The study protocol was approved by the Human Subject

Committee EPZ015938 of National Taiwan College of Physical Education. Study design This study used a double-blind, randomized cross-over design. The procedure of exercise tests and blood and gas samplings is shown in Figure 1. Each subject completed 3 trials in a random order according to their order of admission to this study. Each trial was separated by at least 2 weeks. The same food was provided in the lunch and dinner on the day before, and the breakfast on the day of each trial. The lunch and dinner were meal boxes purchased from a local restaurant. The 2 meals combined to provide approximately 1434 kcal, with 49.7% energy from carbohydrate, 30.1% from fat, and 20.2% from protein. The diet analysis was performed by a dietitian using Taiwanese food exchange table [26]. The breakfast contained white bread 1.2 g/kg, jam 0.1 g/kg, butter 0.l g/kg, and soybean milk 5 ml/kg (6.2 kcal/kg, containing carbohydrate 1.0 g/kg, protein 0.24 g/kg, and fat 0.14 g/kg). For a 70-kg subject, the breakfast contained 434 kcal, including 70 g carbohydrate, 16.8 g protein, and 9.8 g fat. Figure 1 Study protocol.

■: 1-min rest; ↑: supplementation; ▲: blood sampling. Measurement of Vorinostat cardiopulmonary function The cardiopulmonary CRT0066101 function was measured approximately 1-2 weeks prior to the trials using an electrically braked cycle ergometer (ERG 550, Bosch, Stuttgart, Germany). The subjects warmed up at 50 W for 5 min, followed by incremental steps of 25 W every 3 min. The breath-by-breath gas analysis was performed using a gas analyzer (Vmax 29C, Sensormedics, Yorba Linda, CA, USA). The O2max was considered to be achieved if O2 increased by no more than 2 ml/kg/min after increasing the workload or a respiratory exchange ratio was larger than 1.10. Experimental procedure The subjects reported to the laboratory in the early morning after an overnight fast. A cannula was put in the antecubital vein by licensed personnel. After a blood

sample was taken in a fasted state to serve as the baseline, the subjects consumed the standardized breakfast. Phosphatidylethanolamine N-methyltransferase The exercise test started 1 hr after the breakfast was consumed. Each trial contained 3 matches. At the end of the second match, 3 different supplementations were consumed: 1.2 g/kg glucose (CHO trial), 1 g/kg glucose + 0.1 g/kg Arg + 0.1 g/kg BCAA (leucine: isoleucine: valine = 2:1:1, CHO+AA trial), or water (placebo trial). All supplementations were dissolved in 600 ml lemon flavored water to make the tastes similar. The subjects were allowed to drink water ad libtum in the first trial, while the timing and amount of consumption were recorded. The timing and amount of water consumption were repeated in the following trials. Exercise tests The high-intensity intermittent exercise test was designed to mimic the duration of the actual wrestling competition.