NAC (2, 5, and 10 mM). After an addition 24 h of culture, medium was collected and analyzed by ELISA for the cytokines interleukin (IL)-6, IL-8, IL-1a, IL-1b, and TNF-a, as well as prostaglandin E2 (PGE2). N¼4-10 women; each treatment was performed in replicate cultures from the same woman. Results: LTA stimulated release of IL-6, IL-8, TNF-a, and PGE2 from full thickness tissue punches, and these responses were inhibited by NAC. Similarly, live GBS stimulated release of IL-6, IL-8, TNF-a, and PGE2 from choriodecidual punches, and these responses were inhibited by NAC. Notably, the GBS-stimulated release of TNF-a was completely blocked at all concentrations of NAC. In contrast, the GBS-stimulated release of IL-1a and IL-1b was unaffected by NAC treatment (LTA-stimulated release of IL-1a and IL-1b was not measured). Conclusion: The antioxidant NAC effectively blunted release of PGE2 and the proinflammatory cytokines IL-6, IL-8, and TNF-a from human extrap- lacental membranes when challenged in vitro with live GBS or LTA. These results suggest that that GBS elicits at least some of its effects on gesta- tional tissues through reactive oxygen species. Further research is needed to better understand whether this phenomenon can be exploited for therapeutic interventions in clinical practice. P2.49 TROPHOBLAST MITOCHONDRIAL BIOGENESIS AND FUNCTIONALITY IS INCREASED WITH SELENIUM SUPPLEMENTATION Anthony Perkins, Alisha Khera, Olivia Holland, Jessica Vanderlelie. Griffith University, Gold Coast, Queensland, Australia Objectives: Excessive production of reactive oxygen species (ROS) causes cellular damage and has been associated with complication of pregnancy such as preeclampsia. Mitochondria are the major site of reactive oxygen species (ROS) and are susceptible to oxidative stress. In previous studies we have shown that the essential trace element selenium is able protect trophoblast cells and scavenge ROS through the antioxidant enzymes Glutathione Peroxidase (Gpx) and Thioredoxin Reductase (Thx-Red) 1 . The present study was conducted to demonstrate how selenium increases mitochondrial function of trophoblast cells and activates mitochondrial biogenesis. In particular we examined the expression of Selenoprotein-H which activates transcription factors such as nuclear respiratory factor-1 (NRF1) and peroxisome proliferator-activated receptor coactivator -1alpha (PGC-1a) to increase mitochondrial biogenesis 2 . Methods: Trophoblast cells (BeWo, JEG-3 and Swan-71) were treated with Na Selenite (100nM) for 24 hours. Cellular respiration was then measured in an Oxygraph-2k Oroboros high respirometry chamber operated at 37  C. qPCR and citrate synthase assays were done to demonstrate that selenium treated cells have higher mitochondrial number. Western blots were per- formed to determine up-regulation of Selenoprotein H, NRF-1 and PGC-1a. Results: This study revealed that selenium treatment (100nM) for 24 hours effectively stimulates mitochondrial biogenesis, increasing mito- chondrial number in trophoblast cells. The oxygen consumption was higher in the cells treated with NaSe as compared to the non-treated cells. The mitochondrial respiration indicated that oxygen flow was significantly increased in selenium supplemented cells. qPCR data revealed that mito- chondrial gene/nuclear gene ratio was higher in the cells treated with selenium. Selenium supplementation also up-regulated the expression of Selenoprotein H, NRF-1 and PGC-1a 3 , important mediators of mitochon- drial biogenesis. Conclusion: These data demonstrate that supplementation with the trace element selenium increases mitochondrial biogenesis and functionality. This is mediated through enhanced anti-oxidant production, lowering ROS production thus protecting mitochondrial function as well as increased expression of biogenesis activators. This emphasizes the special impor- tance of maintaining adequate selenium during pregnancy and especially in pregnancies complicated by conditions such as preeclampsia. References: (1) Khera, A., Vanderlelie, J.J. & Perkins, A.V. (2013) Selenium Supple- mentation Protects Trophoblast Cells from Mitochondrial Oxidative Stress. Placenta, doi:pii: S0143-4004(13)00184-7. (2) Natalia Mendelev , Suresh L Mehta, Haza Idris, Santosh Kumari, and P Andy Li (2012) Selenite stimulates mitochondrial biogenesis signalling and enhances mitochondrial functional performance in murine Hippocampal Neural Cells. Plos One, 10.1371 0047910. (3) Khera, A., Lan-feng Dong, Holland,O., Vanderlelie, J.J. Pasdar, E.M., Neuzil, J., Perkins, A.V. (2015) Selenium supplementation induces mito- chondrial biogenesis in placental trophoblast cells. Placenta, (8):863-9 P2.50 HYPOXIA AND IRON IMBALANCE IMPAIRS JMJD6-MEDIATED HISTONE DEMETHYLATION OF VHL IN PREECLAMPSIA Sruthi Alahari 1, 2 , Alessandro Rolfo 3 , Isabella Caniggia 1, 2 . 1 Lunenfeld- Tanenbaum Research Institute, Toronto, Ontario, Canada; 2 Departments of Physiology, and Obstetrics & Gynaecology, University of Toronto, Toronto, Ontario, Canada; 3 Department of Surgical Sciences, University of Turin, Turin, Piedmont, Italy Objectives - Chronic hypoxia is a defining feature of preeclampsia. Mounting evidence implicates the O 2 and Fe(II)-dependent Jumonji C domain histone demethylases (JmjDs) as epigenetic mediators of hypoxic gene expression. We recently identified the bifunctional oxygen sensor, JMJD6, as a regulator of VHL (von Hippel Lindau tumour suppressor) protein stability in the human placenta. JMJD6 plays a key role in the histone code by demethylating arginine residues on histones 3 (H3R2me2) and 4 (H4R3me2). Given that VHL is downregulated in preeclamptic placentae, we examined JMJD6 function as a histone demethylase in regulating VHL in preeclampsia. Methods - Placentae were obtained from preeclamptic (PE; n¼25), and normotensive age-matched control (AMC; n¼21) pregnancies. For in vitro demethylation studies, histones isolated from primary cytotrophoblasts or PE and AMC placentae were incubated with recombinant JMJD6 enzyme at 3% or 8% O 2 , followed by Western Blotting. Results - Western blotting revealed significantly elevated H3R2me2 and H4R3me2 expression in PE placentae. JMJD6 enzyme demethylated both histone targets in AMC, but not PE placentae. Placental levels of the Fe(II) ferroxidase, ceruloplasmin were strikingly elevated in PE and, therefore, we hypothesized that reduced Fe(II) availability limits JMJD6 activity in PE. In vitro demethylation of primary cytotrophoblasts revealed decreased expression of both histone targets in 8%, but not in 3% O 2 , indicating optimal demethylase activity in normoxia. Importantly, addition of excess Fe(II) partially rescued JMJD6-mediated demethylation in primary cells in hypoxia and histones from PE placentae, thereby highlighting the impor- tance of iron in mediating JMJD6 function in preeclampsia. Conclusion - This study provides novel evidence of interplay between iron availability and oxygen on JMJD6 demethylase function in the human placenta. The hypoxic environment in PE increases ceruloplasmin, impacting on Fe(II) availability. This, in turn, impairs JMJD6 demethylase activity, thereby contributing to the decreased VHL in PE. (Supported by CIHR) P2.51 VEGF AND SFLT-1 POLYMORPHISMS IN PREECLAMPSIA Pallavi Arora, Renu Dhingra, Neerja Bhatla, Arundhati Sharma. 1 All India Institute of Medical Sciences, New Delhi, India; 2 All India Institute of Medical Sciences, New Delhi, India Objectives: The exact underlying molecular mechanisms for the patho- genesis of preeclampsia are unclear, but are likely to be multifactorial. Various genetic and environmental factors have also been considered to contribute to the pathogenesis of preeclampsia due to a strong familial predisposition observed in individuals of different geographic, socio- economic and racial distribution. Of these imbalance in the proangio- genic (VEGF and PlGF) and antiangiogenic (sFlt-1) factors may serve as a major cause for the most fundamental pathology i.e. shallow tropho- blastic invasion leading to impaired spiral artery remodeling. Thus the aim of the study was to screen for presence of VEGF and sFlt-1 gene polymorphisms and to measure their levels in patients and controls of Indian origin. Methods: 30 each of clinically diagnosed patients and gestational and maternal age matched controls were recruited after taking informed consent. DNA isolated from blood samples was processed for PCR ampli- fication followed by Restriction Fragment Length Polymorphism to screen Abstracts / Placenta 45 (2016) 63e133 118