Each subject participated in a pre-experimental VO2peak test and four identical experimental tests performed one week apart. Samples from two test occasions were used in the primary analysis, while samples from the remaining two test

occasions were used for longitudinal predictions and for that reason, characterized analytically by GC/TOFMS eight months later. The dataset included in total 160 samples, i.e., 96 samples used in the primary analysis (24 subjects at two occasions and two time points) and 64 additional samples characterized eight months later (12 subjects at two additional occasions and two time-points along with 16 analytical www.selleckchem.com/products/carfilzomib-pr-171.html replicates). The Inhibitors,research,lifescience,medical data have been previously used for evaluating physiological variation related to the acute effect of strenuous exercise [51]. Raw data is available upon request. 4.1.1. Pre-Experimental www.selleckchem.com/products/Bortezomib.html Procedures The included subjects performed a pre-experiment incremental test on an Inhibitors,research,lifescience,medical ergometer cycle (Monark 839E) to exhaustion in order to determine the maximum oxygen uptake as a mean of 60 seconds (VO2peak) [52]. At the morning of the experimental test a standardized breakfast in amount related to bodyweight was ingested at 7.30 am, one hour prior to the test. Subjects were instructed to maintain food diaries prior to exercise occasion one and then repeat the same diet prior to exercise occasions two, three Inhibitors,research,lifescience,medical and four. Subjects were

also instructed not to perform any exercise or consume alcohol the day before each exercise occasion and to avoid stress in the morning of the test day. 4.1.2.Experimental Procedure Venous blood samples were Inhibitors,research,lifescience,medical taken after 15 min of bed rest by using a vacutainer system (Becton Dickinson, UK). Thereafter, subjects were equipped with an intravenous catheter (Optiva®2, Medex) in a forearm vein, a transmitter

belt (Polar WearLinkTM31) and a heart frequency monitor Polar S610iTM). Subjects then performed 90 min of ergometer cycling, using an electronically braked bicycle (RodbyTM, RE 829, Enhörna, Inhibitors,research,lifescience,medical Sweden). Each 90 min test session consisted of nine equal 10 minutes sections. The workloads during the sections were loads that corresponded to 40% (2 min), 60% (6 min) and 85% (2 min) of the VO2peak value from the pre-experimental test. 100ml of water was ingested after every 10 min of cycling. Immediately after 90 min completed cycling, blood was collected from the vein catheter into vacutainer tubes. Serum Batimastat was extracted from the collected blood samples following 8 min centrifugation (+4 °C at 3000g) and immediately frozen and stored in -80 °C. Prior to GC/TOFMS analysis, the serum samples were extracted and derivatized according to A et al. [53] The samples were injected in splitless mode by an Agilent 7683 autosampler (Agilent, Atlanta, GA) into an Agilent 6890 gas chromatograph equipped with a 10 m x 0.18 mm i.d. fused silica capillary column with a chemically bonded 0.18 µm DB 5-MS stationary phase (J&W Scientific, Folsom, CA).