The assay was performed in duplicate as per the instructions from DSL and the CV was less than 10%. Xanthine oxidase (XO) was measured because it is involved in free radical production and its elevation contributes to oxidative stress [12, 13]. The XO was assayed in duplicate using a commercially available kit (Invitrogen, Carlsbad, California, USA). Plasma was assayed pre-exercise and immediately post-exercise. The XO stock solution was used to construct a standard curve. The standards
and serum were pipetted into a high binding enzyme immunoassay (Caymen Chemical Co. Ann Arbor, MI USA) 96 well plate. The plasma samples were diluted 1 fold by the placement of a buffer CRT0066101 supplier solution, and the XO reaction was started when a composition of amplex red, horseradish peroxidase, hypoxanthine and buffer solution was added to each well. The plate was incubated at 37°C for 30 min and the absorbance was read at 550 nm using a PolarStar Galaxy plate reader (BMG Laboratory Technologies, Offenburg, H 89 Germany). Statistical analysis A two way repeated measures analysis of variance (ANOVA) was used to evaluate changes over time and condition for power and velocity along with lactate, RPE, GH, CORT and XO. If a significant F value was achieved the Bonferroni post hoc test was performed. The level of significance was set at p ≤ 0.05. All data was analysed using SPSS for Windows version 16. Data are presented as mean ± standard
error of the mean (SEM). Where relevant effect size ratios (ES’r) were calculated using Cohens d[35]. An ES’r of ≥0.5
was considered Succinyl-CoA to display a moderate effect and ≥0.8 a large effect. Results The pre to post HTS, blood lactate concentrations (Blac) increased significantly after both AOX supplementation; 1.23 ± 0.08 to 7.68 ± 3.01 mmol.l−1 (p < 0.05) and placebo supplementation; 1.79 ± 0.30 mmol.l−1 to 8.11 ± 2.98 mmol.l−1 (p < 0.05). Blood lactate continued to be significantly elevated twenty min post-exercise for both groups, but there was no significant difference in Blac levels between the two conditions at any time point (p > 0.05). The RPE was significantly increased in both groups for sets three to six compared to set one. There were however no significant differences in RPE between the AOX and placebo conditions at any point during the HTS (p < 0.05). The concentric mean power and velocity are presented in Figures 1 and 2 respectively. Following AOX supplementation concentric mean power remained consistent across all six sets of the HTS. However, during the placebo trials concentric mean power significantly decreased from sets 1–6. During the placebo trial concentric mean power was significantly lower in comparison to each set in the AOX condition, with sets five and six having the greatest decrease (p < 0.05, ES’r = 0.52). Similarly average velocity during the AOX was higher compared to placebo. Accumulated power output during the AOX HTS was 6746 ± 5.
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