|
| Status |
Public on Jul 01, 2014 |
| Title |
PostExposure_Sputum_141 |
| Sample type |
RNA |
| |
|
| Source name |
Sputum sample, after ozone exposure
|
| Organism |
Homo sapiens |
| Characteristics |
gender: male
age: between 18 and 37 years
exposure: 0.4 ppm ozone for 2 hours
|
| Treatment protocol |
The ozone exposures were conducted in an exposure chamber. Each subject was exposed to 0.4 ppm ozone for two hours while performing four 15-minute sessions of intermittent moderate exercise (expiratory minute ventilation, 30–40 L/min) on a treadmill, separated by 15 minutes of seated rest.
|
| Growth protocol |
Twenty healthy non-asthmatic adult subjects aged 18 – 37 years, with no history of smoking in the past 10 years, completed the study. All subjects underwent a physical examination, a routine blood panel with complete blood cell count, and differential and allergy skin testing for common allergens. Subjects were required to have a negative methacholine challenge test result. Female subjects were required to have a negative urine pregnancy test result before challenge, and all volunteers were required to be free of chronic cardiorespiratory disease and no acute respiratory illnesses within the previous four to six weeks of ozone challenge. All subjects had FEV1 and forced vital capacity (FVC) values of 80% or greater of predicted value and FEV1/FVC ratios of 75% or greater of predicted normal value for height and age. All subjects were screened for their ability to provide an adequate induced sputum sample during their training session 48 hours prior to exposure and this sample acted as their pre-exposure baseline. Pre and post sputum samples were separated by a minimum of 48 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Sputum was obtained 6 hours after exposure. In brief, three 7-minute inhalation periods of nebulized hypertonic saline (3%, 4%, and 5%; UltraNeb 99 ultrasonic nebulizer; DeVilbiss, Jackson, TN) were followed by expectoration of sputum into a sterile specimen cup. Cell-rich “plug” material was selected from the raw sample and treated with a dilute (0.1%) solution of dithiothreitol (Sputolysin; Calbiochem, San Diego, CA) in Dulbecco PBS. The cells were treated with 1 ml of TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA) and frozen at -80 degrees C until RNA extraction. RNA was extracted according to standard TRIzol protocol.
|
| Label |
Cyanine 3-Cytidine bisphosphate (pCp) reagent
|
| Label protocol |
Labeled cRNA were prepared according to the standard Agilent protocol
|
| |
|
| Hybridization protocol |
Following fragmentation, cRNA was hybridized on Agilent Human miRNA Microarray, ver 1
|
| Scan protocol |
Microarrays were scanned using Agilent Microarray Scanner
|
| Description |
Sample from human sputum after ozone inhalation exposure
|
| Data processing |
Data were normalized by quantile using Partek® Genomics SuiteTM software (version 6.5)
|
| |
|
| Submission date |
Jun 14, 2013 |
| Last update date |
Jul 01, 2014 |
| Contact name |
Rebecca Catherine Fry |
| E-mail(s) |
rfry@unc.edu
|
| Organization name |
UNC-Chapel Hill
|
| Department |
Environmental Sciences and Engineering
|
| Street address |
1213 MHRC
|
| City |
Chapel Hill |
| ZIP/Postal code |
27599 |
| Country |
USA |
| |
|
| Platform ID |
GPL6955 |
| Series (1) |
| GSE47977 |
Ozone Exposure Alters MicroRNA Expression Profiles in Humans |
|
