Group size, Data and Statistical analysis
There is no prior study regarding the role of kynurenine pathway metabolites on glutamate receptor mediated neurotransmission in MTLE-HS patients. In the present study, it is not possible to calculate the sample size as there is no prior reports. It is well established that 30% of patients with MTLE-HS are drug resistant (Kwan et al., 2010). Based on this fact, we have collected resected hippocampal samples from 55 drug resistant MTLE-HS patients underwent surgical intervention from seizure freedom over a period of 4 years from 2015 to 2019. Here we have used n=6 as minimum number of repetitions required to have > 95% possibility (p<0.05) to detect. This sample size satisfies the guidelines of the British Journal of Pharmacology for preclinical studies, where n refers to independent values and not replicates (available at: https://bpspubs.onlinelibrary.wiley.com/hub/journal/14765381/author-guidelines.html).
We have complied with the recommendations of the British Journal of Pharmacology on experimental design and analysis in pharmacology (Curtis et al., 2018). Statistical analyses were performed on Sigma Plot 13.0 (RRID: SCR_003210) and GraphPad Prism 6.0 software (RRID: SCR_002798). When normal distribution and equal-variance were valid, statistical significance between two groups were evaluated by two tailed unpaired t-test otherwise Mann Whitney test was used for evaluation and between more than two groups by Kruskal-Wallis non-parametric ANOVA followed by multiple comparison with Dunn’s test was performed. Cumulative distribution of electrophysiological data was analysed by Kolmogorov–Smirnov test. The correlation between duration of seizure and concentration of metabolites was performed using Spearman’s rank correlation test. The least squares method was used to plot the best fit line and to predict the behaviour of dependent variables (KYNA, PLP, and QUIN). The goodness of fit was calculated through R2. The overall significance (P value) was calculated by F-test. Data were presented as a mean ± SEM. P < 0.05 was considered significant.