Unknown,Transcriptomics,Genomics,Proteomics

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Effects of offshore saturation diving on the human peripheral blood transcriptome

ABSTRACT: Illumina HT-12 v4 microarrays were used to measure genome-wide gene expression in peripheral blood from occupational offshore divers, all male and fulfilling the criteria for diving personnel set in the NORSOK U-100 standard for manned underwater operations. Saturation dives lasted 13 - 20 days, with maximal diving depths 100 - 125 mws. Elevated oxygen during deep saturation dives triggers oxidative stress. To test the effects of oral antioxidants on post-dive gene expression, the divers were divided into two groups: one group (13 divers) received daily supplementation of antioxidant vitamins C (500mg/day) and E (200mg/day) during their dives, the other (7 divers) was not.

ORGANISM(S): Homo sapiens

SUBMITTER: Ingrid Eftedal

PROVIDER: E-MTAB-4491 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Decompression sickness (DCS) is a potentially fatal condition usually observed after scuba diving. It involves bubble formation in blood and tissues from dissolved inert gas (usually nitrogen or helium), secondary to decreases in ambient pressure (decompression).To the best of our knowledge, no study has evaluated a DCS-induced transcriptomic signature in humans. In this study, we aim to explore the evolution of leukocyte gene expression in human subjects with DCS compared to closely matched divers after uneventful diving using a hypothesis-free RNA sequencing approach .Recruitment of DCS cases (n = 7) and controls (n = 6) was carried out using specific criteria. For both DCS cases and controls, whole blood was sampled at two time points, T1 - within 8 hours of surfacing from diving and T2 - at 40-44 hours after surfacing. Prior to sampling at T2, subjects were fasted for 10 hours. Specific exclusion criteria included a) age < 18 years b) self-reported consumption of alcohol and/or strenuous physical activity before T2 c) symptoms suggestive of delayed DCS presentation in controls at T2 d) acute life-threatening clinical complications or death within 72 hours of surfacing. All cases received emergency HBO as per United States Navy Treatment Table Six between T1 and T2.For RNA isolation, 2.5mL of whole blood was collected in a PAXgene® Blood RNA Tube (PreAnalytiX, Qiagen/BD) from DCS cases and controls at both T1 and T2. The quality of RNA was evaluated by RNA Integrity Number (RIN) determination using the RNA6000 Nano protocol on an Agilent 2100 Bioanalyzer system (Agilent, USA). The RIN values for samples undergoing transcriptome analysis ranged from 7.8 to 9.3. Depletion of alpha and beta globin mRNA was carried out using the GLOBINclear™ kit (ThermoFisher Scientific). To minimize batch effects, all samples were processed simultaneously by the same investigator. RNA samples were submitted for library generation and sequencing by the Beijing Genomics Institute (BGI-Shenzhen). Briefly, poly(A) mRNA was enriched using poly(T) oligo-attached magnetic beads, followed by fragmentation. First strand cDNA synthesis was carried out using random hexamer N6 primers and reverse transcriptase. Following adaptor ligation to cDNA fragments, PCR amplification and purification, single stranded DNA circles were generated in a final library. DNA nanoballs (DNBs) were subsequently generated by rolling circle replication, which underwent paired end sequencing (100bp) on the BGI DNBseq platform.

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Effects of offshore saturation diving on the human peripheral blood transcriptome

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