With this data, the sum of skin-folds, fat mass and skeletal muscle mass, using an anthropometric

method, were estimated. Body mass was measured using a commercial scale (Beurer BF 15, Beurer GmbH, Ulm, Germany) to the nearest 0.1 kg HSP inhibitor after voiding of the urinary bladder. Body height was determined using a stadiometer (Tanita HR 001 Portable Height Measure, Tanita Europe, Amsterdam, Netherlands) to the nearest 1.0 cm. The circumferences and the lengths of the limbs were measured using a non-elastic tape measure (cm) (KaWe CE, Kirchner und Welhelm, Germany) to the nearest 0.1 cm. The circumference of the upper arm was measured at mid-upper arm; the circumference of the thigh was taken at mid-thigh and the circumference of the calf was measured at mid-calf. The skin-fold data were obtained using a skin-fold calliper (GPM-Hautfaltenmessgerät, Siber & Hegner, Zurich, Switzerland) and

recorded to the nearest 0.2 mm. The skin-fold calliper measures with a pressure of 0.1 Mpa ± 5% over the whole measuring range. The skin-fold measurements were taken following the standard of the International Society Napabucasin for the Advancement of Kinanthropometry (ISAK) once for all four skin-folds and then the procedure was repeated twice more by the same investigator; the mean of the three times was then used for the analyses. The timing of the taking of the skin-fold measurements was standardised to ensure reliability. According to Becque et al.[26] readings were performed 4 s after applying the calliper. One trained investigator took all the skin-fold measurements as inter-tester variability is a major I-BET-762 order source of error in skin-fold measurements. An intra-tester reliability check was conducted on 27 male athletes prior to testing [27]. The intra-class correlation (ICC) within the two

measurers was excellent for all anatomical measurement sites, and various summary measurements of skin-fold Methocarbamol thicknesses (ICC >0.9). Agreement tended to be higher within measurers than between measurers but still reached excellent reliability (ICC >0.9) for the summary measurements of skin-fold thicknesses. Fat mass was estimated using the equation following Stewart and Hannan [28] for male athletes: Skeletal muscle mass (kg) was estimated using the anthropometric equation of Lee et al.[29] with skeletal muscle where Ht = height, CAG = skin-fold-corrected upper arm girth, CTG = skin-fold-corrected thigh girth, CCG = skin-fold-corrected calf girth, sex = 1 for male; age is in years; race = 0 for white men and 1 for black men. The volume and the changes of volume of the right arm and the right lower leg were measured using plethysmography. We used a vessel of plexiglass with the internal dimensions of 386 mm length and 234 mm width. These dimensions were chosen so that any foot size of a male runner would fit in the vessel.