804: The elevation of circulating anti-angiogenic factors in preeclampsia occurs independent of markers of neutrophil activation

Abstract

factors in preeclampsia occurs independent of markers of neutrophil activation Wenda Ramma, Irina A. Buhimschi, Guomao Zhao, Antonette T. Dulay, Unzila A. Ali, Catalin S. Buhimschi, Asif Ahmed University of Edinburgh, Queen’s Medical Research Institute, Edinburgh, United Kingdom, Yale University, Ob/Gyn & Reprod Sci., New Haven, CT OBJECTIVE: The imbalance among proand anti-angiogenic factors and a heightened state of maternal intra-vascular inflammation are proposed as central pathophysiological features of preeclampsia. The objective of this study was to investigate the relationship between levels of anti-angiogenic factors and prototype markers of neutrophil activation in preeclampsia. STUDY DESIGN: We analyzed blood samples of 88 women stratified as follows: severe preeclampsia (sPE, n 45, GA: 30 [27-32 weeks]), maternal systemic inflammatory response (SIR, n 16, GA: 30 [28-33 weeks]) secondary to chorioamnionitis, pyelonephritis, appendicitis, and normotensive controls (CRL, n 27, GA: 29 [26-32 weeks]). All samples were obtained prior to steroids or magnesium. Soluble Fmslike tyrosine kinase receptor-1 (sFlt-1), placenta growth factor (PlGF), soluble Endoglin (sEng), interleukin-6 (IL-6), -defensins, and calprotectin levels were analyzed in serum or plasma as appropriate by ELISA. RESULTS: Women with sPE had elevated levels of serum sEng (Fig. A) and sFlt-1 (Fig. B) (P .001 for both) compared to SIR and CRLs. Women with SIR had sFlt-1 and sEng levels similar to CRLs. Women with sPE had higher IL-6 levels versus CRLs (Fig. C), but lower compared to SIR cases. Compared to CRLs, both sPE and SIR had higher levels of -defensins and calprotectin (P .05 for both). In sPE, there was no correlation between anti-angiogenic factors and the sought markers of neutrophil activation [sEng & -defensins (P .376) and sEng & calprotectin (P .133)] or levels of IL-6 [sEng & IL-6 (P .597) and sFlt-1 & IL-6 (P .423)]. CONCLUSION: We present evidence that activation of the anti-angiogenic and inflammatory systems are associated with development of sPE, but the degree of pathologic dysregulation does not seem to evolve in parallel. 805 Utilization of maternal blood on Guthrie card for first trimester screen for noninvasive fetal sex determination and RhD genotyping Yali Xiong, Indhu M Prabhakaran, Eliezer J Holtzman, Stacey Jeronis, Dan A Liebermann, Barbara Hoffman, Ossie Geifman-Holtzman Temple University School of Medicine, Fels Institute, Philadelphia, PA, Temple University, Fels Institute, Philadelphia, PA, Sheba Medical Center, Tel-Aviv University, Nephrology and Hypertension Institute, Tel Hashomer, Israel, Temple University Dept. OBGYN, OBGYN, Philadelphia, PA, Temple University School of Medicine, Fels Institute, Philadlephia, PA, Drexel University School of Medicine, OBGYN, Philadelphia, PA OBJECTIVE: To evaluate the use of maternal blood on Guthrie card obtained at the first trimester screen as a common source for first trimester screen, fetal sex determination and Fetal RhD genotyping. STUDY DESIGN: Guthrie card blood samples and peripheral blood samples were collected from RhD-negative pregnant women in the first trimester. Guthrie cards blood samples were pretreated in 10XTE buffer. Mononuclear layer was isolated from peripheral blood with Ficoll. DNA from both Guthrie cards and peripheral blood mononuclear layer was isolated using Qiagen mini blood kit. Primers and probes specific to fetal gender determination (male DSY14) gene and polymorphism sites (female) were used. Probe specific Realtime PCR was processed including examining the Exon 4, 5, and 6 of RhD gene and fetus RhD genotype was determined. Maternal and paternal cheek swabs were used for their genotyping and follow up confirmation on the babies were obtained after delivery. RESULTS: Male Fetal DNA was confirmed to be present in the amplified samples using DSY14 gene. Samples that were not amplified for DSY14 and were subjected to polymorphic sites amplification (SO6) confirmed the presence of fetal DNA and correctly identified female fetus. The subsequent determination of fetal RhD type using the fetal DNA from Guthrie Card was confirmed using DNA obtained from maternal blood in the second trimester and /or after delivery as well as by standard serology of the baby’s blood. Fetus RhD genotype results were consistent between DNA obtained from Guthrie Card and from the mononuclear layer isolated from peripheral maternal blood. One alloimmunized patient had blood placed on Guthrie card as early as 5 weeks gestation and was incorrectly diagnosed. CONCLUSION: Our study revealed that fetal DNA from maternal blood samples placed on Guthrie card during first trimester screen could also be used for fetal RhD genotyping and fetal sex determination. The availability of this early testing is important clinically not only for diagnosis of X-linked diseases but also for determination of fetal RhD type in RhD-negative pregnant patients.