3 �� 1.36 years; weight 71.3 �� 5.27 kg; height 179.8 �� 5.30 cm; body fat 13.9 �� 2.21 %) volunteered to participate in this study. The participants were informed of the study requirements, benefits and risks. All the participants were male players participating in an elite junior soccer league. They have trained at least for ten years, five to six days per week. selleck chem The study was performed in the middle of the soccer season, when the league was halted for approximately 45 days. Most training sessions at this time of year are devoted to conditioning first and then, specific tactical drills and game skills with the use of balls. The protocol of this study was in accordance with the guidelines of the Ethical Committee of Ostrava University and Declaration of Helsinki.
Procedure The participants performed three tests, separated by at least 48 hours from one other. The tests included the running-based anaerobic sprint test (RAST), the graded treadmill test (GXT) and the multistage fitness test (20mPST), respectively. The participants were asked to refrain from any form of intense physical exercise for the 24-hour period before each testing session. The RAST and 20mPST were performed in the sports hall and the GXT in the laboratory for diagnostics of human movement. None of the participants was taking any food supplements. They were also instructed to not eat for at least 2 hours prior to each test. Before each test, the participants performed an individual 10�C15 minute warm up. Performance Tests Graded Treadmill Test (GXT).
A continuous graded treadmill test was used in order to measure maximal aerobic power (VO2max), following the Harbor protocol (Wassermann et al., 2005), which was modified in order for the total time of each test to last 8�C12 minutes (Astrand et al., 2003). The test consisted of 3 minutes of a warm-up, followed by constant speed and 2 % inclination increments every minute until the participants reached volitional exhaustion. All participants had the same exercise load throughout the test. The test was stopped when a participant was unable to continue running at the actual velocity and slope. Strong verbal encouragement was provided to each participant as they came to the end of the GXT. The test was performed on a Lode Valiant motor-driven treadmill (Groningen, Nederland).
The expired air was continuously monitored for an analysis of O2 and CO2 concentrations during the GXT by using a breath by breath system (ZAN 600 Ergo; Oberthulba, Germany). Prior Cilengitide to each test, the gas analyzers were calibrated using ambient air with a gas mixture of known O2 and CO2 concentrations and the ventilatory membrane calibrated with a 3 L syringe in accordance with the manufacturer��s instructions. The ambient conditions were automatically recorded by ZAN 600 Ergo and maintained by air-condition between 20�C23�� C. The heart rate (HR) was continuously monitored with a chest belt transmission (Polar Electro, Oy, Finland).
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