Statplus

Statistical analyses were performed using Student’s t-test to compare two samples. Statistical analyses comparing more than two groups were performed using a one-way analysis of variance with the StatPlus software program (AnalystSoft Inc., Walnut, CA, USA). Statistical significance was set at P < 0.05 for all tests.

Raw data were analyzed using a statistical software package (StatPlus, AnalystSoft Inc.—statistical analysis program for Mac OS®. Version v6. See http://www.analystsoft.com/en/). To check the normal distribution of the sampling, a Shapiro-Wilk test was applied. Whether this test failed, a non-parametric approach was applied. In consideration of preliminary results on salivary cortisol, hematological and blood chemistry measurements, and energy expenditure calculation, a sample size of 80 participants was selected to ensure a statistical power higher than 0.80 and a type 1 error of <0.05. Intention to treat approach was utilized to analyze the experimental data: missing values (participants dropout) were considered in the statistical analysis. A two-way (group x time) analysis of variance (ANOVA) was used to establish statistical differences among conditions. When necessary, a Bonferroni post-hoc, with Bonferroni correction test, was used to determine specific differences and interactions among variables A Kruskal Wallis ANOVA on ranks was applied to establish statistical differences among conditions presenting a non-normal distribution. Significance level was set at α<0.05. Results are presented as mean ± standard deviation (SD).

The data from three independent experiments on cytotoxicity, anti-proliferative activity, and apoptosis rate obtained by flow cytometry were used and expressed as means ± SD. Statistical analysis was performed using one-way ANOVA test with Stat-Plus software (AnalystSoft VC 2009, Walnut, CA, USA), with the threshold of significance set at p-value < 0.05.