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Status |
Public on Aug 31, 2013 |
Title |
SLEC-CST0-Ba91-Lung-02 |
Sample type |
RNA |
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Source name |
early stage compstatin treated animal's lung
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Organism |
Papio cynocephalus |
Characteristics |
tissue: Lung stress: E.coli (serotype B7-086a:K61) challenge agent: compstatin sepsis stage: early stage protocol: prevention regimen
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Treatment protocol |
Papio cyanocephalus baboons were held for 30 days at the OUHSC animal facility. Only healthy tuberculosis free animals with hemoglobin greater than 10 g/dL and white blood cell (WBC) count less than 12,000 were included in the study. Animals were infused with 1x109 live E. coli (LD50 dose) as described before. The time point at which the infusion was started is further indicated as T0, a time point of n hours thereafter referred to as T+n hours. Compstatin was administered as a 10-mg/kg iv. bolus followed by 60μg/kg/min continuous infusion. Three experimental E. coli groups were studied: (i) E. coli challenge only (n=4); (ii) E. coli plus compstatin treatment from T0 to T+8 (n=4; prevention regimen); (iii) E. coli plus compstatin from T+5 to T+11 (n=4; rescue regimen). The control group comprising three animals received saline infusion only.
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Growth protocol |
Live E. coli organisms (serotype B7-086a:K61; American Type Culture Collection, Rockville, MD), stored in the lyophilized state at 4°C after growth in tryptic soybean agar, were reconstituted and used. To eliminate differences due to E. coli strain variations, all animals were infused with E. coli from this single isolate.
|
Extracted molecule |
total RNA |
Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions, RNA was further purified with the Qiagen DNeasyTissue kit (Qiagen, Valencia, CA) and the contaminant genomic DNA was removed with a Qiagen on-column DNase digestion kit. RNA concentrations are determined on a scanning UV/VIS spectrophotometer Nanodrop ND-1000 Spectrophotometer. RNA Quality Assessment by using Agilent 2100 Bioanalyzer Capillary Gel Electrophoresis System.
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Label |
Biotin
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Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 10 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
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Hybridization protocol |
Following fragmentation, 20 ug of cRNA were hybridized using Model 640 hybridization oven for 16 hr at 45C on GeneChip Human Genome Array U133A 2.0. GeneChips were washed and stained in the Affymetrix GeneChip® 450 fluidics station.
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Scan protocol |
GeneChips were scanned using GeneChip® 3000 7G scanner with autoloader (0.51 micron resolution)
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Description |
Gene expression data from early stage compstatin treated E.coli challenged baboon lung
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Data processing |
RMA
|
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Submission date |
Aug 12, 2010 |
Last update date |
Aug 31, 2013 |
Contact name |
florea lupu |
E-mail(s) |
[email protected]
|
Phone |
405-271-7483
|
Fax |
405-271-7417
|
Organization name |
Oklahoma Medical Research Foundation
|
Street address |
825 NE 13th Street
|
City |
Oklahoma City |
ZIP/Postal code |
73104 |
Country |
USA |
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|
Platform ID |
GPL571 |
Series (1) |
GSE23590 |
Gene expression data from compstatin treated E.coli-Induced primate sepsis model |
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