Controls consisted of PBS/0.25 μM H2O2 in the absence (control) or presence of diluted plasma. Data were calculated Temsirolimus supplier as a change in fluorescence over time (900 s) minus the fluorescence observed at time zero (ΔFI). Results are calculated as ΔFI after 300 sec and shown as % change from pre-LY2603618 cost damage values. Plasma interleukin (IL)-6. Plasma (100 μL), collected pre and 12, 36 and 60 hours post damage was measured for IL-6 using a sandwich ELISA, purchased from R&D Systems, (Minneapolis, MN, USA). Plasma antioxidant capacity. The capacity to reduce ferric ions was determined using the ferric reducing antioxidant power (FRAP) assay as described by Benzie and Strain . Briefly,
an aliquot of 8.5 μL of normal (non- deproteinized) serum was added to 275 μL of diluted FRAP reagent (pre-warmed to 37°C) using a microplate and the plates were incubated at 37°C for 30 mins before measuring
the absorbance at 595 nm using a plate reader (ELX 808 Ultra Microplate Reader (Bio-tek Instruments. Inc, USA)). The working FRAP reagent was prepared by mixing 10 volumes of 300 mmol/L acetate buffer, pH 3.6, with 1 volume of 10 mmol/L TPTZ (2,4,6-tripyridyl-s-triazine) in 40 mmol/L hydrochloric acid and with 1 volume of 20 mmol/L ferric chloride. A standard curve was prepared using different concentrations (200–2000 μmol/L) of FeSO4.7H2O. FRAP was calculated and expressed as either μmol/L or % of pre-treatment values. Statistical analyses Data were analyzed using Statistical Analysis Software (SAS) 9.1 for Windows (version 5.1.2600). Using a repeated measures analysis of variance (ANOVA), comparison between conditions (blueberries MK-0457 research buy and DCLK1 control) over time for each
measure (independent variable) were determined, providing levels of significance for Trial effect, Treatment effect, and interaction effect between Treatment and Trial. Where significance permitted, post-hoc tests were performed to identify significant differences at each time point. Represented values are means ± standard deviation (or standard error) for n = 10 at a 95% significance level (p = 0.05). Paired t-tests were used to determine order effects for performance measures and effort during the 300 maximal eccentric contractions of the quadriceps. Pearson’s Product Moment Correlation Coefficient’s were determined using SPSS 15.0 for Windows. This allowed us to investigate any relationships between certain variables (i.e. antioxidant activity with muscle performance measures) by giving an r-value between 0.0 and 1.00 (or −0.0 and −1.00). Results Intervention diet All subjects completed the study and there were no reported adverse effects from the dietary intervention. Performance (muscle function) Overall changes in volunteer’s physical performance following a strenuous exercise designed to cause muscle damage were evaluated by measuring the torque generated during a series of isometric, eccentric and concentric exercises over a 60 hour recovery period (Table 2).