[11]. Resting metabolic rate Resting metabolic rate (RMR) was assessed by using a portable indirect calorimeter for 25 minutes (Ulixertinib supplier Cosmed K4b2, Cosmed, Italy). A face mask (Hans Rudolph, Kansas City, MO) covering the mouth and nose of the participant was attached to a bidirectional digital turbine flow-meter and fastened to the participant using a mesh hairnet with Velcro straps. To guarantee an airtight seal, a disposable gel seal (Hans Rudolph) was positioned between the inside of the face mask and the skin. The Cosmed K4b2

system was calibrated prior to each individual test according to the manufacturer’s guidelines. Breath-by-breath O2 and CO2 gas exchange was measured and recorded in the portable unit’s computer system. On completion of each test, the stored data were transferred to the Cosmed K4b2 version 6 computer software running on a Windows-based CH5183284 laptop computer. The data were then averaged over 15 second intervals and transferred to Microsoft Excel for further analysis. The morning before the RMR measurements, the Cosmed K4b2 was calibrated with a calibration gas mixture (16% O2, 5% CO2). The test was carried out

with the participant in a comfortable supine position, at an environmental temperature of 21–22°C. All measurements were done in the morning (between 6 and 9 a.m.) following a 12 hours fast and a minimum of 8 hours of rest. The results find more of the RMR measurement were compared with the RMR predicted by the Harris-Benedict equation [12] and the RMR(kcal)/FFM(kg) ratio was also calculated. Energy and nutrients intake Seven consecutive days of dietary records were obtained under the supervision of dieticians. Athletes had a regularly contact with registered dietitian who teach them and control how to record nutrition intake. All meals (including recipes and item masses), nonmeal foods, beverages, and fluids Phosphoribosylglycinamide formyltransferase were recorded in diary form using a photographic album of dishes [13]. The daily diets were analyzed for their energy and nutrient levels (fat, protein, carbohydrate, dietary fiber, calcium, phosphorus, iron, zinc, vitamins A, D, B1, B2,

niacin, B6, B12, foliate and vitamin C) using the Dietician computer software package, based on Polish food composition tables [14]. Total energy expenditure and energy availability For three days, each subject wore a heart-rate monitor (HR) (Polar Sport Tester, RS 400, Finland) in order to estimate total energy expenditure (TEE). For each subject, the relationship between HR and VO2 was established. The measurements were carried out two or more hours after meals, and after the subject had rested for 30 min, having arriving at the laboratory. Results were obtained by simultaneous measurement of HR and VO2 for the following activities carried out sequentially: lying in supine position, sitting quietly, standing quietly, and continuous graded exercise on a cycle ergometer.