Spss statistics software v 25

Statistical analysis was performed by the Olink Biostatistics unit. Four different statistical methods were used. T-test was used to identify differences in protein concentrations between healthy controls and tumour patients. Regression analysis was conducted to search for correlation between protein concentrations and Ki-67 index in tumour patients. Survival analysis was used to investigate associations between protein concentration and survival in tumour patients. Finally, a one-way ANOVA test was performed to compare protein concentrations in different treatment response groups. All p values were adjusted for multiple testing within each test using the Benjamini–Hochberg approach^{51 (link)}.
A chi-square test was performed to study correlations between FASLG expression and tumour differentiation (IBM SPSS statistics software, v25, USA).

All measurements were performed in triplicate, and results were expressed as mean value ± SD, except in in vivo study where SEM was used instead of SD. After checking the normality of distribution, the statistical analysis involved Student’s t-test for two sets of results or ANOVA with Turkey HSD as a post hoc test for three results groups. Statistical analysis was performed using IBM SPSS Statistics software (v. 25). p < 0.05 was considered as statistically significant.

A statistical analysis of Salmonella spp. culture, qPCR, and histomorphometry was performed using Mann–Whitney and Fisher exact tests following IBM SPSS Statistics Software v25 (IBM, Armonk, NY, USA). The level for statistical significance was set at p < 0.05.

The clinical variables chosen to be investigated in the statistical analyses included gender, age, Ki-67 index, C-reactive protein (CRP), CgA, Syn, therapeutic response (evaluated according to the RECIST 1.0 criteria), survival and small cell/large cell morphology. The Chi-2 test was used for calculating correlations of categorical variables e.g. Ki-67 (<55% and >55%), CgA (positive and negative), Syn (positive and negative), sex (female and male), and morphology (small cell and large cell). Spearman’s correlation test was used for correlations between continuous variables.
Survival was evaluated through the Kaplan-Meier analysis. Mann-Whitney test was used to compare PFS and OS between PD-L1-non-immunoreactive patients and PD-L1- immunoreactive patients. PFS was defined as the time from first treatment to progression or death of any cause. OS was defined as survival time calculated from the date of diagnosis to date of death of any cause.
All statistical analyses were performed using IBM SPSS statistics software (v25, USA).

Chi-square test was used to test the associations between possible risk factors and the presence of antibodies against L.infantum. When associations were significant, odds ratios (OR) and the 95% confidence interval (CI) were estimated by univariable logistic regressions. The variables with a bivariate p value ≤0.25 were included in a multivariable stepwise logistic model [29 ]. The likelihood of the final model was evaluated by Log likelihood chi square test (LRT). A p value <0.05 (two-tailed) was considered statistically significant.
The seroprevalence was calculated as the proportion between the number of seropositive dogs and the total number of dogs tested in the period 2010–2014. The apparent prevalence (AP; i.e. the proportion of a population that tests positive using a diagnostic method), and the 95% CI were calculated using a binomial distribution. The true prevalence (TP; estimated from AP adjusting for the diagnostic test Se and Sp), was calculated using the following Rogan and Gladen equation [30 (link)]: TP = (AP + Sp– 1)/(Se + Sp– 1).
TP and Blaker’s 95% CI were calculated using Epitools epidemiological calculators [31 ]. Analyses were performed by SPSS Statistics Software v.25 (IBM SPSS Statistics).

The standard statistical analyses, the Student’s t-test, followed by the Bonferroni correction and ANOVA, followed by post hoc Tukey’s test, were used to analyze the data except those specially mentioned below. The log transformation for the relative expression levels of genes after RNAi was used before the statistical analysis because the data did not meet the normal distribution. The survival rates of larvae with knocked down CmGMC10 or CmGMC38 after heat exposure were analyzed using the repeated-measures ANOVA in the GLM model, and the survival rates measured at 0, 2, 4, and 6 d after exposure were considered as repeated measurements. All analyses were performed in IBM SPSS Statistics software V25.