Inc., Santa Clara, California, USA) by using a 27-plex Bioplex Human Cytokine kit, which includes IL-1��, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, eotaxin, basic FGF, G-CSF, GM-CSF, IFN-��, IP-10, MCP-1 (MCAF), MIP-1��, MIP-1��, PDGF-BB, RANTES, TNF-��, and selleck VEGF (Bio-Rad Laboratories), according to the manufacturer’s instructions. In 50 children with bacterial meningitis, in whom sufficient CSF existed for analysis, CSF growth factors were determined on admission. Plasma Ang-1 and Ang-2 were determined by using a commercial ELISA assay (R&D Systems Europe, Ltd., Abingdon, UK). We have previously reported the analysis of chemokines and pro-and antiinflammatory cytokines in this cohort [33,35].

HIV determinationHIV status was assessed in children 18 months or older by using at least two of the following tests; Unigold and Serocard (Trinity Biotech, Wicklow, Ireland), or Determine-HIV (Abbott Laboratories, Springfield, IL, USA). At least two tests were required to be positive for a subject to be classified as HIV infected. In children younger than 18 months, and in those with discordant antibody tests, HIV status was determined by using Amplicor HIV-1 DNA Test version 1.5 (Roche Diagnostics, South San Francisco, CA, USA).Statistical analysisThe growth factors and angiopoietins determined were summarized by using geometric means and interquartile ranges (IQRs). Two-sample t tests were used to compare growth-factor concentrations between groups, by using log-transformed data.

Multiway analyses of variance were used to obtain adjusted comparisons for each factor of interest (main effects: SBI/NBI, pneumonia/meningitis, HIV status, survivor/nonsurvivor, and gram positive/negative infection). Correlations between growth factors and other variables were estimated by using Spearman’s rho correlation coefficient. Fisher’s Exact test was used to compare proportions. Univariable and multivariable logistic regression analyses were performed to develop a prognostic model of the influence of confounding factors (HIV status, age, sex, diagnosis, and previous antibiotics) on the primary outcome measure, inpatient mortality. CSF and plasma growth factors in children with bacterial meningitis were analyzed by using Wilcoxon’s Signed Ranks test. Adjusted odds ratios (ORs) were obtained by using logistic regression. All tests were two-tailed, and a P value of < 0.05 was considered significant.ResultsPatient characteristicsWe studied 293 children (57% boys), of whom 64 (22%) died. The median age was 2.4 years, and the IQR, 0.7 to Drug_discovery 6.0 years.