Total distance (km) was recorded and average speed (km.h-1) was calculated. Total distance (unknown by the subject) was considered as physical performance. Protocol 2: Standardized exercise A 20 μL blood sample was collected from the earlobe for the assessment of resting glucose and lactate concentrations. As in protocol 1, 15 min before the test and just before their gentle warm-up subjects

drank 250 mL of PLA or SPD. Thereafter, the subjects exercised for 2 hours at 95% of their individual lowest average speed sustained in PLA or SPD during protocol 1; 250 mL of beverage was provided every 15 min. During exercise, Dabrafenib order , , Respiratory Exchange Ratio (RER: / ), HR and Rate of Perceived Exertion (RPE) were measured and/or recorded every 20 min. Central and peripheral fatigue was evaluated before and immediately after exercise. Material and procedures All exercises were performed on the same treadmill (EF 1800, HEF Tecmachine, Andrezieux-Boutheon, France). Blood lactate and glucose concentrations were determined enzymatically using a YSI 2300 (Yellow Spring Instrument, USA). and were measured as described above (see paragraph Preliminary testing). RPE was determined using the 6 – 20 point Borg scale [31]. Central and peripheral fatigue measurements Tests were performed on the knee extensors. The subjects were seated in the frame of a Cybex II (Ronkonkoma, NY) and Velcro

straps were used to limit lateral and frontal displacements. The subjects were instructed to grip the seat during the voluntary contractions to drug discovery acetylcholine stabilize the pelvis. The knee extensor muscles’ mechanical response was recorded with a strain gauge (SBB 200 Kg, Tempo Technologies, Taipei, Taiwan). All measurements

were taken from the subject’s right leg, with the knee and hip flexed at 90 degrees from full extension. The isometric contractions performed during the experiment included 3-4-s maximal voluntary contractions and electrically evoked contractions. During the 4 MVCs, the subjects were strongly encouraged. Femoral nerve electrical stimulation was performed using a cathode electrode (10-mm diameter, Ag-AgCl, Type 0601000402, Contrôle Graphique Medical, Brie-Comte-Robert, France) pressed over the femoral nerve in the femoral triangle, 3-5 cm below the inguinal ligament with the anode (10.2 cm × 5.2 cm, Compex, SA, Ecublens, Switzerland) placed over the gluteal fold. Electrical impulses (single, square-wave, 1-ms duration) were delivered with a constant current, high-voltage (maximal voltage 400 V) stimulator (Digitimer, DS7A, Hertfordshire, UK). For all stimulus modalities, stimulation intensity corresponded to ~120% of optimal intensity, i.e. the stimulus intensity at which the maximal amplitude of both twitch force and the concomitant vastus lateralis (VL) M wave (see below) were reached.