Prizm 6

The IC₅₀ values from the MTT assay were evaluated with a non-linear regression analysis (Curve fit, GraphPad Prizm 6.01 software, GraphPad Software Inc., San Diego, CA, USA) using sigmoidal concentration–response curves. The cell viability is presented as a percentage of the untreated control. At least four wells were seeded for every concentration. Three independent assays were performed (n = 3). The maximum tolerated concentration (MTC) for the cell lines was determined from three independent assays according to ISO 10993-5 as the maximum concentration at which at least 70% of the cells were viable (nonlin fit, range, GraphPad Prizm 6.01 software) GraphPad Software Inc., San Diego, CA, USA). To compare the MTC values of the solvents for each cell line, the ordinary One-way ANOVA statistical analysis based on Tukey’s multiple comparisons test with a single pooled variance (GraphPad Prizm 6.01 software) GraphPad Software Inc., San Diego, CA, USA) was utilized, and p ≤ 0.05 was set as a significance level. The respiratory activity of the bacterial cultures was calculated as a percentage against the untreated control.

All statistical analyses were performed using statistics program package SAS^®, version 9.2 (SAS Institute, Cary, NC, USA). IL-1β data did not follow a normal distribution. Therefore, nonparametric statistical tests were used and differences between all the groups were evaluated using Kruskal–Wallis test and Bonferroni’s post hoc correction. First, all groups of the animals were compared to healthy dogs and to each other. Afterwards all epileptic dogs together were compared to TBI and healthy group. Mean values with standard deviation were calculated for each evaluated group. The graphical presentation of the results was done using GraphPad Prizm 6 (GraphPad Software, Inc., La Jolla, USA).
To associate the IL-1β concentrations in serum to the important parameters in epilepsy, linear regression analysis was performed for the following variables: duration of the disease, seizure frequency, time point between sample collection and the last seizure event. The relationship was described with the coefficient of determination, R-squared (R²). The levels of IL-1β were compared between groups with different seizure types respectively seizure severity (single, cluster seizures or status epilepticus) applying one-way ANOVA test.