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Decreased placental protein expression of PHOSPHO-AKT correlates with a decrease in expression of eNOS protein at mid-gestion in an ovine model of intrauterine growth restricition (IUGR)
SMFM Abstracts S185 607 DECREASED PLACENTAL PROTEIN EXPRESSION OF PHOSPHO-AKT CORRELATES WITH A DECREASE IN EXPRESSION OF ENOS PROTEIN AT MID-GESTION IN AN OVINE MODEL OF INTRAUTERINE GROWTH RESTRICITION (IUGR) JUAN A. ARROYO1, RUSSELL V. ANTHONY2, THOMAS A. PARKER2, GATES ROE3, HENRY 1 1 L. GALAN , University of Colorado Health Sciences Center, Obstetrics and Gynecology, Denver, Colorado, 2University of Colorado Health Sciences Center, Dept. of Pediatrics, Aurora, Colorado, 3University of Colorado Health Sciences Center, Dept. of Pediatrics, Denver, Colorado OBJECTIVE: Endothelial nitric oxidase synthase (eNOS) and NO are important regulators of blood flow in the fetal-placental circulation and are modulated by shear stress. eNOS activity is regulated by the phosphorylation (p) of ERK and AKT proteins in various tissues. In an established hyperthermic (HT) ovine model of placental and fetal growth restriction (FGR), we hypothesize that eNOS, pERK and pAKT protein levels would be decreased in the placental tissues in 95 day gestational age (dGA; term=147dGA) animals. STUDY DESIGN: 4 ewes were exposed to HT conditions for 55 days to induce FGR and 4 were used as controls. Placentomes were separated into caruncle and cotyledon components, and frozen for Western blot analysis with an antibody against eNOS, pERK, ERK, pAKT and AKT . Actin westerns on the same blots were used to account for inter-lane loading variations. RESULTS: HT animals showed, 1) smaller placentae (440G50 v. 186G18; p!0.004) while fetus weights were similar, 2) decreased O2 saturation (5.5G.06% v. 3.8G0.5% p!0.04) and pO2 levels (48.6G15.8 mmHg v. 23.2G2.34 mmHg; p!0.02), 3) a 2.5-fold decrease in caruncle eNOS protein and a 1.9-fold decrease in cotyledon eNOS protein (p!0.03), 4) a 1.5-fold increase in caruncle pERK with a 1.4-fold decrease in pAKT, 5) no differences in cotyledon pERK with a 2-fold decrease in pAKT protein (p!0.02).
has no recognized infertility predisposition. Thus, dissemination of information about pregnancy and delivery in dwarf women would be useful to LPs and their healthcare providers. STUDY DESIGN: A retrospective cohort study of LPs with R one prior conception (regardless of outcome), pertaining to her pregnancy and delivery experience, maternal physical status pre- and post-delivery, and general opinions about pregnancy in LPs. LP women were recruited via a genderand age-specific mailing to members of the Little People of America, Inc. following Medical Advisory Board and IRB approval. RESULTS: To date, 45 women representing 94 total pregnancies with 12 different skeletal dysplasia diagnoses are enrolled, including achondroplasia (26), pseudoachondroplasia (4), hypochondroplasia (3), diastrophic dysplasia (2), other dysplasias (10). 72 liveborn infants were delivered via cesarean section (39 general anesthesia-GA, 33 spinal and/or epidural), 16 miscarriages, and 1 termination. 4 liveborn infants were delivered vaginally; 2 without anesthesia to a mother with hypochondroplasia, 2 with epidural to a mother with femoral focal hypoplasia. One pregnancy is ongoing. CONCLUSION: Thus far, there is no apparent increased rate of pregnancy loss or fetal complications related to maternal dwarf condition. Most LP women require cesarean section due to cephaloplevic disproportion, yet vaginal delivery is noted in skeletal dysplasias with minimal truncal or pelvic disproportion. Many LPs have carried successful pregnancies and delivered with adequate pain control, under both regional and GA. We neither promote nor discourage childbearing in LP women, yet there is little medical literature regarding pregnancy and delivery in dwarfs. We present practical experiences with obstetrics and obstetrical anesthesia from the dwarf and healthcare provider perspective for others who choose to have children and those who provide their medical care. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.661
CONCLUSION: We conclude that decreased placental eNOS protein correlates with a decrease placental expression of pAKT protien in our ovine model of FGR . These findings support a vasculopathic state that may in part account for the abnormal umbilical velocimetry and fetal hypertension seen in the HT model of FGR. (Supported by NIH grant R01 HL071990-01A1). 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.659
608 OBSTETRICS AND OBSTETRICAL ANESTHESIA ISSUES IN WOMEN WITH DWARFISM JULIE HOOVER-FONG1, GRETCHEN OSWALD2, DEE MILLER2, JENNIFER LEADROOT2, HILLARY BARNES2, DONALD PENNING2, IVOR BERKOWITZ2, DEBORAH KRAKOW3, JUDITH ROSSITER4, 1Johns Hopkins University, Pediatrics, Baltimore, Maryland, 2Johns Hopkins University, Baltimore, Maryland, 3Cedars-Sinai Medical Center, Los Angeles, California, 4St. Joseph Medical Center, Gynecology and Obstetrics, Towson, Maryland OBJECTIVE: There is a paucity of medical literature regarding pregnancies and obstetrical care in dwarf women (aka Little People, LPs). This population
609 INTRAUTERINE E.COLI INOCULATION ALTERS PURKINJE CELL MORPHOLOGY, TUMOR NECROSIS FACTOR- AND CALBINDIN EXPRESSION IN THE DEVELOPING BRAIN KEDRA WALLACE1, SRUTHI VEERISETTY2, JOSEPHINE WYATT-ASHMEAD3, JOSe´ MIGUEL-HIDALGO4, WILLIAM BENNETT1, 1University of Mississippi Medical Center, Obstetrics and Gynecology, Jackson, Mississippi, 2Emory University, Atlanta, Georgia, 3Hammersmith NHS, London, United Kingdom, 4University of Mississippi Medical Center, Psychiatry and Human Behavior, Jackson, Mississippi OBJECTIVE: The cerebellum is involved in the control of motor functions. Purkinje axons are the only output from the cerebellum. Purkinje cells are important targets for toxic substances and are vulnerable to prenatal insults. Functional or morphological abnormalities in Purkinje cells could explain some of the problems seen in children with cerebral palsy (CP) or periventricular leukomalacia (PVL). Using an established animal model of PVL we examined Purkinje cell number and morphology in the brains of pups born to normal and experimentally infected dams. In addition to this, we also looked at the presence of pro-inflammatory cytokine tumor necrosis factor alpha (TNF-a) and calbindin, a calcium binding protein which is also a known marker of Purkinje cells. Intrauterine inoculation will lead to alterations in Purkinje cell morphology and number as well as changes in TNF-aand calbindin expression. STUDY DESIGN: Pregnant rats were inoculated with Escherichia coli (1x105CFU) or sterile saline at ED17. At PN16, brains were extracted and processed for immunhistochemistry (IHC). H&E staining was done to examine the general morphology and IHC was done to detect the presence of TNFaand calbindin. RESULTS: H&E staining demonstrated a change in Purkinje cell morphology. Purkinje cells from rats exposed to infection in-utero were oblong and greater gaps were noted between cells when compared to controls. The axon fibers were also less linear than controls. Decreased expression of TNF-a and calbindin were also noted in the E. coli infected group. CONCLUSION: In this model of PVL, we observed changes in Purkinje cell morphology which could result in alterations in motor capability and function. The decrease in TNF-a and calbindin may be due to a decrease of function in Purkinje cells. This evidence further supports the validity of this model for examining some of the underlying problems children with CP and PVL face with regards to motor coordination. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.662
has no recognized infertility predisposition. Thus, dissemination of information about pregnancy and delivery in dwarf women would be useful to LPs and their healthcare providers. STUDY DESIGN: A retrospective cohort study of LPs with R one prior conception (regardless of outcome), pertaining to her pregnancy and delivery experience, maternal physical status pre- and post-delivery, and general opinions about pregnancy in LPs. LP women were recruited via a genderand age-specific mailing to members of the Little People of America, Inc. following Medical Advisory Board and IRB approval. RESULTS: To date, 45 women representing 94 total pregnancies with 12 different skeletal dysplasia diagnoses are enrolled, including achondroplasia (26), pseudoachondroplasia (4), hypochondroplasia (3), diastrophic dysplasia (2), other dysplasias (10). 72 liveborn infants were delivered via cesarean section (39 general anesthesia-GA, 33 spinal and/or epidural), 16 miscarriages, and 1 termination. 4 liveborn infants were delivered vaginally; 2 without anesthesia to a mother with hypochondroplasia, 2 with epidural to a mother with femoral focal hypoplasia. One pregnancy is ongoing. CONCLUSION: Thus far, there is no apparent increased rate of pregnancy loss or fetal complications related to maternal dwarf condition. Most LP women require cesarean section due to cephaloplevic disproportion, yet vaginal delivery is noted in skeletal dysplasias with minimal truncal or pelvic disproportion. Many LPs have carried successful pregnancies and delivered with adequate pain control, under both regional and GA. We neither promote nor discourage childbearing in LP women, yet there is little medical literature regarding pregnancy and delivery in dwarfs. We present practical experiences with obstetrics and obstetrical anesthesia from the dwarf and healthcare provider perspective for others who choose to have children and those who provide their medical care. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.661
CONCLUSION: We conclude that decreased placental eNOS protein correlates with a decrease placental expression of pAKT protien in our ovine model of FGR . These findings support a vasculopathic state that may in part account for the abnormal umbilical velocimetry and fetal hypertension seen in the HT model of FGR. (Supported by NIH grant R01 HL071990-01A1). 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.659
608 OBSTETRICS AND OBSTETRICAL ANESTHESIA ISSUES IN WOMEN WITH DWARFISM JULIE HOOVER-FONG1, GRETCHEN OSWALD2, DEE MILLER2, JENNIFER LEADROOT2, HILLARY BARNES2, DONALD PENNING2, IVOR BERKOWITZ2, DEBORAH KRAKOW3, JUDITH ROSSITER4, 1Johns Hopkins University, Pediatrics, Baltimore, Maryland, 2Johns Hopkins University, Baltimore, Maryland, 3Cedars-Sinai Medical Center, Los Angeles, California, 4St. Joseph Medical Center, Gynecology and Obstetrics, Towson, Maryland OBJECTIVE: There is a paucity of medical literature regarding pregnancies and obstetrical care in dwarf women (aka Little People, LPs). This population
609 INTRAUTERINE E.COLI INOCULATION ALTERS PURKINJE CELL MORPHOLOGY, TUMOR NECROSIS FACTOR- AND CALBINDIN EXPRESSION IN THE DEVELOPING BRAIN KEDRA WALLACE1, SRUTHI VEERISETTY2, JOSEPHINE WYATT-ASHMEAD3, JOSe´ MIGUEL-HIDALGO4, WILLIAM BENNETT1, 1University of Mississippi Medical Center, Obstetrics and Gynecology, Jackson, Mississippi, 2Emory University, Atlanta, Georgia, 3Hammersmith NHS, London, United Kingdom, 4University of Mississippi Medical Center, Psychiatry and Human Behavior, Jackson, Mississippi OBJECTIVE: The cerebellum is involved in the control of motor functions. Purkinje axons are the only output from the cerebellum. Purkinje cells are important targets for toxic substances and are vulnerable to prenatal insults. Functional or morphological abnormalities in Purkinje cells could explain some of the problems seen in children with cerebral palsy (CP) or periventricular leukomalacia (PVL). Using an established animal model of PVL we examined Purkinje cell number and morphology in the brains of pups born to normal and experimentally infected dams. In addition to this, we also looked at the presence of pro-inflammatory cytokine tumor necrosis factor alpha (TNF-a) and calbindin, a calcium binding protein which is also a known marker of Purkinje cells. Intrauterine inoculation will lead to alterations in Purkinje cell morphology and number as well as changes in TNF-aand calbindin expression. STUDY DESIGN: Pregnant rats were inoculated with Escherichia coli (1x105CFU) or sterile saline at ED17. At PN16, brains were extracted and processed for immunhistochemistry (IHC). H&E staining was done to examine the general morphology and IHC was done to detect the presence of TNFaand calbindin. RESULTS: H&E staining demonstrated a change in Purkinje cell morphology. Purkinje cells from rats exposed to infection in-utero were oblong and greater gaps were noted between cells when compared to controls. The axon fibers were also less linear than controls. Decreased expression of TNF-a and calbindin were also noted in the E. coli infected group. CONCLUSION: In this model of PVL, we observed changes in Purkinje cell morphology which could result in alterations in motor capability and function. The decrease in TNF-a and calbindin may be due to a decrease of function in Purkinje cells. This evidence further supports the validity of this model for examining some of the underlying problems children with CP and PVL face with regards to motor coordination. 0002-9378/$ - see front matter doi:10.1016/j.ajog.2006.10.662