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Severe discordancy in dichorionic diamniotic twin potentially shows preterm labor and poor perinatal outcome
Abstracts / Placenta 36 (2015) A1eA60 2 Department of Obstetrics and Gynecology, Ochsner Baptist Hospital, New Orleans, Louisiana, USA
Background: Obesity during pregnancy is associated with a proinflammatory state affecting the placenta's role in releasing molecules (e.g. exosomes) that can directly affect the maternal metabolism and fetal development. The aim of this study was to test the hypothesis that placenta-derived exosomes from obese women induce higher TNF-a releases from endothelial cells. Methods: Women were recruited, with informed, written consent, by research midwives from the Ochsner Baptist Medical Center (New Orleans, USA). Groups were classified according to the body max index (BMI) in two categories: lean (n ¼ 6, BMI 18.5e24.9 Kg/m2) and obese (n ¼ 6, BMI >30 Kg/m2). Exosomes were isolated from plasma by differential and buoyant density centrifugation. The total number of exosome vesicles and specific placenta-derived exosomes were determined by quantifying immunoreactive exosomal CD63 and PLAP using ELISA kit and nanoparticle tracking analysis (NanoSight NS500). The effect of exosomes on TNF-a cytokine release from endothelial cells was established using protein solution array analysis (Bioplex). Results: Exosome plasma concentration was 2-fold greater in obese than lean women (p < 0.05). In obese women, the number of exosomes present in maternal plasma (9.34 108 ± 2.0 108 particles per ml plasma) was significantly higher compared with lean women (4.14 108 ± 1.3 108 particles per ml plasma). The concentration of placenta-derived exosomes in maternal plasma (as indicated by exosomal PLAP) is higher in obese compared to lean women (1020 ± 75 pg/ml and 634 ± 56 pg/ml, respectively). Exosomes isolated from lean and obese women increased TNF-a releases from endothelial cell by 1.4 ± 0.1 and 2.6 ± 0.1-fold, respectively, compared to the control in the absence of exosomes. Conclusions: Proinflammatory state is associated with obesity and leads to the releases of proinflammatory cytokines into the circulation. Placental exosomes could contribute to this phenomenon by promoting cytokines releases from endothelial cells.
P2.18. SEVERE DISCORDANCY IN DICHORIONIC DIAMNIOTIC TWIN POTENTIALLY SHOWS PRETERM LABOR AND POOR PERINATAL OUTCOME Michihisa Shiro, Naoyuki Iwahashi, Tomoko Noguchi, Tamaki Yahata, Madoka Yamamoto, Yuko Tanizaki, Aya Kobayashi, Nami Ota, Shigetaka Yagi, Sawako Minami, Kazuhiko Ino. Wakayama Medical University, Wakayama, Japan Objectives: In dichorionic diamniotic (DD) twin, there is generically no placental anastomoses and each fetus has a own placenta. In the same environment of uterus, sometimes discordant twin occurs in DD twin. We investigated maternal factors and neonatal outcome of small for gestational age (SGA)at least one twin in DD twin pregnancy. Methods: This was a retrospective study of 113 patients who delivered DD-Twin, between January 2006 and December 2014 in our hospital. Small for gestational age (SGA) group was defined as a weight below the 10th percentile for the gestational age at least one baby. Especially, severe SGA group was defined as a weight below -2.0SD (standard deviation), Mild SGA group was defined as excepting severe SGA in all SGA cases. Results: There was no significant difference in maternal factors and neonatal outcome between SGA group and non-SGA group. Between severe SGA group and mild SGA group, there were statistically significant differences in gestational age (33.7 ± 4.4 weeks vs. 36.5 ± 1.1 weeks, p < 0.001), discordancy rate (37.4 [7.2e58.1] % vs. 14.9 [1.1e36.6] %, p < 0.001). In all SGA group, there were no significant differences in the rate of velamentous or marginal insertion of the umbilical cord in smaller and larger groups. The rate of not intact survival in severe SGA was statistically higher (p < 0.01) than in mild SGA. Conclusion: Severe SGA group is delivered earlier than mild SGA group and intact survival rate is lower in small gestational age twin of severe SGA group. Almost all cases of severe SGA group are delivered earlier because of fetal dysfunction. In smaller twin of severe SGA group, immaturity and fetal growth restriction may affect their poor prognosis.
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P2.19. GROWTH RESTRICTION ALTERS EXPRESSION OF PLACENTAL LEPTIN SIGNALLING TARGETS IN A SEX-SPECIFIC MANNER Deanne Hryciw 1, Jessica Briffa 1, Lisa Jedwab 1, Tania Romano 2, Mary Wlodek 1. 1 Department of Physiology, University of Melbourne, Parkville, Australia; 2 Department of Human Biosciences, LaTrobe University, Bundoora, Australia In the Western world, uteroplacental insufficiency is the leading cause of growth restriction. Consequently, being born small alters development in the offspring, which increases the risk of developing disease in adulthood. The adipokine leptin regulates development and is transferred from the maternal circulation to the offspring via the placenta. The aim of our study was to investigate if growth restriction due to uteroplacental insufficiency alters specific targets involved in leptin transport and signalling in the placenta and whether there are sex specific effects. Female WKY rats were mated and at day 18 of pregnancy (E18, term ¼ E22) underwent sham surgery or bilateral ligation of the uterine vessels to induce uteroplacental insufficiency. At E20, offspring and maternal blood was collected, offspring were sacrificed and the placenta was separated into the basalis and labyrinth regions. RNA was then extracted, reverse transcribed into cDNA and key signalling targets in the leptin pathways were analysed by a RT2 profiler PCR-array (Qiagen). Growth restriction reduced offspring body weight, and resulted in a reduction in plasma leptin concentrations, despite no changes in maternal and amniotic fluid leptin concentrations. In the basalis, growth restriction significantly reduced SOCS3 and significantly increased AMPKa. There were sex specific differences in the expression of STAT5a, Akt3, PI3K and AMPKb. Megalin, a leptin transport protein, was significantly increased in the Restricted males but significantly decreased in the Restricted females. In the labyrinth, growth restriction significantly increased JAK2, STAT5a, STAT3, megalin and AMPKa but significantly decreased PI3K. There was a sex specific difference in the expression of megalin. Thus we have demonstrated that growth restriction induced by uteroplacental insufficiency alters key targets in the leptin signalling pathway in the placenta, which may provide a mechanism for the altered placental growth and development in the growth restricted offspring.
P2.20. MATERNAL DIETARY NITRATE SUPPLEMENTATION ALTERS FETOPLACENTAL VASCULAR FUNCTION IN THE ENDOTHELIAL NITRIC OXIDE SYNTHASE KNOCKOUT (ENOS¡/¡) MOUSE Elizabeth Cottrell 1, Mark Wareing 1, Elizabeth Cowley 1, Sarah FinnSell 1, Susan Greenwood 1, Phillip Baker 2, Colin Sibley 1. 1 Maternal and Fetal Health Research Centre; University of Manchester, Manchester, UK; 2 Liggins Institute; University of Auckland, Auckland, New Zealand Objectives: In non-pregnant individuals, supplementation with dietary nitrate, providing nitrite, a source of nitric oxide-generating potential, improves blood flow and cardiovascular function. Previously, we have shown that maternal dietary nitrate supplementation, via beetroot juice, increased fetal growth and improved ex vivo uterine artery function in a model of fetal growth restriction, the eNOS-/- mouse. Here, we aimed to assess whether this maternal treatment alters umbilical or fetal artery function. Methods: Pregnant C57Bl/6J (wildtype; WT) and eNOS/ mice received drinking water supplemented with beetroot juice from embryonic day (E) 12.5-E18.5. Treatments were a nitrate-replete beetroot juice (delivering ~1 mmol/kg/day nitrate) or a nitrate-depleted control juice, at equivalent dose. At E18.5, umbilical arteries (UmbA) and fetal aortas (FA) were isolated and vascular function assessed by wire myography. Results: In eNOS-/- mice, both UmbA and FA vessel diameters were significantly smaller (by ~25%; p < 0.01) and exhibited an increased maximal constriction to the thromboxane mimetic U46619 compared with WT vessels. Significant endothelial-dependent vasodilation (to acetylcholine; ACH) was observed in pre-constricted FA of WT but not eNOS-/mice. In pre-constricted UmbA, ACH induced a further constriction, with this response being greater in eNOS-/- compared with WT UmbA (p<0.05).
Background: Obesity during pregnancy is associated with a proinflammatory state affecting the placenta's role in releasing molecules (e.g. exosomes) that can directly affect the maternal metabolism and fetal development. The aim of this study was to test the hypothesis that placenta-derived exosomes from obese women induce higher TNF-a releases from endothelial cells. Methods: Women were recruited, with informed, written consent, by research midwives from the Ochsner Baptist Medical Center (New Orleans, USA). Groups were classified according to the body max index (BMI) in two categories: lean (n ¼ 6, BMI 18.5e24.9 Kg/m2) and obese (n ¼ 6, BMI >30 Kg/m2). Exosomes were isolated from plasma by differential and buoyant density centrifugation. The total number of exosome vesicles and specific placenta-derived exosomes were determined by quantifying immunoreactive exosomal CD63 and PLAP using ELISA kit and nanoparticle tracking analysis (NanoSight NS500). The effect of exosomes on TNF-a cytokine release from endothelial cells was established using protein solution array analysis (Bioplex). Results: Exosome plasma concentration was 2-fold greater in obese than lean women (p < 0.05). In obese women, the number of exosomes present in maternal plasma (9.34 108 ± 2.0 108 particles per ml plasma) was significantly higher compared with lean women (4.14 108 ± 1.3 108 particles per ml plasma). The concentration of placenta-derived exosomes in maternal plasma (as indicated by exosomal PLAP) is higher in obese compared to lean women (1020 ± 75 pg/ml and 634 ± 56 pg/ml, respectively). Exosomes isolated from lean and obese women increased TNF-a releases from endothelial cell by 1.4 ± 0.1 and 2.6 ± 0.1-fold, respectively, compared to the control in the absence of exosomes. Conclusions: Proinflammatory state is associated with obesity and leads to the releases of proinflammatory cytokines into the circulation. Placental exosomes could contribute to this phenomenon by promoting cytokines releases from endothelial cells.
P2.18. SEVERE DISCORDANCY IN DICHORIONIC DIAMNIOTIC TWIN POTENTIALLY SHOWS PRETERM LABOR AND POOR PERINATAL OUTCOME Michihisa Shiro, Naoyuki Iwahashi, Tomoko Noguchi, Tamaki Yahata, Madoka Yamamoto, Yuko Tanizaki, Aya Kobayashi, Nami Ota, Shigetaka Yagi, Sawako Minami, Kazuhiko Ino. Wakayama Medical University, Wakayama, Japan Objectives: In dichorionic diamniotic (DD) twin, there is generically no placental anastomoses and each fetus has a own placenta. In the same environment of uterus, sometimes discordant twin occurs in DD twin. We investigated maternal factors and neonatal outcome of small for gestational age (SGA)at least one twin in DD twin pregnancy. Methods: This was a retrospective study of 113 patients who delivered DD-Twin, between January 2006 and December 2014 in our hospital. Small for gestational age (SGA) group was defined as a weight below the 10th percentile for the gestational age at least one baby. Especially, severe SGA group was defined as a weight below -2.0SD (standard deviation), Mild SGA group was defined as excepting severe SGA in all SGA cases. Results: There was no significant difference in maternal factors and neonatal outcome between SGA group and non-SGA group. Between severe SGA group and mild SGA group, there were statistically significant differences in gestational age (33.7 ± 4.4 weeks vs. 36.5 ± 1.1 weeks, p < 0.001), discordancy rate (37.4 [7.2e58.1] % vs. 14.9 [1.1e36.6] %, p < 0.001). In all SGA group, there were no significant differences in the rate of velamentous or marginal insertion of the umbilical cord in smaller and larger groups. The rate of not intact survival in severe SGA was statistically higher (p < 0.01) than in mild SGA. Conclusion: Severe SGA group is delivered earlier than mild SGA group and intact survival rate is lower in small gestational age twin of severe SGA group. Almost all cases of severe SGA group are delivered earlier because of fetal dysfunction. In smaller twin of severe SGA group, immaturity and fetal growth restriction may affect their poor prognosis.
A43
P2.19. GROWTH RESTRICTION ALTERS EXPRESSION OF PLACENTAL LEPTIN SIGNALLING TARGETS IN A SEX-SPECIFIC MANNER Deanne Hryciw 1, Jessica Briffa 1, Lisa Jedwab 1, Tania Romano 2, Mary Wlodek 1. 1 Department of Physiology, University of Melbourne, Parkville, Australia; 2 Department of Human Biosciences, LaTrobe University, Bundoora, Australia In the Western world, uteroplacental insufficiency is the leading cause of growth restriction. Consequently, being born small alters development in the offspring, which increases the risk of developing disease in adulthood. The adipokine leptin regulates development and is transferred from the maternal circulation to the offspring via the placenta. The aim of our study was to investigate if growth restriction due to uteroplacental insufficiency alters specific targets involved in leptin transport and signalling in the placenta and whether there are sex specific effects. Female WKY rats were mated and at day 18 of pregnancy (E18, term ¼ E22) underwent sham surgery or bilateral ligation of the uterine vessels to induce uteroplacental insufficiency. At E20, offspring and maternal blood was collected, offspring were sacrificed and the placenta was separated into the basalis and labyrinth regions. RNA was then extracted, reverse transcribed into cDNA and key signalling targets in the leptin pathways were analysed by a RT2 profiler PCR-array (Qiagen). Growth restriction reduced offspring body weight, and resulted in a reduction in plasma leptin concentrations, despite no changes in maternal and amniotic fluid leptin concentrations. In the basalis, growth restriction significantly reduced SOCS3 and significantly increased AMPKa. There were sex specific differences in the expression of STAT5a, Akt3, PI3K and AMPKb. Megalin, a leptin transport protein, was significantly increased in the Restricted males but significantly decreased in the Restricted females. In the labyrinth, growth restriction significantly increased JAK2, STAT5a, STAT3, megalin and AMPKa but significantly decreased PI3K. There was a sex specific difference in the expression of megalin. Thus we have demonstrated that growth restriction induced by uteroplacental insufficiency alters key targets in the leptin signalling pathway in the placenta, which may provide a mechanism for the altered placental growth and development in the growth restricted offspring.
P2.20. MATERNAL DIETARY NITRATE SUPPLEMENTATION ALTERS FETOPLACENTAL VASCULAR FUNCTION IN THE ENDOTHELIAL NITRIC OXIDE SYNTHASE KNOCKOUT (ENOS¡/¡) MOUSE Elizabeth Cottrell 1, Mark Wareing 1, Elizabeth Cowley 1, Sarah FinnSell 1, Susan Greenwood 1, Phillip Baker 2, Colin Sibley 1. 1 Maternal and Fetal Health Research Centre; University of Manchester, Manchester, UK; 2 Liggins Institute; University of Auckland, Auckland, New Zealand Objectives: In non-pregnant individuals, supplementation with dietary nitrate, providing nitrite, a source of nitric oxide-generating potential, improves blood flow and cardiovascular function. Previously, we have shown that maternal dietary nitrate supplementation, via beetroot juice, increased fetal growth and improved ex vivo uterine artery function in a model of fetal growth restriction, the eNOS-/- mouse. Here, we aimed to assess whether this maternal treatment alters umbilical or fetal artery function. Methods: Pregnant C57Bl/6J (wildtype; WT) and eNOS/ mice received drinking water supplemented with beetroot juice from embryonic day (E) 12.5-E18.5. Treatments were a nitrate-replete beetroot juice (delivering ~1 mmol/kg/day nitrate) or a nitrate-depleted control juice, at equivalent dose. At E18.5, umbilical arteries (UmbA) and fetal aortas (FA) were isolated and vascular function assessed by wire myography. Results: In eNOS-/- mice, both UmbA and FA vessel diameters were significantly smaller (by ~25%; p < 0.01) and exhibited an increased maximal constriction to the thromboxane mimetic U46619 compared with WT vessels. Significant endothelial-dependent vasodilation (to acetylcholine; ACH) was observed in pre-constricted FA of WT but not eNOS-/mice. In pre-constricted UmbA, ACH induced a further constriction, with this response being greater in eNOS-/- compared with WT UmbA (p<0.05).