The load was set according to each subject’s mass . The test was a 30-second WAnT followed by 5 min of rest and then eight 10-s intervals of all-out cycling. Each interval was separated by 2 min of rest. The resistance for the WAnT and intervals was set at 0.10 kP/kg body mass. Performance Measures Peak power during the WAnT was defined as the highest mechanical power output elicited during each 30 s test. Mean power was calculated based on the average mechanical power Selleckchem OICR-9429 produced during the test. Additionally, average peak power output and average mean power output were both calculated across the WAnT and all 8 intervals.
Temsirolimus datasheet Biochemical Measures Capillary blood samples (5 μL) were taken from the fingertip during the baseline resting blood draw and at 0, 5, and 10 min post-exercise buy LY2603618 in order to determine peak blood lactate values and clearance. The Lactate Pro (Arkray, Japan) portable analyzer was used to determine whole blood lactate content. Before (t0), immediately after (t1), 30 min post (t2), and 60 min post (t3) each WAnT + interval session, blood samples were collected via an indwelling cannula inserted into an antecubital
vein using a vacutainer system (Becton Dickinson, Rutherford, NJ). Approximately 10 mL were collected in a serum separator tube and 10 mL in an EDTA coated tube. After removing a 1 mL aliquot of whole blood for hemoglobin and hematocrit analysis to account for plasma volume changes, an additional 300 μL aliquot (2 × 100 μL for GSSG; 2 × 50 μL for GSH) was obtained for GSH/GSSG analysis. 1-methyl-2-vinylpyridium (M2VP) was added to the tubes containing samples for GSSG analysis. Plasma for 8-isoprostane assay was obtained by centrifugation
of whole blood in the EDTA tubes at 3000 × g 10 min at 4°C with 1 mL aliquots placed in microvials pre-coated with 200-μg of butylatedhydroxytoluene (BHT). The serum separator tubes were left to stand for 30 min to facilitate clotting before being centrifuged at 3500 × g for 15 min at 4°C in order to obtain serum for IL-6 and CORT analysis. Aliquots of blood, serum, and plasma were stored at -80°C until analysis of the dependent measures. All assays were performed in duplicate and assays for each measure were run in one batch. Thiamet G Total and oxidized glutathione were analyzed using a commercially-available EIA kit (Bioxytech® GSH/GSSG-412, OxisResearch, Portland, OR). The within assay coefficient of variation (CV) for GSH was ± 7.3% and for GSSG was ± 8.6%. Similarly, IL-6 was determined via ELISA using commercial kits (IBL, Hamburg, Germany). Within assay CV for IL-6 was ± 6.9%. Serum CORT was analyzed using RIA (MP Biomedicals, Irvine, CA), and the within assay CV was ± 6.2%. In order to analyze plasma free 8-iso PGF2α, plasma from the EDTA tubes was first purified by diluting the sample in a 1:5 ratio with Eicosanoid Affinity Column Buffer (Cayman Chemical, Ann Arbor, MI).