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.