cted for each patient were aliquoted and stored at 270uC until used for analysis. p MDT HCV Mono-infected 193.8000 und .8000 MST 26 2 Cytokine Analysis Plasma samples were assayed using the Luminex 200 IS System according to manufacturer’s guidelines at the Human Immune Monitoring Center. All samples were assayed for the following 50 proinflammatory plasma immune markers: CXCL5, CCL11, FGF, G-CSF, GM-CSF, CXCL1, HGF, IFN-a, IFN-b, IFN-c, IL-10, IL-12p40, IL-12p70, IL-13, 16483784 IL-15, IL-17A, IL-17F, IL-18, IL-1a, IL-1b, IL-1RA, IL-2, IL-4, IL-5, IL-6, IL-7, CXCL8, CXCL10, leptin, LIF, CCL2, CCL7, M-CSF, MIG, CCL3, CCL4, NGF, PAI-1, PDGF-bb, CCL5, resistin, SCF, sFasL, sICAM-1, sVCAM-1, TGF-a, TGF-b, TNFa, TNF-b, and VEGF. All samples were run in duplicate, and the average of duplicate measures was used for analysis. Values were reported in picograms per milliliter. 0.17 247 CDT 83 .37000 365 0.27 0.02 11 16 514 48 409 C-NR und 16035 HIV/HCV Co-infected C-SVR 501 15 1 11 0.36 p 48 353 Statistical Analysis For participants whose HCV viral load fell to undetectable levels at FU, the midpoint between 0 and the lowest detectable limit was imputed. For participants whose cytokine concentrations were below the quantifiable limit, a new value of was imputed in order to carry out the 12504917 analyses. To compare clinical and demographic variables, non-parametric statistical tests were conducted for non-normally distributed data, and parametric tests were conducted for normally distributed data. Specifically the Kruskal-Wallis test was used for cross-sectional analyses to compare across groups at BL and FU. For the longitudinal analyses, the signed-rank test compared BL to FU values within groups. ANOVA was used when testing means across three groups, and the Student’s t-test was used for comparing means across two groups. HIV viral load at both BL and FU was categorized into detectable versus non-detectable levels. For categorical data, the general association for comparison of proportions was calculated. For variables where there were no observations in a given cell, Fisher’s exact p-value was calculated. Overall, we conducted tests of 50 biomarkers for each of 17 different comparisons or scientific hypotheses for a total of 850 Median HCV treatment duration, weeks Median CD4+ cell count, cells/mL Characteristic Undetectable HIV RNA level Follow-up Follow-up Baseline Baseline 14 10 Biomarkers in HCV and HIV Infection 5 Biomarkers in HCV and HIV Infection tests. Simultaneous testing has the potential to inflate Type 1 error. However, multiple testing corrections necessarily inflate Type 2 error, increasing the purchase GLPG-0634 chance of false negative conclusions. To balance the risks of Type 1 and Type 2 error, we applied a withincomparison Bonferroni correction, using an a = 0.05/50 = 0.001 significance level for all biomarker analyses. Whether to correct across distinct scientific hypotheses is a matter of controversy. We note that the 0.001 significance level guarantees that, within a 50-test comparison, the probability of zero false positives is greater than 95%. 
Furthermore, we expect at most one false positive in 1,000 tests. For the total 850 tests conducted, this amounts to less than one false positive. Using a more stringent significance level of 0.001/17 = 0.0006 to correct across comparisons would have reduced the overall expected number of false positives to at most 0.05, while increasing the expected number of false negatives by an unknown number. For those biom
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