Prism statistical

Data are expressed as means ± standard deviation. We used a linear mixed-effects model to evaluate the slope of change in BW over time as well as the interaction between time (weeks) and the combinations of juvenile-period and adulthood protein source (casein or REP). In addition, two-way analysis of variance (ANOVA) for unpaired repeated measures was performed to determine the interaction between the effects of protein source during the juvenile period and adulthood as independent variables on parameters measured at 22 weeks of age as dependent variables. This analysis was followed by multiple comparison testing using Tukey’s correction as post-hoc analysis to identify differences. The results of the mice fed an HFD with amino acids were analyzed by Student’s t-test. All statistical analyses were performed using GraphPad Prism statistical software version 5 (GraphPad Software, Inc., San Diego, CA, USA) and SPSS statistical software version 22 (IBM Japan, Tokyo, Japan). Statistical significance was set at p < 0.05.

All statistical analyses were performed using GraphPad Prism statistical software (versions 6 & 7; GraphPad Software, La Jolla, CA, United States). The D’Agostino and Pearson omnibus test was used to determine whether data were normally distributed prior to correlation or group analyses and to test residuals after performing linear regression analyses. Depending on the results from normality testing and the relationship being tested, results are reported as either r-values for parametric Pearson correlations or as ρ-values for non-parametric Spearman correlations. The co-efficient of determination (r-squared, R²) is reported for linear regression analyses; residuals were normally distributed unless otherwise noted. P-values are indicated for each correlation or linear regression correlation. Two-tailed paired t-tests were used to compare two groups of normally distributed data and Sidak’s multiple comparisons test during one-way ANOVA for testing between multiple groups. Non-parametric groups were compared using Dunn’s multiple Mann–Whitney test. For multiple comparisons, the adjusted p-value is reported.

The titer of the blood/virus suspension, and the mosquito bodies and saliva expectorates was calculated using the method of Reed and Meunch [18 ] and expressed as TCID₅₀/ml. Differences in PCV body titer between the three mosquito species was analysed using a Kruskill-Wallis test, with a Dunn’s post hoc multiple comparisons test. WNV infection, dissemination and transmission rates in PCV infected and non-infected Cx. annulirostris were compared using Fisher’s exact tests. Differences in WNV titer within bodies and saliva expectorates of PCV infected and non-infected Cx. annulirostris were analyzed using Mann-Whitney U tests. Replication of PCV and WNV_KUNMRM16 in the C6/36 and RML-12 cells was compared using a two-way ANOVA. All statistical tests were performed using Graphpad Prism statistical software Version 6 (GraphPad Software, Inc, San Diego, USA).

A PARISS^® hyperspectral imaging system (LightForm, Inc., Asheville, NC) was used to provide spatial mapping of the spectral output from the samples when normally illuminated with white light. Each sample was imaged without a coverslip for structure and spectral mapping. For structure 20% of the light output was imaged using a monochrome QIClick camera (QImaging); for spectral reflectance measurements, 80% of the light output was collected using a 100× 0.9 N.A. air objective (giving ≤ 0.5 µm spatial resolution) on a Nikon Eclipse 80i microscope with a PARISS^® spectrometer utilizing a Retiga 2000DC CCD camera (QImaging). A 50 µm slit was used for window collection, and radiometric calibration was done with a Hg⁺/Ar⁺ lamp (LightForm Inc.), with spectral resolution measured better than 2 nm. A silver mirror (Thorlabs) was used as a reflectance reference for all measurements. Hyperspectral mapping was performed using a library of selected spectra with a minimum correlation coefficient (MCC) of 0.99 used as a discrimination factor to identify and map common spectra. All spectra were smoothed, normalized, and plotted using GraphPad Prism statistical software (GraphPad Software, Inc., La Jolla, CA, USA). The colours of the curves were estimated based on the “spec2rgb” function in R script “pavo”⁴⁹ .

Concentration–response curves were compared by two-way analysis of variance (ANOVA) for repeated measurements. For each concentration, a Tukey test was used to identify differences between groups. Responses to U46619, bradykinin, and SNP were presented as relative change in vessel diameter from baseline. For the analysis of resting arteriole diameters, HPLC-based assays, PCR, DHE and immunostainings, one-way analysis of variance (ANOVA), and Tukey’s test were used to identify differences between groups. Data are expressed as mean ± SE. The significance level was set at 0.05. All data were analyzed using GraphPad PRISM statistical software (Version 9, GraphPad Inc., San Diego, CA, USA).