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See the Supplemental Experimental Procedures for primer sequences. In preparation for microCT PD-0332991 manufacturer scanning a radio-opaque silicone rubber contrast agent (Microfil; Flow Technology, Carver, MA) was perfused via the umbilical artery via methods previously described (Rennie et?al., 2011). See the Supplemental Experimental Procedures for details of perfusion protocol and how quantification of vascular measurements was obtained. Vascular resistance was calculated with standard formulas for resistance in parallel and in series (Yang et?al., 2010) and the viscosity of blood in small vessels (Pries and Secomb, 2003). The distribution of pressure and flow in the tree was calculated assuming (1) Posieuille��s law for flow of fluid through a pipe-like structure, (2) equal pressure at each terminal vessel, and (3) a diameter dependent blood viscosity correction affecting small vessels as previously described (Pries and Secomb, 2003). Analysis was performed on placentas from wild-type?(nuninfected?= selleck chemical 7 from four litters; ninfected?= 7 from four litters) and C5aR?/? (nuninfected?= 5 from three litters; ninfected?= 8 from three litters) mice. One data set (C5aR+/+ uninfected) for which the umbilical vessel was not present due to the umbilical cord being tied off too close to the chorionic plate during the perfusion process was eliminated from hemodynamic modeling analyses. Continuous variables were analyzed by Mann-Whitney U test. Categorical data were analyzed by Pearson��s chi-square test or Fisher��s exact test. Adjusted odds ratios were obtained via logistic regression (SPSS). In order to test our hypothesis within a multivariate framework, we employed structural equation modeling (AMOS) to simultaneously examine the relationships between multiple dependent and independent variables (Calis et?al., 2008), as well as latent concepts and their multiple indicators. Risk factors for adverse outcomes in pregnancy were included in exploratory models and, through a process of trimming, were subsequently removed if insignificant (p > 0.05; e.g., number of antenatal visits, number of antimalarial doses). Inflammatory cytokines were not measured, so their relative impact on angiogenesis and birth outcomes was not assessed in this structural equation model. Model estimates were generated via maximum-likelihood estimation, and fitness was assessed Megestrol Acetate via the likelihood ratio test statistic (Byrne, 2010), expressed as chi-square and rmsea, respectively. A low rmsea (