, 2004). Volunteers evaluated each item in four domains (physical, psychological, social-relational, and environmental), using a five-point Likert scale and scoring from 1 (very dissatisfied/very poor) to 5 (very satisfied/very good). Summing across these four domains, we calculated an overall quality of life; with a potential score ranging from 24 to 120, and a high number indicating Selleckchem Tacrolimus a good quality of life. The peak aerobic
power ( V˙O2peak) was measured using a modified Bruce treadmill test protocol (American College of Sports Medicine, 2006). Subjects walked on an ATL-10200 treadmill (Inbramed, Porto Alegre, RS, BRA) with continuous monitoring of a 12-lead electrocardiogram, blood pressure, and metabolic response (CPX/D metabolic cart, Medgraphics, St Paul, MN, calibrated by gases of known composition immediately Obeticholic Acid in vivo before each stress test). After collecting three minutes of resting data with the subject standing on the treadmill, walking began at 2.6 km h−1, 5% grade, and thereafter the speed and grade were increased every
three minutes to volitional fatigue. Criteria of V˙O2peak were: (i) RER > 1.10; (ii) attainment of maximal age-predicted heart rate; and (iii) volitional fatigue. Muscle strength was determined as the one repetition maximal (1RM) effort attained in a leg press exercise; it reflected the maximum load (N) that a subject could lift just once, using the required technique (applying the force via the specified muscle groups, without assistance from momentum or changes in body position). Three familiarization sessions each comprised three sets of eight to 12 repetitions of the leg press exercise preceded the definitive test. Subjects avoided solid or liquid Olopatadine foods containing caffeine, chocolate, or cola-based products, and moderate or vigorous physical activity for 48 h prior to collection of blood samples. They came to the laboratory at 7:00 a.m., having fasted overnight,
and ante-cubital blood samples were collected after 30 min of seated rest. Blood in non-heparinized syringes was dispensed into evacuated tubes coated with ethylene diamine tetra-acetic acid (EDTA) and kept refrigerated until analysis later on the same day, when differential cell counts were made using a Cell-Dyn 3500 cell analysis system (Coulter Corp., Miami, FL). Proliferative responses and natural killer cell activity (NKCA) were tested on samples collected in heparinized syringes after an interval of no more than 4 h. Two hundred microliters of whole blood was incubated for one-, two-, or three-color immunophenotyping, using appropriate combinations of monoclonal antibodies (Becton–Dickinson, Miami, FL) conjugated to fluorescein isothiocyanate (FITC (CD25, CD45RA, CD95)), phycoerythrin (PE (CD19, CD28, CD45RO, CD69, HLA-DR)), or phycoerythrin-cyanine (PE-Cy-5 or PCy-5 (CD3, CD4, CD8, CD56)).