All body composition measurements were carried out at the same time
each morning after urination and defecation. Height was measured to the nearest selleckchem 0.1 cm with a stadiometer. Body weight was measured using a calibrated balance beam scale (Shinko Denshi Vibra Co., Ltd., Tokyo, Japan) to the nearest 0.01 kg, with the subjects wearing only light undergarments. Hydrostatic weighing and stable isotope dilution method estimated body density and total body water. Subjects were administered these stable isotopes using the following protocol: 2H2O, H218O, and 2H2O for BL1st, BL2nd, and OF measurements, respectively. Our previous study provides details regarding the evaluation of body composition using the three-component model [12]. Physical activity and energy intake Daily AEE was evaluated using a triaxial accelerometer (Panasonic Electric Works Co., Ltd., Osaka, Japan) [13], which was attached to the waist for about 1 month until the end of the study (from 1 week before BL1st until the postintervention observation period finished). Subjects were instructed to refrain from vigorous exercise and to maintain their lifestyle for about 1 month. The data of baseline PA were obtained for 7 days with the exception of the first 3 days since attaching the triaxial accelerometer. Subjects strictly maintained baseline PA by checking
levels of PA using the triaxial accelerometer during the overfeeding period (between BL2nd and OF measurement). If the non-wear activity time of the accelerometer exceeded 3 h in a day, with the exception of the time for taking a bath and sleeping, that day was excluded from the analysis. All foods and beverages were weighed using a portable digital scale (KS-232; Dretec Co. Ltd., Saitama, Japan) during the BL2nd and OF measurement periods (3 days). Furthermore, a survey of food intake was conducted using both
self-reporting methods and visual records obtained using a digital camera or a mobile phone with a camera. A well-trained registered dietitian checked calculated nutrients from the diet records with the photographs. EI was measured daily from a week before the BL1st until the OF measurement. All diet records were analyzed using a computerized nutrient analysis program (Excel Eiyoukun Ver. 4.5; Kenpakusha, Tokyo, Japan). Statistical analysis The results are presented as means±standard deviations. Comparisons between two groups (BL1st versus BL2nd and BL2nd versus OF) were made with the paired AV-951 t-test using Microsoft Excel 2010 from Microsoft Office 2010 (Microsoft Corp., Redmond, WA, USA). The intraclass correlation coefficient (ICC) and the coefficient of variation (CV) were used to test the reproducibility of body weight, % fat, FM, FFDS and TBW measured by the three-component models. Values of ICC above 0.7 were considered as having excellent reproducibility. An alpha of 0.05 was used to denote statistical significance.