so that the cuff pressure gauge is mounted on the upper arm and inflated above the systolic pressure value. The test is positive during 60 seconds occurs pain and/or paresthesia in the innervation area of median nerve. + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 8810 + Alongside this symptom, electrical neurophysiological diagnostic is the"gold standard" in the diagnosis of carpal tunnel syndrome, in order to determine the exact location of nerve compression, and in order to set the indication for surgical treatment (Figure 3, 4, 5). + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + FIG + AIM-22-333-g003.jpg + F3 + fig_caption + 9084 + EMNG median nerve recording procedure + + + FIG + AIM-22-333-g004.jpg + F4 + fig_caption + 9122 + EMNG: Motor Conduction Velocity of nervus medianus + + + FIG + AIM-22-333-g005.jpg + F5 + fig_caption + 9173 + EMNG: Sensory Conduction Velocity of nervus medianus + + + INTRO + paragraph + 9226 + In the case of known primary disease that caused Carpal tunnel syndrome the primary disease is treated first. The conservative methods of treatment include immobilization, local application of corticosteroids and avoidance of chronic, repeated trauma. If the cause of the syndrome is mechanical compression, the treatment is surgical (cutting of retinaculum flexorum). + + MESH:D014947 + Disease + + trauma + + + MESH:D002349 + Disease + + Carpal tunnel syndrome the primary disease + + + + INTRO + paragraph + 9595 + In Turkey was conducted a study that suggests that treatment with vitamin B6 improves clinical symptoms as well as sensory and electro diagnostic results in patients with carpal tunnel syndrome, and therefore it is advisable to treat carpal tunnel syndrome with vitamin B6. + + 9103 + Species + + Turkey + + + 9606 + Species + + patients + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + MESH:D025101 + Chemical + + vitamin B6 + + + MESH:D025101 + Chemical + + vitamin B6 + + + + INTRO + paragraph + 9869 + Another study in Turkey was carried out at the Institute of Neurology in cooperation with the Clinic for Physiotherapy in which patients were included with carpal tunnel syndrome. They are divided into two groups, one in the treatment arm used immobilization and massages, while the other control group used only immobilized arm. + + 9103 + Species + + Turkey + + + 9606 + Species + + patients + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + INTRO + paragraph + 10199 + The study results showed that the massage of the painful area only in patients with carpal tunnel syndrome is sufficient for mild cases. + + 9606 + Species + + patients + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + INTRO + paragraph + 10336 + This was also one of the biggest studies with massage treatment in patients with carpal tunnel syndrome. The researchers recommend self-administration of massage methods, and this would reduce the time to go to the doctor and rehabilitation centers, and thereby save money, both for health funds, and patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + INTRO + paragraph + 10647 + Manipulation (effleurage): Thirty seconds smoothing the skin's surface, from distal to proximal forearm. Friction: sixty seconds, massaging the deep tissues from the distal to proximal. + + + INTRO + title_1 + 10834 + 3. CERVICAL PAIN SYNDROME + + MESH:D000699 + Disease + + PAIN + + + MESH:D002575 + Disease + + CERVICAL + + + + INTRO + paragraph + 10860 + Cervical pain syndrome (CPS) refers to a range of disorders caused by changes in the cervical spine and soft tissue surrounding it, with pain as the predominant symptom. Neck pain is a common problem for a large portion of today's population. + + MESH:D019547 + Disease + + Neck pain + + + MESH:D020165 + Disease + + CPS + + + MESH:D010146 + Disease + + pain syndrome + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 11103 + Factors contributing to this problem are the modern way of life, prolonged sitting and inadequate, fixed or artificial positions. The root of these problems is found in the mechanical disorders of the cervical spine, poor posture and quick movements of the body. + + + INTRO + paragraph + 11366 + Anatomical specificity of the cervical spinal column are: a small side vertebral joints that cervical spine provide great mobility, transversaria foramina through which the vertebral artery (C6-C2) and the spinal cord enters in the vertebral canal. + + OMIM:217000 + Disease + + C6-C2 + + + MESH:D007592 + Disease + + small side vertebral joints + + + + INTRO + paragraph + 11615 + Cervical radiculopathy causes symptoms that radiate from the neck. Although the problem is in the spine, the symptoms can be felt in the shoulder or arm. Symptoms will be felt in the area where the nerve is in its path. + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D004194 + Disease + + radiate + + + + INTRO + paragraph + 11835 + By clinical examination of the spine the specialist can usually determine which nerve is involved. Symptoms include pain, stiffness and weakness. Reflexes on the upper arm may be affected.. + + MESH:D010146 + Disease + + pain + + + MESH:D018908 + Disease + + stiffness and weakness + + + + INTRO + paragraph + 12025 + Risk factors for radiculopathy are activities that put excessive or repetitive load on the spine. Patients involved in heavy work or are in contact sports, are more prone to the development of those with radiculopathy more sedentary. A family history of radiculopathy or other spine disorders also increase the risk of developing radiculopathy (Figure 6). + + 9606 + Species + + Patients + + + MESH:D016135 + Disease + + spine disorders + + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D011843 + Disease + + radiculopathy + + + + FIG + AIM-22-333-g006.jpg + F6 + fig_caption + 12381 + Example of cervical syndrome caused by disc herniation with reduction of liquor space (Source: I.M.) + + MESH:D002575 + Disease + + cervical syndrome + + + + INTRO + paragraph + 12482 + In the younger population, cervical radiculopathy is the result of disc herniation or acute injury. In the older population, cervical radiculopathy is often the result of foramen narrowing and the formation of osteophytes then reduced the amount of disk and degenerative changes in the joints. + + MESH:D058186 + Disease + + acute injury + + + MESH:D011843 + Disease + + cervical radiculopathy + + + MESH:D011843 + Disease + + radiculopathy + + + + INTRO + paragraph + 12776 + Cervical syndrome classification: + + MESH:D002575 + Disease + + Cervical syndrome + + + + INTRO + paragraph + 12810 + CERVICAL SYNDROME patients complain of pain in the neck that occurs gradually or abruptly, after taking a certain position, rapid or uncontrolled movements of the neck or cooling. + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + MESH:D002575 + Disease + + CERVICAL + + + + INTRO + paragraph + 12990 + CERVICOCEPHALIC SYNDROME usually develops as a result of irritation or compression of the nerve roots C1-C3. The main symptom is headache of varying character and intensity. Headache is localized in the neck with a tendency to spread to the shoulders, the face, orbital or auricular region. + + MESH:D006261 + Disease + + Headache + + + MESH:D006261 + Disease + + headache + + + MESH:D001523 + Disease + + irritation + + + + INTRO + paragraph + 13281 + CERVOCOBRACHIAL SYNDROME result is irritation or compression of the nerve roots C4-C8, mostly due to prolapse of the intervertebral discus, severe degenerative changes or injury. + + MESH:D001523 + Disease + + irritation + + + Disease + + CERVOCOBRACHIAL + + + MESH:D058186 + Disease + + injury + + + + INTRO + paragraph + 13460 + VERTEBROBASILAR SYNDROME occurs as a result of irritation of the last cervical sympathetic with fiber mesh wrap vertebral artery, causing reflex vascular disorders in the vertebrobasilar basin. + + MESH:D001523 + Disease + + irritation + + + MESH:D000783 + Disease + + vascular disorders + + + + INTRO + paragraph + 13654 + Irritation commonly caused by degenerative changes in the cervical spine: spondylosis (osteophytes), uncarthrosis, spondylosis. Atherosclerosis can worsen symptoms. + + MESH:D050197 + Disease + + Atherosclerosis + + + MESH:D055009 + Disease + + spondylosis + + + Disease + + uncarthrosis + + + MESH:D055009 + Disease + + spondylosis + + + + INTRO + paragraph + 13819 + The typical clinical presentation of patients with cervical syndrome is characterized by the presence of pain and sensitivity in the muscles back of the neck with the spread of pain in the back of the head, shoulders or scapular region (Figure 6). The sensitivity of the muscle can occur during execution of one or more movements, and the headache is a common symptom. + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D006261 + Disease + + headache + + + MESH:D002575 + Disease + + cervical syndrome + + + + INTRO + paragraph + 14188 + Symptoms and signs are: a) pain in the neck, which may be sharp or blunt; b) the tension in the neck; c) painful and limited mobility; d) inability to perform daily duties due to stiffness in the neck; e) pain in the shoulders and arms associated with pain in the neck; f) fainting; g) dizziness; h) tinnitus; i) blurred vision; j) headache; k) diplopia; l) weakness; m) feeling of heaviness, tenderness and paresthesia in the upper extremities; n) impaired concentration and memory, etc.. + + MESH:D010146 + Disease + + pain + + + Disease + + tenderness + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D008595 + Disease + + heaviness + + + MESH:D004244 + Disease + + dizziness + + + MESH:D004172 + Disease + + diplopia + + + MESH:D014786 + Disease + + blurred vision + + + MESH:D018908 + Disease + + weakness + + + MESH:D014012 + Disease + + tinnitus + + + MESH:D006261 + Disease + + headache + + + + INTRO + paragraph + 14678 + Diagnostic imaging such as magnetic resonance imaging, computerized tomography, or myelography should be used as a strategy for assessment. Electromyography is useful in differentiating between the various entities when distinguishing clinical diagnosis. Treatment of this disorder has not been studied systematically in a controlled manner. However, using a variety of treatments, radiculopathy usually improves without the need for surgery. Indications for surgery were persistent pain, increased weakness, or new or progressive process. Future studies evaluating different treatment options will be helpful in guiding practitioners towards optimal economic evaluation. + + MESH:D018908 + Disease + + weakness + + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 15350 + The goal of treatment is absolutely the same as for any other illness. Always seeks the same goal, or cure, and if this is not possible, at least alleviate symptoms and apply palliative care. Since we are talking about a technological disease that affects more and more to older and younger people should therefore work on the prevention of disease. It is very important to advise patients about lifestyle, how to maintain mobility and that despite the difficult situation improve the quality of life. + + 9606 + Species + + people + + + 9606 + Species + + patients + + + MESH:D002908 + Disease + + illness + + + + INTRO + paragraph + 15852 + Treatment without medicationshould be: a) massage hot/cold; b) rest; c) exercises with light stretching (gently stretch your neck to one, then the other aside and keep 30 seconds); d) exercises for neck recommended by physiologists; e) transcutaneous electrical stimulation (TENS); f) short-term immobilization; g) surgical treatment; h) acupuncture; i) traction (enlargement of the intervertebral space), j) medicamentous treatment: + + 137872 + Gene + 5865 + + hot + + + + INTRO + paragraph + 16286 + Early and adequate treatment with analgesics, anti-inflammatory drugs, muscle relaxants and help with sleep problems are carriers of pharmacological treatment in patients with neck pain. + + 9606 + Species + + patients + + + MESH:D019547 + Disease + + neck pain + + + + INTRO + paragraph + 16473 + Performed is a study on how the steroid injections affect the reduction of pain in patients with cervical syndrome, who are waiting for surgery discus, and whether the possible suspension of surgical treatment. + + 9606 + Species + + patients + + + MESH:D002575 + Disease + + cervical syndrome + + + MESH:D013256 + Chemical + + steroid + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 16684 + Transforaminal injection of steroids has gained popularity with the explanation that the inflammation of nerve roots causing radicular pain, and because steroids are placed on the local level should relieve symptoms. + + MESH:D013256 + Chemical + + steroids + + + MESH:D007249 + Disease + + inflammation + + + MESH:D010146 + Disease + + radicular pain + + + MESH:D013256 + Chemical + + steroids + + + + INTRO + paragraph + 16901 + Prospective-cohort study indicates a decrease in the need for surgical treatment because of treatment-injection steroid injections. The clinical effect is measurable and statistically significant improvement registered radicular pain. + + MESH:D010146 + Disease + + pain + + + MESH:D013256 + Chemical + + steroid + + + + INTRO + paragraph + 17138 + Another in a series of studies on the treatment of cervical syndrome confirmed the effectiveness of acupuncture in the cervical region of patients with radicular symptoms. Favorable results have been seen in almost 90% of cases. These results indicate that treatment of acupuncture cervical region can be effective as the conservative therapy for the treatment of cervical radiculopathy. + + 9606 + Species + + patients + + + MESH:D002575 + Disease + + cervical syndrome + + + MESH:D011843 + Disease + + cervical radiculopathy + + + + INTRO + title_1 + 17526 + 4. MOUSE SHOULDER + + 10090 + Species + + MOUSE + + + + INTRO + paragraph + 17544 + Computers are considered as an integral part of everyday life. They are used in education, health and medicine, science, banking, recreation and entertainment, politically engaged people. Approximately 75% of jobs are dependent on the work on the computer. + + 9606 + Species + + people + + + + INTRO + paragraph + 17801 + Chen and colleagues examined the effect of five computer mice made at different angles in 12 respondents employed in the business sector in Taiwan. They concluded that too frequent use of non ergonomic computer mice causes extreme discomfort in the muscle and tendon system of the hand along with various manifest symptoms in the shoulder and forearm. The use of ergonomic mice custom hand shape, thanks to the different corners of the obliquity have less adverse effect on the activity of the forearm and shoulder. + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + 10090 + Species + + mice + + + + FIG + AIM-22-333-g007.jpg + F7 + fig_caption + 18318 + Recommended proper position of the neck, torso, arms and legs when working at a computer + + + INTRO + paragraph + 18407 + The shoulder belt is common, even among the most frequent localization of musculoskeletal pain after pain in the back and knees. + + MESH:D059352 + Disease + + musculoskeletal pain + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 18536 + The long-term work at the computer leads to inflammation of tendons and exchanges shoulder joint, and in severe cases can lead to tearing of tendons and muscles "rotator cuff". If inflammation persists, the capsule and ligaments of the shoulder joint becomes stiffer and limited mobility. Inappropriate placement of computers may have adverse effects on posture in children. + + 9606 + Species + + children + + + MESH:D052256 + Disease + + tendons + + + MESH:D007249 + Disease + + inflammation + + + MESH:D007249 + Disease + + inflammation of tendons + + + + INTRO + paragraph + 18915 + Limited and painful mobility is especially pronounced when raising your arms above your head or behind your back. Long-term pain becomes stronger and leads to the development of muscle weakness areas of the shoulder belt through a series of hands, and preventing further activities. + + MESH:D010146 + Disease + + pain + + + MESH:D018908 + Disease + + muscle weakness + + + + INTRO + paragraph + 19198 + Symptoms of pain in the shoulder differently vary from intense that it can spread to other parts of the body, to moderate that lingers on the shoulder belt, but still hinder the function of the hinge, and can even lead to sleep disorders. + + MESH:D010146 + Disease + + pain + + + MESH:D012893 + Disease + + sleep disorders + + + + INTRO + paragraph + 19437 + The anatomy of the shoulder is a specific because the shoulder joint consists of primary, secondary and ancillary wrist joints. The anatomical structure makes the most complex joint of the body. Joints shoulders: glenohumeral joint (shoulder joint primary), Scapulothoracic joint (secondary shoulder joint), and sternoclavicular joint (extra joints). + + MESH:D007592 + Disease + + sternoclavicular joint + + + + INTRO + paragraph + 19788 + Muscle groups that may be a potential source of pain are rotator cuff (responsible for balance of the glenohumeral joint), stabilizing the scapula (responsible for the position of the scapula) the primary drivers (responsible for the strong movements). + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 20041 + For the diagnosis of mouse shoulder, the most important is detailed history with regard to occupation and profession of the patient and physical review. We should not forget to examine whether the patient has in the past had a shoulder injury, or some localized inflammation in the same. + + 10090 + Species + + mouse + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + MESH:D007249 + Disease + + inflammation + + + MESH:D058186 + Disease + + injury + + + + INTRO + paragraph + 20329 + Information about the character, pain intensity and directions of its expansion are of great importance. In fact it is necessary to establish that the primary cause of shoulder pain, and that the anatomical structure of the shoulder is affected. It is known that the pain can be transmitted from the adjacent region to the shoulders, for example. The cervical region, or by visceral fibers of the heart. + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 20733 + In equivocal cases, x-rays of the shoulder joint may help in differentiating existing inflammation of the joints and structure surrounding joint of a possible slight shoulder dislocation. + + 255564 + Species + + rays + + + MESH:D007249 + Disease + + inflammation of the joints + + + + INTRO + paragraph + 20921 + The primary method for the treatment of mouse shoulder in a timely manner to reduce the strain on shoulders, mostly working with computer mouse. + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + + INTRO + paragraph + 21066 + If the patient is in question, and cannot make a break, they definitely need the to replace old with new ergonomic computer mouse that is adapted to grip and allows the natural position of the hand during mouse use. + + 9606 + Species + + patient + + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + + INTRO + paragraph + 21282 + NSAIDs are the first drugs of choice. In the long-term intractable pain states and may prescribe therapy application of corticosteroid injections in the shoulder joint. An indispensable aspect of therapy is physical therapy. Physical therapy is appropriate in the chronic phase of the disease, which aims to stabilize and strengthen the muscles of the shoulder belt, and the resultant reduction in pain. + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 21686 + Jan C. Winters and colleagues came to the conclusion that only 50% of patients with newly diagnosed painful condition experience a full recovery within the first 6 months. This percentage rises to 60% in the first year of the onset of pain. They also proved that speaking to the long-term treatment, at the end of treatment, there was no difference in effect between the corticosteroid and physical therapy. + + 9606 + Species + + patients + + + MESH:D010146 + Disease + + pain + + + + INTRO + paragraph + 22094 + Onyebeke LC investigated the effects of props during the use of the mouse on the palm, forearm and upper extremities, and came to the result that 90% of respondents who have used the support arm had less pain in the shoulder girdles of those who are not used to. The use of any support results in less tension, and less applied force to a computer mouse during operation, and therefore less hassle and pain in the shoulder. + + 10090 + Species + + mouse + + + 10090 + Species + + mouse + + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + INTRO + title_1 + 22518 + 5. PHYSICAL REHABILITATION AND PREVENTION OF TECHNOLOGICAL DISEASES + + + INTRO + title_2 + 22586 + 5.1. The role and organization of the Centers for Physical Rehabilitation (CBR) + + + INTRO + paragraph + 22666 + If the patient required outpatient treatment, family doctor referred him Physiotherapy in CBR (Community based rehabilitation) i.e. the center for rehabilitation in the community. CBR's are organized within the framework of primary health care and are located within the health centers. CBR-rehabilitation in the community-the definition of a Joint Paper, ILO, UNESCO, WHO, in 1994. + + 9606 + Species + + patient + + + 9606 + Species + + outpatient + + + + INTRO + paragraph + 23053 + Community rehabilitation strategy based on community development for the rehabilitation, equalization of opportunities and social integration of people with disabilities, their families and communities and the appropriate health of educational, professional and social services. + + 9606 + Species + + people + + + + INTRO + paragraph + 23332 + Community rehabilitation allows people with disabilities to take initiative and improve their own lives, and to contribute to the community, and not just use the funds and resources that are available. Thus, the entire community and all its members are winners. + + 9606 + Species + + people + + + + INTRO + paragraph + 23594 + CBR became the backbone of the carrier non-institutional organization of the program in physical medicine and rehabilitation of persons with disabilities as well as a large number of chronically ill patients in the prevention of disability. + + 9606 + Species + + persons + + + 9606 + Species + + patients + + + + INTRO + title_2 + 23835 + 5.2. The forms of work in the CBR + + + INTRO + paragraph + 23869 + Rehabilitation system reaches into every community rehabilitation team becomes more flexible and actively participate in all aspects of rehabilitation. Rehabilitation includes all age groups. + + + INTRO + paragraph + 24061 + Treatment in the community increases the accessibility and quality of services for people with disabilities and their families. + + 9606 + Species + + people + + + + INTRO + paragraph + 24189 + Services include primary treatment damage and disability in the center and at home, as well as patient education, health promotion and prevention of disease and disability. + + 9606 + Species + + patient + + + + INTRO + paragraph + 24362 + CBR Team composition: a) 1 medical doctor specialist in physical medicine and rehabilitation; b) 1 physiotherapist; c) 1 occupational therapist; d) 2 medical technicians-physiotherapist; e) Medical technician-general direction; f) 0.5 Social Worker; g) 0.4 special education teacher-speech therapist. + + + INTRO + title_2 + 24667 + 5.3. The most common reasons for referral of patients in the CBR + + 9606 + Species + + patients + + + + INTRO + paragraph + 24732 + The most common reasons due to which family physician refers patients to the ambulance treatment in rehabilitation centers in the community are: + + 9606 + Species + + patients + + + + INTRO + paragraph + 24877 + Adult patients: a) Cervical pain syndrome; b) Lumbar pain syndrome; c) Rheumatic diseases; d) Rehabilitation after stroke. Children and young people: a) Neurorisk children; b) Deformities of the spinal column; c) Deformities of the lower extremities; d) Rehabilitation after injuries of the locomotor apparatus + + 9606 + Species + + patients + + + 9606 + Species + + Children + + + 9606 + Species + + people + + + 9606 + Species + + children + + + MESH:D010146 + Disease + + pain syndrome + + + MESH:D020521 + Disease + + stroke + + + MESH:D010146 + Disease + + pain + + + MESH:D012216 + Disease + + Rheumatic diseases + + + + INTRO + title_1 + 25188 + 6. ACCREDITATION STANDARDS RELATED TO PHYSICAL REHABILITATION + + + INTRO + paragraph + 25250 + Standard 5I: Center/Service for Physical Rehabilitation + + + INTRO + paragraph + 25306 + Health Center promotes the concept of active participation of disabled people and their families in identifying needs and resources in rehabilitation develop a shared vision of their life in society, the implementation of the vision and the monitoring and evaluation of implementation. + + 9606 + Species + + people + + + + INTRO + paragraph + 25592 + Criteria + + + INTRO + paragraph + 25601 + 5I.295 Within the health center there is a center for the rehabilitation of disabled persons (CBR) with support groups and self-help. + + 9606 + Species + + persons + + + + INTRO + paragraph + 25735 + 5I.296 center is managed by the appointed specialist in physical medicine and rehabilitation. + + + INTRO + paragraph + 25829 + 5I.297 Depending on the systematization of jobs, the Centre employed physiotherapy technicians, occupational therapists, nurses, general direction. + + + INTRO + paragraph + 25977 + 5I.298 Physical therapists adhere to the Code Company physiotherapists with permission to work in practice. + + + INTRO + paragraph + 26085 + 5I.299 specialist in physical medicine and rehabilitation therapists are members professional organizations that provide guidance on continuing professional development. + + + INTRO + paragraph + 26255 + 5I.300 All employed staff in service has a documented plan for ongoing personal professional development that includes measurable goals for learning. + + + INTRO + paragraph + 26405 + 5I.301 There is evidence in writing of actions taken on the permanent professional development that reflects the plan. + + + INTRO + paragraph + 26524 + 5I.302 There are dated, documented criteria for referring patients to physical therapy. The criteria are written/reviewed in the past three years. + + 9606 + Species + + patients + + + + INTRO + paragraph + 26671 + 5I.303 There are dated, documented procedures on granting new cases to team members for physical therapy. The procedure is written/rewritten in the past three years. Instructions: Cases should be awarded according to the skills and experience required for the indicated treatment, together with the need for equitable distribution of the number of cases. + + + INTRO + paragraph + 27026 + 5I.304 There are documented procedures on the regulation of proper completion of the treatment of patients having implemented the rehabilitation plan. The procedure is written/rewritten in the past three years. + + 9606 + Species + + patients + + + + INTRO + paragraph + 27237 + 5I.305 estimated needs of the patient/user for the physical treatment by physical examination which receives measurable data for evaluation. + + 9606 + Species + + patient + + + + INTRO + paragraph + 27378 + 5I.306 Each patient was referred to physical therapy has an individual rehabilitation plan for the implementation of physical therapy. + + 9606 + Species + + patient + + + + INTRO + paragraph + 27513 + 5I.307 Before each procedure/treatment is carried out risk assessments. Note: This includes risk assessment, contraindications to treatment and precautions. Also, include the verification of hazards such as wet floors, and the provision of suitable clothing and shoes worn by therapists and patients/users. + + 9606 + Species + + patients + + + + INTRO + paragraph + 27820 + 5I.308 rehabilitation program undertaken only after the results of the risk assessment. + + + INTRO + paragraph + 27908 + 5I.309 risks associated with the use of electrical equipment reduces the use of safety switches. + + + INTRO + paragraph + 28005 + 5I.310 Before using the apparatus and its application to a patient, is made visual and physical verification of security equipment. + + 9606 + Species + + patient + + + + INTRO + paragraph + 28137 + 5I.311 There is plenty of space and are available partitions/curtains in the room/rooms for physiotherapy treatment that patients provide visual privacy when using the equipment. + + 9606 + Species + + patients + + + + INTRO + paragraph + 28316 + 5I.312 equipment is kept in a place where not to interfere with access to fire exits, entrances, hallways and other equipment. + + + INTRO + paragraph + 28443 + 5I.313 Health Centre organizes regular meetings for people with disabilities and their family members, performs basic assessment of their needs and implements early education and basic rehabilitation programs under the guidance of an appropriate therapist. + + 9606 + Species + + people + + + + INTRO + paragraph + 28700 + 5I.314 health center, or CBR, is developing a multi-sectoral cooperation and contracts with appropriate institutions for labor and social policy, education, institutes Employment, sports institutions, etc. Panel evidence-based clinical practice guidelines on musculoskeletal rehabilitation interventions, + + MESH:D048949 + Disease + + labor + + + + DISCUSS + title_1 + 29005 + 7. DISCUSSION AND RECCOMENDATIONS ON THERAPEUTIC MODALITIES + + + DISCUSS + paragraph + 29065 + Several studies have reported possitive or negative association between computer use and musculosceletal symptoms. The prevalence of Computer Carpal Syndrome (CTS) among computer professionals based on clinical signs and sympoms is approximately 13,1% and almost 1 out of every computer professional suffers from this condition. Very few data published about this topic - studies about CTS prevalence based on clinical signs and symptoms, according to Ali KM and Sathiyasekaran BWC, ranging from 3 to 6 %b only, but we think real situation is much worse. Postural stress due to inadeqauate workstation ergonomics (inappropriate location of monitor, keyboard or mouse) discussed as cause of all mentioned technological diseases in this text (Figure 7). Aydan Oral et al. Had written paper about Evidence Based Physical Medicine and Rehabilitation strategies for patients with cervical radiculopathy due to disc herniation. They reported that neck pain affected 4,82% of the world population in 2010, ranks second after low back pain among musculoskeletal disorders as one of the leading causes of years lived with disability, with contribution of 33,6 million years. Physical medicine and rehabilitation interventions in radicular neck pain and their evidence based are: a) educational interventions; b) exercise; c) workplace interventions/ergonomics; d) phyasical agents (TENS, therapeutic ultrasound, low-level laser therapy) pulsed electromagnetic field therapy, non-invasive brain stimulation techniques, etc); e) injection therapy (epidural steroids, botulinum toxin, ozone); f) cervical collars; g) traction; manual therapies (massage, manipulation and mobilization (commonly used in the management of discogenic neck pain); complementary and alternative medicine treatments (acupuncture, herbal medicine, etc.). Much research efforts have been spent and several risk factors such as heavy lifting, lifestyle, psichosocial factors identified, but the etiology of technological diseases are still or not enough unclear. Recurrences and functional limitations can be minimized limitations with appropriate conservative management, including medications, physical therapy, exercise and patient education. But, these subgroup of developing chronic and disabling symptoms generating large social costs. + + 7276 + Gene + 317 + + CTS + + + 7276 + Gene + 317 + + CTS + + + 10090 + Species + + mouse + + + 9606 + Species + + patients + + + 1407750 + Species + + herbal medicine + + + 9606 + Species + + patient + + + MESH:D010146 + Disease + + pain + + + MESH:D002349 + Disease + + Computer Carpal Syndrome + + + MESH:D009140 + Disease + + musculoskeletal disorders + + + MESH:D019547 + Disease + + neck pain + + + MESH:D001416 + Disease + + back pain + + + MESH:D019547 + Disease + + radicular neck pain + + + MESH:D011843 + Disease + + cervical radiculopathy + + + + DISCUSS + paragraph + 31371 + According to the Panel evidence-based clinical practice guidelines on musculoskeletal rehabilitation interventions the key points to clinicians are (Table 1 and 2): + + + TABLE + T1.xml + T1 + table_caption + 31536 + Summary grid of neck pain guidelines. *Adapted from the Philadelphia Panel Members and Ottawa Methods Group.2. A, benefit demonstrated; C, no benefit demonstrated; EMG, electromyographic; ID, insufficient or no data; TENS, transcutaneous electrical nerve stimulation + + MESH:D000309 + Disease + + insufficient + + + MESH:D010146 + Disease + + pain + + + + TABLE + T2.xml + T2 + table_caption + 31803 + Summary grid of shoulder pain guidelines. *Adapted from the Philadelphia Panel Members and Ottawa Methods Group.1A, benefit demonstrated; C, no benefit demonstrated; EMG, electromyographic; ID, insufficient or no data; TENS, transcutaneous electrical nerve stimulation. + + MESH:D000309 + Disease + + insufficient + + + MESH:D010146 + Disease + + pain + + + + DISCUSS + paragraph + 32073 + The Philadelphia Panel recommends continued normal activities for acute, uncomplicated low back pain and therapeutic exercise for chronic, subacute, and postsurgical low back pain. + + MESH:D017116 + Disease + + low back pain + + + MESH:D017116 + Disease + + low back pain + + + + DISCUSS + paragraph + 32254 + The Philadelphia Panel also recommends transcutaneous electrical nerve stimulation and + + + DISCUSS + paragraph + 32341 + exercise for knee osteoarthritis. + + MESH:D010003 + Disease + + osteoarthritis + + + + DISCUSS + paragraph + 32375 + For chronic neck pain, the Philadelphia Panel recommends proprioceptive and thera-peutic exercise. + + MESH:D019547 + Disease + + neck pain + + + + DISCUSS + paragraph + 32474 + The Philadelphia Panel found evidence to sup-port the use of therapeutic ultrasound in thetreatment of calcific tendonitis of the shoulder. + + MESH:D052256 + Disease + + calcific tendonitis + + + + DISCUSS + paragraph + 32614 + The main difficulty in determining the effec-tiveness of rehabilitation interventions is the lack of well-designed, prospective, random-ized, controlled trials. + + + DISCUSS + paragraph + 32775 + Acute neck pain is often associated with injury or accident, whereas chronic neck pain is related to repetitive injury. Neck pain is commonly managed with analgesics and rest, but referrals to rehabilitation are increasing. The Philadelphia Panel sought to improve the appropriate use of rehabilitation interventions for neck pain by providing evidence-based guidelines. A summary of the Panel's recommendations can be found in Table 2. + + MESH:D019547 + Disease + + Neck pain + + + MESH:D019547 + Disease + + neck pain + + + MESH:D000081084 + Disease + + injury or accident + + + MESH:D019547 + Disease + + neck pain + + + MESH:D012090 + Disease + + repetitive injury + + + MESH:D019547 + Disease + + neck pain + + + + DISCUSS + paragraph + 33212 + Rehabilitation specialists offer several conservative interventions for the management of shoulder pain. There are few published guidelines for the management of shoulder pain. + + MESH:D010146 + Disease + + pain + + + MESH:D010146 + Disease + + pain + + + + CONCL + title_1 + 33389 + 8. CONCLUSION + + + CONCL + paragraph + 33403 + Thanks to the high use of computers, resulting diseases of the modern era, known as the"Technological disease." Under this term we mean the diseases caused by the harmful influence of the job in the first place, the working position in which it is involved excessive work for a computer, such as the banking sector, the health sector and many others who are directly or indirectly connected with the work on the computer and overuse keyboards and computer mice, which are in a number of cases of non-ergonomic, and inappropriate for a handful. Establishing the diagnosis of diseases of technology is an interdisciplinary process that requires special knowledge in medicine and related fields related to health and safety at work. Determining the causes and diagnosis of occupational diseases is carried out according to the criteria of modern medicine work. + + 10090 + Species + + mice + + + MESH:D009784 + Disease + + occupational diseases + + + + CONCL + paragraph + 34265 + First, we identify the clinical picture of the disease on the one hand and identification in the working process on the other side, and their immediate connections. Medical history is the gold standard, because without it, it would be possible to find information on working conditions and the duration and intensity of exposure to a particular hazard from the workplace. The intensity and length of exposure to harmful factor must be that level that is known and proven scientific research that can damage health. Prevention would involve, educate employees by computers to ensure better and more regular position of the spine and shoulder while working at the computer, as well as the use of stylish ergonomic mice in hand. Of course, adequate work breaks are required, in order to avoid fatigue and exhaustion syndrome, but today it is difficult to appreciate the extent and speed of work. No less important and efficient are exercises to strengthen the shoulder belt, as well as exercises for posture. + + 10090 + Species + + mice + + + MESH:D005221 + Disease + + fatigue + + + MESH:D006359 + Disease + + exhaustion syndrome + + + + CONCL + footnote + 35271 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 35308 + REFERENCES + + + 3 + surname:Ashalatha;given-names:STL + surname:Sebastian;given-names:A + REF + surname:Pereira;given-names:M + 73 + 11 + 2013 + IJHSR + ref + 79 + 35319 + Body Mechanics and Perceived Physical Health Problems among Computer Users + + + surname:Ganos;given-names:D + surname:Janevski;given-names:P + surname:Holsbeeck;given-names:M + surname:Lee;given-names:D + 859 + 4 + 10442084 + 1999 + Radiologic Clinics of North America + ref + 37 + REF + 872 + surname:Darian;given-names:V + surname:Ditmars;given-names:D + 35394 + Diagnosis of carpal tunnel syndrome + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + surname:Golub;given-names:R + 306 + surname:Robert;given-names:M + surname:Lynm;given-names:C + REF + surname:Torpy;given-names:JM + 2283 + 20 + 22110111 + 2011 + JAMA + ref + 35430 + Carpal Tunnel Syndrome + + + 155 + surname:Demartini;given-names:FE + REF + surname:Grokoest;given-names:AW + 635 + 7 + 1954 + JAMA + ref + 637 + 35453 + Sistemic disease and the carpal tunnel syndrome + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + 38 + surname:Kolundzic;given-names:R + surname:Bilic;given-names:R + REF + surname:Crnkovic;given-names:T + 3-4 + 2008 + Medica Jadertina + ref + 35501 + Syndrome carpal tunnel + + + surname:Meholjic;given-names:A + REF + surname:Vavra-Hadziahmetovic;given-names:N + 64 + 2011 + Medicinski fakultet Univerziteta u Sarajevu + ref + 65 + 35524 + Osnove klinickog pregleda u fizijatriji + + + surname:Ayromlou;given-names:H + surname:Bakhti;given-names:S + surname:Andalib;given-names:S + surname:Talebi;given-names:M + 283 + 2 + 2013 + Neurosciences Research Center + ref + 3 + REF + 288 + surname:Talebi;given-names:A + surname:Aghili;given-names:A + 35565 + Effect of vitamin B6 on clinical symptoms and electrodiagnostic results of patients with carpal tunnel sndrome + + 9606 + Species + + patients + + + MESH:D025101 + Chemical + + vitamin B6 + + + + surname:Mustafa;given-names:Yilmaz + surname:Koca;given-names:I + surname:Altindag;given-names:O + surname:Madenci;given-names:E + 3171 + 10 + 21953301 + 2011 + Rheumatology International + ref + 32 + REF + 3179 + surname:Gur;given-names:A + 35676 + Reliability and efficacy of the new massage technique on the treatment in the patients with carpal tunnel syndrome + + 9606 + Species + + patients + + + MESH:D002349 + Disease + + carpal tunnel syndrome + + + + surname:Osmanovic;given-names:J + 67 + surname:Gorcevic;given-names:S + surname:Gorcevic;given-names:E + REF + surname:Kasumovic;given-names:M + 414 + 6 + 2013 + Med Arh + ref + 417 + 35791 + Cervical Syndrome - the Effectiveness of Physical Therapy Interventions + + + surname:Klinicka;given-names:neurologija + REF + surname:Kantardzic;given-names:Dz + 460 + 2001 + Sarajevo + ref + 461 + 35863 + IP "Svjetlost" d.d., Zavod za udzbenike i nastavna sredstva + + + surname:Vavra-Hadziahmetovic;given-names:N + surname:Masic;given-names:I + surname:Talic;given-names:A + surname:Karic-Skrijelj;given-names:M + 25 + 1 + 24109153 + 2008 + Acta Inform Med + ref + 16 + REF + doi: 10.5455/aim.2008.16.25-28 + 28 + surname:Suljic-Mehmedika;given-names:E + surname:Pandza;given-names:H + 35928 + Cervical Pain Syndrome as Consequence of Computer Use in Daily Practice + + MESH:D010146 + Disease + + Pain + + + + REF + ref + 36000 + + + REF + ref + 36001 + + + surname:Treanor;given-names:WJ + surname:Honet;given-names:JC + REF + surname:Ellenberg;given-names:MR + MI 48235 + 1994 + ref + 2899 + 36002 + Cervical radiculopathy + + MESH:D011843 + Disease + + radiculopathy + + + + 147 + surname:Nygaard;given-names:OP + surname:Leivseth;given-names:G + REF + surname:Kolstad;given-names:F + 1065 + 10 + 2005 + A prospective outcome study. Acta Neurochir (Wien) + ref + 1070 + 36025 + Transforaminal steroid injections in the treatment of cervical radiculopathy + + MESH:D011843 + Disease + + radiculopathy + + + MESH:D013256 + Chemical + + steroid + + + + surname:Kitakoji;given-names:H + 31 + surname:Itoi;given-names:M + surname:Inoue;given-names:M + REF + surname:Nakajima;given-names:M + 364 + 23965301 + 2013 + Acupunct Med + ref + 367 + 36102 + Clinical effect of acupuncture on cervical spondylotic radiculopathy: results of a case series + + MESH:D011843 + Disease + + spondylotic radiculopathy + + + + REF + ref + 36197 + + + REF + ref + 36198 + + + 22 + surname:Leung;given-names:CT + REF + surname:Chen;given-names:HM + 518 + 5 + 2007 + Clin Biomech (Bristol, Avon) + ref + 523 + 36199 + The effect on forearm and shoulder muscle activity in using different slanted computer mice + + + REF + ref + 36291 + + + 102 + surname:Schouten;given-names:JS + REF + surname:Picavet;given-names:HS + 167 + 12620608 + 2003 + Pain + ref + 178 + 36292 + Musculoskeletal pain in the Netherlands: prevalences, consequences and risk groups, the DMC(3)-study + + MESH:C023025 + Chemical + + DMC + + + MESH:D059352 + Disease + + Musculoskeletal pain + + + + surname:Arendzen;given-names:HJ + surname:Groenier;given-names:KH + surname:Sobel;given-names:JS + surname:Winters;given-names:JC + 1320 + 9158469 + 1997 + BMJ + ref + 314 + REF + 1325 + surname:Meyboom-de Jong;given-names:B + 36393 + Comparison of physiotherapy, manipulation, and corticosteroid injection for treating shoulder complaints in general practice: randomised, single blind study + + + surname:Chain;given-names:IS + surname:Sze;given-names:WW + surname:Tsui;given-names:MM + REF + surname:Sheto;given-names:GP + available on. + DRC + ref + surname:Lee;given-names:FW + surname:Chung;given-names:CC + 36550 + Issues about home computer workstations and primary school children in Hong Kong: A pilot study + + 9606 + Species + + children + + + + 60 + surname:Ketenci;given-names:A + surname:Sindel;given-names:D + REF + surname:Oral;given-names:A + 47 + 2014 + Doi: 10.5152/tftrd.2014.76735 + Turk J Phys Med Rehab + ref + 53 + 36646 + Evidence-Based Physical Medicine and Rehabilitatio Strategies for Patients with Cervical Radiculopathy Due to Disc Herniation + + 9606 + Species + + Patients + + + MESH:D011843 + Disease + + Cervical Radiculopathy + + + + 12 + surname:Sathiyasekaran;given-names:BWC + REF + surname:Ali;given-names:Mohamad K + 319 + 3 + 16984790 + 2006 + JOSE + ref + 325 + 36772 + Computer Professionals and Carpal Tunel Syndrome (CTS) + + 7276 + Gene + 317 + + CTS + + + + surname:Mustafa;given-names:A + surname:Haxhiu;given-names:B + surname:Murtezani;given-names:A + surname:Ibraimi;given-names:Z + 410 + 6 + 2013 + Med Arch + ref + 67 + REF + Doi. 10.5455/medarh.2013.67.419-413 + 413 + surname:Martinaj;given-names:M + 36827 + Prevalence and Pharmacologic Treatment of Patients with Low Back Pain Treated at Kosovo Energetic Corporation + + 9606 + Species + + Patients + + + MESH:D017116 + Disease + + Low Back Pain + + + + + 4272840 + + surname:Genadieva-Stavric;given-names:Sonja + surname:Balkanov;given-names:Trajan + surname:Sotirova;given-names:Tatjana + 25568637 + surname:Krstevska;given-names:Svetlana B. + 348 + 5 + 2014 + 4272840 + MSM-26-348 + 10.5455/msm.2014.26.348-351 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 348-51. doi:10.5455/msm.2014.26.348-351 + multiple myeloma treatment transplant noneligible patients + 351 + Krstevska SB, Sotirova T, Balkanov T, Genadieva-Stavric S + 0 + Tretatment Approach of Nontransplant Patients with Multiple Myeloma + + 9606 + Species + + Patients + + + MESH:D009101 + Disease + + Multiple Myeloma + + + + ABSTRACT + abstract + 68 + Multiple myeloma is still an incurable disease with pattern of regression and remission followed by multiple relapses raising from the residual myeloma cells surviving even in the patients who achieve complete clinical response to treatment. In recent years there is a huge improvement in treatment of patients with multiple myeloma. The milestones of these improvement are: autologous transplantation and high-dose melphalan, imunomodulating drugs (thalidomide, lenalidomide), proteosom inhibitors (bortesomib, carfilzomib). The most significant improvement in overall survival has been achieved in the patients younger than 65 years. So, the major challenge for hematologist is to translate this improvement in the elderly patients with multiple myeloma. Today, physicians are able to offer wider variety of treatment options for elderly patients with multiple myeloma. Therapeutic options should be tailored and personalized according to patient's characteristics by balancing efficacy and toxicity of each drug which is especially important for elderly patients. In the mode of sequencing treatment for elderly patients with multiple myeloma, our goal is to achieve and maintain maximal response while limiting treatment -related toxicities as much as possible. Second-generation novel agent, such as carfilzomib, pomalidomide, elotuzumab, bendamustine are currently being evaluated as an option to improve treatment outcome in elderly patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + - + Chemical + + bortesomib + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D000069461 + Chemical + + bendamustine + + + MESH:D064420 + Disease + + toxicity + + + MESH:D009101 + Disease + + Multiple myeloma + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:C524865 + Chemical + + carfilzomib + + + MESH:C467566 + Chemical + + pomalidomide + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D064420 + Disease + + toxicities + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + myeloma + + + MESH:C546027 + Chemical + + elotuzumab + + + MESH:C524865 + Chemical + + carfilzomib + + + + INTRO + paragraph + 1518 + Myeloma multiplex is a malignant disorder first recognized in the 19th century and at that time point was described as "mollities ossium" accompanied by the presence of Bence Jones protein in urine. At that time there was no effective treatment and median overall survival was only short: a few months. Over the time, especially over the past decade, many advances in myeloma treatment have been made and improvement of the median overall survival have been achieved. But, myeloma multiplex is still considered incurable disease. + + MESH:D009101 + Disease + + Myeloma multiplex + + + MESH:D009101 + Disease + + myeloma + + + MESH:D009101 + Disease + + myeloma + + + MESH:D009369 + Disease + + malignant disorder + + + + INTRO + paragraph + 2052 + Myeloma multiplex is a malignant disorder that arises from the malignant proliferation of plasma cell and is characterized by the presence of at least 10% clonal bone marrow plasma cells and serum and/or urinary monoclonal protein. Myeloma multiplex accounts for 1% of all types of cancers and is the second most common hematologic neoplasm, approximately 13% for all hematologic malignances. Myeloma multiplex is considered as a disease of elderly reflected by the average age at diagnosis 70 years, with 37% of patients younger than 65 years, 26% aged 65 to 74 years, and 37% older than 75years. + + 9606 + Species + + patients + + + MESH:D009369 + Disease + + cancers + + + MESH:D009101 + Disease + + Myeloma + + + MESH:D019337 + Disease + + hematologic neoplasm + + + MESH:D009369 + Disease + + malignant disorder + + + MESH:D009101 + Disease + + Myeloma multiplex + + + MESH:D009101 + Disease + + Myeloma + + + + INTRO + paragraph + 2650 + Patient with myeloma multiplex could be diagnosed as symptomatic or asymptomatic disease. Patients with a symptomatic disease should be treated immediately, and the mainstay of asymptomatic disease is still an observation. Symptomatic disease could be defined with so called CRAB features: C-hypercalcemia (>11.5mg/dl (2.65mmol/l); R-renal failure (serum creatinin >mg/dl; 1.73mmol/l); A-anemia (hemoglobin <10g/dl; 12.5 mmol/l) or 2 g/dl; 1.25mmol/l below the lower limit of normal; and B-bone disease (lytic lesions, severe osteopenia, or pathologic fractures). Patients are stratified into three risk groups according to the International Staging system (ISS). This system defines three risk groups based on serum beta 2 micro globulin and albumin levels at diagnosis. High-risk disease and poor prognosis are defined with the presence of high levels of serum beta2 micro globulin (stage III). + + 10242 + Gene + 4257 + + beta 2 + + + 10242 + Gene + 4257 + + beta2 + + + 9606 + Species + + Patient + + + 9606 + Species + + Patients + + + 9606 + Species + + Patients + + + MESH:D009101 + Disease + + myeloma + + + + INTRO + paragraph + 3547 + Nowadays, chromosomal abnormalities t(4;14), t(14;16) and t(14;20) chomosome 1 abnormalities and del17 detected by fluorescent in situ hybridization (FISH) are associated with poor prognosis. Hiperdiplpdy, t (11;14), t, are associated with good prognosis and can be considered "standard risk". + + MESH:D002869 + Disease + + chromosomal abnormalities + + + Disease + + Hiperdiplpdy + + + + INTRO + paragraph + 3845 + Initial therapy for multiple myeloma depends to a certain extent on patients characteristics such as: eligibility for autologous stem cell transplantation per se; age and co-morbidities. The role of induction therapy is to induce remission, but patient's characteristics have a significant role in the initial treatment approach. Goals of treatment are: to eradicate the tumor clone, including cancer stem cell, to search for an appropriate balance between efficacy and toxicity with three different but complementary aims: quality of life, survival prolongation and eventually the dream of cure. This can be achieved if we use appropriate tools to evaluate treatment efficacy. Achieving the lowest level of minimal residual disease can be an important goal of therapy, a step in the path to cure. + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D009369 + Disease + + tumor + + + MESH:D009369 + Disease + + cancer + + + MESH:D064420 + Disease + + toxicity + + + MESH:D009101 + Disease + + multiple myeloma depends + + + + INTRO + paragraph + 4643 + Today, treatment goals in myeloma patients are shifting and the goal of therapy in elderly patients is to achieve and maintain maximal response. Many believe that multiple myeloma can be converted into a chronic disease and that a functional cure maybe a realistic goal. Attainment of complete remission at any time point during treatment is associated with improvement outcome; so it is likely to be established as a goal of therapy. The increasing number of treatment possibilities as an optimal therapeutic strategy improve patient outcome. Physicians have the opportunity to choose the best treatment regimen according to patient characteristics, while limiting treatment - related toxicities as much as possible. In elderly patients optimal treatment should be always balance efficacy and toxicity. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + MESH:D064420 + Disease + + toxicities + + + MESH:D064420 + Disease + + toxicity + + + MESH:D002908 + Disease + + chronic disease + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + myeloma + + + + INTRO + paragraph + 5449 + Initial therapy for multiple myeloma depends on eligibility for high-dose therapy and autologous stem cell transplant (HDT-ASCT). In many European countries, elderly patients (older than 65) are generally considered ineligible for autologous stem cell transplantation (ASCT). In past times in practice, patients were stratified to those who are transplant eligible and consolidated with high-dose melphalan/SCT and those who were transplant ineligible and received oral melphalan with prednisolone (MP). This stratification has been made in the era of conventional therapy where response rates to the induction therapy were poor and the goal was achievement of partial response and prevention of organ damage. So, in contrast, today we considered that biological age and chronological age do not always correspond, and a greater emphasis should be placed on the former rather than the latter. CR is a good surrogate end point for survival in transplant-ineligible patients and novel drugs have been incorporated into the treatment of non-transplant candidates. Patients are generally considered eligible for ASCT if they have good performance status, no-comorbidities, and normal cardiac, pulmonary, liver and renal function. Patients older than 75 years or vulnerable ones are more susceptible to adverse events and in this setting we are looking for less toxic regimens and appropriate dose reductions should be adopted. Even, ASCT with a reduced melpahlan conditioning dose is well tolerated by the selected population of patients up to the age of 75 in good clinical conditions. Until novel agents were introduced, for more than 40 years the combination of melpahalan and prednisone (MP) was considered the standard approach for transplant -ineligible patients, with PFS of approximately 18 months and 2-3 years (at best) overall survival in the population treated with MP. A meta-analysis including 27 randomized studies compared MP with other chemotherapy-containing regimens. The introduction of novel agents has challenged this combination and new and more effective combination is available. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D011241 + Chemical + + prednisone + + + - + Chemical + + MP + + + MESH:D011239 + Chemical + + prednisolone + + + - + Chemical + + melpahalan + + + MESH:D009101 + Disease + + myeloma depends + + + - + Chemical + + MP + + + + INTRO + paragraph + 7552 + Six randomized phase III studies have shown that the combination melphalan-prednisone-thalidomide (MPT) is superior to MP in terms of response and progression-free survival, but it was translated into improved survival in the 2 IFM studies. An efficacy meta-analysis of the six MPT trials including 1685 patients was conducted and has showed that the addition of thalidomide to MP significantly prolonged both PFS and extended OS by 20%-39 months compared to MP-33 months. The meta-analysis further confirmed the progression-free survival advantage achieved with MPT. So, MPT is therefore regarded as a new standard of care in transplant ineligible patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D013792 + Chemical + + thalidomide + + + - + Chemical + + MPT + + + + INTRO + paragraph + 8215 + For selected elderly patients, particularly for standard risk patients with favorable FISH, CTD is a feasible approach. A phase III assessed the role of thalidomide in combination with different alkylating agent and steroid than those commonly used, respectively cyclophosphamide and dexamethasone with an attenuated schedule (CTDa). Despite a deeper response, no differences were noted in median PFS and OS between patients treated with CTDa and MP. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D013792 + Chemical + + thalidomide + + + - + Chemical + + CTDa + + + MESH:D013256 + Chemical + + steroid + + + - + Chemical + + CTDa + + + MESH:D003907 + Chemical + + dexamethasone + + + MESH:D003520 + Chemical + + cyclophosphamide + + + + INTRO + paragraph + 8666 + Lenalidomide is an immunomodulatory drug with higher potency than its analogue thalidomide and without sedative or neurotoxic adverse effects. Lenalidomide showed to be safe and effective in relapsed/refractory multiple myeloma patients, as well as in newly diagnosed multiple myeloma patients. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000077269 + Chemical + + Lenalidomide + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D009101 + Disease + + myeloma + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000077269 + Chemical + + Lenalidomide + + + MESH:D020258 + Disease + + neurotoxic + + + + INTRO + paragraph + 8961 + The phase III trial RD (lenalidomide plus high-dose dexamethason) versus Rd (lenalidomide plus low-dose dexamethason) included newly diagnosed multiple myeloma patients both eligible and ineligible for autologus stem cell transplantation. Rd seems preferable for elderly patients because has significantly longer 1-year overall survival, particularly evident in patients older than 65 years of age and can be considered a valid therapeutic option for elderly newly diagnosed myeloma patients. RD remains a good option for patients with renal failure, hypercalcemia, pain or spinal cord compression because more adverse events occurred when RD was given compared with Rd. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D006934 + Disease + + hypercalcemia + + + MESH:D009101 + Disease + + myeloma + + + MESH:D010146 + Disease + + pain + + + MESH:D051437 + Disease + + renal failure + + + MESH:D003907 + Chemical + + dexamethason + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D003907 + Chemical + + dexamethason + + + MESH:D009101 + Disease + + multiple myeloma + + + + INTRO + paragraph + 9632 + The role of lenalidomide was assessed in a recent phase III study that compared melphalan-prednisone-lenalidomide followed by lenalidomide maintenance (MPR-R), with MPR and MP. MPR-R significantly improved median progression-free survival compared with MPR and MP (31 months versus 14 months vs. 13 months; p<0.001), which means that MPR-R reduced the risk of progression by 51% and 60% compared with MPR and MP. Lenalidomide maintenance was well tolerated with few reported incidence of grade 3/4 adverse events and second cancer. However the benefit associated with MPR-R outweigh the increased risk of second primary malignancies. + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D009369 + Disease + + cancer + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D000077269 + Chemical + + Lenalidomide + + + MESH:D009369 + Disease + + malignancies + + + + INTRO + paragraph + 10266 + VISTA study compared MP versus VMP and proved that addition of bortezomib to MP is rational, because responses, time to progression and survival were significantly higher with VMP. + + 64115 + Gene + 81923 + + VISTA + + + MESH:D000069286 + Chemical + + bortezomib + + + + INTRO + paragraph + 10447 + In the Spanish PETHEMA trial a cohort of 260 patients older than 65, transplant ineligible, were randomized to receive induction treatment with 6 cycles of bortezomib-thalidomeide-prednisone VTP, or VMP considered as more gentle approach where melphalan is used instead of thalidomide. Both regimens led to high ORR rate (81%and 80%), but there were more adverse events among patients with VDT. VMP with once-weekly schedule of botezomib should be preferred to VTP as induction for elderly multiple myeloma patients. One-weekly bortezomib was scheduled instead of standard twice-weekly administration to reduce adverse events, especially neuropathy associated with bortezomib administration. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D011241 + Chemical + + prednisone + + + MESH:C562515 + Disease + + especially neuropathy + + + MESH:D013792 + Chemical + + thalidomide + + + - + Chemical + + botezomib + + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D008558 + Chemical + + melphalan + + + - + Chemical + + VTP + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000069286 + Chemical + + bortezomib + + + - + Chemical + + thalidomeide + + + Disease + + VDT + + + + INTRO + paragraph + 11139 + The combination of bortezomib-lenalidomide-dexamethasone (VRD) was evaluated in both young and elderly patients with multiple myeloma. + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + - + Chemical + + VRD + + + - + Chemical + + bortezomib-lenalidomide-dexamethasone + + + + INTRO + paragraph + 11274 + The addition of thalidomide to the new treatment standard with VMP followed by bortezomib-thalidomide maintenance is valid alternative with 3-years overall survival 565 with VMP-VT compared with 41% with VMP. VMP-VT with once weekly bortezomib seems to be a valid alternative for elderly patients particularly those younger than 75 years, where there is the same efficacy, but without additional toxicity, particular peripheral neuropathy. + + 9606 + Species + + patients + + + MESH:D010523 + Disease + + particular peripheral neuropathy + + + MESH:D017180 + Disease + + VT + + + MESH:D000069286 + Chemical + + bortezomib + + + - + Chemical + + bortezomib-thalidomide + + + MESH:D064420 + Disease + + toxicity + + + MESH:D017180 + Disease + + VT + + + MESH:D013792 + Chemical + + thalidomide + + + + INTRO + paragraph + 11714 + A sequential approach consisting of an induction regimen associated with a high rate of complete response, followed by consolidation, maintenance therapy, induce a profound cytoreduction and delays relapse, thus improving survival as a therapy approach that minimized toxicity maximized quality of life and emphasized patient's preference. This choice of treatment strategy improve outcome and prevent the occurrence of relapse with a continuous treatment keeping residual disease under control. The international MM015 phase III study assessed the role of lenalidomide given in maintenance. Having this in mind MPR followed by lenalidomide maintenance emerges as a new standard treatment option for elderly multiple myeloma patients. Another treatment option for elderly patients with impressive result is more intensive regimen VMPT followed by VT maintenance. Bortezomib showed same efficacy with decreased toxicity administrated in a new schedule from twice to once weekly. + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D064420 + Disease + + toxicity + + + MESH:D000069286 + Chemical + + Bortezomib + + + MESH:D017180 + Disease + + VT + + + MESH:D064420 + Disease + + toxicity + + + MESH:D000077269 + Chemical + + lenalidomide + + + + INTRO + paragraph + 12692 + As we already emphasized, the choice among these regimens should be based on the patient's characteristics. For elderly patients with renal impairment bortezomib or thalidomide containing regiment is considered the best treatment option, because lenalidomide is excreted by the kidneys. In such a case dose of lenalidomide should be reduced according to creatinin clearance. Despite that, for patients with peripheral neuropathy lenalidomide is considered as the best treatment option because of the reduced neurological toxicity. In elderly patients there is an increased risk of treatment-related adverse events in response to novel agent-containing regimens. Clinicians should be aware and proposed guidelines for dose reduction of novel agents and protocol modification in elderly should be followed in clinical practice for elderly patients with multiple myeloma to optimize outcome. So, there is an attempt to personalize the therapy in multiple myeloma according to patient age and vulnerability proposed by European Myeloma Network (EMN). + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D009101 + Disease + + Myeloma + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D010523 + Disease + + peripheral neuropathy + + + MESH:D020258 + Disease + + neurological toxicity + + + MESH:D007674 + Disease + + renal impairment bortezomib + + + MESH:D009101 + Disease + + multiple myeloma + + + + INTRO + paragraph + 13739 + Multiple myeloma is still an incurable disease with pattern of regression and remission followed by multiple relapses raising from the residual myeloma cells surviving even in the patients who achieve complete clinical response to treatment. New anti-myeloma drugs change treatment paradigm providing both tumor reduction and tumor suppression. There is so much progress, but still many unsolved questions. Today, physicians are able to offer wider variety of treatment options for both young and elderly patients with multiple myeloma. Therapeutic options should be tailored and personalized according to patient's characteristics by balancing efficacy and toxicity of each drug which is especially important for elderly patients. In the mode of sequencing treatment for elderly patients with multiple myeloma, our goal is to achieve and maintain maximal response while limiting treatment -related toxicities as much as possible. Second-generation novel agent, such as carfilzomib, pomalidomide, elotuzumab, bendamustine are currently being evaluated as an option to improve treatment outcome in elderly pateints. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D064420 + Disease + + toxicities + + + MESH:D064420 + Disease + + toxicity + + + MESH:C467566 + Chemical + + pomalidomide + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + myeloma + + + MESH:D000069461 + Chemical + + bendamustine + + + MESH:D009101 + Disease + + Multiple myeloma + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:C524865 + Chemical + + carfilzomib + + + MESH:D009369 + Disease + + tumor reduction and tumor + + + MESH:C546027 + Chemical + + elotuzumab + + + MESH:D009101 + Disease + + myeloma + + + + INTRO + footnote + 14854 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 14891 + REFERENCES + + + 111 + surname:Rajkumar;given-names:SV + REF + surname:Kyle;given-names:RA + 2962 + 18332230 + 2008 + Blood + ref + 2972 + 14902 + Multiple myeloma + + MESH:D009101 + Disease + + Multiple myeloma + + + + 15 + surname:Palumbo;given-names:A + surname:Cavallo;given-names:F + REF + surname:Genadieva;given-names:Stavric S + 157 + 2 + 24578203 + 2014 + Curr Treat Options Oncol + ref + 170 + 14919 + New approaches to management of multiple myeloma + + MESH:D009101 + Disease + + multiple myeloma + + + + 364 + surname:Anderson;given-names:K + REF + surname:Palumbo;given-names:A + 1046 + 21410373 + 2011 + N Engl J Med + ref + 1060 + 14968 + Multiple myeloma + + MESH:D009101 + Disease + + Multiple myeloma + + + + 23 + surname:Durie;given-names:BGM + surname:San Miguel;given-names:J + REF + surname:Greipp;given-names:PR + 3412 + 2005 + J Clin Oncol Off J Am Soc Clin Oncol + ref + 3420 + 14985 + International staging system for multiple myeloma + + MESH:D009101 + Disease + + multiple myeloma + + + + 23 + surname:Rajkumar;given-names:SV + REF + surname:Kyle;given-names:RA + 3 + 2009 + Leuk Off J Leuk Soc Am Leuk Res Fund UK + ref + 9 + 15035 + Criteria for diagnosis, staging, risk stratification and response assessment of multiple myeloma + + MESH:D009101 + Disease + + myeloma + + + + 43 + REF + surname:Suzuki;given-names:K + 116 + 2 + 23293370 + 2013 + Jpn J Clin Oncol + ref + 124 + 15132 + Current Threpautic Starategy for Multiple myeloma + + MESH:D009101 + Disease + + Multiple myeloma + + + + 23 + surname:Drach;given-names:J + surname:Bergsagel;given-names:PL + REF + surname:Fonseca;given-names:R + 2210 + 19798094 + 2009 + Leukemia + ref + 2221 + 15182 + International Myeloma Working Group molecular classification of multiple myeloma: spotlight review + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + Myeloma Working + + + + 27 + surname:Cavo;given-names:M + surname:Durie;given-names:BGM + REF + surname:Avet-Loiseau;given-names:H + 711 + 23032723 + 2013 + Leukemia + ref + 717 + 15281 + Combining fluorescent in situ hybridization data with ISS staging improves risk assessment in myeloma: an International Myeloma Working Group collaborative project + + MESH:D009101 + Disease + + myeloma + + + MESH:D009101 + Disease + + Myeloma Working + + + + 32 + surname:San Miguel;given-names:F + surname:Rajkumar;given-names:V + REF + surname:Palumbo;given-names:A + 587 + 24419113 + 2014 + J Clin Oncol + ref + 600 + 15445 + International Myeloma Working Group Consensus Stateet for the Managmnent, treatment, and Supportive Care of Patients with myeloma not eligible for standard Autologous Stem-Cell transplantation + + 9606 + Species + + Patients + + + MESH:D009101 + Disease + + myeloma + + + MESH:D009101 + Disease + + Myeloma Working + + + + 92 + surname:Kleber;given-names:M + surname:Trepos;given-names:E + REF + surname:Engelhardt;given-names:M + 232 + 2 + 24497560 + 2014 + Haematologica + ref + 242 + 15638 + European myeloma network recommendations on the evaluation and treatment of newly diagnosed patients with multiple myeloma + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + myeloma + + + + surname:San Miguel;given-names:JF + REF + surname:Mateos;given-names:MV + 488 + 24319223 + 2013 + Hematology + ref + 495 + 15762 + How should we treat newly diagnosed multple myeloma patients? + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + myeloma + + + + surname:Gay;given-names:F + REF + surname:Palumbo;given-names:A + 566 + 20008241 + 2009 + Hematology + ref + 577 + 15824 + How to treat elderly patients with multiple myeloma: combination of therapy or sequencing + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + + 28 + surname:Cerrato;given-names:C + REF + surname:Palumbo;given-names:A + 263 + 23682217 + 2013 + Korean J Intern Med + ref + 273 + 15914 + Diagnosis and therapy of multiple myeloma + + MESH:D009101 + Disease + + myeloma + + + + 28 + surname:Anderson;given-names:KC + REF + surname:Lional;given-names:S + 258 + 23868105 + 2014 + Leukemia + ref + 268 + 15956 + Association of response endpoints with survival outomes in multiple myeloma + + MESH:D009101 + Disease + + myeloma + + + + 111 + surname:Dispenzieri;given-names:A + surname:Rajkumar;given-names:SV + REF + surname:Kumar;given-names:SK + 2516 + 17975015 + 2008 + Blood + ref + 2520 + 16032 + Improved survival in multiple myeloma and the impact of novel therapies + + MESH:D009101 + Disease + + myeloma + + + + 118 + surname:Ludwig;given-names:H + surname:Bringhen;given-names:S + REF + surname:Palumbo;given-names:A + 4519 + 17 + 21841166 + 2011 + Blood + ref + 4529 + 16104 + Personalized therapy in multiple myeloma according to p[atient age and vulnerability: a report of the Eouropean Myeloma Network (EMN) + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D009101 + Disease + + Myeloma + + + + 99 + surname:Bringhen;given-names:S + surname:Palumbo;given-names:A + REF + surname:Zweegman;given-names:S + 1133 + 7 + 24986875 + 2014 + Haematologica + ref + 1137 + 16238 + Age and aging in blood disoreders: multiple myeloma + + MESH:D009101 + Disease + + myeloma + + + MESH:D006402 + Disease + + blood disoreders + + + + 25 + surname:Palumbo;given-names:A + REF + surname:Gay;given-names:F + 65 + 21295387 + 2011 + Blood Reviews + ref + 73 + 16290 + Management of older patients with multiple myeloma + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + + 124 + surname:Dingli;given-names:D + REF + surname:Gertz;given-names:MA + 882 + 6 + 24973360 + 2014 + Blood + ref + 890 + 16341 + How we manage autologous stem cell transplantation for patients with multiple myeloma + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + + 40 + surname:Laubach;given-names:J + surname:Jasielec;given-names:J + REF + surname:Rosenbaum;given-names:C + 592 + 5 + 24135404 + 2013 + Seminars in Oncology + ref + 601 + 16427 + Evolving Starategies in the Initial Treatment of multiple myeloma + + MESH:D009101 + Disease + + multiple myeloma + + + + 40 + surname:Palumbo;given-names:A + REF + surname:Cerrato;given-names:C + 577 + 5 + 24135402 + 2013 + Seminars in Oncology + ref + 584 + 16493 + Initial treatment of nontransplant patients With Multiple myeloma + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + Multiple myeloma + + + + 16 + REF + 3832 + 9850028 + 1998 + J Clin Oncol + ref + 3842 + 16559 + Myeloma trialist Collaborative Group. Combination chemotherapy versus melphalan plus prednisone as treatment for myltiple myeloma: an overview of 6,633 patients from 27 randomized trials + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + myltiple myeloma + + + MESH:D009101 + Disease + + Myeloma + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + + 370 + surname:Hulin;given-names:C + surname:Mary;given-names:JY + REF + surname:Facon;given-names:T + 1209 + 17920916 + 2007 + Lancet + ref + 1218 + 16746 + Melphalan and prednisone plus thalidomide versus melphalan and prednisone alone or reduced-intensity autologous stem cell transplantation in elderly patients with multiple myeloma (IFM 99-06): a randomized trial + + 9606 + Species + + patients + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + Melphalan + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D013792 + Chemical + + thalidomide + + + + 27 + surname:Rodon;given-names:P + surname:Facon;given-names:T + REF + surname:Hulin;given-names:C + 3664 + 19451428 + 2009 + J Clin Oncol + ref + 3670 + 16958 + Efficacy of melphalan and prednisone plus thalidomide in patients older than 75 years with newly diagnosed multiple myeloma: IFM01/01 trial + + 9606 + Species + + patients + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D011241 + Chemical + + prednisone + + + + 86 + surname:Firatli-Tuglular;given-names:T + surname:Haznedar;given-names:R + REF + surname:Beksac;given-names:M + 16 + 2011 + Eur J Hematol + ref + 22 + 17098 + Addition of thalidomide to oral melphalan/prednisone in patients with multiple myeloma not eligible for transplantation: result of a randomized trial from tha Turkish Myeloma Study Group + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D009101 + Disease + + Myeloma + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D008558 + Chemical + + melphalan + + + + 116 + surname:Fayers;given-names:P + surname:Gimsing;given-names:P + REF + surname:Waage;given-names:A + 1405 + 20448107 + 2010 + Blood + ref + 1412 + 17285 + Melphalan and prednisone plus thalidomide or placebo in elderly patients with multiple myeloma + + 9606 + Species + + patients + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D008558 + Chemical + + Melphalan + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D011241 + Chemical + + prednisone + + + + 28 + surname:Termorshuizzen;given-names:F + surname:Schaafsma;given-names:M + REF + surname:Wijermans;given-names:P + 3160 + 20516439 + 2010 + J Clin Oncol + ref + 3166 + 17380 + Phase III study of the value of thalidomide added to melphalan plus prednisone in elderly patients with newly diagnosed multiple myeloma: the HOVON 49 Study + + 9606 + Species + + patients + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D013792 + Chemical + + thalidomide + + + + 112 + surname:Liberati;given-names:AM + surname:Bringen;given-names:S + REF + surname:Palumbo;given-names:A + 3107 + 18505783 + 2008 + Blood + ref + 3114 + 17537 + Oral melphalan, prednisone, and thalidomide in elderly patients with multiple myeloma: update results oa a randomized controlled trial + + 9606 + Species + + patients + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D013792 + Chemical + + thalidomide + + + + 98 + surname:Hulin;given-names:C + surname:Waage;given-names:A + REF + surname:Palumbo;given-names:A + 87 + 2013 + Hematologica + ref + 94 + 17672 + Safety of thalidomide in newly diagnosed elderly myeloma patients: a mata-analysis of data from individual patients in six randomized trials + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D009101 + Disease + + myeloma + + + MESH:D013792 + Chemical + + thalidomide + + + + 118 + surname:Gregory;given-names:WM + surname:Davies;given-names:FE + REF + surname:Morgan;given-names:GJ + 1231 + 21652683 + 2011 + Blood + ref + 1238 + 17813 + Cyclophosphamide, thalidomide and dexamathasone (CTD) as initial therapy for patients with multiple myeloma unsuitable for autologous transplantation + + 9606 + Species + + patients + + + - + Chemical + + CTD + + + MESH:D003520 + Chemical + + Cyclophosphamide + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D009101 + Disease + + multiple myeloma + + + - + Chemical + + dexamathasone + + + + 11 + surname:Callander;given-names:NS + surname:Jacobus;given-names:S + REF + surname:Rajkumar;given-names:SV + 29 + 19853510 + 2010 + Lancet.Oncol + ref + 37 + 17963 + lenalidomide plus high-dose dexamathasone versus lenalidomide plus low-dose dexamethasone as initial therapy for newly diagnosed multiple myeloma : an open - lablel randomized controlled trial + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D003907 + Chemical + + dexamethasone + + + - + Chemical + + dexamathasone + + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D000077269 + Chemical + + lenalidomide + + + + 366 + surname:Delforge;given-names:M + surname:Hajek;given-names:R + REF + surname:Palumbo;given-names:A + 1759 + 19 + 22571200 + 2012 + N Engl J Med + ref + 1769 + 18158 + Continuos lenalidomide treatment for newly diagnosed multiple myeloma + + MESH:D000077269 + Chemical + + lenalidomide + + + MESH:D009101 + Disease + + myeloma + + + + 371 + surname:Gay;given-names:F + surname:Cavallo;given-names:F + REF + surname:Palumbo;given-names:A + 895 + 25184862 + 2014 + N Engl J Med + ref + 905 + 18228 + Autologous Transplantation and Maintenance Therapy in Multiple Myeloma + + MESH:D009101 + Disease + + Multiple Myeloma + + + + 359 + surname:Khuageva;given-names:NK + surname:Schlag;given-names:R + REF + surname:San-Miguel;given-names:JF + 906 + 18753647 + 2008 + N Engl J Med + ref + 917 + 18299 + Bortezomib plus melphalan and prednisone for initial treatment of multiple myeloma + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D007625 + Disease + + Bortezomib plus melphalan + + + + 28 + surname:Schlag;given-names:R + surname:Richardson;given-names:PG + REF + surname:Mateos;given-names:MV + 2259 + 20368561 + 2010 + J Clin Oncol + ref + 2266 + 18382 + Bortezomib plus melphalan and prednisone compared with melphalan and prednisone in previously untretated multiple myeloma: update follow-up and impact of subsequent therapy in phase III VISTA trial + + 64115 + Gene + 81923 + + VISTA + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D007625 + Disease + + Bortezomib plus melphalan + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D009101 + Disease + + untretated multiple myeloma + + + MESH:D008558 + Chemical + + melphalan + + + + 11 + surname:Martinez-Lopez;given-names:J + surname:Oriol;given-names:A + REF + surname:Mateos;given-names:MV + 934 + 20739218 + 2010 + Lancet Oncol + ref + 941 + 18580 + Bortezomib, melphalan, and prednisone versuis bortezomib, thalidomide, and prednisone as induction therapy followed by maintenance treatment with bortezomib and thalidomide versus bortezomib and prednisone in elderly patients with untreated multiple myeloma: a randomized trial + + 9606 + Species + + patients + + + MESH:D000069286 + Chemical + + Bortezomib + + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D013792 + Chemical + + thalidomide + + + MESH:D000069286 + Chemical + + bortezomib + + + MESH:D011241 + Chemical + + prednisone + + + + 28 + surname:Rossi;given-names:D + surname:Bringhen;given-names:S + REF + surname:Palumbo;given-names:A + 5101 + 20940200 + 2010 + J Clin Oncol + ref + 5109 + 18858 + Bortezomib-melphalan-prednisone-thalidomide followed by maintenance with bortezomib-thalidomide compared with bortezomib-melphalan-prednisone for initial treatment of multiple myeloma: a randomized controlled trial + + - + Chemical + + melphalan-prednisone-thalidomide + + + MESH:D011241 + Chemical + + prednisone + + + MESH:D008558 + Chemical + + melphalan + + + - + Chemical + + bortezomib-thalidomide + + + MESH:D000069286 + Chemical + + Bortezomib + + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000069286 + Chemical + + bortezomib + + + + 116 + surname:Rossi;given-names:D + surname:Larocca;given-names:A + REF + surname:Bringhen;given-names:S + 4745 + 20807892 + 2010 + Blood + ref + 4753 + 19073 + Efficacy and safety of once- weekly bortezomib in multiple myeloma patients + + MESH:D009101 + Disease + + multiple myeloma + + + MESH:D000069286 + Chemical + + bortezomib + + + + 98 + surname:Zweegman;given-names:S + surname:Victoria;given-names:M + REF + surname:Bringhen;given-names:S + 6 + 2013 + Haematologica + ref + 19149 + Age and organ damage correlate with poor survival in myeloma patients: meta - analysis of 1435 individual patient data from 4 randomiuzed trials + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + MESH:D009101 + Disease + + myeloma + + + + + 4272841 + + 25568587 + surname:Masic;given-names:Izet + 350 + 5 + 2014 + 4272841 + AIM-55-350 + 10.5455/aim.2014.22.350-350 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 350. doi:10.5455/aim.2014.22.350-350 + 350 + Masic I + 0 + The Most Influential Scientists in the Development of the Medical Informatics (2): Morris F. Collen + + + INTRO + title_1 + 100 + MORRIS F. COLLEN (1913-2014) + + + INTRO + paragraph + 129 + Morris F. Collen was born in St. Paul, Minnesota. He attended the University of Minnesota, where he earned a bachelor's degree in electrical engineering in 1935. In 1938 he earned his MD "with distinction" from the School of Medicine and completed a residency in internal medicine at USC/Los Angeles County General Hospital. + + + INTRO + paragraph + 458 + Dr. Morris Collen has had a profound influence, not only on the creation of the field of informatics, but also on health-care delivery and the creation of new models of payment and prevention. Dr. Collen's remarkable career began in 1942 when he was selected by Dr. Sidney Garfield, a surgeon, to join him as an internist in a California group practice. Drs Garfield and Collen subsequently worked with the industrialist Henry Kaiser, who is credited with creating one of the first comprehensive prepaid health plans for both office and hospital care. This led to the establishment of Kaiser Perma-nente in the post-World War II period plus a comprehensive infrastructure of hospitals in the Bay Area near San Francisco and near Portland, Oregon. In the subsequent decades, the Kaiser organization grew to become a nationwide healthcare provider with millions of enrollees. + + 8406 + Gene + 4598 + + Drs + + + + INTRO + paragraph + 1332 + Collen became a nationally recognized authority on the treatment of pneumonia during World War II. His gift for research showed early in his published studies in The Permanente Foundation Medical Bulletin of which he was long-time editor. After two decades as an internist with Kaiser Permanente, his career took a turn into early medical information technology. Morris Collen and his team set to work to automate the 10-year-old multiphasic health screening exam to develop a prototype electronic health record. Within a decade, Dr. Collen accumulated several millions of health checkup data sets on more than a million subjects, creating in the process not only a prototype electronic health record, but also a phenomenal and unique basis for research, and this despite the immaturity of the technology available in the fifties and sixties. For the pursuit of the scientific aspects of his work, Dr. Collen founded the Medical Methods Research Division within Kaiser Permanente in Oakland, to which he added the Division of Technology Assessment in 1979 that he directed until his retirement in 1983, at age 70. + + MESH:D011014 + Disease + + pneumonia + + + + INTRO + paragraph + 2446 + He was elected to membership in the Institute of Medicine of the National Academy of Sciences (1971), and has served in many capacities on many committees of the National Library of Medicine. + + + INTRO + paragraph + 2638 + By the time of his retirement that year, Dr. Collen listed some 150 publications in his scientific output and had held appointments at multiple first-class universities, including Johns Hopkins and Stanford. His work ,,Hospital Information Systems" and ,,Multiphasic Health Testing Services", both became classics. The Morris F. Collen Award is given each year, when appropriate, to pioneers in the field of medical informatics who best exemplify the teaching and practice of Morrie Collen. + + + INTRO + footnote + 3133 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 3170 + REFERENCES + + + REF + surname:Masic;given-names:I + 236 + 2014 + ref + 237 + 3181 + Contributions to the History of Medical Informatics + + + 52 + surname:Morris;given-names:F + surname:Ball;given-names:MJ + REF + surname:Lindberg;given-names:DA + 371 + 3 + 24114266 + 2013 + Methods Inf Med + ref + 373 + 3233 + Collen at 100: A Trib ute to "The Father of Medical Informatics" + + 26150 + Gene + 9230 + + Trib + + + + surname:Masic;given-names:I + surname:Lindberg;given-names:D + surname:Ball;given-names:M + 4 + 1 + 24648612 + 2014 + Acta Inform Med + ref + 22 + REF + doi: 10.5455/aim.2014.22.4-5 + 5 + 3303 + Special Tribute on Morris F. Collen: Charismatic Leader of Medical Informatics + + + + 4272842 + + surname:Cai;given-names:Liuhong + surname:Xing;given-names:Weijie + surname:Tao;given-names:Xin + 25568631 + surname:Zhang;given-names:Erhong + 321 + 5 + 2014 + 4272842 + MSM-26-321 + 10.5455/msm.2014.26.321-323 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 321-3. doi:10.5455/msm.2014.26.321-323 + Intrauterine insemination Male factor infertility Semen analysis + 323 + surname:Zhang;given-names:Bin + Zhang E, Tao X, Xing W, Cai L, Zhang B + 0 + Effect of Sperm Count on Success of Intrauterine Insemination in Couples Diagnosed with Male Factor Infertility + + MESH:D007248 + Disease + + Male Factor Infertility + + + + ABSTRACT + abstract_title_1 + 112 + Objective: + + + ABSTRACT + abstract + 123 + To exam semen parameters in predicting intrauterine insemination (IUI) outcomes in couples with male factor. + + + ABSTRACT + abstract_title_1 + 232 + Study design: + + + ABSTRACT + abstract + 246 + This retrospective study was performed at department of infertility and sexual medicine from September 2007 to February 2014. 307 couples with male factor infertility were included and 672 IUI cycles were analyzed. + + MESH:D007247 + Disease + + infertility + + + MESH:D007247 + Disease + + infertility + + + + ABSTRACT + abstract_title_1 + 461 + Results: + + + ABSTRACT + abstract + 470 + From 672 inseminations performed on 307 couples, there are 27.36% couples get pregnancy (84 out of 307) and the overall pregnancy rate was 12.95% (87 out of 672) of IUI. With the increase of post total progressive sperm count, the clinical pregnancy rate increased. When the initial progressive sperm count was lower than 5*106, there was no pregnant in the IUI cycle. At the end of the third cycle, 85 clinical pregnancies had been achieved (97.70%). + + + ABSTRACT + abstract_title_1 + 922 + Conclusions: + + + ABSTRACT + abstract + 935 + The initial total progressive sperm count lower than 5*106 means the poor outcome of IUI in the infertile couples with male factor. If the infertile couples with male factor don't get pregnancy after three IUI cycles, the couples should receive re-assessment or other artificial reproductive technology. + + + INTRO + title_1 + 1241 + 1. INTRODUCTION + + + INTRO + paragraph + 1257 + Intrauterine insemination (IUI) is a widely used fertility treatment for couples with infertility and is a simple, non-invasive, and a cost-effective technique. IUI is often suggested to infertile couples in which the woman has at least one permeable fallopian tube. This method is indicated in cases of cervical infertility, male factor infertility, anovulation, endometriosis, and unexplained infertility. + + 9606 + Species + + woman + + + MESH:D007247 + Disease + + cervical infertility + + + MESH:D004715 + Disease + + endometriosis + + + MESH:D007247 + Disease + + unexplained infertility + + + MESH:D007247 + Disease + + infertility + + + MESH:D007247 + Disease + + infertility + + + + INTRO + paragraph + 1665 + There are certain variables that are currently known to be predictive of IUI success, most of which relate to the female partner including follicle number, endometrial thickness, duration of infertility, and sperm motility. Pregnancy rate after IUI differ between studies according to patient selection criteria, the presence of various infertility factors, ovarian stimulation methods, number of cycles performed, different sperm parameters and preparation technique. + + 9606 + Species + + patient + + + MESH:D007247 + Disease + + infertility + + + MESH:D007247 + Disease + + infertility factors + + + + INTRO + paragraph + 2134 + Semen analysis is the first step to accurately diagnose male infertility. Sperm count, sperm motility and the percentage of sperm with normal morphology are the main criteria for the quality of semen and number of motile sperm and normal morphology have been shown to correlate with IUI outcome. Others found that male factors were not found to correlate with the treatment outcome. The aim of this retrospective study is to exam the use of semen parameters in predicting IUI outcomes in couples diagnosed with male factor infertility. + + MESH:D007247 + Disease + + infertility + + + MESH:D007248 + Disease + + male infertility + + + + METHODS + title_1 + 2670 + 2. MATERIAL AND METHODS + + + METHODS + paragraph + 2694 + In this retrospective cohort study, all couples undergoing intrauterine insemination at department of infertility and sexual medicine from September 2007 to February 2014 were retrospectively enrolled into our database for evaluation of a diagnosis of male factor infertility. Before each course of treatment, infertile couples underwent the following tests: hysterosalpingography, hormone concentrations, semen analysis and postcoital test. Male factor infertility was diagnosed if the male had at least two pre-treatment semen analyses which were abnormal. In this analysis 672 IUI were performed in couples which fit the criteria listed above for male factor infertility. Clinical features of couples, pre and post processing semen analysis parameters at the time of IUI were included in the analyses. + + MESH:D007247 + Disease + + infertility + + + MESH:D007248 + Disease + + male factor infertility + + + MESH:D007247 + Disease + + infertility + + + MESH:D007247 + Disease + + infertility + + + + METHODS + paragraph + 3499 + Patients who underwent natural cycles have regular menstrual cycle and ovulation. Clomiphene citrate (50 or 100 mg daily) and letrozole (2.5 mg or 5 mg daily) were administered orally for 5 days stating on cycle day 3 to 5. Gonadotropin injections were performed daily starting on cycle day 3 to 5 and titrated to develop 2 to 3 mature follicles. When the lead follicle reached at least 16 mm (usually 18 mm) in diameter 10000IU of human chorionic gonadotropin (hCG, livzon pharmaceutical Co.Ltd, China) subcutaneously to induce ovulation. Washed sperm was inseminated 24-36 hours after hCG injection according the LH level on HCG day. All patients were monitored with either ultrasound or both serum LH, estradiol and progesterone levels. + + 93659 + Gene + 37338 + + hCG + + + 93659 + Gene + 37338 + + HCG + + + 93659 + Gene + 37338 + + hCG + + + 9606 + Species + + Patients + + + 9606 + Species + + human + + + 9606 + Species + + patients + + + MESH:D011374 + Chemical + + progesterone + + + MESH:D002996 + Chemical + + Clomiphene citrate + + + MESH:D000077289 + Chemical + + letrozole + + + MESH:D004958 + Chemical + + estradiol + + + MESH:D007986 + Chemical + + LH + + + + METHODS + paragraph + 4239 + For the purpose of semen collection, individuals were asked to refrain from ejaculating for two to four days prior to collection of the specimen. Specimens were produced with masturbation in a collection room adjacent to the laboratory. Freshly ejaculated sperm was allowed to liquefy before initial semen analysis. Liquefied semen was thoroughly mixed before an aliquot was placed on a standard count slide for the pre-processing analysis. The samples were assessed by CASA system (Sperm Class Analyzer, V4.0.0, Spain) and at least four random fields were evaluated for each analysis. + + 1446 + Gene + 48054 + + CASA + + + + METHODS + paragraph + 4825 + After place semen sample in a discontinuous density (40%-80%) gradient column (SpermGrad TM, SpermRinse, Vitrolife Sweden AB, Sweden), the gradient was centrifuged for 20 min at 400g and subsequently, the 40% layer and the seminal plasma fraction were removed from the test tube, leaving the 80% layer undisturbed. Approximately 6-8 ml of sperm-washing medium was added to the 80% layer and centrifuged for 5 min at 400g. The sperm pellet was then reconstituted to approximately 0.5 ml. The analysis of an aliquot of the processed sample was performed as previously. The insemination was performed in a sterile fashion, using a flexible plastic catheter with the patient in the dorsal lithotomy position. The patient did not assume a prone position for at least thirty minutes after the end of the insemination. + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + + METHODS + paragraph + 5639 + Serum beta-human chorionic gonadotropin (beta-hCG) levels were analyzed 14 to 17 days after IUI to determine pregnancy status. A level greater than 5 mIU/ml was considered positive for pregnancy including biochemical pregnancy and clinical pregnancy. Clinical pregnancies were defined as those with a gestational sac on ultrasound. In this article, we use clinical pregnancy rate to analyze. + + 9606 + Species + + human + + + 93659 + Gene + 37338 + + hCG + + + + METHODS + paragraph + 6031 + Statistical methods + + + METHODS + paragraph + 6051 + All statistical analyses were done using the statistical package for social sciences 13.0 (SPSS, Inc, Chicago, IL). Results are reported as mean value+-standard deviation (SD). Statistical significance was accepted as a two-sided P<0.05. + + + RESULTS + title_1 + 6289 + 3. RESULTS + + + RESULTS + paragraph + 6300 + From 672 IUI cycles performed on 307 couples, there are 27.36% couples get pregnancy (84 out of 307) and the overall pregnancy rate was 12.95% (87 out of 672) of inseminations. At the primary infertile group pregnancy rate was found as 14.29% (68 out of 475). In a group of patients with secondary infertility pregnancy rate was observed as 9.69% (19 out of 197). There were more dominant preovulatory follicles (>= 16mm) in pregnant group than that in not pregnant groups (1.72+-0.89 vs 1.50+-0.77, P<0.05). The mean age of couple, duration of infertility were compared between two groups. When comparing the semen analysis parameters between the two outcome groups, we found that the initial motile and progressive sperm count and post motile and progressive sperm count was higher in the pregnant and not pregnant groups, whereas the post motile sperm count has no significant difference between two groups (Table 1). + + 9606 + Species + + patients + + + MESH:D007247 + Disease + + infertility + + + MESH:D007247 + Disease + + infertility + + + + TABLE + T1.xml + T1 + table_caption + 7222 + Comparison of the baseline characteristics and sperm parameters of subjects + + + RESULTS + paragraph + 7298 + Table 2 shows that with the increase of post total progressive sperm count, the clinical pregnancy rate increased. There was no pregnant in the cycles which the pre total progressive sperm count lower than 5*106. The clinical pregnancy rate per cycle was 12.95%. At the end of the third cycle, 85 clinical pregnancies had been achieved (97.70%). There was no clinical pregnancies occurred in these cases after five attempts (Table 3). + + + TABLE + T2.xml + T2 + table_caption + 7733 + Initial and post total progressive sperm count with pregnancy rate + + + TABLE + T3.xml + T3 + table_caption + 7800 + Pregnancy rate in different number of IUI cycles + + + DISCUSS + title_1 + 7849 + 4. DISCUSSION + + + DISCUSS + paragraph + 7863 + Intrauterine insemination using the husband's sperm is commonly performed to overcome male factor problems, as well as to enhance the probability of conception in various other infertility conditions. The pregnancy rate depends on sperm parameters, female factors, and the number of dominant preovulatory follicles. In Stephanie M. Luco's study, the total pregnancy rate observed is 5.3% in 356 IUI cycles. In Ahmed Badawy's study, Seventy-nine clinical pregnancies followed 714 IUI cycles, for a clinical pregnancy rate per cycle of 11.06% and a clinical pregnancy rate per couple of 20.1%. The total clinical pregnancy rate observed in this study, 12.95% per cycle, is considerably higher than Stephanie M. Luco's study and is comparable with Ahmed Badawy's study. + + MESH:D007247 + Disease + + infertility conditions + + + + DISCUSS + paragraph + 8630 + Miller et al. reported a pregnancy rate per cycle of 12.4% when the total number of collected motile spermatozoa (TMS) is over 20 million, compared with 7.4% when it is between 10 and 20 million. For a value under 10 million, he suggests steering the couple towards IVF. In the same sense, Dickey et al. and Van Voorhis et al. reported the best pregnancy rates when more than 10 million motile spermatozoa were selected. It has been suggested that the number of motile spermatozoa inseminated is a potential predictive factor. In our study, when comparing the semen analysis parameters between the two outcome groups, we found that the initial and post progressive sperm count and the initial motile sperm count was higher in the pregnant group than not pregnant group. These results mean that the semen parameters were related to the outcome of IUI in the current study. In this study, when initial total progressive sperm count was >=5x106, the pregnancy rate was higher than in the group with initial total progressive sperm count <5x106 and the pregnancy rate was compared among the groups with initial total progressive sperm count 5-9.9x106, 10-19.9x106 and >=20x106 (12.73%, 11.11%, and 13.68%, respectively (Table 2). There was no pregnant in the cycles which the initial total progressive sperm count was lower than 5*106, so we should told these infertile couples the poor outcome of IUI and suggested them receive in vitro fertilization (IVF). + + MESH:C537182 + Disease + + IVF + + + Disease + + TMS + + + MESH:D000072660 + Disease + + motile spermatozoa + + + MESH:D000072660 + Disease + + motile spermatozoa + + + MESH:D000072660 + Disease + + motile spermatozoa + + + + DISCUSS + paragraph + 10096 + The findings in Stephanie Marticle indicated that post processing semen analysis parameters are not more predictive of pregnancy than are preprocessing semen analysis results in couples with male factor infertility. Tan et al. found that post washed total progressively motile sperm count (TPMSC) is an independent predictor of pregnancy test result and TPMSC of half million or greater is adequate to achieve statistically similar pregnancy test results after non-donor IUI cycles. The predicted odd of positive pregnancy result is statistically significantly higher when TPMSC is >0.51 million compared to the TPMSC of <0.51 million (OR = 1.68, 95 % CI: 1.04-2.71). Ok EK found that an average TPMSC of 10x106 may be a useful threshold value for IUI success. In this study, there was no pregnancy occurred when TPMSC was <4 million (0/20). Our results also showed that with the increase of post total progressive sperm count, the clinical pregnancy rate increased, similar with the results of those articles (Table 2). + + 9606 + Species + + donor + + + MESH:D007247 + Disease + + infertility + + + + DISCUSS + paragraph + 11118 + In the present study, 85 clinical pregnancies had been achieved (97.70%) at the end of the third cycle (Table 3). Plosker and Amato advised infertile couples to received IVF after three unsuccessful IUI. Nuoja-Huttunen et al noted that the highest pregnancy rate in 811 cycles (18%) was seen during the first cycle and that 97% of all pregnancies result from the first four cycles. In cases of unexplained infertility, Aboulghar et al found a cumulative pregnancy rate of 39.2% after three cycles and 48.5% after six cycles in a study of 1112 IUI cycles (16.4% per cycle). At present, it is generally admitted that IUI should be limited to four or six cycles and that IVF should be performed. Our results supported the opinion that three inseminations prior to IVF treatments have been usually performed in couples where the cause of infertility was unexplained, due to unilateral tubal factor, mild endometriosis or mild male factor infertility. + + MESH:D007247 + Disease + + infertility + + + MESH:C537182 + Disease + + IVF + + + MESH:D007247 + Disease + + infertility + + + MESH:C537182 + Disease + + IVF + + + MESH:D004715 + Disease + + endometriosis + + + MESH:D007247 + Disease + + unexplained infertility + + + MESH:C537182 + Disease + + IVF + + + + CONCL + title_1 + 12065 + 5. CONCLUSION + + + CONCL + paragraph + 12079 + IUI is a powerful method to deal with infertile couple with male factors. The initial total progressive sperm count lower than 5*106 means the poor outcome of IUI in the infertile couples with male factor. If the couples with male factor infertility don't get pregnancy after three IUI cycles, the couple should receive re-assessment or other artificial reproductive technology. + + MESH:D007247 + Disease + + infertility + + + + CONCL + footnote + 12460 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 12497 + REFERENCES + + + surname:Al-Inany;given-names:H + surname:Cohlen;given-names:BJ + REF + surname:Cantineau;given-names:AE + CD001502 + 15266448 + 2004 + Cochrane Database Syst Rev + ref + 12508 + Intrauterine insemination versus fallopian tube sperm perfusion for non tubal infertility + + MESH:D005184 + Disease + + Intrauterine insemination versus fallopian tube + + + MESH:D007247 + Disease + + tubal infertility + + + + surname:Kasraie;given-names:JL + surname:Thompson;given-names:KA + surname:Amissah-Arthur;given-names:JB + surname:Tomlison;given-names:MJ + 1892 + 9 + 8921060 + 1996 + Hum Reprod + ref + 11 + REF + 1896 + surname:Bentick;given-names:B + 12598 + Prognostic indicators for intrauterine insemination (IUI): statistical model for IUI success + + + surname:Hassiakos;given-names:D + surname:Makrakis;given-names:E + surname:Pantos;given-names:K + surname:Grigoriou;given-names:O + 773 + 15749516 + 2005 + Fertil Steril + ref + 83 + REF + 775 + surname:Creatsas;given-names:G + surname:Konidaris;given-names:S + 12691 + Impact of isolated teratozoospermia on the outcome of intrauterine insemination + + MESH:D000072660 + Disease + + teratozoospermia + + + + 91 + surname:Eltotongy;given-names:M + surname:Elnashar;given-names:A + REF + surname:Badawy;given-names:A + 777 + 18304534 + 2009 + Fertil Steril + ref + 781 + 12771 + Effect of sperm morphology and number on success of intrauterine insemination + + + 93 + surname:Grenier;given-names:N + surname:Heraud;given-names:MH + REF + surname:Merviel;given-names:P + 79 + 1 + 18996517 + 2010 + Fertil Steril + ref + 88 + 12849 + Predictive factors for pregnancy after intrauterine insemination (IUI): an analysis of 1038 cycles and a review of the literature + + + 45 + surname:Amiri;given-names:I + REF + surname:Farimani;given-names:M + 101 + 2 + 2012 + Acta Med Iran + ref + 106 + 12979 + Analysis of prognostic factors for successful outcome in patients undergoing intrauterine insemination + + 9606 + Species + + patients + + + + 19 + surname:Dorion;given-names:A + surname:Albert;given-names:M + REF + surname:Wainer;given-names:R + 2060 + 15243004 + 2004 + Hum Reprod + ref + 2065 + 13082 + Influence of the number of motile spermatozoa inseminated and of their morphology on the success of intrauterine insemination + + MESH:D000072660 + Disease + + motile spermatozoa + + + + surname:Barry;given-names:Behr + 179 + surname:Chioma;given-names:Agbo + surname:Luco + REF + surname:Stephanie;given-names:M + 159 + 24965998 + 2014 + European Journal of Obstetrics & Gynecology and Reproductive Biology + ref + 162 + 13208 + The evaluation of pre and post processing semen analysis parameters at the time of intrauterine insemination in couples diagnosed with male factor infertility and pregnancy rates based on stimulation agent. A retrospective cohort study + + MESH:D007247 + Disease + + infertility + + + + 60 + surname:Randolph;given-names:JF + surname:Smith;given-names:GD + REF + surname:Miller;given-names:DC + 497 + 12350496 + 2002 + Urology + ref + 501 + 13444 + Processed total motile sperm count correlates with pregnancy outcome afterintra-uterine insemination + + + 71 + surname:Taylor;given-names:SN + surname:Lu;given-names:PY + REF + surname:Dickey;given-names:RP + 684 + 10202879 + 1999 + Fertil Steril + ref + 689 + 13545 + Comparison of the sperm qualitynecessary for successful intra-uterine insemination with World HealthOrganization threshold values for normal sperm + + + 75 + surname:Sparks;given-names:AE + surname:Barnett;given-names:M + REF + surname:Van Voorhis;given-names:BJ + 661 + 11287015 + 2001 + Fertil Steril + ref + 668 + 13692 + Effect of the total motile sperm count on the efficacy and cost-effectiveness of intrauterine insemination and in vitro fertilization + + MESH:D005317 + Disease + + cost-effectiveness of intrauterine insemination + + + + 31 + surname:Carr;given-names:BR + surname:Ha;given-names:T + REF + surname:Tan;given-names:O + 1147 + 9 + 25106939 + 2014 + J Assist Reprod Genet + ref + 1153 + 13826 + Predictive value of postwashed total progressively motile sperm count using CASA estimates in 6871 non-donor intrauterine insemination cycles + + 1446 + Gene + 48054 + + CASA + + + 9606 + Species + + donor + + + + 14 + surname:Okyay;given-names:RE + surname:Dogan;given-names:OE + REF + surname:Ok;given-names:EK + 142 + 3 + 24592093 + 2013 + J Turk Ger Gynecol Assoc + ref + 145 + 13968 + The effect of post-wash total progressive motile sperm count and semen volume on pregnancy outcomes in intrauterine insemination cycles: a retrospective study + + + 9 + surname:Amato;given-names:P + REF + surname:Plosker;given-names:S + 2014 + 7868666 + 1994 + Hum Reprod + ref + 2021 + 14127 + Predicting and optimizing success in an intra-uterine stimulation program + + + surname:Tuomivaara;given-names:L + surname:Bloigu;given-names:R + surname:Tomas;given-names:C + surname:Nuojua-Huttunen;given-names:S + 698 + 10221698 + 1999 + Hum Reprod + ref + 14 + REF + 703 + surname:Martikainen;given-names:H + 14201 + Intrauterine insemination treatment in subfertility: an analysis of factors affecting outcome + + + 75 + surname:Serour;given-names:G + surname:Mansour;given-names:R + REF + surname:Aboulghar;given-names:M + 88 + 11163821 + 2001 + Fertil Steril + ref + 91 + 14295 + Controlled ovarian hyperstimulation and intrauterine insemination for treatment of unexplained infertility should be limited to a maximum of three trials + + MESH:D007247 + Disease + + unexplained infertility + + + + 15 + REF + 265 + 3 + 19240042 + 2009 + Hum Reprod Update + ref + 277 + 14449 + The ESHRE Capri Workshop Group. Intrauterine insemination + + + + 4272843 + + surname:Karami;given-names:Mahtab + 25568582 + surname:Barzekar;given-names:Hosein + 325 + 5 + 2014 + 4272843 + AIM-22-325 + 10.5455/aim.2014.22.325-328 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 325-8. doi:10.5455/aim.2014.22.325-328 + Organizational factors Information technology Managers Implementation + 328 + Barzekar H, Karami M + 0 + Organizational Factors that Affect the Implementation of Information Technology: Perspectives of Middle Managers in Iran + + + ABSTRACT + abstract_title_1 + 121 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 130 + Objective: + + + ABSTRACT + abstract + 141 + to examine the organizational factors affecting the application of information technology in hospitals. Since the organizational factors are one of the most important determinants of successful projects, by understanding their impact and identifying them it can help planning a systematic IT implementation. + + + ABSTRACT + abstract_title_1 + 449 + Methods: + + + ABSTRACT + abstract + 458 + In this cross-sectional descriptive study 110 middle managers were chosen from teaching hospitals. Structured questionnaire was used for the data collection. + + + ABSTRACT + abstract_title_1 + 616 + Results: + + + ABSTRACT + abstract + 625 + There was a significant relationship between organization resource, organizational knowledge, process, management structure and values and goals with implementation of information technology. + + + ABSTRACT + abstract_title_1 + 817 + Conclusion: + + + ABSTRACT + abstract + 829 + Findings showed that organizational factors had a considerable impact on implementation of information technology. Top managers must consider the important aspects of effective organizational factors. + + + INTRO + title_1 + 1030 + 1. INTRODUCTION + + + INTRO + paragraph + 1046 + Nowadays, organizations perform in an environment which the economy, politics and society constantly change. With such aggressive perspective, organizations should be adaptable in order to maintain the competitive market conditions, increase productivity and re-configure due to changing workforce, the global business environment and e-commerce development. Studies indicate that one of the most important reasons that differentiated organizations from each other is the degree of information technology application in their organizational activities. Accordingly, the organizations that use widespread and optimized information technology have sustainable competitive advantages, and in the view of stakeholders are more distinguished than the other organizations. Based on a research conducted between 100 companies between 1980 and 1995 the average cost to implement the changes has been estimated to be a million dollars. Since after 1980, almost %50 of the investments within the organizations are spent on development and deployment of information technology. It is believed that IT can increase the capabilities of organizations. Furthermore, Health system is not excluded from these changes, and many countries utilize Information Technology in order to enhance the health level and to improve the outcomes. Research shows that application of information technology in different countries brings various benefits to the health systems such as improving service delivery, reduction of medical errors, it supports the healthcare personnel in inpatient and outpatient units, enhances the effectiveness of healthcare (by reduction of waiting times for the patients and improving patient care). + + 9606 + Species + + outpatient + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + + INTRO + paragraph + 2745 + Successful implementation and use of IT in health care systems and any other system is influenced by various factors like economic, political, social, cultural, etc. Identifying the factors resulting in increased use of information technology leads to enhanced accuracy and it accelerates the application of this technology. Attention to organizational factors is one factor in the application of information technology that plays an important role in health systems. Nowadays organizations live in an active and dynamic environment which is influenced by internal and external factors of the organization. Some external factors affecting the performance of organizations include economic, political, social, cultural, etc. Implementation of Information Technology in Health Care systems often requires changes in work duties and processes either simultaneously or before application of the new technology, However, studies show that almost %70 of IT implementation projects failed. Mostly, failure to utilize and incompatibility with the needs of IT users were the reasons that IT projects failed. The present study sought to examine the organizational factors affecting the application of information technology in hospitals. Since the organizational factors are one of the most important determinants of successful projects, by understanding their impact and identifying them it can help planning a systematic IT implementation. + + + METHODS + title_1 + 4177 + 2. METHODS + + + METHODS + paragraph + 4188 + This descriptive-analytic study is a cross-sectional kind and it is done in the second half of 2012. The study population included middle managers of the teaching hospitals of Tehran University of Medical Sciences (hospital administrator, nursing services director, staffing and recruiting office, finance, medical records, laboratory and pathology, radiology and pharmacy). Total statistical population was 110 middle managers that 89 people answered to the questionnaire. The data collection was done by a questionnaire that its validity was confirmed by the experts and its reliability was examined by Cronbach's alpha coefficient of 20 samples that were selected randomly from the population. The questionnaire included an introduction at the beginning in order to show the goals of the research. The first section of the questionnaire was about demographic information of the population like age, sex, name of hospital, length of occupation, educational degree and organizational position. The second part consists of nine questions related to computer use. In the third part there are questions related to organizational factors and is composed of 5 parts: the first section includes organizational resources. The second section is regarding the organizational knowledge. The third section is about organizational processes. The fourth one is about managerial structure and the last section is about values and goal of the organization. The scores given to the responses are according to a five-option Likert-type ranking number one as "totally disagree" to number five as "totally agree". Overall, the degree of importance for each item was made and calculated. In order to determine a relationship between each of the variables with information technology the test of Pearson's correlation coefficient, independent T and analysis of one-way Variance was used and then multiple regressions was used to study all factors' effect on information technology usage. Also the data were analyzed using SPSS 16 software. At the beginning of the research the ethical consideration authorization was obtained from the Deputy office of research in the university. Afterwards, the goal of the research was explained and people were assured that their information stays confidential and it is only for the sake of doing the research. + + 9606 + Species + + people + + + 9606 + Species + + people + + + + RESULTS + title_1 + 6526 + 3. RESULTS + + + RESULTS + paragraph + 6537 + From 110 questionnaires distributed, 89 questionnaires were returned that %37.1 was woman and %62.9 was men. The average age of the people participating in the study was 42+-15.6 years old. %56.2 of the participants had a bachelor's degree, %12.4 had master's degree, %6.7 had a PhD and %24.7 was physicians. The average length of their occupation was 15+-5.91 and more than %83 of them had more than 5 years working record. %82 of the participants used computer several times a day. + + 9606 + Species + + woman + + + 9606 + Species + + men + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + + RESULTS + paragraph + 7021 + As shown in Table 1 there is a meaningful difference regarding the sex of the participants in case of average usage of computer (t= -2.815, p<0.05) average scores gained from the usage of computer by men (33.58) and women (37.45) indicates this difference. Nevertheless, Variance analysis test shows that between the age of the participants with usage of computer (F=0.718, and education with usage of computer (F=1.613, there is no meaningful difference. + + 9606 + Species + + participants + + + 9606 + Species + + men + + + 9606 + Species + + women + + + 9606 + Species + + participants + + + + TABLE + T1.xml + T1 + table_caption + 7477 + Middle managers use of computer according to age, sex and education + + + RESULTS + paragraph + 7545 + In table 2 the results of Pearson correlation test is provided about the relationship of resources of the organizations, organizational knowledge, processes, managerial structure, values and goals with use of computer. It can be seen that between each of these variables and use of computer there is a meaningful relationship: resources of the organizations (p<0.01, r=0.297), organizational knowledge (p<0.01, r=0.297), processes (p<0.01, r=0.409), managerial structure (p<0.01, r=0.789), values and goals (p<0.05, r=0.246). + + + TABLE + T2.xml + T2 + table_caption + 8071 + Correlation coefficient of middle managers' attitude toward each of the variables + + + RESULTS + paragraph + 8155 + In table 3 Linear Regression was used for mass observation of the organizational factors about Information technology application. The results of the regression test showed a meaningful relationship between managerial structure and attitude (p<0.001, beta= 0.760), resources of the organizations and attitude (p<0.05, beta=0.153); in this regression model the affect of the variables decreased and the only variables remained are managerial structure and the resources of the organization. + + + TABLE + T3.xml + T3 + table_caption + 8645 + Linear Regression for mass observation of the organizational factors associated with Information technology application + + + DISCUSS + title_1 + 8765 + 4. DISCUSSION + + + DISCUSS + paragraph + 8779 + Information Technology application in health systems is associated with a variety of factors. Identifying these factors will result in faster implementation of the technology since they play a significant role in information technology application in health systems. Some factors to name are economic, social and cultural factors. Our goal in this study is to find and review the organizational factors affecting the information technology implementation in the teaching hospitals of Tehran University of Medical Sciences. + + + DISCUSS + paragraph + 9302 + This research shows that there is a significant and meaningful relationship between the sexes of users and using computer systems; thus, female middle manager show more utilization of the computer systems than male managers and this finding is consistent with the finding of Al-Gahtani, 2004 and Meade, et al., 2009. In this research between the age and the education level of the participants with computer usage system did not show any meaningful relationship, while in the studies of Marchewka and Al-Gahtani it showed a significant relationship. + + 9606 + Species + + participants + + + + DISCUSS + paragraph + 9852 + In a Level of %95 between organizational resources and IT implementation there is a significant relationship which showed a correlation intensity of r= 0.297. Regarding the relationship between these two variables it can be said that due to some facts like accessible financial and human resources, enough budget for the application of new technologies, strategic planning of the organizations' resources that can lead the managers to realize the mission and targets of the organizational resources, the attitude of the users might be influenced and it is concluded that the more available resources is provided for the users, the more tendency they show in application of Information technology. + + 9606 + Species + + human + + + + DISCUSS + paragraph + 10551 + The above theory is confirmed due to a significant relationship between the users' attitude toward IT application and organizational knowledge with a level of %99 and a correlation coefficient of (r= 0.297). Therefore, it can be inferred however the organizational knowledge is not in a technology oriented category, information technology application is a necessity for knowledge management projects to succeed. Hence some organizations seek knowledge management systems for executing their organizational knowledge management projects. In this way they might become confused because of lack of deep and correct understanding of capabilities and abilities of the organizations and susceptible areas for knowledge technology products. Considering the main three factors of knowledge management (individuals, contents, processes) an organization should identify the proper technology for itself. Accordingly, the tools for knowledge management should be determined based on the needs of the organization. Thus, it can be generally stated that when organizational knowledge in an organizations is used properly, people tend to use more of information technology because it facilitates the access and management of this knowledge. These findings are in agreement with previous researches like Armenakis&Bernerth in 2007 and Bartunek& Rousseau in 2006. + + 9606 + Species + + people + + + + DISCUSS + paragraph + 11902 + The relationship between processes and the attitude toward application of information technology is confirmed with a level of %99 and a correlation coefficient of (r= 0.409) that approves the above theory. Therefore, it can be concluded that in organizations in which processes are shorter and easier beside shorter organizational communication and fewer organizational levels, they show more flexibility in case of facing with environmental and technological changes. In other words, staff in organizations with horizontal structure can communicate with managers easier and they participate in decision makings more effectively. So, organizations with the least division of responsibilities and informal processes have an organic structure in which people tend to use information technology more. This finding is also in consistent with Armenakis Schraeder & Tsiknakis &Kouroubali, 2009 and Yu li H, P, 2009. + + 9606 + Species + + people + + + + DISCUSS + paragraph + 12812 + Between managerial structure and attitude toward using IT there is a correlation of (r=0.789) with a level of 99%. However management is similar to leadership, on the whole in any organization they have similar strategies and they can be different in some specifications. It is clear that although all managers cope with leading a small or large group for reaching determined goals, this trend can be performed through various methods which each of them counts as a specific kind of management style. Results indicate that the managerial structure and leadership method that top managers practice can greatly effective in the level of using IT as one of high technologies of nowadays. In turn, the results of this study is compatible with the previous studies of MarchewkaLiu 2007 and Jimmieson 2008. + + + DISCUSS + paragraph + 13613 + The relationship between values and goals and the attitude toward application of information technology is confirmed with a level of %95 and a correlation coefficient of (r= 0.246) in this regard, it can be stated that goals and strategies are as the main framework of planning. Any plan requires a goal which explains the philosophy of existence and it also elaborates the needs, the objectives and the final results in order to support the subordinate goals of the organization. In fact, the goals create a kind of network or a hierarchy system. In addition, just like the need to know the tradition for a better understanding of organizational behavior, noting the values would be another aspect that can affect people's behavior in the organizations. Values have great influence on career decision makings, directions, behaviors, relationships and the attitudes of the staffs. According to the findings in this research, when implementation and application of information technology becomes a priority in the organizations' goals, there should be a strategy and it also ought to be valued in order to be relatively effective in application of information technology. This is in accordance with Simpson & Dansereau, 2007. + + 9606 + Species + + people + + + + DISCUSS + paragraph + 14840 + Among the variables included in the equation, managerial structure (beta= 0.760) and then human resources (beta=0.153) have the strongest affect on information technology application, so it is concluded that senior managers should focus more attention to these variables since they are the main important variables. + + 9606 + Species + + human + + + + CONCL + title_1 + 15156 + 5. CONCLUSION + + + CONCL + paragraph + 15170 + In order to improve the measures to successful implementation of information technology in health systems, identification of effective and various factors such as social factors, humanitarian factors, organizational culture, simplification of occupational relationships, enhancing the communication and reduction of bureaucracy, designing a strategic plan for implementation of information technology in the organization for its correct application is of a great importance. + + + CONCL + footnote + 15645 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 15682 + REFERENCES + + + surname:van de Wijngaert;given-names:L + REF + surname:Bouwman;given-names:H + 2005 + ref + 15693 + Information and communication technology in organizations: adoption, implementation, use and effects + + + 78 + surname:Kouroubali;given-names:A + REF + surname:Tsiknakis;given-names:M + 39 + 1 + 18723389 + 2009 + International Journal of Medical Informatics + ref + 52 + 15794 + Organizational factors affecting successful adoption of innovative eHealth services: A case study employing the FITT framework + + MESH:C043685 + Chemical + + FITT + + + + REF + surname:Arlington;given-names:V + 2008 + The global information economy + ref + 15921 + World Information Technology and Service Alliance + + + 45 + REF + surname:Benamati;given-names:J + 833 + 4 + 2008 + Decision Support Systems + ref + 844 + 15971 + Decision support systems unfrastructure: The root problems of the management of changing IT + + + surname:Maglione;given-names:M + surname:Wu;given-names:S + surname:Wang;given-names:J + surname:Chaudhry;given-names:B + 742 + 10 + 16702590 + 2006 + Annals of internal medicine + ref + 144 + REF + surname:Roth;given-names:E + surname:Mojica;given-names:W + 16063 + Systematic review: impact of health information technology on quality, efficiency, and costs of medical care + + + surname:George;given-names:B + surname:Placide;given-names:PN + surname:Claude;given-names:S + REF + surname:Guy;given-names:P + 6 + Implementation Science + ref + 16172 + Clinicians' perceptions of organizational readiness for change in the context of clinical information system projects: insights from two cross-sectional surveys + + + surname:Harris;given-names:S + 43 + surname:Feild;given-names:H + surname:Armenakis;given-names:A + REF + surname:Holt;given-names:D + 232 + 2 + 2007 + The Journal of applied behavioral science + ref + 16335 + Readiness for organizational change: The systematic development of a scale + + + 78 + surname:Speedie;given-names:SM + surname:Pannarunothai;given-names:S + REF + surname:Kijsanayotin;given-names:B + 404 + 6 + 19196548 + 2009 + International Journal of Medical Informatics + ref + 416 + 16410 + Factors influencing health information technology adoption in Thailand's community health centers: applying the UTAUT model + + + 15 + REF + surname:Oreg;given-names:S + 73 + 1 + 2006 + European Journal of Work and Organizational Psychology + ref + 101 + 16534 + Personality, context, and resistance to organizational change + + + 76 + REF + surname:Anderson;given-names:J + 480 + 5 + 17064955 + 2007 + International Journal of Medical Informatics + ref + 483 + 16596 + Social, ethical and legal barriers to e-health + + + 7 + REF + surname:Al-Gahtani;given-names:S + 5 + 1 + 2004 + Journal of Global Information Technology Management + ref + 29 + 16643 + Computer technology acceptance success factors in Saudi Arabia: an exploratory study + + + 78 + surname:Boland;given-names:M + surname:Buckley;given-names:D + REF + surname:Meade;given-names:B + 551 + 8 + 19375381 + 2009 + International Journal of Medical Informatics + ref + 558 + 16728 + What factors affect the use of electronic patient records by Irish GPs? + + 9606 + Species + + patient + + + + 8 + surname:Morris;given-names:M + surname:Davis;given-names:F + REF + surname:Venkatesh;given-names:V + 267 + 4 + 2007 + Journal of the Association for Information Systems + ref + 286 + 16800 + Dead or alive? 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Only those lateral cephalograms were selected where Point A was easily identified. Landmarks: Sella (S), Nasion (N), Point A and three substitution points Y, L, X were traced. Angles formed by SN with Point A (Angle SNA) and three substitution points (Angle SNY, SNX, SNL) were measured. Correlation of angle SNA with angles SNY, SNX and SNL were derived. Statistical analysis used + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + + ABSTRACT + abstract_title_1 + 981 + Results: + + + ABSTRACT + abstract + 990 + Mean and standard deviation for Angles SNA, SNY, SNL and SNX were calculated individually for males and females. 'T' Test was applied to determine statistical significance for all the parameters i.e Age, Angles SNA, SNY, SNL and SNX respectively. Karl Pearson correlation coefficient was carried out to determine the statistical significant correlation for Angle SNA with SNY, SNL and SNX. Results: A mean value of 82.8 +-1.9 , 83.1 +-1.8 , 78.3 +-2.9 and 78.7 +-2.7 for Angle's SNA, SNY, SNL and SNX respectively was observed. A statistically significant correlation was observed between angles SNA & SNY, SNL, SNX & strong positive correlation was observed with angle SNY. Conclusions: We conclude that Point Y is the most nearing maxillary apical base landmark to Point A. Hence maxillary apical base landmark can be substituted by Point Y where identification of point A is not obvious. + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + INTRO + title_1 + 1899 + 1. INTRODUCTION + + + INTRO + paragraph + 1915 + Cephalometric assessment of patients is an essential adjunct to achieve an accurate orthodontic diagnosis aiding for comprehensive orthodontic treatment planning. The extreme range or radiolucency between bone and soft tissues makes it impossible to locate consistently all landmarks on routine radiographs. Studies regarding the reliability of cephalometric landmarks have been differentiated by (a) differences between two films of the same subject, (b) observed differences in locating the points, and (c) variations in measuring the distance between two marked points. The factors influencing accurate identification were quoted as distinctness of structural detail, noise from adjacent structures due to superimposition of conflicting anatomic details, and conceptual judgment, a factor which is largely based on the past experience and radiological knowledge of the observer. In spite of improved techniques, occasionally certain landmarks are still difficult to locate, among them Point A or Subspinale is one such landmark. Point A is a midline point whose relationship to the anterior teeth in a lateral head film may be influenced by head position. Almost all cephalometric analysis such as Steiner's, Down's, Wit's Appraisal, Mc Namara to name a few use point A or the NA plane as a reference point or plane. Because of difficulty in locating point A various authors i.e Van der Linden, Jarabak and Fizzel, Jacobson R. and Jacobson A have given different substitutions for Point A. The aim of this study was to identify the most nearing alternate maxillary apical base landmark from pre-existing cephalometric points given by different authors. + + 9606 + Species + + patients + + + + METHODS + title_1 + 3571 + 2. MATERIAL AND METHODS + + + METHODS + paragraph + 3595 + The study was started after obtaining the approval from the Ethical Committee. A cross-sectional study was conducted on 30 (12 males; 18 females) pretreatment good quality lateral cephalograms from patients visiting to Department of Orthodontics and Dentofacial Orthopedics. The lateral cephalograms were selected such that the Point A could be accurately located. A purposive convenience sampling technique was utilized for our study. Patients of age more than 16 yrs only were included in the sample. Patients with tooth agenesis or supernumeraries, any developmental anomalies, traumatic injuries or fractured upper and lower incisors and molars, complex craniofacial deformities or syndromes, Patients who have undergone any orthodontic treatment were excluded from our study. The lateral cephalograms (Kodak 8000C Digital Panoramic and Cephalometric Systems) utilized in our study were of true size (1:1) and any faulty radiographs with image distortion were excluded. The exposure time ranged from 12.8 to 13.9 seconds with kV 69-71 and m/A 10-12. All the cephalograms were manually traced for landmark by one investigator. + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + Patients + + + 9606 + Species + + Patients + + + MESH:D050723 + Disease + + traumatic injuries or fractured upper + + + MESH:D000848 + Disease + + tooth agenesis + + + MESH:D019465 + Disease + + craniofacial deformities + + + MESH:D000014 + Disease + + developmental anomalies + + + + METHODS + paragraph + 4725 + Identification of landmarks + + + METHODS + paragraph + 4753 + After categorization of the cephalograms, on an acetate tracing paper the following landmarks were traced, Sella (S), Nasion (N), Point A, Point B, Point Y, Point L and Point X (Table 1, Figure 1) were located and the angles SNA, SNY, SNL and SNX were measured (Figure 2). + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + MESH:D000085 + Chemical + + acetate + + + + TABLE + T1.xml + T1 + table_caption + 5026 + Various landmarks utilized in the study. + + + FIG + AIM-22-347-g002.jpg + F1 + fig_caption + 5067 + Various substitutions given by diff erent authors + + + FIG + AIM-22-347-g003.jpg + F2 + fig_caption + 5117 + Various angles formed a: Angle SNA, b: Angle SNY, c: Angle SNL, d: Angle SNX + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + + METHODS + paragraph + 5194 + Satistical analysis + + + METHODS + paragraph + 5214 + All the statistical tests were conducted using SPSS (version 19.0) Mean and standard deviation for Angles SNA, SNY, SNL and SNX were calculated individually for males and females. 'T' Test was applied to determine statistical significance for all the parameters i.e Age, Angles SNA, SNY, SNL and SNX respectively. Karl Pearson correlation coefficient was carried out to determine the statistical significant correlation for Angle SNA with SNY, SNL and SNX. + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + RESULTS + title_1 + 5675 + 3. RESULTS + + + RESULTS + paragraph + 5686 + Males: A mean value of 22.1 +-4.3 years for Age was observed. A mean value of 82.80 +-1.950 for Angle SNA was observed. A mean value of 83.10 +-1.80 was observed with Angle SNY. A mean value of 78.30 +-2.90 was observed with Angle SNL. A mean value of 78.70 +-2.70 was observed with Angle SNX (Table 2). + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + + TABLE + T2.xml + T2 + table_caption + 5990 + Measured angles of SNA, SNY, SNL and SNX in our study + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + RESULTS + paragraph + 6044 + Females: A mean value of 19.8 +-3.2 years of Age was observed. A mean value of 80.90 +-2.40 was observed for Angle SNA. A mean value of 81.10 +-2.50 was observed with Angle SNY. A mean value of 77.30 +-3.40 was observed with Angle SNL. A mean value of 77.90 +-3.40 was observed with Angle SNX (Table 2). T test was carried out to determine significance amongst all the parameter Age, SNA, SNY, SNL and SNX. Angles SNA (P<0.033) and SNY (P<0.023) revealed statistical significance (Table 3). + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + TABLE + T3.xml + T3 + table_caption + 6535 + Determination of statistical signifi cance for parameters SNA, SNY, SNL, SNX and Age + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + RESULTS + paragraph + 6620 + A correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. SNA revealed a strong statistically significant correlation with SNY>SNX>SNL (Table 4). + + 6615 + Gene + 4363 + + SNA + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + + TABLE + T4.xml + T4 + table_caption + 6853 + Determination of overall correlation coeffi cient for parameters SNA, SNY, SNL, SNX and Age + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + + RESULTS + paragraph + 6945 + In males correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. Only SNA revealed a strong statistically significant correlation with SNY only. (Table 5). + + 6615 + Gene + 4363 + + SNA + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + + TABLE + T5.xml + T5 + table_caption + 7188 + Determination of correlation coefficient for parameters SNA, SNY, SNL, SNX and Age for females. + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + + RESULTS + paragraph + 7284 + In females correlation of age, SNA, SNY, SNL and SNX was calculated. Age had a non significant correlation with all the parameters utilized in the study. SNA revealed a strong statistically significant correlation with SNY, SNL and SNX (Table 6). + + 6615 + Gene + 4363 + + SNA + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + + TABLE + T6.xml + T6 + table_caption + 7531 + Determination of correlation coefficient for parameters SNA, SNY, SNL, SNX and Age for males + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 310 + Gene + 36149 + + SNX + + + + DISCUSS + title_1 + 7624 + 4. DISCUSSION + + + DISCUSS + paragraph + 7638 + Apical base of maxilla and mandible help in determining the spatial relation of both maxilla and mandible to the cranial base. It also determines the limit of placement of incisors in the anteroposterior position. Numerous controversies exist in landmarks which are difficult to identify. Among those points, Point A is the most common point which encounters difficulty in identification. The cephalometric landmark, Point A, was investigated with regard to definition, location and usefulness in cephalometric analysis. Point A or Subspinale represents the maxillary apical base; the projection of cheeks frequently obscures this landmark in lateral cephalogram. Due to shortcomings of Point A various substitute landmarks have been sought by different authors by keeping the root apex of maxillary central incisor as a stable landmark. Van der Linden, suggested the use of point L, which is located on the anterior surface of the image of the labial lamella at the region of the apex of the maxillary incisors Jarabak and Fizzel identified a Point X 2mm ahead of the root apex as a redefinition of point A. Another Point Y plotted 3mm labial to a point between upper and lower two-thirds of the long axis of the root of the maxillary central incisor as suggested by Jacobson R. and Jacobson A. Comparison of mean values of Angle SNA with Angles SNY, SNL and SNX When the mean values of angles SNA, SNL, SNX and SNY were compared we observed the mean value of SNY was more in comparison to SNA, SNL and SNX. The mean values of SNL and SNX was less in comparison to SNA. Probable cause for such an observation could be attributed the variation in the definition of those cephalometric landmarks. When T test was carried out to determine significance amongst all the parameters individually like Age, SNA, SNY, SNL and SNX, Angles SNA and SNY revealed statistical significance (Table 3). This suggested that angles SNA and SNY were more specific for all the samples included in the study. On an overall SNA and SNY; SNX and SNL revealed strong statistical correlation between each other. Clear visibility of maxillary central incisor would act as a guide for marking the landmarks as require for Point Y could be attributed for such an observation. In males there was high correlation of angle SNA with angle SNY. In females, angle SNA correlated with Angle SNY, SNL and SNX, as we had observed that all the angles were less in comparison to males, which resulted in such an observation. However a further study has to be conducted on females to ascertain the probable cause for such an observation. + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6615 + Gene + 4363 + + SNA + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6624 + Gene + 48164 + + SNL + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 310 + Gene + 36149 + + SNX + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + + DISCUSS + paragraph + 10237 + In almost all cases the angles SNA and SNY were equal and showed high statistical significant correlation. Abdwani et al stated that the effects of incisal inclination changes, due to orthodontic treatment, are of no clinical relevance to the position of Point A and B, even though they may be statistically significant. However Kazem et al reported that the position of Point A is affected by local bone remodeling associated with proclination of the upper incisor in Class II division 2 malocclusion, but this minor change does not significantly affect the SNA angle. According to Jacobson a point closer to the center of the root of a tooth is less vulnerable to displacement than, say, a point close to the root apex during crown tipping procedures. Point Y represents closer to center of the tooth root hence point Y can be used more precisely as a substitute for point A. Whereas Point L and Point X are located in relation to the root apex, which can change if the tooth is proclined or retroclined. + + 6615 + Gene + 4363 + + SNA + + + 6615 + Gene + 4363 + + SNA + + + + CONCL + title_1 + 11244 + 5. CONCLUSION + + + CONCL + paragraph + 11258 + The situation of Point A is rather complex, and its location depend on a number of variables. Thus from the present study the following conclusions can be drawn: Age had no significance on identification of maxillary apical base; Overall Point; A showed strong statistically significant correlation with Point Y.; In males Point A and Point Y showed strong statistically significant correlation.; In females, Point A showed statistical significant correlation with Point Y, L and X. + + + CONCL + footnote + 11741 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 11778 + REFERENCES + + + 59 + REF + surname:der;given-names:Linden V + 111 + 5276196 + 1971 + Am J Orthod + ref + 125 + 11789 + A study of roentgenographic bony landmarks + + + 60 + surname:Frantz;given-names:RC + REF + surname:Baumrind;given-names:S + 111 + 5283996 + 1971 + Am J Orthod + ref + 11832 + The reliability of head film measurements. 1. Landmark identification + + + REF + surname:Fizzel;given-names:JR + 146 + 1963 + ref + 11902 + Technique and Treatment with the Light wire appliance + + + 77 + REF + surname:Rick;given-names:L + 1 + 1980 + Am J Orthod + ref + 11956 + Jacobson and Alex Jacobson, Point A revisited + + + REF + surname:Jacobson;given-names:A + 2006 + ref + 12002 + Radiographic cephalometry + + + 34 + REF + surname:Downs;given-names:WB + 812 + 18882558 + 1948 + Am J Orthod + ref + 840 + 12028 + Variations in facial relationship. Their significance in treatment and prognosis + + + 38 + surname:Kronman;given-names:JH + REF + surname:Kalafa;given-names:JA + 225 + 3 + 5242882 + 1968 + The Angle Orthodontist + ref + 230 + 12109 + A Critical Evaluation of Cephalometric "A" Point and Proposal of a More Significant Landmark + + + 79 + REF + surname:Al-Abdwani;given-names:R + 462 + 19413385 + 2009 + Angle Orthod + ref + 467 + 12206 + Change of Incisor Inclination Effects on Points A and B + + + 79 + surname:Al-Nimri + REF + surname:Kazem;given-names:S + 880 + 19705940 + 2009 + Angle Orthod + ref + 884 + 12262 + Maxillary Incisor Proclination Effect on the Position of Point A in Class II division 2 Malocclusion + + + + 4272845 + + surname:Mirzaeian;given-names:Razieh + surname:Mobasheri;given-names:Mahmoud + 25568630 + surname:Jafari;given-names:Fatemeh + 318 + 5 + 2014 + 4272845 + MSM-26-318 + 10.5455/msm.2014.26.318-320 + front + 26 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Mater Sociomed; 2014 Oct; 26(5) 318-20. doi:10.5455/msm.2014.26.318-320 + Hemodialysis Diet Education Blood Pressure Change Weight + 320 + Jafari F, Mobasheri M, Mirzaeian R + 0 + Effect of Diet Education on Blood Pressure Changes and Interdialytic Weight in Hemodialysis Patients Admitted in Hajar Hospital in Shahrekord + + 9606 + Species + + Patients + + + + ABSTRACT + abstract_title_1 + 142 + Background and aim: + + + ABSTRACT + abstract + 162 + Nutrition is a key factor in the treatment of patients with chronic kidney disease because kidney burden decrease causes uremic reduction and its side effects. The aim of this research is to examine the effect of diet education on blood pressure changes and interdialytic weight in Hemodialysis patients admitted to Hemodialysis ward of Hajar hospital in Shahrekord. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D051437 + Disease + + kidney burden decrease causes uremic reduction + + + MESH:D051436 + Disease + + chronic kidney disease + + + + ABSTRACT + abstract_title_1 + 529 + Methods: + + + ABSTRACT + abstract + 538 + This quasi-experimental and interventional study of 100 dialysis patients referred to Hemodialysis ward of Hajar hospital was performed in a pre-test and post-test in 2011. Diet education, including face to face training with instruction booklets, were conducted in the two sessions. Having carried out the educational program, blood pressure and interdialytic weight gain were measured and recorded one month before and during three stages and after the educational program by researcher-designed checklists. The data were analyzed through Spss16 software by Paired t-test and ANOVA. + + 9606 + Species + + patients + + + MESH:D015430 + Disease + + interdialytic weight gain + + + + ABSTRACT + abstract_title_1 + 1123 + Results: + + + ABSTRACT + abstract + 1132 + The results showed that mean of primary weight of the patients increase from 66.15+-15.10 to 64.43+-14.67. Mean of Systolic and diastolic blood pressure in patients in three stages were reduced to 6.65+-1.51 mmg 2.24+-1.82 mmg respectively. There was a significant difference between the creatinine amount in patients before and after of training (p<=0.01) but no meaningful difference was observed between the BUN amount before and after of training (p<=0.031). + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D003404 + Chemical + + creatinine + + + + ABSTRACT + abstract_title_1 + 1597 + Conclusion: + + + ABSTRACT + abstract + 1609 + Training to patients underwent hemodialysis in order to observe diet and its effects on improvement in treatment are of significant importance. + + 9606 + Species + + patients + + + + INTRO + title_1 + 1753 + 1. INTRODUCTION + + + INTRO + paragraph + 1769 + Advanced chronic renal failure is a progressive and irreversible impairment where the kidney's ability to filter metabolic waste products from the blood, and maintain water and electrolyte balance disappears, hence resulting in uremia. In patients with advanced chronic renal failure, it is impossible for them to live without adequate alternative treatment. Nowadays, the best alternative treatment is hemodialysis. Renal failure is highly prevalent in the world and in Iran. Nowadays, 242 out of 1000 people in the world suffer from renal failure; besides, about 8% are added to this figure annually. Although hemodialyses has improved renal disease and prolong patients' lives, without the proper diet and medication, it is not highly efficient. Following diet, fluid restriction and drugs are essential in care and health maintenance of renal dialysis patients. For these patients, the purpose of diet is the reduction of kidney burden, prevention of renal complications, maintaining proper nutrition, and prevention of uremia and its complications. Evidence indicates that many patients undergoing hemodialysis do not adhere to proper diet and fluid intake restrictions. It seems that many patients are not aware of diet and fluid restriction. Dialysis alone cannot provide the health and survival to the patients in long term. Besides hemodialysis, therefore, diet, fluid restriction, and drug therapies are essential treatments for chronic renal failure. Poor diet, and lack of fluid restriction can have serious consequences for patients, including impaired physical abilities, depression, acute pulmonary edema, congestive heart failure, and premature death. A poor dietary habit is associated with low quality of life, and morbidity and mortality of dialysis patients. Further, malnutrition and inflammation increase cardiovascular risk and mortality in hemodialysis patients, while following the diet and restricting fluid intake decrease the risk of complications and increase quality of life. Yet, many patients do not follow their diet. Previous studies on the role of nurses in the renal failure patients' diet plan show that the most important objectives in hemodialysis patient care are preventing malnutrition and maintaining good nutrition, edema control, delaying renal osteodystrophy by controlling phosphorus and calcium levels, and finally encouraging the patient to consume tolerable foods which are in accordance with lifestyle changes. Lack of awareness regarding the effects of diet on blood pressure made hemodialysis patients always suffer from its complications. Since this matter is of vital importance, the present study aims to determine the effect of diet on the interdialytic blood pressure and weight in patients undergoing hemodialysis treatment. + + 9606 + Species + + patients + + + 9606 + Species + + people + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D014867 + Chemical + + water + + + MESH:D000275 + Disease + + depression + + + MESH:D014511 + Disease + + uremia + + + MESH:D051437 + Disease + + renal failure + + + MESH:D051437 + Disease + + Renal failure + + + MESH:D007676 + Disease + + chronic renal failure + + + MESH:D051437 + Disease + + renal failure + + + MESH:D007676 + Disease + + chronic renal failure + + + MESH:D004487 + Disease + + edema + + + MESH:D007674 + Disease + + renal complications + + + MESH:D006333 + Disease + + congestive heart failure + + + MESH:D010758 + Chemical + + phosphorus + + + MESH:D012080 + Disease + + delaying renal osteodystrophy + + + MESH:D002118 + Chemical + + calcium + + + MESH:D007676 + Disease + + chronic renal failure + + + MESH:D014511 + Disease + + uremia + + + MESH:D003643 + Disease + + premature death + + + MESH:D002318 + Disease + + inflammation increase cardiovascular + + + MESH:D007674 + Disease + + renal disease + + + MESH:D011654 + Disease + + acute pulmonary edema + + + + METHODS + title_1 + 4557 + 2. MATERIAL AND METHOD + + + METHODS + paragraph + 4580 + This quasi-experimental and interventional study of 100 dialysis patients admitted in Hemodialysis ward of Hajar hospital was conducted in 2011 where a pretest as well as a post test took place. Patients were enrolled after giving informed consent. The patients who intended to undergo a kidney transplant during the educational program, or leave the place of dialysis, or those who are frequently ill and unable to afford a full time hemodialysis or those who suffer from another known disease besides advanced chronic kidney failure were excluded from the study. The instrument used for data collection was a two-part researcher-designed checklist which consisted of questions on demographic information, knowledge of consumption of food, systolic and diastolic hypertension control, as well as the weight control of the patients. The reliability of this checklist was determined by nutritionists. Before starting the educational program, data on weight, blood pressure and the results of Cr and BUN tests were recorded. The educational program was conducted during two months for the patients. Carried out through face to face interview, bulletins, pamphlets and educational CD, this program was concerned with the control of salt and fluid intake, and the right way of food consumption. Two weeks after beginning of the educational program, weight and blood pressure changes were measured, then at the end of the first month of program, a questionnaire was given to patients through which demographic data were recorded. Their knowledge of proper diet and nutrition was evaluated, as well. Two weeks later and at the end of the second month, weight and blood pressure were measured, and again the patients were given the questionnaire, then data obtained from the questionnaires, including weight, blood pressure and other variables were recorded. The data were entered into SPSS 16, and then the results were analyzed by using descriptive statistics, paired t-test and ANOVA. + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D012492 + Chemical + + salt + + + MESH:C563897 + Disease + + diastolic hypertension + + + MESH:D007676 + Disease + + chronic kidney failure + + + + RESULTS + title_1 + 6561 + 3. RESULTS + + + RESULTS + paragraph + 6572 + Results showed that patients were between 13-80 years old with a mean age of 56.2+- 15.14. Of them, 69% were male and the rest 31% were female. In terms of education, the majority of the subjects (81.4%) were illiterate or educated below the 9th grade, and 18.6% were educated beyond this grade. The mean duration of dialysis was 40.7+- 36.40. The maximum and minimum of duration for the subjects undergoing dialysis were 20 years and 4 month, respectively. Further, the mean frequency of dialysis was 22.40+- 2.9 times a week. + + 9606 + Species + + patients + + + + RESULTS + paragraph + 7100 + According to the patient's medical record, which was filled by the physician regarding muscle mass and cardiovascular problems each month, the initial mean weight of the patients was 66.15 +- 15.10. The lightest patient was 31.5 kg and the heaviest one 105 kg. After the first stage of educational program, the mean of weight become 64.43 +- 14.67. + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + + RESULTS + paragraph + 7449 + Results showed that the mean of BUN and Cr for patients before training were 77.98 +- 21.74 and 9.47 +- 2.80 respectively, which become 66.14 +- 12.13 and 8.48 +- 2.36 after the first stage of training. There was a significant difference between the Cr of the patients before and after training (P <= 0/01). However, the BUN of the patients were not significantly different before and after the training (P <= 0/31). + + 5714 + Gene + 37686 + + P <= 0/31 + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D002857 + Chemical + + Cr + + + + RESULTS + paragraph + 7868 + Moreover, the findings represented that the mean systolic and diastolic blood pressure in the three stages of the research reduced to 6.65 +- 1.51 mmHg and 2.24 +- 1.82 mmHg, respectively. The mean systolic blood pressure before and after the test was significantly different (P <= 0/05). However, there was no significant difference between the mean diastolic blood pressure before and after the test (P <= 0/20) (Table 1). + + 51673 + Gene + 32659 + + P <= 0/20 + + + 106479023 + Gene + + P <= 0/05 + + + + TABLE + T1.xml + T1 + table_caption + 8295 + Weight, creatinine, urea, and systolic and diastolic blood pressure in stages of study + + MESH:D003404 + Chemical + + creatinine + + + MESH:D014508 + Chemical + + urea + + + + TABLE + T2.xml + T2 + table_caption + 8382 + Comparison of laboratory indices in two genders + + + RESULTS + paragraph + 8430 + Findings on the determination and comparison of the problems of the patients undergoing hemodialysis (with regard to the systolic and diastolic blood pressure, weight gain, and the changes before and after training including the changes in Urea, creatinine, blood pressure and weight) are important indicators and have a huge role in creating problems and reducing the quality of life. These findings indicated that there was no significant difference in the Cr means of males and females after diet education in the experimental investigation (P=0.2). The relationship between gender and BUN showed a difference in males and females after training (P=0.026). Further, males and females significantly differed from each other in terms of interdialytic weight gain and systolic blood pressure. (P=0. 003), but no significant difference was observed between diastolic blood pressure (P=0.4). + + 9606 + Species + + patients + + + MESH:D015430 + Disease + + interdialytic weight gain + + + MESH:D015430 + Disease + + weight gain + + + MESH:D003404 + Chemical + + creatinine + + + MESH:D014508 + Chemical + + Urea + + + + DISCUSS + title_1 + 9320 + 4. DISCUSSION + + + DISCUSS + paragraph + 9334 + The findings represented that educating patients with a special emphasis on diet can have a positive effect on their attitude. Interdialytic weight loss indicated that training had an effect on patients and enabled them to make adjustments in their diet. Toolabi has investigated the effect of diet education on the knowledge and practice of Hemodialysis Patients in Lorestan, which indicated a significant increase in knowledge of patients after training. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + Patients + + + 9606 + Species + + patients + + + MESH:D015431 + Disease + + weight loss + + + + DISCUSS + paragraph + 9791 + One of the common problems to these patients is Uremia that can cause poor appetite, memory loss, drowsiness, misjudgment and poor concentration. Interdialytic weight gain increases blood pressure by 3 mm Hg per kg of body weight. Results of nutritional evaluation of 99 hemodialysis patients in Tehran represented a significant positive correlation between protein intake through the correct way of nutrition, and body weight. Therefore, nutrition education is an important variable in increasing awareness and health status of hemodialysis patients. Previous research showed that it is necessary to educate patients with high blood pressure to follow a good diet and restrict the use of fat and sodium. Lack of attention to the patient education regarding drug use, and food and fluid restriction has adverse consequences. Considering the persistent problems as well as their kinds, inefficiency of dialysis in complete resolution of these problems, and the need for continuous care, education can provide an informed and active participation of the patient to care for themselves. The subjects in this study had a low level of awareness before training. This was due to the insufficient diet education which is consistent with the study carried out by Toulabi entitled Impact of education on knowledge and practice of dialysis patients in Lorestan. In this study, there was a significant relationship between education and weight after training which was consistent with Alipour's findings on knowledge, attitude and practice of renal dialysis patients' nutrition. In the study entitled the effect of diet education on dialysis efficacy which is conducted in Zanjan, Talebipour in three stages of training showed that after training, there was an increase in the tendency of these patients to follow an efficient diet. The results of Talebipour's study is similar to those of the current research. Shab Bidar, et al. in their study of dialysis adequacy and prevalence of malnutrition in hemodialysis patients have suggested that in 23% of patients albumin level was below 3 (severe malnutrition), and in 32% of them albumin was in the range of 3-3.5 (medium malnutrition). Moreover, 75 to 80% of hemodialysis patients have high blood pressure, and despite using various drugs, the major cause of hypertension in these patients is the increase in blood volume, which is due to water and salt retention in the body; therefore, withdrawing fluid in a short period from these patients has had a significant effect on blood pressure. Durose, et al have shown that educating patients about diet and alternative consumption can lead to the fluid intake restriction and subsequently to weight loss. Based on the findings, there was a significant association between the weight of the subjects and their level of education that is consistent with the findings of the study conducted by Alipour representing a positive correlation between the knowledge and practice of the subjects, and their level of education. The main achievement in this study was that education is positively effective since the goal of education is to enable us to establish an assuring relationship between patients and nurses:considering education and support of the patients. This is implemented if once patients are allowed to actively participate in decision-making and planning for themselves. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patient + + + 9606 + Species + + patient + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D000309 + Disease + + insufficient + + + MESH:D012492 + Chemical + + salt + + + MESH:D015430 + Disease + + weight gain increases blood + + + MESH:D014511 + Disease + + Uremia + + + MESH:D015431 + Disease + + weight loss + + + MESH:D008569 + Disease + + memory loss + + + MESH:D014867 + Chemical + + water + + + MESH:D012964 + Chemical + + sodium + + + MESH:D006970 + Disease + + drowsiness + + + MESH:D006973 + Disease + + hypertension + + + + CONCL + title_1 + 13165 + 5. CONCLUSION + + + CONCL + paragraph + 13179 + Diet and drug therapy besides hemodialysis can improve health condition and therapeutic effectiveness for patients with renal failure. However, it seems that many patients are not fully aware of diet and fluid intake restriction. This study showed that periodic educational programs can play a significant role in the health of patients and their attitude and knowledge of nutrition. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + MESH:D051437 + Disease + + renal failure + + + + CONCL + footnote + 13563 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 13600 + REFERENCES + + + surname:Bare;given-names:BG + REF + surname:Smeltezer;given-names:SC + 1326 + 2004 + Textbook of medical - Surgical nursing + ref + 1334 + 13611 + Brunner and Suddarth S + + + surname:Bare;given-names:BG + REF + surname:Smeltzer;given-names:SC + 1110 + 2006 + Text book of medical-surgical nursing + ref + 1150 + 13634 + Brunner and Suddarth S + + + surname:Braunwald;given-names:E + surname:Brenner;given-names:BM + surname:Green;given-names:J + surname:Skorecki;given-names:K + 1653 + 2005 + Harrison principle of internal medicine + ref + REF + surname:Jameson;given-names:JL + surname:Hauser;given-names:SL + 1667 + surname:Longo;given-names:DL + surname:Kasper;given-names:D + surname:Fauci;given-names:AS + 13657 + Chronic renal failure + + MESH:D007676 + Disease + + Chronic renal failure + + + + 20 + REF + surname:Vinpound;given-names:GM + 161 + 9 + 1991 + Dialysis and Transplantation + ref + 163 + 13679 + Intervention on Strategies for improving fluid restriction compliance in chronic hemodialysis patients + + + surname:Stump;given-names:SE + REF + surname:Kattleen;given-names:ML + 2000 + Philadelphia + ref + 13782 + Food, nutrition & diet therapy + + + surname:Ing;given-names:TS + REF + surname:Daugirdas;given-names:JT + 1989 + Handbook of Hemodialysis + ref + 13813 + + + 20 + REF + surname:Jones;given-names:K + 67 + 2 + 1996 + ANNA Journal + ref + 68 + 13814 + Are hemodialysis patient population at risk + + 9606 + Species + + patient + + + + 30 + REF + surname:Wells;given-names:C + 637 + 6 + 14730784 + 2003 + Nephrology Nursing Journal + ref + 646 + 13858 + Optimizing nutrition in patients with chronic kidney disease + + 9606 + Species + + patients + + + MESH:D051436 + Disease + + chronic kidney disease + + + + surname:Bartucci;given-names:MR + surname:Morgan;given-names:J + surname:Marek;given-names:JF + surname:Marantides;given-names:DK + 1465 + 2003 + Medical surgical nursing concept & clinical practice + ref + REF + 1477 + surname:Marker;given-names:J + surname:Sands;given-names:J + surname:Phipps;given-names:W + 13919 + Management of persons with renal failure + + 9606 + Species + + persons + + + MESH:D051437 + Disease + + renal failure + + + + surname:Rizza;given-names:GM + surname:Morelli;given-names:E + surname:D'Alessandro;given-names:C + surname:Cupisti;given-names:A + 127 + 3 + 2004 + Journal of Nutrition + ref + 14 + REF + 133 + surname:Franzoni;given-names:F + surname:Galetta;given-names:F + 13960 + Nutritional status and dietary manipulation in predialysis chronic renal failure patients + + MESH:D007676 + Disease + + chronic renal failure + + + + surname:Boeschoten;given-names:EW + surname:Dekker;given-names:FW + surname:Jager;given-names:KJ + surname:Merkus;given-names:MP + 584 + 4 + 9100049 + 1997 + American Journal of Kidney Diseases + ref + 29 + REF + 592 + surname:Krediet;given-names:RT + surname:Stevens;given-names:P + 14050 + Quality of life in patients on chronic dialysis: self-assessment 3 months after the start of treatment + + 9606 + Species + + patients + + + + surname:Stump;given-names:PSE + REF + surname:Kattreen;given-names:ML + 46 + 2000 + Nutrition + ref + 49 + 14153 + Diet Therapy & Food + + + 1 + REF + surname:Toulabi;given-names:T + 36 + 2 + 2001 + Lorestan University of Medical Sciences + ref + 47 + 14173 + Response of patients undergoing Hemodialysis + + 9606 + Species + + patients + + + + 21 + surname:Heyka;given-names:RJ + surname:Yan;given-names:G + REF + surname:Rocco;given-names:MV + 280 + 11509799 + 2001 + Am J Nephrol + ref + 288 + 14218 + Risk factors for hypertension in chronichemodialysis patients: baseline data from the HEMO study + + 9606 + Species + + patients + + + MESH:D006973 + Disease + + hypertension + + + + REF + surname:Borazjani;given-names:A + 2000 + ref + 14315 + Evaluating the hemodialysis patients based on nutrition + + 9606 + Species + + patients + + + + 1 + REF + surname:Bashardoost;given-names:B + 13 + 7 + 2007 + Ardabil Journal of Ardabil University of Medical Sciences + ref + 16 + 14371 + Evaluatin the relationship between the weight during the hemodialys session in hemodialysis patients and hypertension increasing in Booali hospital + + 9606 + Species + + patients + + + MESH:D006973 + Disease + + hypertension + + + + surname:Alipour;given-names:S + 56 + surname:Alipour;given-names:fard A + surname:Koshavar;given-names:H + REF + surname:Alipour;given-names:B + 57 + 2002 + Medical Journal of Tabriz University of Medical Sciences + ref + 62 + 14519 + Study of knowledge, attitude and practice of nutrition for kidney patients undergoing hemodialysis + + 9606 + Species + + patients + + + + 36 + REF + surname:Talebipoor;given-names:B + 28 + 1 + 2001 + Gynecologist Journal of Medical Sciences + ref + 36 + 14618 + Establishment of dietary education in hemodialysis efficacy in dialysis patients in Dr. Reisi hospital + + 9606 + Species + + patients + + + + REF + 2005 + ref + 14721 + Estimating the efficience of dialysis and malnutrition prevalence in hemodialysis patients. Razi training and treatment center of Rasht + + 9606 + Species + + patients + + + + surname:Przygrodzka;given-names:F + 104 + surname:Watson;given-names:V + surname:Holdsworth;given-names:M + REF + surname:Durose;given-names:CL + 35 + 1 + 2004 + Journal of American Diet Association + ref + 42 + 14857 + Knowledge of dietary restrictions and the medical consequences of noncompliance by patients on Hemodialysis are not predictive of dietary compliance + + 9606 + Species + + patients + + + + + 4272846 + + surname:Jaafreh;given-names:Amany + surname:Kawar;given-names:Jane + surname:Ghuweri;given-names:Ali AL + 25568576 + surname:Alkhawaldeh;given-names:Khaled + 297 + 5 + 2014 + 4272846 + AIM-22-297 + 10.5455/aim.2014.22.297-301 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 297-301. doi:10.5455/aim.2014.22.297-301 + Bone scan back pain pediatric 99mTc-MDP + 301 + Alkhawaldeh K, Ghuweri AA, Kawar J, Jaafreh A + 0 + Back Pain in Children and Diagnostic Value of 99mTc MDP Bone Scintigraphy + + 1800 + Gene + 80192 + + MDP + + + 9606 + Species + + Children + + + MESH:D010146 + Disease + + Pain + + + + ABSTRACT + abstract_title_1 + 74 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 83 + Aim: + + + ABSTRACT + abstract + 88 + The aim of our study is to assess the diagnostic value of Technituim-99m-Methyle diphosphonate (99mTc-MDP) Bone scintigraphy in the assessment of children with back pain. + + 1800 + Gene + 80192 + + MDP + + + 9606 + Species + + children + + + - + Chemical + + Technituim-99m-Methyle diphosphonate + + + MESH:D001416 + Disease + + back pain + + + + ABSTRACT + abstract_title_1 + 259 + Methods: + + + ABSTRACT + abstract + 268 + Included in this retrospective study were 68 child referred to us complaining of back pain (mean age of 13+ 2). There were 45 boys and 23 girls. All children have been investigated with conventional x-ray which revealed normal or inconclusive result. All underwent bone scintigraphy after the injection of 99mTc-MDP with calculated doses according to there body weights. + + 1800 + Gene + 80192 + + MDP + + + 9606 + Species + + child + + + 9606 + Species + + boys + + + 9606 + Species + + girls + + + 9606 + Species + + children + + + MESH:D001416 + Disease + + back pain + + + + ABSTRACT + abstract_title_1 + 639 + Results: + + + ABSTRACT + abstract + 648 + Bone scintigraphy revealed 17 (25%) abnormal scans in 11 boys and 6 girls. Scans findings were suggestive of spondylolysis (n=4); malignancy including primary tumors and metastases (n=3); infection including osteomyelitis and discitis (n=3); sacroiliitis (n=2); benign tumors (n=2); pseudo fractures in ribs (n=1); necrosis in femoral head epiphysis(n=1) and nonskeletal-renal retention due to hydronephrosis (n=1). Sensitivity, specificity and accuracy of bone scan in detecting gross skeletal abnormality as a cause for back pain were 94% and 100% and 99% respectively. + + 9606 + Species + + boys + + + 9606 + Species + + girls + + + MESH:D007239 + Disease + + infection + + + MESH:C538496 + Disease + + skeletal abnormality + + + MESH:D009362 + Disease + + metastases + + + MESH:D001416 + Disease + + back pain + + + MESH:D007674 + Disease + + nonskeletal-renal retention + + + MESH:D015299 + Disease + + osteomyelitis and discitis + + + MESH:D009369 + Disease + + malignancy + + + MESH:D058566 + Disease + + sacroiliitis + + + MESH:D009369 + Disease + + benign tumors + + + MESH:D009369 + Disease + + tumors + + + MESH:D006869 + Disease + + hydronephrosis + + + MESH:D050723 + Disease + + fractures + + + MESH:D009336 + Disease + + necrosis + + + + ABSTRACT + abstract_title_1 + 1220 + Conclusion: + + + ABSTRACT + abstract + 1232 + Bone isotope scan is a sensitive imaging modality in the assessment of pediatric patients with back pain. It is a reliable modality to detect and role out most benign and aggressive serious etiologies. + + 9606 + Species + + patients + + + MESH:D001416 + Disease + + back pain + + + + INTRO + title_1 + 1434 + 1. INTRODUCTION + + + INTRO + paragraph + 1450 + Back pain in relatively less common in children than in adults. Most cases of back pain in children clear up in a few days without medical attention. In few cases the symptoms will persist for a longer time which may indicate underlying serious problem more than just a simple backache. Causes can range from serious malignant causes to just benign lesions. Therefore, it is important to start with sensitive and cost effective imaging modality that is capable of distinguishing simple back pain from serious underlying diseases. + + 9606 + Species + + children + + + 9606 + Species + + children + + + MESH:D010146 + Disease + + pain + + + MESH:D001416 + Disease + + back pain + + + MESH:D001416 + Disease + + back pain + + + + INTRO + paragraph + 1980 + A variety of imaging techniques has been used to confirm the cause of back pain. A conventional x-ray is often the first imaging technique which has been used. Unfortunately; this technique suffers from low sensitivity, and can miss most causes of back pain including serious pathologies. On the other hand, advanced imaging modalities like computerized tomography (CT scan) and magnetic resonance imaging (MRI) can be ordered. However, such modalities are not cost effective, and not used routinely to image the whole body. + + MESH:D001416 + Disease + + back pain + + + MESH:D001416 + Disease + + back pain + + + + INTRO + paragraph + 2505 + Bone scintigraphy (BS) has been used in the diagnostic work-up of unexplained bone pain. The technique is sensitive and allows visualization of the whole skeleton in a short time. This modality is more sensitive than x-rays, since early or non-aggressive lesions generally cannot be seen on radiographs until significant bone mineral matrix has been lost. + + 255564 + Species + + rays + + + MESH:D010146 + Disease + + bone pain + + + + INTRO + paragraph + 2861 + Our study will assess the diagnostic value of BS in children presented with back pain. We will try to emphasize on efficiency of this modality in localizing many pathologies that would have been missed by conventional x-ray study. Also, we will make an overview for scan findings in some lesions encountered during our study. + + 9606 + Species + + children + + + MESH:D001416 + Disease + + back pain + + + + METHODS + title_1 + 3187 + 2. PATIENTS AND METHODS + + 9606 + Species + + PATIENTS + + + + METHODS + paragraph + 3211 + Included in this retrospective study were 68 children with ages ranging from 4 to 15 years (mean age of 11+- 3). There were 45 boys and 23 girls. All children presented with back pain of 7 to 88 days duration with a mean value of 18+-5 days. All children underwent BS for assessment of back pain. There initial laboratories and conventional radiological investigation failed to define or localize any possible etiology. All patients underwent 99mTc MDP BS which has been carried out two hours after intravenous administration of radioisotope tracer with calculated dose according to body weights by the formula: Pediatric dose = age (in years)/age+12 x adult dose (20 mCi). + + 9606 + Species + + children + + + 9606 + Species + + boys + + + 9606 + Species + + girls + + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + patients + + + 1800 + Gene + 80192 + + MDP + + + MESH:D001416 + Disease + + back pain + + + MESH:D001416 + Disease + + back pain + + + + METHODS + paragraph + 3885 + Whole body BS was performed, using G.E Millennium dual head gamma camera with low energy high resolution collimators. Matrix set for 128 x 128 and time set for 30 minutes. For better assessment of any suspected or equivocal abnormality in the whole body scans, all children had additional static views by 500,000 counts performed after whole body scans, and additional SPECT images for the spine. + + 9606 + Species + + children + + + + METHODS + paragraph + 4282 + All scans were reported by two nuclear medicine specialists. The abnormal scan finding was defined as either single or multiple foci of increase or reduced tracer uptake in the spine or pelvic area; or any extra skeletal soft issue activity within chest or abdomen that can explain the etiology of back pain. + + MESH:D001416 + Disease + + back pain + + + + METHODS + paragraph + 4591 + Final diagnosis was established using histopathology and other imaging modalities (MRI and CT scan). Patients with normal scans had clinical follow up for 6 months to 1 year, or until relieve of symptoms. + + 9606 + Species + + Patients + + + + RESULTS + title_1 + 4796 + 3. RESULTS + + + RESULTS + paragraph + 4807 + Bone scintigraphy revealed 17 (25%) abnormal scans in 11 boys and 6 girls. Abnormal scans were described according to anatomical sites which were congruent with the site of pathology in all cases. Table 1 shows anatomical distribution of scan abnormalities in 17 abnormal scans. Scans findings were suggestive of spondylolysis (n=4); malignancy including primary tumors and metastases (n=3); sacroiliitis (n=2); infection including osteomyelitis and discitis (n=3); benign tumors (n=2); pseudofracture (ribs) (n=1); necrosis in femoral head epiphysis (n=1) and nonskeletal (renal) (n=1). Table 2 shows BS findings and diagnosis in 17 abnormal scans encountered during our study. BS failed to detect one case of disc herniation. + + 9606 + Species + + boys + + + 9606 + Species + + girls + + + MESH:D009336 + Disease + + necrosis + + + MESH:D009369 + Disease + + benign tumors + + + MESH:D007239 + Disease + + infection + + + MESH:D058566 + Disease + + sacroiliitis + + + MESH:D009369 + Disease + + tumors + + + MESH:D009362 + Disease + + metastases + + + MESH:D009369 + Disease + + malignancy + + + MESH:D015299 + Disease + + osteomyelitis and discitis + + + + TABLE + T1.xml + T1 + table_caption + 5535 + Anatomical distribution of bone scans abnormalities + + + TABLE + T2.xml + T2 + table_caption + 5587 + Bone scintigraphy fi ndings and diagnosis in 17 abnormal scans + + + RESULTS + paragraph + 5650 + Sensitivity, specificity and accuracy of bone scan in detecting gross skeletal abnormality as a cause for back pain were 94% and 100% and 99% respectively. Positive predictive value (PPV) and negative predictive value (NPV) were found to be 100% and 98% respectively. + + MESH:C538496 + Disease + + skeletal abnormality + + + MESH:D001416 + Disease + + back pain + + + + DISCUSS + title_1 + 5918 + 4. DISCUSSION + + + DISCUSS + paragraph + 5932 + Back pain is uncommon in children younger than 3 years, increasing its frequency with age until reaching a plateau during early adolescence. Most often, back pain is benign and self-limited. However, it is occasionally the presenting symptom of systemic diseases as cancer or infection. Thus, the major diagnostic task is to distinguish the patients with simple back pain from those with serious underlying diseases. + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D001416 + Disease + + back pain + + + MESH:D034721 + Disease + + systemic diseases + + + MESH:D007239 + Disease + + infection + + + MESH:D001416 + Disease + + back pain + + + MESH:D010146 + Disease + + pain + + + MESH:D009369 + Disease + + cancer + + + + DISCUSS + paragraph + 6349 + Low cost and availability make plain radiography the most common spinal imaging test. Conventional x-ray is often the first imaging technique which has been used. Unfortunately, the interpretation of spine X-ray in children is more difficult than in adults due to the presence of epiphyseal and apophyseal growth center. Also conventional x-ray is lack the sensitivity and would miss most causes of back pain including serious pathologies. The limitation of plain radiography was obvious in our study group, in which most x-ray studies were negative or inconclusive. + + 9606 + Species + + children + + + MESH:D001416 + Disease + + back pain + + + + DISCUSS + paragraph + 6916 + In our study, bone scintigraphy was able to detect many benign and malignant conditions in 25% of children (Table 2). Our results for scintigraphy with 99mTc-MDP showed an overall sensitivity, specificity values of 94%, 100% respectively. + + 1800 + Gene + 80192 + + MDP + + + 9606 + Species + + children + + + + DISCUSS + paragraph + 7155 + Skeletal scintigraphy is highly sensitive for detecting stress-induced changes in bone and is an important part in the evaluation of the young athlete with low back pain. Spondylolysis is an osseous defect of the pars interarticularis, thought to be a developmental or acquired stress fracture secondary to chronic low-grade trauma. Most patients with spondylolysis present with either acute or chronic pain. It can be found in as many as 47% of adolescent athletes with low back pain. Most of pars defects (90%) occur at the L5 level and unilateral defect found in 30% of cases. Spondylolysis will show focal tracer accumulation in the region of the pars interarticularis during delayed BS (Figure 1). SPECT BS is more sensitive than planar images and can detect abnormalities in about one third of individuals with normal planar exams. + + 9606 + Species + + patients + + + MESH:D015775 + Disease + + stress fracture + + + MESH:D017116 + Disease + + low back pain + + + MESH:D015868 + Disease + + pars defects + + + MESH:D014947 + Disease + + trauma + + + MESH:D015868 + Disease + + osseous defect of the pars interarticularis + + + MESH:D010146 + Disease + + pain + + + MESH:D017116 + Disease + + low back pain + + + + FIG + AIM-22-297-g003.jpg + F1 + fig_caption + 7993 + Whole body scan in 14 years old male boy with LBP of 3 weeks duration; scan showed focus with abnormal activity at L5 level (arrow); finding was suggestive of spondylolysis. + + 3929 + Gene + 3055 + + LBP + + + 9606 + Species + + boy + + + + DISCUSS + paragraph + 8167 + Also, BS is a very sensitive exam for the detection of acute bone fractures, and it does have increased sensitivity for the detection of rib fractures compared to plain radiographs. BS can be the modality of choice in the assessment of suspected child abuse cases. Also, findings can sometimes draw attention to possible underlying metabolic bone disease (Figure 2). + + 9606 + Species + + child + + + MESH:D001851 + Disease + + metabolic bone disease + + + MESH:D050723 + Disease + + fractures + + + MESH:D050723 + Disease + + bone fractures + + + + FIG + AIM-22-297-g004.jpg + F2 + fig_caption + 8534 + 10 years old male child with multiple pseudofractures invoving multiple ribs and lower tibii. + + 9606 + Species + + child + + + + DISCUSS + paragraph + 8628 + Other causes of low back pain in children like Spinal process apophysitis, Lumbosacral transitional vertebra, Facet Syndrome, Schuermann's Disease and disc herniation can have nonspecific findings during BS. For instant; spinal process apophysitis produces uptake of tracer along the lumbar spinous process and is due to a hyperextension overuse injury. A lumbosacral transitional vertebra is a common anomaly characterized by a large transverse process that follows the contour of the sacral ala and forms a non-ossified articulation with the sacrum. Bertolotti syndrome refers to increased tracer uptake along the transverse process pseudoarthrosis of a transitional vertebrae which is indicative of stress change. This finding can be seen in about 80% of patients with a transitional vertebra and low back pain. + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D010300 + Disease + + Schuermann's Disease + + + MESH:D000014 + Disease + + anomaly + + + MESH:D012090 + Disease + + hyperextension overuse injury + + + MESH:D017116 + Disease + + low back pain + + + MESH:D017116 + Disease + + low back pain + + + MESH:D011542 + Disease + + pseudoarthrosis + + + MESH:D013577 + Disease + + Bertolotti syndrome + + + + DISCUSS + paragraph + 9443 + Facet Syndrome can produce both local and radiating pain. SPECT BS will usually detect an abnormality (sensitivity 100%), but with poor specificity (70%). However, a normal exam can rule out facet syndrome. + + MESH:D010146 + Disease + + pain + + + + DISCUSS + paragraph + 9650 + Schuermann's Disease is relatively uncommon disorder, occurring in 3 to 5% of adolescents. The diagnosis is established when the radiographs demonstrate anterior wedging greater than 5 degrees with involvement of 3 or more contiguous vertebral bodies and a structural kyphosis of greater than 40 degrees plate. + + MESH:D007738 + Disease + + kyphosis + + + MESH:D010300 + Disease + + Schuermann's Disease + + + MESH:D025063 + Disease + + contiguous vertebral bodies + + + + DISCUSS + paragraph + 9961 + An acute herniated nucleus pulposus may not have any changes on BS until there has been excessive stress placed on the adjacent vertebrae. When this occurs, mild increased activity can be seen in the end-plates of the vertebral bodies on either side of the disc. In our study BS failed to detect any abnormalities in one case of disc herniation. + + + DISCUSS + paragraph + 10307 + Early stage of Perth's disease has a characteristic appearance during BS with photopenic defect within femoral epiphysis. On the other hand; slipped Capital Femoral Epiphysis in the absence of avascular necrosis the bone scan findings is nonspecific and consist of mildly increased activity about the hip and widening and blurring of the growth plate activity. + + MESH:D010020 + Disease + + avascular necrosis + + + MESH:D007873 + Disease + + Perth's disease + + + MESH:D001929 + Disease + + photopenic defect within femoral epiphysis + + + + DISCUSS + paragraph + 10668 + 99mTc-MDP BS can detect osteomyelitis one to two weeks before radiologic changes. For the detection of osteomyelitis in non-violated bone, the 3 phase bone scan has a reported sensitivity of 90-100% and a specificity of 70-95%. Osteomyelitis has characteristic appearance in 3 phase BS with abnormal increase in flow, blood pool and delayed radiotracer uptake. + + 1800 + Gene + 80192 + + MDP + + + MESH:D010019 + Disease + + Osteomyelitis + + + MESH:D010019 + Disease + + osteomyelitis + + + MESH:D010019 + Disease + + osteomyelitis + + + + DISCUSS + paragraph + 11029 + There is a higher incidence of discitis in children with peaks seen between the ages of 1 to 4, and 12 to 15 years. In young children, the higher incidence is felt to be related to the vascular supply of the spine. In children subchondral vessels are present until age of 8 and supply the disc, and may persist until teenage years. During BS delayed images; discitis will show increased tracer activity in the disc space and the contiguous vertebral bodies (Figure 3). + + 9606 + Species + + children + + + 9606 + Species + + children + + + 9606 + Species + + children + + + MESH:D025063 + Disease + + contiguous vertebral bodies + + + MESH:D015299 + Disease + + discitis + + + MESH:D015299 + Disease + + discitis + + + + FIG + AIM-22-297-g005.jpg + F3 + fig_caption + 11498 + 14 years old girl with back pain; bone scan revealed intervertebral disc activity at level of T12-L1 extending to adjacent vertebral end plates (arrow). Finding was consistent with discitis. + + 9606 + Species + + girl + + + MESH:D001416 + Disease + + back pain + + + MESH:D015299 + Disease + + discitis + + + + DISCUSS + paragraph + 11689 + 99mTc-MDP BS can detect synovial inflammations of rheumatoid arthritis and ankylosing spondylitis. Sacroiliitis can have prominent unilateral or bilateral uptake within sacroiliac joints (Figure 4). However; bilateral sacroiliitis, can be difficult to detect due to the normal increased activity within the SI joints. SI joint to sacrum (SIJ index) of greater than 1.35 can have value in this case. + + 1800 + Gene + 80192 + + MDP + + + MESH:C563037 + Disease + + sacroiliac joints + + + MESH:D001172 + Disease + + inflammations of rheumatoid arthritis + + + MESH:D013167 + Disease + + ankylosing spondylitis + + + Disease + + SIJ + + + MESH:D007592 + Disease + + SI joint + + + MESH:D058566 + Disease + + Sacroiliitis + + + MESH:D007592 + Disease + + SI joints + + + MESH:D058566 + Disease + + bilateral sacroiliitis + + + + FIG + AIM-22-297-g006.jpg + F4 + fig_caption + 12088 + 15 years old boy with low back pain of one month duration. Bone scan revealed prominent activity in both SIJs which is more prominent in the left side. Finding was consistent with sacroiliitis. + + 9606 + Species + + boy + + + MESH:D058566 + Disease + + sacroiliitis + + + MESH:D017116 + Disease + + low back pain + + + Disease + + SIJs + + + + DISCUSS + paragraph + 12282 + 99mTc-MDP BS is 50 to 80% more sensitive than radiographs in detecting skeletal metastases. This is probably because about 30-75% of the bone mineral content must be lost before a metastasis is evident on a radiograph. In contrast, as little as a 5-10% change is required to detect an abnormality on BS. The sensitivity for BS in detecting bone metastases is between 62%-100%. About 90% of patients with skeletal metastases present with multiple lesions which increase its specificity of this modality. Nearly 80% of all metastatic lesions are in the axial skeleton. Multiplicity of active lesions can help to define metastatic bone lesions and tracer uptake involving both the vertebral body and pedicles is usually indicative of metastatic disease. + + 9606 + Species + + patients + + + 1800 + Gene + 80192 + + MDP + + + MESH:D009362 + Disease + + bone metastases + + + MESH:D009362 + Disease + + metastases + + + MESH:D009362 + Disease + + metastases + + + MESH:C538445 + Disease + + metastatic disease + + + MESH:C536543 + Disease + + vertebral body + + + + DISCUSS + paragraph + 13035 + Primary bone tumors also can present with back pain in children since more than 80% of the Ewing's sarcomas occur in patients under 20 years of age. The tumor usually arises in the diaphysis of a long bone, but 20% arise in the pelvis. Bone scan can show gross abnormality during the three phases of bone scan; and hardly differentiated from osteomyelitis in some cases (Figure 5). + + 9606 + Species + + children + + + 9606 + Species + + patients + + + MESH:D010019 + Disease + + osteomyelitis + + + MESH:C563168 + Disease + + Ewing's sarcomas + + + MESH:D009369 + Disease + + tumor + + + MESH:D001859 + Disease + + Primary bone tumors + + + MESH:D001416 + Disease + + back pain + + + + FIG + AIM-22-297-g007.jpg + F5 + fig_caption + 13417 + 13 years old child with low back pain of 2 months duration; bone scan revealed prominent activity within left iliac bone suggestive of aggressive bone pathology. Biopsy revealed primary malignant Ewing sarcoma. + + 9606 + Species + + child + + + MESH:C563168 + Disease + + malignant Ewing sarcoma + + + MESH:D017116 + Disease + + low back pain + + + + DISCUSS + paragraph + 13628 + Benign lesions like osteoblastomas and osteoid osteoma can involve the posterior elements of the spine and they have characteristic appearance during 3 phase bone scan, osteoblastomas and ostioid osteomas will be hot on both blood pool and delayed images. Osteoblastomas are typically larger than 2 cm and possess both sclerotic and lytic features. Osteoid osteoma may occur in the spine (10-20% of cases), it most commonly involves the lumbar spine, nearly always occurs in the posterior elements (50%); only 10% of lesions involve the vertebral body, and is frequently associated with a painful scoliosis with the convexity of the curve oriented away from the side of the lesion. + + MESH:D010016 + Disease + + ostioid osteomas + + + MESH:C538213 + Disease + + sclerotic + + + MESH:D018215 + Disease + + osteoblastomas + + + MESH:D018215 + Disease + + Benign lesions like osteoblastomas and osteoid osteoma + + + MESH:D010017 + Disease + + Osteoid osteoma + + + MESH:C536543 + Disease + + vertebral body + + + MESH:D018215 + Disease + + Osteoblastomas + + + MESH:D010146 + Disease + + painful scoliosis + + + + DISCUSS + paragraph + 14310 + 99mTc-MDP B.S showed nonskeletal abnormalities that would have been missed by conventional imaging. Retention of tracer in the kidney due to Hydronephrosis is frequently encountered, which can draw attention to presence of obstructive uropathy as a possible etiology (Figure 6). + + 1800 + Gene + 80192 + + MDP + + + MESH:D006869 + Disease + + Hydronephrosis + + + MESH:C536483 + Disease + + obstructive uropathy + + + + FIG + AIM-22-297-g008.jpg + F6 + fig_caption + 14589 + 6 years old girl with back pain; scan revealed radiotracer retention within left kidney. Finding was suggestive of hydronephrosis-PUJ stenosis. + + 9606 + Species + + girl + + + MESH:D001416 + Disease + + back pain + + + MESH:D006869 + Disease + + hydronephrosis-PUJ stenosis + + + + DISCUSS + paragraph + 14733 + Finally we believe that plain x-ray can miss serious pathologies in pediatric patients with back pain, causing delay and unnecessary radiation exposure to this group of patients. Bone isotope scan has high negative predictive value in this study (98%), and consequently it can be safe to proceed into clinical follow up and maybe symptomatic management without the need to do any advanced investigations. We do not recommend the use bone isotope scan in each child with back pain, but when we have clinical data suggestive of an underlying bony pathology, bone isotope scan should be the first line of radiological investigation, before proceeding to CT or MRI. Also, Bone scintigraphy SPECT/CT which includes optimized CT as part of the study, provide improved diagnostic accuracy for back pain and oncology compared to routine BS, which may decrease the number of studies an individual child may need. This imaging modality, however, must be tailored on an individual basis as the addition of the CT study can increase exposure to the child and should only be performed after appropriate justification and with adherence to optimized low dose pediatric protocols. + + 9606 + Species + + patients + + + 9606 + Species + + patients + + + 9606 + Species + + child + + + 9606 + Species + + child + + + 9606 + Species + + child + + + MESH:D001416 + Disease + + back pain + + + MESH:D001847 + Disease + + bone isotope + + + MESH:D001416 + Disease + + back pain + + + MESH:D001416 + Disease + + back pain + + + MESH:D001847 + Disease + + bone isotope + + + + CONCL + title_1 + 15899 + 5. CONCLUSION + + + CONCL + paragraph + 15913 + Bone isotope scan is a non-invasive, highly sensitive and reliable imaging modality in the assessment of pediatric patients with back pain and in detecting most possible bony pathologies. We recommend this investigation as first imaging procedure to avoid any delay in the diagnosis and to avoid unnecessary investigations. + + 9606 + Species + + patients + + + MESH:D001416 + Disease + + back pain + + + + CONCL + footnote + 16237 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 16274 + REFERENCES + + + 46 + surname:McGrath;given-names:PJ + REF + surname:Goodman;given-names:JE + 247 + 1758709 + 1991 + Pain + ref + 264 + 16285 + The epidemiology of pain in children and adolescents: a review + + 9606 + Species + + children + + + MESH:D010146 + Disease + + pain + + + + 20 + surname:Waldburger;given-names:M + surname:Dutoit;given-names:G + REF + surname:Balague;given-names:F + 175 + 2976526 + 1988 + Scand J Rehabil Med + ref + 179 + 16348 + Low back pain in school children. 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Roentgenography;gallium, indium-labeled white blood cell, gammaglobulin, bone scintigraphy;and MRI + + MESH:D007204 + Chemical + + indium + + + MESH:D005708 + Chemical + + gallium + + + MESH:D009140 + Disease + + musculoskeletal infection + + + + 155 + REF + surname:Al-Sheikh;given-names:W + 501 + 3157204 + 1985 + Radiology + ref + 506 + 17721 + Subacute and chronic bone infections: Diagnosis using In-111, Ga-67 and Tc-99m MDP bone scintigraphy, and radiography + + 1800 + Gene + 80192 + + MDP + + + MESH:D001847 + Disease + + bone infections + + + MESH:D005708 + Chemical + + Ga + + + + 21 + surname:Luk;given-names:KD + surname:Ho;given-names:WY + REF + surname:Peh;given-names:WC + 54 + 9117933 + 1997 + Clin Imaging + ref + 62 + 17839 + Applications of bone scintigraphy in ankylosing spondylitis + + MESH:D013167 + Disease + + ankylosing spondylitis + + + + 46 + REF + surname:Even-Sapir;given-names:E + 1356 + 16085595 + 2005 + J Nucl Med + ref + 1367 + 17899 + Imaging of malignant bone involvement by morphologic, scintigraphic, and hybrid modalities + + + 23 + REF + surname:Love;given-names:C + 341 + 12640151 + 2003 + Radiographics + ref + 358 + 17990 + Radionuclide bone imaging: an illustrative review + + + 33 + REF + surname:Murphey;given-names:MD + 803 + 23674776 + 2013 + Radiographics + ref + 831 + 18040 + From the radiologic pathology archives. Ewing sarcoma family of tumors: radiologic-pathologic correlation + + MESH:D009369 + Disease + + tumors + + + MESH:C563168 + Disease + + Ewing sarcoma + + + + 1 + surname:Khan;given-names:A + surname:Epstein;given-names:JA + REF + surname:Janin;given-names:Y + 31 + 7207770 + 1981 + Neurosurgery + ref + 38 + 18146 + Osteoid osteomas and osteoblastoma of the spine + + MESH:D010017 + Disease + + Osteoid osteomas + + + MESH:D018215 + Disease + + osteoblastoma + + + + 198 + REF + surname:Iyer;given-names:RS + 1039 + 22528893 + 2012 + AJR + ref + 1052 + 18194 + Pediatric bone imaging: diagnostic imaging of osteoid osteoma + + MESH:D010017 + Disease + + osteoid osteoma + + + + 29 + surname:Konerding;given-names:KF + surname:Katz;given-names:PG + REF + surname:Haden;given-names:HT + 1781 + 11 + 3183746 + 1988 + J Nucl Med + ref + 1785 + 18256 + Detection of obstructive uropathy by bone scintigraphy + + MESH:C536483 + Disease + + obstructive uropathy + + + + + 4272847 + + surname:Roupa;given-names:Zoe + surname:Hatzoglou;given-names:Chryssi + surname:Gourgoulianis;given-names:Konstantinos + 25568580 + surname:Koulouri;given-names:Agoritsa + 315 + 5 + 2014 + 4272847 + AIM-22-315 + 10.5455/aim.2014.22.315-319 + front + 22 + This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-Share Alike 3.0 Unported, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. + TITLE + Acta Inform Med; 2014 Oct; 22(5) 315-9. doi:10.5455/aim.2014.22.315-319 + Chronic Obstructive Pulmonary Disease Quality of life Telemetry + 319 + Koulouri A, Gourgoulianis K, Hatzoglou C, Roupa Z + 0 + Telemetric Detection of Chronic Obstructive Pulmonary Disease and Investigation of Quality of Life for People Working in Shipbuilding Industry + + MESH:D006212 + Disease + + People Working + + + MESH:D029424 + Disease + + Chronic Obstructive Pulmonary Disease + + + + ABSTRACT + abstract_title_1 + 143 + ABSTRACT + + + ABSTRACT + abstract_title_1 + 152 + Introduction: + + + ABSTRACT + abstract + 166 + Chronic Obstructive Pulmonary Disease (COPD) has a significant impact on quality of life-related health. + + 260431 + Gene + + COPD + + + MESH:D029424 + Disease + + Chronic Obstructive Pulmonary Disease + + + + ABSTRACT + abstract_title_1 + 271 + Aim: + + + ABSTRACT + abstract + 276 + It was the detection of Chronic Obstructive Pulmonary Disease by using telemetric methods and the investigation of the quality of life for people working in Shipbuilding Industry compared with a control group. + + 9606 + Species + + people + + + MESH:D029424 + Disease + + Chronic Obstructive Pulmonary Disease + + + + ABSTRACT + abstract_title_1 + 486 + Methods: + + + ABSTRACT + abstract + 495 + A group of one hundred men working in the shipbuilding industry aged 51.8 +- 8.2 years old and a control group of one hundred men of the general population aged 51.1 +- 6.4 years were studied. All participants completed the General Health Questionnaire - 28, the Fagerstrom test and a form with demographic characteristics. Pulmonary function test results were electronically sent to a specialist for evaluation. + + 9606 + Species + + men + + + 9606 + Species + + men + + + 9606 + Species + + participants + + + + ABSTRACT + abstract_title_1 + 910 + Results: + + + ABSTRACT + abstract + 919 + People working in the shipbuilding zone had significantly lower values (p<0.001) in FVC, FEV1 and FEV1/FVC compared with the general population participants. Worse social functionality was exhibited by workers in the shipbuilding zone, people with elementary education, unemployed and by those suffering from comorbidities (p <0.001). + + 9606 + Species + + People + + + 9606 + Species + + participants + + + 9606 + Species + + people + + + + ABSTRACT + abstract_title_1 + 1254 + Conclusions: + + + ABSTRACT + abstract + 1267 + Health level and its individual dimensions are both associated with health self-assessment and occupational and economic status. The coexistence of chronic diseases and smoking dependence affects emotion and social functioning of individuals. + + MESH:D002908 + Disease + + chronic diseases + + + + INTRO + title_1 + 1510 + 1. INTRODUCTION + + + INTRO + paragraph + 1526 + Chronic Obstructive Pulmonary Disease (COPD) is a life-threatening disease characterized by airflow limitation, which is not fully reversible. The World Health Organization (WHO) estimates that 210 million people worldwide suffer from COPD. Despite prevention efforts, the disease appears to be increasing and predictions show that in 2030 it will be the third cause of death. COPD is responsible for a large number of visits to doctors, outpatient departments and hospitals. It has been recorded as the 8th leading cause of years lived with disability in men and 7th in women in the United States of America. Reduction in expiratory flow is slow, progressive and associated with an abnormal inflammatory response of the lungs to irritating particles or gases. + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 9606 + Species + + people + + + 9606 + Species + + outpatient + + + 9606 + Species + + men + + + 9606 + Species + + women + + + MESH:D029424 + Disease + + Chronic Obstructive Pulmonary Disease + + + MESH:D003643 + Disease + + death + + + + INTRO + paragraph + 2287 + COPD affects the entire body and nowadays is considered a multi systemic disease, since it is one of the leading causes of death in the world with high levels of hospitalization and significant financial cost. Smoking is the main causative factor of COPD, while other factors deemed to act aggravatingly and contribute to its occurrence is air pollution and occupational exposure to industrial pollutants (dust, chemicals, fumes, irritant gases, smoke). Hnizdo E. et al., found that COPD percentage caused by the working environment was calculated as 19.2% overall and 31.1% among those who never smoked. The diagnosis of COPD is done by spirometry which is a simple but basic examination in the context of respiratory function testing. The use of spirometry with telemetric data transmission may allow early diagnosis of the disease and facilitate the initiation of medication. In patients with chronic diseases such as asthma, diabetes and COPD, the use of telehealth methods can reduce hospital admissions without increasing mortality, but with increasing people's quality of life. + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 9606 + Species + + patients + + + 9606 + Species + + people + + + MESH:D001249 + Disease + + asthma + + + MESH:C564969 + Disease + + multi systemic disease + + + MESH:D003643 + Disease + + death + + + MESH:D003920 + Disease + + diabetes + + + MESH:D002908 + Disease + + chronic diseases + + + + INTRO + paragraph + 3372 + Quality of life is a highly individualized concept associated with health and formed mainly by health status (physical and psychological), functionality, knowledge of the disease, health habits, use of services, social support system and by the individual's adaptive capacity, financial situation and educational level. We can quantify it by using measurement tools, such as questionnaires, and find the correlation that probably exists between the objective laboratory data on the health status of people with COPD and the subjective perception that affects one's personality. The purpose of this study was the detection of Chronic Obstructive Pulmonary Disease by using telemetric methods and investigation of the quality of life for people working in Shipbuilding Industry by a control group. + + 260431 + Gene + + COPD + + + 9606 + Species + + people + + + 9606 + Species + + people + + + MESH:D029424 + Disease + + Chronic Obstructive Pulmonary Disease + + + + METHODS + title_1 + 4168 + 2. METHODS + + + METHODS + paragraph + 4179 + Sample - Participants + + + METHODS + paragraph + 4203 + A group of one hundred people working in the Shipbuilding Industry (SI) was studied as well as a control group of one hundred people of the general population. All were middle aged men and constituted a convenience sample. + + 9606 + Species + + people + + + 9606 + Species + + people + + + 9606 + Species + + men + + + Disease + + SI + + + + METHODS + paragraph + 4426 + Data were collected on demographic characteristics, education, marital status, smoking dependence and personal medical history. Additionally, data were collected on employment status, self-assessment of health and use of health services. Independence in everyday life activities and quality of life were assessed by using St. George's Respiratory Questionnaire and smoking habit by using Fagerstrom test. St. George's Respiratory Questionnaire is a questionnaire designed to assess the relationship of pulmonary disease with quality of life associated with health and wellbeing. The average time to fill it out ranges from 10 to 15 minutes depending on age and educational level of the individual. The questionnaire provides a sum total and scores for individual symptoms (cough, sputum, wheezing, dyspnea, their frequency and duration), activities (which cause or are limited by dyspnea) and impacts (in a wide range of social functioning and mental disorders as a result of airway obstruction). The lowest score represents a better quality of life. + + MESH:D003371 + Disease + + cough + + + MESH:D004417 + Disease + + dyspnea + + + MESH:D001523 + Disease + + mental disorders + + + MESH:D012135 + Disease + + wheezing + + + MESH:D004417 + Disease + + dyspnea + + + MESH:D008171 + Disease + + pulmonary disease + + + + METHODS + paragraph + 5477 + The questionnaire assessing the degree of nicotine dependence (Fagerstrom test) was used to provide a meaningful picture of smoking habit. It is a questionnaire easy to fill out, with its composite score of 10 points indicating the maximum nicotine dependence. + + MESH:D009538 + Chemical + + nicotine + + + MESH:D009538 + Chemical + + nicotine + + + + METHODS + paragraph + 5738 + Study Design -Research Tools + + + METHODS + paragraph + 5769 + All individuals underwent spirometry and wireless medical devices were used in conjunction with means of web access and software for biomedical parameters telemetry service. The telemetry system used is a web application based on the new generation telemedical devices of automatic measurement and wireless transmission of biological signals. + + + METHODS + paragraph + 6112 + Data were stored in an electronic page which was accessed by the specialist physician who recorded the COPD diagnosis. Diagnosis was based on symptoms and spirometry findings and airway obstruction was staged according to GOLD guidelines. Participants were asked to make at least three attempts and the highest values of FEV1 and FVC were used to classify the severity of the degree of airway obstruction. + + 260431 + Gene + + COPD + + + 9606 + Species + + Participants + + + + METHODS + paragraph + 6518 + Statistical analysis + + + METHODS + paragraph + 6539 + Descriptive and inferential analysis was performed Depending on indications, Student's t-test or Mann-Whitney nonparametric test was used to compare quantitative variables between two groups. Respectively, parametric analysis of variance (ANOVA) or Kruskal-Wallis nonparametric test was used to compare quantitative variables between more than two groups. Bonferroni correction was used to counteract type I error Pearson's chi-squared test or Fisher's exact test was used to compare proportions where necessary. Spearman's or Pearson's (r) correlation coefficient was used to control the relationship between two quantitative variables. The level of statistical significance was set at 0.05 and SPSS 17.0 statistical software was used in the analysis. + + MESH:C536353 + Disease + + type I error Pearson + + + + RESULTS + title_1 + 7292 + 3. RESULTS + + + RESULTS + paragraph + 7303 + Characteristics of Participants + + + RESULTS + paragraph + 7335 + The mean age of workers and general population group was 51.8 +- 8.2 and 51.1 +- 6.4 years with 25.1 +- 11.5 and 25.46 +- 10 years duration of smoking, respectively. There was a significantly higher percentage of workers in the general population (75%) compared with the corresponding percentage in the group of people who worked in the shipbuilding zone (62%). The working years in the two groups were 10 +- 3 and 25.4 +- 8.7 respectively. + + 9606 + Species + + people + + + + RESULTS + paragraph + 7776 + Fagerstrom score of participants did not differ between the two groups, as well as their smoking dependence level. + + 9606 + Species + + participants + + + + RESULTS + paragraph + 7891 + The following diseases were found in the general population in order of frequency: arterial hypertension 12%, COPD 10%, heart diseases 9%, orthopedic problems 2%, diabetes mellitus 6% and other 7%. Respectively, in the shipbuilding industry workers were found: arterial hypertension 7%, COPD 13%, heart diseases 4%, orthopedic problems 7%, diabetes mellitus 4% and other 7%. + + 544442 + Gene + + COPD 10 + + + 544417 + Gene + + COPD 13 + + + MESH:D006331 + Disease + + heart diseases + + + MESH:D006331 + Disease + + heart diseases + + + MESH:D006973 + Disease + + hypertension + + + MESH:D003920 + Disease + + diabetes mellitus + + + MESH:D003920 + Disease + + diabetes mellitus + + + MESH:D006973 + Disease + + hypertension + + + + RESULTS + paragraph + 8266 + No participant was found in stage IV in the general population group and 3 persons respectively were found in the group of people working in the shipbuilding industry, according to GOLD guidelines. The workers in the shipbuilding zone had significantly lower values (p < 0.001) in FVC, FEV1 and FEV1/FVC compared with the participants of the general population. There was no significant difference in COPD stage of participants depending on their workplace, although most workers in the zone had advanced-stage COPD compared with the general population (Table 1). + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 9606 + Species + + participant + + + 9606 + Species + + persons + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + + TABLE + T1.xml + T1 + table_caption + 8830 + Spirometric findings in cases and controls. *Student's t-test **Pearson's x test +Fisher's exact test + + + RESULTS + paragraph + 8932 + Quality of life + + + RESULTS + paragraph + 8948 + Significantly higher score in "Symptoms and Activity" dimensions was found in the workers in the shipbuilding industry, those with basic education and non-actively working during this period. Health evaluation as moderate/poor and comorbidity are also associated with high score in these particular dimensions of life quality (p < 0.001). Also, people who were hospitalized during the last year and those diagnosed with III-IV COPD stage had significantly higher values in Symptoms and Activity score, compared to participants with I-II stage. There was a significant difference in Symptoms score depending on smoking dependence. Participants with low smoking dependence had significantly lower scores compared with participants with high smoking dependence (p = 0.016). Participants who visited a general practitioner more than 3 times during the last year had significantly higher values in Activity score compared to participants who visited him/her 1-2 times and to participants who did not visit him/her at all (p = 0.002 and p = 0.008 respectively) (Table 2). + + 260431 + Gene + + COPD + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + Participants + + + 9606 + Species + + participants + + + 9606 + Species + + Participants + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + + TABLE + T2.xml + T2 + table_caption + 10018 + Symptoms score, Activity score, self-reported health and use of health services indicators. *Kruskal-Wallis test + + + RESULTS + paragraph + 10131 + In "Impact score" dimension, a significantly higher score (p < 0.001), corresponding to more problems of social conciliation, was exhibited by those who considered their health as moderate/poor, suffered from a chronic disease and were diagnosed with III-IV COPD stage. In "Total score", participants who had visited a general practitioner more than three times during the last year had significantly higher values in Total score compared both to participants who had visited him/her 1-2 times and to those who had not visited him/her at all (p = 0.004 and p = 0.001 respectively) (Table 3). A significant positive correlation was observed between the score of participants in dimensions under study of quality of life with age, years of smoking and values of FVC, FEV1, FEV1/FVC and Fagerstrom score (p < 0.001) (Table 4). + + 260431 + Gene + + COPD + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + 9606 + Species + + participants + + + MESH:D002908 + Disease + + chronic disease + + + + TABLE + T3.xml + T3 + table_caption + 10963 + Impact score, Total score, self-reported health and use of health services indicators. *Kruskal-Wallis test + + + TABLE + T4.xml + T4 + table_caption + 11071 + Demograhic and spirometric correlations + + + DISCUSS + title_1 + 11111 + 4. DISCUSSION + + + DISCUSS + paragraph + 11125 + Health professionals have recognized the need for early detection of COPD and introduced new ways to facilitate its diagnosis by using modern telehealth methods such as telemetric data transfer. Masa et al. argue that, if high quality spirometry could be performed online through a connection of the specialist to the Pulmonary Laboratory and the patient in a PHC structure by qualified personnel and high quality spirometer, possibly the problem of performing a reliable and qualitative spirometry would be resolved. + + 260431 + Gene + + COPD + + + 9606 + Species + + patient + + + + DISCUSS + paragraph + 11643 + Spirometry with telemetric data transmission method was used in the present study and it was found that the prevalence of COPD in the general population was 10%. Similarly, in their study Tzanakis et al., found a prevalence of 11.6% in men and in an epidemiological study conducted by Sichleridis et al the prevalence of COPD was also estimated at 8.2% among men. + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 9606 + Species + + men + + + 9606 + Species + + men + + + + DISCUSS + paragraph + 12007 + The results of an epidemiological study conducted in Ireland from 1999 to 2001 on a sample of the general population aged 40-69 years showed that the prevalence of obstructive lung disease was 6.3%. In order to estimate the prevalence of COPD in this study, medical history, spirometry, a special respiratory symptoms questionnaire as in ours and dermal tests were used. Also, the BOLD (Burden of Obstructive Lung Disease) study that attempted to identify the prevalence of COPD by using population sample from 12 different countries found that the prevalence of COPD was 11.8 +- 7.9% in men. It is noted that in our study the prevalence of COPD was estimated at 10% in the general population and 13% in people working in the shipbuilding industry and no statistically significant difference was observed between the two groups. It is well known that there are many factors responsible for COPD such as environmental pollution, occupation, place of residence and respiratory infections during childhood. In this study, it was not possible to study the effects of each of these factors, but probably the highest prevalence of COPD in the group of people working in the shipbuilding industry is due to overburdened environment because of occupational exposure. Although it is known that air pollution has a serious impact on respiratory symptoms, the relationship between pollution and COPD remains to be elucidated. + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 260431 + Gene + + COPD + + + 9606 + Species + + men + + + 9606 + Species + + people + + + 9606 + Species + + people + + + MESH:D012131 + Disease + + respiratory infections + + + MESH:D008173 + Disease + + obstructive lung disease + + + + DISCUSS + paragraph + 13422 + The results of our research showed that there was a significant positive correlation of the participants' score in the SGRQ scales with age, years of smoking and Fagerstrom Score. Regarding the presence of respiratory symptoms, limitation of activities and their impacts which affect the field of social functioning, results were similar in a research of Xydeas-Kikemenis et al. in a sample of 203 people working in the construction industry. It was observed that age and duration of working activity were strong determinants of mental problems incidence affected by the workload and the negative characteristics of the workplace. Pierrakos et al., in their work concerning the quality of life related to smokers and non-smokers health, find that high smoking dependence negatively affects the quality of life which is exacerbated when tobacco consumption increases. + + 9606 + Species + + participants + + + 9606 + Species + + people + + + 4097 + Species + + tobacco + + + + DISCUSS + paragraph + 14291 + In contrast, SGRQ scales showed a high negative correlation with spirometric results (FEV1%, FVC% and FEV1/FVC) and thus, as values of the above measurements of the participants increase, the respective score implying better quality of life decreases. The studies of Stahl . and Ketelaars CA., in patients with COPD also demonstrate that the quality of life is positively correlated with good respiratory function while it is burdened with advancing age and disease. A moderate correlation of FEV1 values with health level and life quality emerges from the studies of Aibar Arregui et al., and Pereira et al.. + + 260431 + Gene + + COPD + + + 9606 + Species + + participants + + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 14905 + In our study, comorbidities showed a significant positive correlation with all subscales of quality of life, as supported by many researchers, to the extent that the disease has physical and psychosocial impacts on the overall situation of the individual and chronic patient is observed living under conditions acting bindingly as to his/her quality of life. Overall, the coexistence of chronic diseases burdens the person mainly with organic and functional problems, frequent hospitalizations, continuous medical visits and examinations and secondarily with social, psychological and occupational problems that affect his/her quality of life. + + 9606 + Species + + patient + + + 9606 + Species + + person + + + MESH:D002908 + Disease + + chronic diseases burdens + + + MESH:C535569 + Disease + + psychosocial + + + + DISCUSS + paragraph + 15549 + Educational level and occupational status have significant positive correlation with all SGRQ scales. Participants with basic educational level had higher scores in scales of symptoms, impacts, activities and overall, as in the study of Dimitropoulos et al., in which people with lower educational level had a more aggravated symptoms scale compared to the more educated participants. Based on that, people with low educational level tend to occupy lower-paid jobs in the labor market and often those associated with exposure to harmful for health factors. + + 9606 + Species + + Participants + + + 9606 + Species + + people + + + 9606 + Species + + participants + + + 9606 + Species + + people + + + + DISCUSS + paragraph + 16106 + In our study, admission to hospital during the last year is significantly correlated with the quality of life scales and is partially consistent with the studies of Dimitropoulos et al., and Miravitlles et al., where patients with more than two admissions to hospital during the last year had worse scores in all scales, but without statistical correlation. + + 9606 + Species + + patients + + + + DISCUSS + paragraph + 16464 + The contact frequency of participants (> 3 times) with a general practitioner in the last 6 to 12 months was positively correlated with the quality of life scales and is consistent with the results of the study of Tountas et al., of whom 37% of the sample under study had visited a doctor >1 time in the previous month. + + 9606 + Species + + participants + + + + DISCUSS + paragraph + 16784 + The loss of employment (unemployment) and self-assessment of health as moderate/poor in our study were statistically significantly associated with higher score in almost all scales of the quality of life questionnaire. Our findings are consistent with those of Zimmer et al., who argue that active employment is a determinant of subjective evaluation of health status. + + + CONCL + title_1 + 17153 + 5. CONCLUSION + + + CONCL + paragraph + 17167 + Educational level, occupational status and presence of chronic diseases affect the quality of life of individuals, while "low" self-assessment of health, increased frequency of visits to a general practitioner and repeated admissions to hospital are associated with social dysfunction and reduced quality of life. Telemetric diagnosis of COPD can become a method of choice in a community environment, but requires enhancement of education and training of health professionals that are going to perform it. + + 260431 + Gene + + COPD + + + MESH:D000067404 + Disease + + social dysfunction + + + MESH:D002908 + Disease + + chronic diseases + + + + CONCL + footnote + 17677 + CONFLICT OF INTEREST: NONE DECLARED. + + + REF + title + 17714 + REFERENCES + + + surname:Kilgert;given-names:K + surname:Lippert;given-names:B + surname:Berger;given-names:K + surname:Nowak;given-names:D + 381 + 16336024 + 2005 + Treat Respir Med + ref + 4 + REF + 395 + surname:Sandtmann;given-names:R + surname:Caeser;given-names:M + 17725 + Epidemiology and health economics of COPD across Europe: a critical analysis + + 260431 + Gene + + COPD + + + + REF + 2004 + ref + 17802 + GOLD, Global Initiative for Chronic Obstructive Lung Disease. 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surname:Rubio;given-names:M + 918 + surname:Zamorano;given-names:J + surname:Corral;given-names:J + surname:Riesco;given-names:JA + 18548 + Validity of spirometry performed online + + + 125 + surname:Filaditaki;given-names:V + surname:Anagnostopoulou;given-names:U + REF + surname:Tzanakis;given-names:Nu + 892 + 15006947 + 2010 + Chest + ref + 900 + 18588 + Prevalence of COPD in Greece + + 260431 + Gene + + COPD + + + + surname:Kottakis;given-names:I + surname:Tsiotsios;given-names:I + surname:Chloros;given-names:D + surname:Sichletidis;given-names:LT + 92 + 16701767 + 2006 + Prim Care Respir J + ref + 15 + REF + surname:Posporelis;given-names:S + surname:Kalkanis;given-names:D + 97 + surname:Karamanlidis;given-names:A + surname:Kaouri;given-names:S + surname:Kaiafa;given-names:O + 18617 + High prevalence of smoking in Northern Greece + + + surname:Shepherd;given-names:R + surname:Gingles;given-names:J + surname:Heaney;given-names:L + surname:Murtagh;given-names:E + 443 + 16080593 + 2005 + Eur J Epidemiol + ref + 20 + REF + 453 + surname:MacMahon;given-names:J + surname:Patterson;given-names:C + surname:Kee;given-names:F + 18663 + Prevalence of obstructive lung disease in a general population sample: the NICECOPD study + + MESH:D008173 + Disease + + obstructive lung disease + + + + surname:Elias;given-names:JA + REF + surname:Fishman;given-names:AP + 2008 + ref + 18753 + Fishman's Pulmonary Diseases and Disorders + + MESH:C535736 + Disease + + Fishman's Pulmonary Diseases + + + + REF + Available at: + BOLD Study + ref + 18796 + BOLD Study + + + 277 + surname:Weinmann;given-names:GG + REF + surname:Petty;given-names:TL + 246 + 9005275 + 1997 + JAMA + ref + 253 + 18807 + Building a national strategy for the prevention and management of and research in chronic obstructive pulmonary disease: National Heart, Lung, and Blood Institute workshop summary + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + + surname:Ktenas;given-names:E + surname:George;given-names:G + surname:Aloumanis;given-names:P + REF + surname:Xydea-Kikemeni;given-names:A + 2003 + Greek Medical Journal + ref + 18987 + Workers' in the construction sector morbidity + + + surname:Ganas;given-names:A + 50 + surname:Kondili;given-names:I + surname:Michopoulou;given-names:E + REF + surname:Pierrakos;given-names:G + 203 + 2011 + NOSILEFTIKI + ref + 213 + surname:Sarris;given-names:M + 19035 + Health-related Quality of Life of Smokers and Non-Smokers + + + surname:Ronmark;given-names:E + surname:Jansson;given-names:SA + surname:Lindberg;given-names:Alpha + surname:Stahl;given-names:Epsilon + 56 + 16153294 + 2005 + Health and Quality of Life Outcomes + ref + 3 + REF + surname:Lundback;given-names:B + surname:Lofdahl;given-names:CG + surname:Andersson;given-names:F + surname:Svensson;given-names:K + 19093 + Health-related quality of life is related to COPD disease severity + + MESH:D029424 + Disease + + COPD disease + + + + surname:Huyer Abu-Saad;given-names:H + surname:Mostert;given-names:R + surname:Schlosser;given-names:MA + surname:Ketelaars;given-names:CA + 39 + 8658367 + 1996 + Thorax + ref + 51 + REF + 43 + surname:Wouters;given-names:EF + surname:Halfens;given-names:RJ + 19160 + Determinants of health-related quality of life in patients with chronic obstructive pulmonary disease + + 9606 + Species + + patients + + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + + 24 + surname:Conget;given-names:Lopez F + surname:Laborda;given-names:Ezquerra K + REF + surname:Aibar;given-names:Arregui MA + 473 + 18271650 + 2007 + An Med Interna + ref + 477 + 19262 + Relationship between objective data and health-related quality of life in COPD patients + + 260431 + Gene + + COPD + + + + surname:Medeiros;given-names:M + surname:Alcantara;given-names:M + surname:Pinto;given-names:R + surname:Pereira;given-names:ED + 730 + 19750324 + 2009 + J Bras Pneumol + ref + 35 + REF + 736 + surname:Mota;given-names:RM + 19350 + Influence of respiratory function parameters on the quality of life of COPD patients + + 260431 + Gene + + COPD + + + + 11 + surname:Simmons;given-names:R + REF + surname:Dew;given-names:HA + 23 + 1990 + Scand J Urol Nephrol + ref + 30 + 19435 + The advantage of multiple measures of quality of life + + + surname:Skopelitou;given-names:M + 8 + surname:Sotiropoulou;given-names:P + surname:Roupa;given-names:Z + REF + surname:Koulouri;given-names:A + 255 + 2009 + Rostrum of Asclepius + ref + 269 + 19489 + Mood disorders and handling of daily routine in patients with diabetes mellitus type II + + 9606 + Species + + patients + + + MESH:D003924 + Disease + + diabetes mellitus type II + + + MESH:D019964 + Disease + + Mood disorders + + + + surname:Nikolaou;given-names:A + surname:Kopitopoulou;given-names:A + surname:Theodorakopoulou;given-names:E + surname:Dimitropoulos;given-names:C + 127 + 2013 + PNEUMON + ref + 2 + REF + surname:Kosmas;given-names:E + 151 + surname:Ntanos;given-names:I + surname:Koulouris;given-names:N + surname:Tsapas;given-names:C + 19577 + The health related quality of life of Greekpatients with chronic obstructive pulmonary disease + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + + surname:Scambler;given-names:G + surname:Blane;given-names:D + REF + surname:Scambler;given-names:G + 107 + 2003 + Sociology as applied to medicine + ref + 123 + 19672 + Inequality and social class + + + 59 + surname:Pont;given-names:A + surname:Ferrer;given-names:M + REF + surname:Miravitlles;given-names:M + 387 + 15115864 + 2004 + Thorax + ref + 395 + 19700 + Effect of exacerbations on quality of life in patients with chronic obstructive pulmonary disease: a 2 year follow up study + + 9606 + Species + + patients + + + MESH:D029424 + Disease + + chronic obstructive pulmonary disease + + + + REF + surname:Tountas;given-names:Y + 2008 + ref + 19824 + Health services in Greece (1996-2006) Athens: Center for Health Services Research + + + surname:Chayovan;given-names:N + 41 + surname:Lin;given-names:H + surname:Natividad;given-names:J + REF + surname:Zimmer;given-names:Z + 465 + 11198569 + 2000 + Journal of health and Social Behavior + ref + 481 + 19906 + A cross-national examination of the determinants of self-assessed health + + + diff --git a/data/temp/batch_log.tsv b/data/temp/batch_log.tsv new file mode 100644 index 0000000..1753c29 --- /dev/null +++ b/data/temp/batch_log.tsv @@ -0,0 +1,133 @@ +batch pmcid +0 PMC2289422 +0 PMC2289423 +0 PMC2289424 +0 PMC2289425 +0 PMC2289426 +0 PMC2289427 +0 PMC2289428 +0 PMC2289429 +0 PMC2289430 +0 PMC2289434 +0 PMC2289435 +0 PMC2289436 +0 PMC2289437 +0 PMC2289438 +0 PMC2289439 +0 PMC2289440 +0 PMC2289441 +0 PMC2289442 +0 PMC2289443 +0 PMC2289444 +0 PMC2289445 +0 PMC2289446 +0 PMC2289447 +0 PMC2289448 +0 PMC2289449 +0 PMC2289450 +0 PMC2289451 +0 PMC2289452 +0 PMC2289453 +0 PMC2289454 +0 PMC2289455 +0 PMC2289456 +0 PMC2289457 +0 PMC2289458 +0 PMC2289459 +0 PMC2289460 +0 PMC2289461 +0 PMC2289462 +0 PMC2289463 +0 PMC2289464 +0 PMC2289465 +0 PMC2289466 +0 PMC2289467 +0 PMC2289468 +0 PMC2289469 +0 PMC2289470 +0 PMC2289471 +0 PMC2289472 +0 PMC2289473 +0 PMC2289474 +0 PMC2289475 +0 PMC2289476 +0 PMC2289477 +0 PMC2289478 +0 PMC2289479 +0 PMC2289480 +0 PMC2289481 +0 PMC2289482 +0 PMC2289483 +0 PMC2289484 +0 PMC2289485 +0 PMC2289486 +0 PMC2289487 +0 PMC2289488 +0 PMC2289489 +0 PMC2289490 +0 PMC2289491 +0 PMC2289492 +0 PMC2289493 +0 PMC2289494 +0 PMC2289495 +0 PMC2289496 +0 PMC2289497 +0 PMC2289498 +0 PMC2289499 +0 PMC2289500 +0 PMC2289501 +0 PMC2289502 +0 PMC2289503 +0 PMC2289504 +0 PMC2289505 +0 PMC2289506 +0 PMC2289507 +0 PMC2289508 +0 PMC2289509 +0 PMC2289510 +0 PMC2289512 +0 PMC2289513 +0 PMC2289514 +0 PMC2289515 +0 PMC2289517 +0 PMC2289518 +0 PMC2289519 +0 PMC2289520 +0 PMC2289521 +0 PMC2289522 +0 PMC2289524 +0 PMC2289525 +0 PMC4272812 +0 PMC4272813 +1 PMC4272814 +1 PMC4272815 +1 PMC4272816 +1 PMC4272818 +1 PMC4272820 +1 PMC4272821 +1 PMC4272822 +1 PMC4272823 +1 PMC4272824 +1 PMC4272825 +1 PMC4272826 +1 PMC4272827 +1 PMC4272828 +1 PMC4272829 +1 PMC4272830 +1 PMC4272831 +1 PMC4272832 +1 PMC4272833 +1 PMC4272834 +1 PMC4272835 +1 PMC4272836 +1 PMC4272837 +1 PMC4272838 +1 PMC4272839 +1 PMC4272840 +1 PMC4272841 +1 PMC4272842 +1 PMC4272843 +1 PMC4272844 +1 PMC4272845 +1 PMC4272846 +1 PMC4272847 diff --git a/environment.yml b/environment.yml deleted file mode 100644 index 5d0b0b9..0000000 --- a/environment.yml +++ /dev/null @@ -1,11 +0,0 @@ -name: pubtator -dependencies: -- python=3.8 -- lxml=4.4 -- tqdm -- requests -- pandas=0.25.3 -- pip -- pip: - - git+https://github.com/en-dash/PyBioC@a21bd181ce3b89de791db4bb7db6ca1c48167648 - - ratelimit \ No newline at end of file diff --git a/execute.py b/execute.py new file mode 100644 index 0000000..462944c --- /dev/null +++ b/execute.py @@ -0,0 +1,136 @@ +import argparse +from pathlib import Path +import json + +import wget + +from scripts.download_full_text import download_full_text, merge_full_text +from scripts.extract_tags import extract_annotations +from scripts.hetnet_id_extractor import filter_tags +from scripts.map_ids import map_ids +from scripts.pubtator_to_xml import convert_pubtator + +parser = argparse.ArgumentParser( + description='Runs Pubtator/Pubtator Central Parser Pipeline' +) + +parser.add_argument( + "--config", help="The config file for the extractor.", + default="config_files/pubtator_central_config.json" +) + +args = parser.parse_args() + +configuration = json.load(open(args.config, "r")) + +if ( + "repository_download" in configuration and + not configuration["repository_download"]["skip"] +): + print("Executing repository_download...") + wget.download( + configuration["repository_download"]['url'], + out=configuration["repository_download"]['download_folder'] + ) + + del configuration["repository_download"] + +if ( + "pubtator_to_xml" in configuration and + not configuration["pubtator_to_xml"]["skip"] +): + print("Executing pubtator_to_xml...") + convert_pubtator( + configuration["pubtator_to_xml"]['documents'], + configuration["pubtator_to_xml"]['output'] + ) + + del configuration["pubtator_to_xml"] + +if ( + "extract_tags" in configuration and + not configuration["extract_tags"]["skip"] +): + print("Executing extract_tags...") + extract_annotations( + configuration["extract_tags"]["input"], + configuration["extract_tags"]["output"] + ) + + del configuration["extract_tags"] + +if ( + "hetnet_id_extractor" in configuration and + not configuration["hetnet_id_extractor"]["skip"] +): + print("Executing hetnet_id_extractor...") + filter_tags( + configuration["hetnet_id_extractor"]["input"], + configuration["hetnet_id_extractor"]["output"] + ) + + del configuration["hetnet_id_extractor"] + +if ( + "map_pmids_to_pmcids" in configuration and + not configuration["map_pmids_to_pmcids"]["skip"] +): + print("Executing map_pmids_to_pmcids...") + map_ids( + configuration["map_pmids_to_pmcids"]["input"], + configuration["map_pmids_to_pmcids"]["output"], + configuration["map_pmids_to_pmcids"]["debug"] + ) + + del configuration["map_pmids_to_pmcids"] + +if ( + "download_full_text" in configuration and + not configuration["download_full_text"]["skip"] +): + print("Executing download_full_text...") + download_full_text( + configuration["download_full_text"]["input"], + configuration["download_full_text"]["document_batch"], + configuration["download_full_text"]["temp_dir"], + configuration["download_full_text"]["log_file"] + ) + + merge_full_text( + configuration["download_full_text"]["temp_dir"], + configuration["download_full_text"]["output"] + ) + + del configuration["download_full_text"] + +if ( + "extract_full_text_tags" in configuration and + not configuration["extract_full_text_tags"]["skip"] +): + print("Executing extract_full_text_tags...") + extract_annotations( + configuration["extract_full_text_tags"]["input"], + configuration["extract_full_text_tags"]["output"] + ) + + del configuration["extract_full_text_tags"] + +if ( + "hetnet_id_extractor_full_text" in configuration and + not configuration["hetnet_id_extractor_full_text"]["skip"] +): + print("Executing hetnet_id_extractor_full_text...") + filter_tags( + configuration["hetnet_id_extractor_full_text"]["input"], + configuration["hetnet_id_extractor_full_text"]["output"] + ) + + del configuration["hetnet_id_extractor_full_text"] + +if len(configuration) > 0: + not_executed_commands = '\n'.join(configuration.keys()) + print( + "\n" + "Commands not executed:\n" + f"{not_executed_commands}" + ) diff --git a/execute.sh b/execute.sh deleted file mode 100644 index 76c46fd..0000000 --- a/execute.sh +++ /dev/null @@ -1,76 +0,0 @@ -# Exit on an error -set -o errexit - -if [ "$1" == "--pubtator" ]; then - # PubTator FTP download - FTP_URL=ftp://ftp.ncbi.nlm.nih.gov/pub/lu/PubTator; - - wget \ - --timestamping \ - --directory-prefix=download \ - --output-file=download/bioconcepts2pubtator_offsets.gz.log \ - $FTP_URL/bioconcepts2pubtator_offsets.gz - - # Convert pubtator format to BioC XML - python scripts/pubtator_to_xml.py \ - --documents download/bioconcepts2pubtator_offsets.gz \ - --output data/pubtator-docs.xml.xz - - # Extract tags from the BioC XML to a TSV - python scripts/extract_tags.py \ - --input data/pubtator-docs.xml.xz \ - --output data/pubtator-tags.tsv.xz - - # Extract hetnet tags from the pubtator tags - python scripts/hetnet_id_extractor.py \ - --input data/pubtator-tags.tsv.xz \ - --output data/pubtator-hetnet-tags.tsv.xz - -else - # PubTator Central FTP download - FTP_URL=ftp://ftp.ncbi.nlm.nih.gov/pub/lu/PubTatorCentral; - - wget \ - --timestamping \ - --directory-prefix=download \ - --output-file=download/bioconcepts2pubtatorcentral_offset.gz.log \ - $FTP_URL/bioconcepts2pubtatorcentral.offset.gz - - # Convert pubtator format to BioC XML - python scripts/pubtator_to_xml.py \ - --documents download/bioconcepts2pubtatorcentral.offset.gz \ - --output data/pubtator-central-docs.xml.xz - - # Extract tags from the BioC XML to a TSV - python scripts/extract_tags.py \ - --input data/pubtator-central-docs.xml.xz \ - --output data/pubtator-central-tags.tsv.xz - - # Extract hetnet tags from the pubtator tags - python scripts/hetnet_id_extractor.py \ - --input data/pubtator-central-tags.tsv.xz \ - --output data/pubtator-central-hetnet-tags.tsv.xz - - # Grab ids with full text - python scripts/map_ids.py \ - --input data/pubtator-central-tags.tsv.xz \ - --output data/pubtator-pmids-to-pmcids.tsv - - # Download full text from pubtator central - python scripts/download_full_text.py \ - --input data/pubtator-pmids-to-pmcids.tsv \ - --document_batch 100 \ - --output data/pubtator-central-full-text.xml \ - --temp_dir data/temp - - # Extract tags from the BioC XML - python scripts/extract_tags.py \ - --input data/pubtator-central-full-text.xml \ - --output data/pubtator-central-full-text-tags.tsv.xz - - # Filter-Convert tags to Hetnet IDs - python scripts/hetnet_id_extractor.py \ - --input data/pubtator-central-full-text-tags.tsv.xz \ - --output data/pubtator-central-full-hetnet-tags.tsv.xz - -fi diff --git a/requirements.txt b/requirements.txt new file mode 100644 index 0000000..38230cc --- /dev/null +++ b/requirements.txt @@ -0,0 +1,7 @@ +lxml==4.5.2 +pandas==0.25.3 +ratelimit==2.2.1 +requests==2.22.0 +tqdm==4.11.2 +wget==3.2 +git+https://github.com/en-dash/PyBioC@a21bd181ce3b89de791db4bb7db6ca1c48167648 \ No newline at end of file diff --git a/scripts/download_full_text.py b/scripts/download_full_text.py index 2fa3d66..efc86a2 100644 --- a/scripts/download_full_text.py +++ b/scripts/download_full_text.py @@ -3,6 +3,7 @@ import requests import time import os +import sys import lxml.etree as ET from lxml.etree import XMLSyntaxError @@ -33,7 +34,7 @@ def read_id_chunk(id_file, batch_size): for id_batch in pd.read_csv(id_file, sep="\t", chunksize=batch_size): yield id_batch -def download_full_text(ids_file, document_batch, temp_dir): +def download_full_text(ids_file, document_batch, temp_dir, log_file="batch_log.tsv"): """ Download full text from Pubtator Central. This section involves heavy api calls and subject to get blocked. @@ -51,22 +52,70 @@ def download_full_text(ids_file, document_batch, temp_dir): # Create the directory if it doesn't exist Path(f"{temp_dir}").mkdir(parents=True, exist_ok=True) + + # Load a logger to track which ids have been queried + log_file_path = Path(f"{temp_dir}/{log_file}") + if not log_file_path.exists(): + log = pd.DataFrame([], columns=["batch", "pmcid"]) + log.to_csv( + log_file_path.resolve(), + sep="\t", index=False + ) + + else: + log = pd.read_csv( + log_file_path.resolve(), + sep="\t" + ) + # test each query against Pubtator's API for idx, pmcid_batch_df in tqdm.tqdm(enumerate(read_id_chunk(ids_file, document_batch))): - query = f"{pubtator_central_api}pmcids={','.join(pmcid_batch_df.PMCID.values)}" - response = call_api(query) + # Measure the ids that haven't been seen by the logger + already_seen = ( + set(pmcid_batch_df.PMCID.values) + .difference(set(log.pmcid.values)) + ) + + # If all ids have been processed skip batch + if len(already_seen) == 0: + continue - with open(f"{temp_dir}/batch_{idx}.xml", "wb") as xml_file: + query = f"{pubtator_central_api}pmcids={','.join(pmcid_batch_df.PMCID.values)}" + response = call_api(query) + + batch_file_path = Path(f"{temp_dir}/batch_{idx}.xml") + with open(batch_file_path.resolve(), "wb") as xml_file: try: root = ET.fromstring( bytes(response.text, encoding="utf8") ) xml_file.write(ET.tostring(root, pretty_print=True)) - except Exception as e: - print(query) + + # Save to the logger each pmcid batch + ( + log + .assign( + batch=[idx]*pmcid_batch_df.shape[0], + pmcid=pmcid_batch_df.PMCID.values.tolist() + ) + .to_csv( + f"{temp_dir}/{log_file}", sep="\t", + mode="a", header=False, index=False + ) + ) + except Exception as e: + sys.stderr.write( + f"There is an error processing batch {idx}." + ) + + sys.stderr.write( + f"Here is the query:{query}." + ) + + def merge_full_text(temp_dir, output): """ Download full text from Pubtator Central. @@ -108,8 +157,13 @@ def merge_full_text(temp_dir, output): xml_file.write(ET.tostring(document, pretty_print=True)) except XMLSyntaxError as e: - print(f"Please check {file}! There is an error at: {e}") - print("I am skipping this file.") + sys.stderr.write( + f"Please check {file}! There is an error at: {e}", + ) + + sys.stderr.write( + "I am skipping this file.", + ) xml_file.write(b"\n") @@ -118,8 +172,9 @@ def merge_full_text(temp_dir, output): parser.add_argument("--input", help="a tsv file that contains pmids to query for full text", required=True) parser.add_argument("--document_batch", help="the number of documents to query at once", default=100, type=int) parser.add_argument("--temp_dir", help="The directory to store the temporary files", required=True, type=str) + parser.add_argument("--log_file", help="The directory to store the temporary files", default="batch_log.tsv", type=str) parser.add_argument("--output", help="the name of the outputfile containing full text documents", default="pubtator_full_text.xml") args = parser.parse_args() - download_full_text(args.input, args.document_batch, args.temp_dir) + download_full_text(args.input, args.document_batch, args.temp_dir, args.log_file) merge_full_text(args.temp_dir, args.output) diff --git a/scripts/extract_tags.py b/scripts/extract_tags.py index 519c6e8..7c23c4f 100644 --- a/scripts/extract_tags.py +++ b/scripts/extract_tags.py @@ -5,7 +5,7 @@ from lxml import etree as ET import tqdm -import utilities +import scripts.utilities as utilities diff --git a/scripts/hetnet_id_extractor.py b/scripts/hetnet_id_extractor.py index 316ea34..98c5bf1 100644 --- a/scripts/hetnet_id_extractor.py +++ b/scripts/hetnet_id_extractor.py @@ -3,7 +3,7 @@ import pandas as pd import tqdm -import utilities +import scripts.utilities as utilities def filter_tags(infile, outfile): """ This method filters pubtator tags to consist of only diff --git a/scripts/pubtator_to_xml.py b/scripts/pubtator_to_xml.py index 1b33ba1..410139a 100644 --- a/scripts/pubtator_to_xml.py +++ b/scripts/pubtator_to_xml.py @@ -11,7 +11,7 @@ from lxml.etree import tostring import tqdm -import utilities +import scripts.utilities as utilities def bioconcepts2pubtator_annotations(tag, index): diff --git a/tests.sh b/tests.sh index 6574e3a..26bf4ac 100644 --- a/tests.sh +++ b/tests.sh @@ -1,36 +1,4 @@ set -o errexit -# Convert pubtator format to BioC XML -python scripts/pubtator_to_xml.py \ - --documents data/example/1-sample-annotations.txt \ - --output data/example/2-sample-docs.xml - -# Extract tags from the BioC XML -python scripts/extract_tags.py \ - --input data/example/2-sample-docs.xml \ - --output data/example/3-sample-tags.tsv - -# Filter-Convert tags to Hetnet IDs -python scripts/hetnet_id_extractor.py \ - --input data/example/3-sample-tags.tsv \ - --output data/example/4-hetnet-tags.tsv - -python scripts/map_ids.py \ - --input data/example/3-sample-tags.tsv \ - --output data/example/5-sample-pmids-to-pmcids.tsv \ - --debug - -python scripts/download_full_text.py \ - --input data/example/5-sample-pmids-to-pmcids.tsv \ - --output data/example/6-sample-full-text.xml \ - --temp_dir data/temp - -# Extract tags from the BioC XML -python scripts/extract_tags.py \ - --input data/example/6-sample-full-text.xml \ - --output data/example/7-sample-full-text-tags.tsv - -# Filter-Convert tags to Hetnet IDs -python scripts/hetnet_id_extractor.py \ - --input data/example/7-sample-full-text-tags.tsv \ - --output data/example/8-hetnet-full-text-tags.tsv +python execute.py \ +--config config_files/tests_config.json