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Comparisons were made using Mann-Whitney U tests (nonnormally distributed data, between-group comparisons) or repeated measures two-way ANOVA (normally distributed data, where the time factor represents differences across both groups over time and the interaction factor represents between-group differences over time). Ex vivo cytokine data were log-transformed to obtain a normal distribution. A P value Crenolanib at the vaccination site in 9 out 10 subjects (median (range) size of 6 (1�C9)?mm). In the placebo group, 1 subject developed a small scar (1?mm). BCG vaccination did not result in fever or other clinical symptoms Lapatinib and no serious adverse events occurred during the trial. Table 1 Baseline characteristics. 3.2. Hemodynamic Parameters and Symptoms LPS administration resulted in a typical increase in heart rate and flu-like symptoms and a decrease in mean arterial blood pressure (MAP) in all subjects, with no differences between groups (Figure 2). Figure 2 Blood pressure, heart rate, and symptoms during experimental endotoxemia in subjects vaccinated with gamma-irradiated BCG or placebo. Data are expressed as mean �� SEM (n = 10 per group). P values calculated using repeated measures two-way analysis ... 3.3. Plasma Cytokines and Circulating Leukocyte Counts Liothyronine Sodium BCG vaccination did not result in increased plasma levels of any of the measured cytokines in the days following vaccination (Figure 3). As expected, administration of LPS resulted in a sharp increase in plasma levels of the proinflammatory cytokines TNF-��, IL-6, IL-8, and MCP-1, as well as the anti-inflammatory cytokines IL-10 and IL-1RA (Figure 3). However, no differences were observed between the BCG and placebo groups. Similar to previous human endotoxemia studies [26], plasma levels of IL-1�� and IFN-�� were below the lower detection limits in the majority of the subjects at most time points. In some subjects and/or time points, very low levels (approximately 10?pg/mL) were found, but no clear patterns over time or differences between groups were observed. Figure 3 Plasma cytokine concentrations in subjects vaccinated with gamma-irradiated BCG or placebo. In the panels (a�Cd), median values of proinflammatory cytokines TNF-��, IL-6, IL-8, and MCP-1 are depicted while in panels (e) and (f) median values ... As described earlier, scar size differed substantially between BCG-vaccinated subjects, which might represent vaccination efficacy, and is associated with nonspecific beneficial effects of BCG [27, 28].