Each training block lasted for seven days, consisting of five exercise training sessions and two rest days. The training sessions were designed to engage both the aerobic and anaerobic energy systems,

and consisted of a variety of training types (e.g. low intensity aerobic, BMN 673 mouse fartlek, and intervals). Participants were assigned to RTB or CTB in a randomised, counter-balanced order. Subsequently, in the week prior to each training block, a familiarisation session consisting of a graded exercise test (GXT) was performed on a motorised treadmill or cycle ergometer to determine each individual’s running and cycling VO2peak, maximum HR (HRmax), and the corresponding velocity (vVO2peak)

or power Selleck HIF inhibitor output (pVO2peak). During each seven day period, exercise training was performed on Day One (D1), Two (D2), Four (D4), Five (D5) and Six (D6), while Days Three (R3) and Seven (R7) were recovery days (Figure 1). After completing their first training block, participants had a seven day recovery period before they started the subsequent condition. In addition, no manual labour or exercise training was performed outside of the experimental protocol, and participants were asked to keep their physical activity levels to a minimum during the seven days of recovery between conditions. Figure 1 Diagrammatic representation of the running and cycling training blocks. For the duration of both conditions, all exercise sessions started between 0700–0800 each day, and cAMP participants were provided with 300 ml of water to be consumed ad-libitum. For RTB

and CTB, baseline venous blood samples were taken on three separate occasions, which included D1, R3 and R7. Finally, urine samples were obtained on arrival (baseline) and 3 h post-exercise on D1, D2 and D6, as well as R3 and R7 (baseline only). All baseline venous and urine samples were obtained between 0700 and 0800 to minimise diurnal variation. Experimental procedures Graded exercise test The running GXT was conducted on a motorised treadmill (VR 3000, NuryTech Inc, Germany) utilising 3 min exercise and 1 min rest periods. The initial speed was 10 km.h−1, with subsequent 1 km.h−1 increments over each exercise period until volitional exhaustion. The cycle GXT was conducted in a similar fashion (3 min exercise: 1 min rest), and performed on a calibrated wind-braked ergometer (Evolution Pty. Ltd., Melbourne, Australia), using customised data collection software (Cyclemax, School of Sport Science, Exercise & Health, The University of Western Australia). The initial workload was 100 W, with increments by 40 W every 3 min. Both HR and ratings of perceived exertion (RPE) were recorded during the final 10 s of each workload.