Statistics
Statistics were carried out using IBM SPSS Statistics Version 25. Graphs were made using GraphPad Prism 6. For parametric assessment of baseline confounders between the groups, as well as plasma leptin levels between time points 0 and 135 min in the ANP group, we used 2-tailed student’s t-test and standard deviation. For comparison of plasma leptin levels, we calculated a generalized linear model using an identity link function under a normal distribution for the leptin delta between the two groups. To assess goodness of fit and model superiority, we used Pearson Chi-Square test and Omnibus test. Wald confidence interval and Wald Chi-square metric were used for variance analysis. To assess dose dependency of plasma leptin decrease on ANP level, we calculated the Pearson correlation using a 1-tailed test of significance. Confidence levels were set at 95% and significance was assumed at α = 0.05. mRNA expression in SGBS cells was assessed using the ΔCt method, with HPRT used as reference gene. We compared mRNA levels and protein densitometry values between the groups using two-sided t-test.