0 and 7.0 pH standards provided by the manufacturer. Physical Activity Monitors (AMs) and Data Processing Algorithm The operating mechanism for the AM used for this study (Actical Monitor; Mini Mitter Company, Inc., Bend, OR USA) will be described briefly since it has been described in detail previously [14]. The AM is the size of a small wristwatch (2.8 × 2.7 × 1.0 cm3), light weight (0.017 kg), water resistant, utilizes a single “”multidirectional”" accelerometer to quantify motion, and has over five weeks of continuous data storage capacity using one-minute recording epochs. The raw AM data are stored in units

of counts/min where a count is proportional to the magnitude and duration of accelerations during the user-specified epoch. When activity monitoring is complete, the raw AM data are downloaded to a computer using an Wortmannin concentration external reader unit and a serial port connection as an ASCII formatted file. A custom Visual Basic (Version 6.0) AZD0156 computer program then transforms the minute-by-minute AM data into units of activity energy expenditure

(AEE, kcals/kg/min) using a previously validated 2R algorithm [14] and post-processing methods [15, 16] previously validated for wrist-worn monitoring in adults. For the present study, AEE was defined as the relative energy expenditure to perform a task above resting metabolism. LY2835219 manufacturer Each subject’s computed AEE data were then summarized into a time-based moderate-to-vigorous PA variable by summing the corresponding one-minute epochs greater than or equal to a moderate intensity about cut point of 0.0310 kcals/kg/min [14]. This cut-point is

the equivalent of the 3 MET cut point commonly used to define the lower boundary of moderate intensity in adults [17]. This processing routine was repeated with each ASCII formatted AM file to compute the 7-day average daily PA (mins/day) for each of the three periods within the Testing Phase. Statistical Analyses Dependent variables for which there was only one value per measurement period (daily PA, SRWC, and all of the diet diary variables) were evaluated using two-factor multivariate repeated measures ANOVA and planned contrasts for post-hoc comparisons within the Control and Experimental group means. Thus, the analytical strategy was to identify changes in the dependent variables within the groups rather than between groups. All other dependent variables (blood and urine osmolality and pH, as well as 24-hour urine volume) were evaluated with a similar two-factor multivariate repeated measures ANOVA model, but Dunnett’s test was used for post-hoc comparisons within the Control and Experimental group means. Dunnett’s test compares the dependent variable means to a control, or reference condition. In the current study, no one measure could truly serve as a reference, so the mean of the pre-treatment values for each subject and each dependent variable was computed for use as this reference value. All ANOVA and post-hoc tests were performed at the 0.05 alpha level.