Progesterone and estrogen receptor expression in labored and unlabored women

SMFM Abstracts S133 421 ALTERATIONS IN P-CREB ACTIVITY: A PATHWAY FOR FAS RELATED NEUROTOXITICITY ROBIN ROBERSON1, IRENE CAMERONI2, LAURA TOSO1, DANIEL ABEBE1, STEPHANIE BISSELL1, CATHERINE SPONG1, 1National Institutes of Health (NIH), Unit on Perinatal and Develomental Neurobiology, NICHD&NIAAA, Bethesda, Maryland, 2University of Milano-Bicocca, Obstetrics and Gynecology, Monza, Milan, Italy OBJECTIVE: Fetal alcohol syndrome (FAS) is the leading cause of a spectrum of preventable non-genetic learning disorders. In the adult, neurogenesis occurs in the hippocampal dentate gyrus, which is thought to be important for learning and memory. Previously we have shown alterations in the NMDA receptor underlie some of the learning abnormalities in FAS. NMDA exerts it action through CREB. CREB is a transcription factor that controls gene expression via phosphorylation promoting neuronal survival. Using a model for FAS, we measured P-CREB levels in hippocampal subregions to evaluate if alterations in P-CREB underlie the FAS learning abnormalities after in utero alcohol exposure. STUDY DESIGN: Pregnant C57Bl6/J mice were treated on gestational day 8 with alcohol, n = 10 or control (saline) n = 5. Adult males were assessed in the Morris water maze for learning on days 50-56. On day 57, 5 males/ treatment were perfused for immunohistochemistry, and stained with SER 144 P-CREB antibody with relative densities of staining assessed using NIH Image software. Statistical analysis ANOVA, P!.05 significant. RESULTS: The hippocampal areas CA1, dentate and CA3 were quantitated. In the control, P-CREB staining was greatest in dentate granule cells and lowest in CA1. In all of the hippocampal subregions, alcohol exposure in utero resulted in significant downregulation of P-CREB (P!.001).

Prenatal alcohol down regulates P-CREB CONCLUSION: In utero alcohol exposure decreased P-CREB activity in all hippocampal subregions of adult mice. The dentate granules cells had the most robust decrease in P-CREB staining, suggesting that in FAS the adult learning deficits may correlate to enhanced dentate granule neurodegeneration. Since the dentate gyrus is the area of adult neurogenesis, these findings may explain in part the learning deficit in FAS.

423 REDUCED LEVELS OF SOLUBLE HLA-G PROTEIN ASSOCIATED WITH SEVERE PREECLAMPSIA AT THE THIRD TRIMESTER RINAT HACKMON1, ARIE KOIFMAN2, HIROHITO HYOBO3, HAGIT GLICKMAN1, EYAL SHEINER4, DAN GERAGHTY3, 1TelAviv University, Ob/Gyn, Tel Aviv, Israel, 2Soroka Medical Center, Ob/ Gyn, Beer-Sheva, Israel, 3Fred Hutchinson Cancer Research Center, Clinical Research Division, Seattle, Washington, 4Soroka Medical Center, BeerSheba, Israel OBJECTIVE: Recently maternal lower plasma HLA-G protein levels in preeclampsia (PE) in the first and second trimesters was reported. Thus we sought to evaluate the levels of HLA-G protein in severe PE during the thirdtrimester. STUDY DESIGN: In this prospective case-control study amniotic fluid and maternal and cord blood samples were aspirated from 50 pregnant women undergoing cesarean section during the third-trimester. The study group included 26 pregnant women diagnosed with severe PE while 24 women without PE served as controls. The control group was matched for gestational age and for nulligravidity vs. multiparity. Exclusion criteria were: multiple gestations, preterm labor, active labor and genetic or congenital malformations. An HLA-G specific ELISA was used to measure protein levels in PE and normal control pregnancies. Capturing antibody used was MEM-G/9 and biotinylated detecting antibody was 16G1, the former HLA-G1 specific and the latter specific for only soluble HLA-G1. Standard protein was diluted serially in human AB serum depleted of HLA-G protein. HLA-G concentrations in samples were determined by four-parameter logistic curve fitting in logarithmic plotting. Statistical analysis included Student’s t-test and p-value !0.05 was considered statistically significant. RESULTS: Serum HLA-G levels in PE pregnancies were found to be significantly lower than from normal pregnancies (10.97G6.55 vs. 36.05G34.53 mg/ml, p=0.003). No statistically significant difference was found between the study and the control groups in cord serum and amniotic fluid (5.39G6.42 vs. 7.19G 10.42 mg/ml, p=0.47 and ÿ4.11G6.59 vs. ÿ1.35G9.86mg/ml, p=0.25; respectively). CONCLUSION: A reduced levels of maternal HLA-G protein is associated with severe preeclampsia during the third-trimester. Based upon this and upon previous findings we conclude that in preeclampsia maternal serum HLA-G levels are lower during all three trimesters of pregnancy. In addition to the potential diagnostic implications, this study suggests an essential role for HLA-G in normal and abnormal pregnancies. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.462

0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.459 422 ASSESSMENT OF FIRST TRIMESTER MATERNAL SERUM PP13 IN EARLY VS. TERM SEVER PREECLAMPSIA ROBERTO ROMERO1, JIMMY ESPINOZA2, SAM EDWIN3, ILANA CHEFETZ4, MAREI SAMMAR5, HAMUTAL MEIRI5, YOSSI TAL6, IDO KUHNREICH6, HOWARD CUCKLE7, 1Perinatology Research Branch, Intramural Division, NICHD, NIH, DHHS, Detroit, Michigan, 2National Institutes of Health (NIH), Perinatology Research Branch, NICHD, NIH, DHHS, Detroit, Michigan, 3National Institute of Child Health and Human Development, Perinatology Research Branch, Detroit, Michigan, 4 Diagnostic Technologies Ltd, Yokneam, Israel, 5Diagnostic Technologies, Yokneam, Israel, 6Technostat, Kfar Saba, Israel, 7University of Leeds, Reproductive Epidmiology, Leeds, United Kingdom OBJECTIVE: To assess placenta protein 13 (PP13) as a first trimester marker for early vs. term severe preeclampsia. STUDY DESIGN: We tested 48 cases of preeclampsia-13 necessitating delivery before gestation week (GA) 37 (6 at GA!34, 11 with IUGR!5%), 21 of term severe preeclampsia (5 with IUGR), and 16 with mild preeclampsia at term, all matched to 250 unaffected cases. Maternal PP13 of stored serum samples GA 8-13 tested for PP13 blinded to outcome. RESULTS: In unaffected and mild term preeclampsia the multiple of the median (MoM) of PP13 was 1.0 and 0.87 (not significantly different), respectively compared to MoM=0.25 in early (GA!37), and MoM=0.69 in term severe preeclampsia and 0.43 in the two combined (p! 0.001). Using receiver operating characteristic (ROC) the area under the curve (AUC) was 0.85C0.07 (95% CI) for early preeclampsia, 0.66C0.10 for term severe preeclampsia and 0.73C0.08 for the two combined. Setting specificity to 80% at MoM cutoff=0.45, the sensitivities were 85% (early preeclampsia), 29% (term severe preeclampsia), and 50% (the two combined). Lower PP13 was found in preeclampsia complicated by IUGR. CONCLUSION: First trimester PP13 may serve to assess the risk for developing early preeclampsia (with or without IUGR), whereas assessment of severe preeclampsia at term may not be effective. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.461

424 PROGESTERONE AND ESTROGEN RECEPTOR EXPRESSION IN LABORED AND UNLABORED WOMEN RICHARD H. LEE1, FRANK STANCZYK1, GLORIA YANG1, JI QING2, ANDREW STOLZ2, CRISTIANE GUBERMAN1, T. MURPHY GOODWIN1, 1 University of Southern California, Obstetrics and Gynecology, Los Angeles, California, 2University of Southern California, Gastroenterology, Los Angeles, California OBJECTIVE: It is hypothesized that changes in progesterone receptor (PR) and estrogen receptor (ER) isoforms convert the uterus from a quiescent state to a contractile state. The purpose of this study is to compare expression levels of PR isoforms (PR-A, PR-B) and ER isoforms (ER-a, ER-b) in the myometrium, membranes, and placentas of term subjects in labor and not in labor. STUDY DESIGN: Samples of myometrium, chorioamniotic membranes, and placentas were obtained from term pregnancies undergoing elective cesarean section not-in-labor (n=13) and from those in spontaneous labor (n=5). Labor was defined as regular painful uterine contractions resulting in cervical dilatation. Quantitative real-time PCR was performed using primers specific for PR isoforms AB and B, as well as, ER isoforms a and b. Statistical analysis was performed using Mann-Whitney U and c2. RESULTS: In the myometrium, there was no significant difference in PR-A, ER-a, ER-b expression. A statistical trend towards higher PR-B expression (2.8 folds) was found in the myometrium of non-laboring subjects compared to laboring subjects (p=0.09). The membranes demonstrated no statistical difference in the expression of PR-B, PR-A, or ER-a. However, the expression of membrane ER-b was 4.5 folds higher in labor (p=0.03). In the placenta, there were no statistical differences in the expression of ER-a or ER-b. Laboring subjects demonstrated statistical trends towards higher expression of PR-B (3.3 fold, p=0.053) and PR-A (12 fold, p=0.07). CONCLUSION: Our data suggest that alterations in the relative expression of PR-A and ER-a do not occur in laboring lower-uterine segment myometrium. The relative increase of myometrial PR-B expression in non-laboring subjects supports the hypothesis that PR-B maintains uterine quiescence. Alterations in PR isoforms do not occur in the membranes. Both PR-A and PR-B are expressed in the placenta. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.463