Centurion

Descriptive data were calculated and a test of variance analysis (ANOVA) for comparing simple averages was applied, verifying the model assumptions; in case of default, the Kruskal Wallis test was used (Navidi, 2006 ). The statistical software Statgraphics Centurion (version 18) was used in this research.

The effects of the independent formulation parameters, including PCL payload (A), TPGS concentration (B), and sonication time (C), were studied using the Box–Behnken factorial design to predict the particle size (Y1), zeta potential (Y2), entrapment efficiency (EE; Y3), and drug release rate (Y4), as shown in Table 1. The PCL payload levels used were 20, 35, and 50 mg, respectively. Additionally, three concentrations of TPGS (0.03%, 0.165%, and 0.3% w/v) and three sonication times (2, 5, and 8 min) were investigated (Table 5 and Table 6). Design of experiment (DoE) was used to optimize different formulations; the software program Statgraphics Centurion (version 17.2.02) was adopted for this purpose, which suggested the 15 formulations that are listed in Table 1.

Statistical calculations were performed using Statgraphics Centurion (version 19, Statgraphics Technologies, Inc., The Plains, VA, USA). Normality of variables was assessed with Shapiro–Wilk test. Statistical significance of the differences for binomial variables was assessed using proportion comparisons with normal approximation. Mann–Whitney test was used to compare BCVA and CRT, respectively, at different visits (COVID-1, COVID 0, COVID/last…). A p value < 0.05 was considered statistically significant.

The statistical data were processed with Excel Microsoft Office and StatPlus2007 (http://www.analystsoft.com, accessed on 20 October 2022) software packs. The sample comparison was based upon a null-hypothesis of the absence of difference between the two samples compared. The null-hypothesis was checked by the Mann–Whitney U-test. Distributions of copy numbers in the groups were compared using the Kolmogorov-Smirnov test (D and α) and the Kruskal-Wallis test (H and p). A difference was deemed significant in case of p < 0.001. The logistic curve was plotted using Statgraphics Centurion (www.statgraphics.com). ROC (Receiver Operating Characteristic) curves were drawn using MedCalc software (https://www.medcalc.org/manual/roc-curves.php, accessed on 20 October 2022). Each point on the curve displays a ratio between sensibility and specificity, which corresponds to a preset threshold value of rDNA copy number. The area under the curve (AUC) shows the difference between the groups analyzed.

Analyses were conducted separately for day 19 (early phase) and days 20–30 (late phase). To be consistent across studies, for the analysis, for day 19 only hourly data collected between 2 and 12 h following dosing was used for the statistical analysis. For both periods, the total number of observed withdrawal symptoms was compared between treatments and placebo, using analysis of variance (ANOVA). Tukey’s tests were used for post hoc comparisons. Differences from placebo were considered statistically significant if p < 0.05.
Trendline data was analyzed using Statgraphics Centurion, version 19. For each treatment group, linear regression models were computed for the daily total number of withdrawal symptoms scores of the late phase, and trendlines were plotted. The equation of the trendlines (y = ax + b) and intercept and slope were determined. Using the corresponding equations, the daily withdrawal sum scores (x) were entered to determine the day at which zero symptoms are expected according to the trendline. Intercepts and slopes of the trendlines of different groups were compared using ANOVA. Differences were considered statistically significant if p < 0.05. Data on body weight and food intake were compared using ANOVA. Tukey’s tests were used for post hoc comparisons. Differences were considered statistically significant if p < 0.05.

Data were summarised as mean and standard deviation. Simple ANOVA analyses were performed to assess the statistical significance of the processes applied to chia (milling, defatting and sprouting) on nutritional composition and particle size; and these processes and the intestinal conditions on proteolysis, lipolysis, calcium and polyphenols bioaccessibility and antioxidant activity. Statgraphics Centurion was used, and the analyses were conducted with at least a significance of 95% (p-value < 0.05).

All assays were conducted in triplicate. The data were processed using the software Statgraphics Centurion version 16.1.18 (Statgraphics.Net, Madrid, Spain). All values were subjected to analysis of variance (ANOVA) with a 95% confidence level. Pearson’s correlation coefficients were also calculated to corroborate relationships among the selected parameters.