05).

Subjects were allowed 60 minutes to consume the entire volume of beverage. Each condition was consumed on a different test day, with a minimum of five days separating mTOR inhibitor test visits. Table 2 Study timeline and outcome measures Time → Variable ↓ Pre Dehydrating Exercise Test Immediately Post Dehydrating Exercise Test 1 Hour Post Dehydrating Exercise Test 2 Hours Post Dehydrating Exercise Test 3 Hours Post Dehydrating Exercise Test† Immediately Post Performance Exercise Test Body Mass†† X X* X** X X   Plasma Osmolality X X     X   Urine Specific Gravity X X     X   Subjective Measures (VAS)   X X X X   Heart Rate X X     X X Blood Pressure X X     X X † The Performance

Exercise Test began following this measurement time (total exercise time was recorded) †† Body Mass was used to calculate fluid retention (as described in the Methods section) * For determination of fluid volume to consume ** For determination of “”baseline”" body mass Performance Exercise Test Three hours after the completion of the dehydrating exercise test (and HMPL-504 two hours after subjects consumed their assigned condition), a test of physical performance was conducted using a treadmill as previously done [20]. Specifically, subjects began walking on a motorized treadmill at a self-selected comfortable speed (0% grade) for five minutes. At the conclusion of the five-minute period,

the actual performance test began. The protocol involved an increase in intensity every three minutes. While the speed of the treadmill remained constant at 4.2 miles per hour throughout the test, the grade increase in the following manner: Rapamycin cost min 1-3, 0%; min 4-6, 2.5%; min 7-9, 5%; min 10-12, 7.5%; min 13-15, 10%; min 16-18, 12.5%; min 19-21, 15%. Subjects exercised until volitional exhaustion and the total exercise time was recorded. This identical protocol was administered at the screening visit (for familiarization) and on each of the four test day visits. Therefore, we do not believe that there was any significant degree of “”learning”" involved with this test. Outcome Measures In addition to the measure of total exercise time obtained in the performance test described above, the following variables were used as outcome measures; some of which have been discussed previously [21]. With regard to hydration status, body mass, fluid retention (based on body mass), plasma osmolality, and urine specific gravity were measured. Specifically, for fluid retention based on body mass, it was expected that the administration of test product at the amount prescribed would bring the subject’s body mass back to very near its P005091 research buy pre-exercise level.