Resistance training protocol Participants engaged Torin 2 ic50 in a 4-day per week resistance-training program split into two upper and two lower extremity workouts per week for a total of seven weeks. The upper body resistance-training program consisted of nine exercises

(bench press, lat pull, shoulder press, seated rows, shoulder shrugs, chest flies, biceps curl, triceps press down, and abdominal curls) twice per week and a seven exercise lower extremity program (leg press or squat, back extension, step ups, leg curls, leg extension, heel raises, and abdominal crunches) performed twice per week. We have previously shown this program to be effective at promoting significant gains in muscle strength and mass [18]. Participants performed

3 sets of 8–10 repetitions with 70–80% 1-RM. Rest periods NVP-BSK805 mw between exercises lasted no longer than three minutes and rest between sets lasted no longer than two minutes. Training sessions were not supervised, but were documented in training logs, and signed off to verify compliance and to monitor progress. Muscle biopsies and venous blood sampling Based on our previously-established guidelines [18], at each of the four testing sessions at days 0, 6, 27, and 48 percutaneous muscle biopsies (50–70 mg) were obtained using a Bergstrom (5 mm) needle. Muscle samples were obtained from the middle portion of the vastus lateralis muscle of the dominant leg at the midpoint between the patella and the greater MEK inhibitor trochanter of the femur, at a depth between one and two cm. For the remaining three biopsies, attempts were made to extract tissue from approximately the same location as the initial biopsy by using the pre-biopsy scar, depth markings on the needle, and a successive incision that was made approximately

0.5 cm to the former from medial to lateral. After removal, the muscle specimens were immediately frozen Fenbendazole in liquid nitrogen and then stored at -80°C for later analysis. At each of the four testing sessions, venous blood samples were obtained from the antecubital vein using a standard Vacutainer apparatus. Once collected, the samples were centrifuged for 15 minutes. The serum was removed and frozen at -80°C for later analysis. An 8-hour fast prior to blood donation was required for the participants before each of the four testing sessions. Muscle and serum creatine analysis Muscle tissue samples were analyzed spectrophotometrically for total creatine by the diacetyl/α-napthtol reaction [19]. Using similar methods, serum samples were measured in duplicate for creatine concentration. Serum samples were immediately ready for creatine analysis, whereas muscle tissue had to first be prepared. For serum creatine analysis, duplicates for all samples yielded a coefficient of variation of 5.4%.