Prism 8.3.0 for macos

Statistical analysis was performed using GraphPad Prism 8.3.0 for MacOS (GraphPad Software, Inc. La Jolla, CA, USA). The distribution of the obtained results was assessed using the Shapiro–Wilk test. Due to the lack of normal distribution, the Mann–Whitney U test was used for quantitative comparisons. Chi-square test with Yates’s modification was used to analyze the differences in the prevalence of qualitative variables. Correlations of the results were assessed using the Spearman rank correlation coefficient. The statistical significance level was set at p < 0.05.
The number of subjects was determined based on our previous experiment, assuming that the power of the test would be equal to 0.9 (ClinCalc sample size calculator).

Statistical analysis was performed using GraphPad Prism 8.3.0 for MacOS (GraphPad Software, Inc. La Jolla, USA). The normality of the distribution was assessed using the Shapiro–Wilk test. For comparison of quantitative variables, the Kruskal–Wallis ANOVA test and Dunn’s test were used. Multiplicity adjusted p-value was also calculated. The relationship between the assessed redox biomarkers was evaluated using the Spearman rank correlation. In order to determine the diagnostic utility of measured parameters, receiver operating characteristic (ROC) curves were drawn, and the area under the curve (AUC) was calculated. The statistical significance level was set at p < 0.05.
The number of subjects was determined based on our previous experiment, assuming that the power of the test would be equal to 0.9.

Blood was drawn from a voluntary 30 y/o female donor (according to permit from Ethical Review Board in Lund: Dnr. 2015/801) in K₂EDTA-coated vacutainers (BD, Franklin Lakes, NJ, USA). The blood was transferred to a Falcon tube and centrifuged (800×g, 10 min) for blood fractionation. The RBCs were washed five times with PBS (pH 7.4) and afterwards diluted in PBS to 1% suspension (v/v). Thereafter, RBCs were incubated in 1.5 ml Eppendorf tubes for spontaneous hemolysis with increasing concentrations of HS6 (3.5, 14, 56, 225 or 1000 μg/ml) with or without recombinant A1M (220 μg/ml) or PBS in control samples. After 5 h of incubation in room temperature (end-over-end rotation, 8 rpm) the tubes were centrifuged (500×g, 5 min) and the supernatants were used for analysis of lactate dehydrogenase (LDH). The LDH release was measured with the CytoTox 96® Non-Radio Cytotoxicity Assay (Promega, Madison, WI, USA) according to instructions from the manufacturer. The absorbance was measured at 490 nm (VICTOR 1420 multilabel reader, PerkinElmer). Results are presented as a protection ratio where addition of only A1M is set to 1. Statistical comparison of differences was performed with a Kruskal-Wallis test with Dunn's multiple comparisons test using GraphPad Prism 8.3.0 for MacOS (GraphPad, Bethesda, MD, USA).