Prism version 7.0b

Cryopreserved plasma aliquots were thawed and prepared following kit manufacturer’s guidelines. MCP-1, sCD163, and IP-10 were measured using a custom multiplex kit (RD Systems, MN, USA), acquired on a Luminex 200™ analyzer (Luminex), and analyzed using Bio-Plex Manager™ software (Bio-Rad). Neopterin and sCD14 were measured by ELISA (Neopterin competitive enzyme immunoassay, ALPCO, NH, USA; Human CD14 Quantikine ELISA kit, RD Systems), optical density read on a microplate spectrophotometer (Bio-Rad), and standard curve interpolation conducted on Prism version 7.0b (Graphpad Software Inc., CA, USA). Samples were run in duplicate and the CV of replicates were less than 10%.
Examination of biomarker distributions revealed two outliers greater than 4 SDs above the biomarker distribution means – one for neopterin and one for MCP-1. Because these values were considered physiologically plausible, we substituted these outlier values with the next most extreme value in the biomarker distribution during statistical analysis, as has been done previously by our group [15 (link)]. The biomarker distributions of sCD163 and IP-10 were log₁₀ transformed prior to statistical analysis.

All data is expressed as arithmetic means ± standard deviation (SD). For the comparison of continuous variables from independent groups we used Student’s t-test for two groups and one-way ANOVA for more than two groups. All values were normally distributed. The statistical details (n-numbers, mean ± SD and tests) are given in the figure legends and were calculated by Prism version 7.0b software (GraphPad).