239 results on '"Chorion enzymology"'
Search Results
2. Reduced Expression of Hydrogen Sulfide-Generating Enzymes Down-Regulates 15-Hydroxyprostaglandin Dehydrogenase in Chorion during Term and Preterm Labor.
- Author
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Sun Q, Chen Z, He P, Li Y, Ding X, Huang Y, Gu H, and Ni X
- Subjects
- Cystathionine gamma-Lyase genetics, Down-Regulation, Female, Humans, Hydrogen Sulfide metabolism, Hydroxyprostaglandin Dehydrogenases genetics, Obstetric Labor, Premature genetics, Pregnancy, Term Birth genetics, Chorion enzymology, Cystathionine beta-Synthase metabolism, Cystathionine gamma-Lyase metabolism, Hydroxyprostaglandin Dehydrogenases metabolism, Obstetric Labor, Premature metabolism, Term Birth metabolism
- Abstract
Chorionic NAD-dependent 15-hydroxyprostaglandin dehydrogenase (PGDH) plays a pivotal role in controlling the amount of prostaglandins in the uterus and has been implicated in the process of labor. Prior studies identified hydrogen sulfide-generating enzymes cystathionine-β-synthetase (CBS) and cystathionine-γ-lyase (CSE) in fetal membranes. We investigated whether hydrogen sulfide is involved in the regulation of PGDH expression in the chorion during labor. The chorionic tissues were obtained from pregnant women at preterm in labor and at term in labor or not in labor at term. Levels of CSE and CBS and hydrogen sulfide production rate were down-regulated in term in labor and preterm in labor groups compared with not in labor at term group. The CBS level correlated to PGDH expression in the chorion. Hydrogen sulfide donor NaHS and precursor l-cysteine dose-dependently stimulated PGDH expression and activity in cultured chorionic trophoblasts. The effect of l-cysteine was blocked by CBS inhibitor and CBS siRNA but not by CSE inhibitor and CSE siRNA. Hydrogen sulfide treatment suppressed miR-26b and miR-199a expression in chorionic trophoblasts. miR-26b and miR-199a mimics blocked hydrogen sulfide upregulation of PGDH expression. Our results indicate that hydrogen sulfide plays pivotal roles in maintenance of PGDH expression in the chorion during human pregnancy. Reduced expression of hydrogen sulfide-generating enzymes contributes to an increased amount of prostaglandins in the uterus during labor., (Copyright © 2018 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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3. Expression of 15-Hydroxyprostaglandin Dehydrogenase in Human Chorion Is Associated with Peroxisome Proliferator-Activated Receptor Isoform Expression in Term Labor.
- Author
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He P, Li Y, Ding X, Sun Q, Huang Y, Gu H, and Ni X
- Subjects
- Adult, Cells, Cultured, Chorion enzymology, Female, Gene Expression Regulation, Enzymologic, Humans, Hydroxyprostaglandin Dehydrogenases genetics, Infant, Newborn, Male, PPAR alpha genetics, Peroxisome Proliferator-Activated Receptors genetics, Pregnancy, Premature Birth, Prostaglandins metabolism, Protein Isoforms, RNA, Messenger genetics, Term Birth, Trophoblasts metabolism, Chorion metabolism, Hydroxyprostaglandin Dehydrogenases metabolism, PPAR alpha metabolism, Peroxisome Proliferator-Activated Receptors metabolism
- Abstract
Chorionic NAD-dependent 15-hydroxy prostaglandin dehydrogenase (PGDH) plays a pivotal role in controlling the amount of prostaglandins in the uterus. Peroxisome proliferator-activated receptors (PPARs) are implicated to be involved in parturition. In this study, we investigated whether PPARs are involved in control of PGDH expression in chorion. The chorionic tissues were collected from the following groups of the women with singleton pregnancy: term no labor (TNL), term labor (TL) and preterm labor (PTL). Chorionic trophoblasts were isolated and cultured in vitro. Immunocytochemistry analysis showed that PPARα, PPARβ, and PPARγ were localized to trophoblasts in chorion. The protein levels of PGDH, PPARβ, and PPARγ were localized to trophoblasts in chorion. The protein levels of PPARα, PPARβ, and PPARγ were reduced in TL tissues compared to that of TNL group. PPARα, PPARβ, and PPARγ expression correlated to PGDH in TNL tissues, whereas only PPARγ expression correlated to PGDH in TL chorion tissues. PGDH expression was decreased in PTL tissues compared with TL group, whereas the expression of PPARs was not significantly different between TL and PTL groups. The agonists of three PPARs dose-dependently stimulated PGDH activity, mRNA, and protein expression in cultured chorionic cells. PPARs did not affect the stability of PGDH mRNA but stimulated the transcriptional activity of HPGD gene. Our results suggest that PPARs play pivotal roles in maintenance of PGDH expression in chorion during human pregnancy., (Copyright © 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2015
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4. The Effect of Progestins on Tumor Necrosis Factor α-Induced Matrix Metalloproteinase-9 Activity and Gene Expression in Human Primary Amnion and Chorion Cells In Vitro.
- Author
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Allen TK, Feng L, Nazzal M, Grotegut CA, Buhimschi IA, and Murtha AP
- Subjects
- 17 alpha-Hydroxyprogesterone Caproate, Amnion cytology, Amnion enzymology, Cells, Cultured, Chorion cytology, Chorion enzymology, Female, Gene Expression Regulation, Enzymologic drug effects, Humans, Matrix Metalloproteinase 9 genetics, Pregnancy, RNA, Messenger biosynthesis, Amnion drug effects, Chorion drug effects, Hydroxyprogesterones pharmacology, Matrix Metalloproteinase 9 metabolism, Medroxyprogesterone Acetate pharmacology, Progesterone pharmacology, Progestins pharmacology, Tumor Necrosis Factor-alpha pharmacology
- Abstract
Background: Current treatment modalities for preventing preterm premature rupture of membranes are limited, but progestins may play a role. Tumor necrosis factor α (TNFα) enhances matrix metalloproteinase-9 (MMP-9) gene expression and activity in fetal membranes, contributing to membrane weakening and rupture. We previously demonstrated that progestins attenuate TNFα-induced MMP-9 activity in a cytotrophoblast cell line. However, whether they have a similar effect in primary amnion and chorion cells of fetal membranes is unknown. In this study, we evaluated the effect of progestins on basal and TNFα-induced MMP-9 activity and gene expression in primary chorion and amnion cells harvested from the fetal membranes of term nonlaboring patients., Methods: Primary amnion and chorion cells were isolated from fetal membranes obtained from term uncomplicated nonlaboring patients following elective cesarean delivery (n = 11). Confluent primary amnion and chorion cell cultures were both pretreated with vehicle (control), progesterone (P4), 17α-hydroxyprogesterone caproate (17P), or medroxyprogesterone acetate (MPA) at 10 M concentration for 6 hours followed by stimulation with TNFα at 10 ng/mL for an additional 24 hours. Cell cultures pretreated with the vehicle only served as the unstimulated control and the vehicle stimulated with TNFα served as the stimulated control. Both controls were assigned a value of 100 units. Cell culture medium was harvested for MMP-9 enzymatic activity quantification using gelatin zymography. Total RNA was extracted for quantifying MMP-9 gene expression using real-time quantitative PCR. Basal MMP-9 activity and gene expression data were normalized to the unstimulated control. TNFα-stimulated MMP-9 activity and gene expression were normalized to the stimulated control. The primary outcome was the effect of progestins on TNFα-induced MMP-9 enzymatic activity in term human primary amnion and chorion cells in vitro. Secondary outcomes included the effect of progestin therapy on TNFα-induced MMP-9 gene expression and on basal MMP-9 activity and gene expression in primary amnion and chorion cells in vitro., Results: Primary cells were harvested from 11 patients. Compared with the unstimulated control, TNFα increased MMP-9 activity (P = 0.005 versus control in primary amnion cells and P < 0.001 versus control in primary chorion cells) and MMP-9 gene expression (P = 0.030 versus control in primary amnion cells, P < 0.001 versus control in primary chorion cells). Compared with the unstimulated controls, MPA, but not P4 or 17P, reduced basal MMP-9 activity [mean difference (95% CI) -49.6 (-81.9, -17.3) units, P = 0.001] and gene expression [mean difference (95% CI) -53.4 (-105.9, -0.9) units, P = 0.045] in primary amnion cells. Compared with the stimulated control, MPA also reduced TNFα-induced MMP-9 activity [mean difference (95% CI) -69.0 (-91.8, -46.3) units, P < 0.001] and gene expression [mean difference (95% CI) -86.0 (-120.7, -51.3) units, P < 0.001] in primary amnion cells. Progestin pretreatment had no significant effect on basal or TNFα-induced MMP-9 activity and gene expression in primary chorion cells., Conclusions: The inhibitory effect of MPA on both basal and TNFα-induced MMP-9 activity and gene expression in primary amnion cells demonstrate a possible mechanism by which progestins may prevent fetal membrane weakening leading to preterm premature rupture of membranes.
- Published
- 2015
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5. Reduced expression of 15-hydroxy prostaglandin dehydrogenase in chorion during labor is associated with decreased PRB and increased PRA and GR expression.
- Author
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Li Y, He P, Sun Q, Liu J, Gao L, You X, Gu H, and Ni X
- Subjects
- Cells, Cultured, Chorion cytology, Chorion drug effects, Dexamethasone pharmacology, Dihydrotestosterone analogs & derivatives, Dihydrotestosterone pharmacology, Extraembryonic Membranes drug effects, Extraembryonic Membranes metabolism, Female, Fluorescent Antibody Technique, Glucocorticoids pharmacology, Humans, Hydroxyprostaglandin Dehydrogenases genetics, Labor, Obstetric drug effects, Pregnancy, Progesterone pharmacology, Proto-Oncogene Proteins c-jun metabolism, Transcription, Genetic drug effects, Trophoblasts cytology, Trophoblasts drug effects, Trophoblasts metabolism, Up-Regulation drug effects, Chorion enzymology, Hydroxyprostaglandin Dehydrogenases metabolism, Labor, Obstetric metabolism, Receptors, Glucocorticoid metabolism, Receptors, Progesterone metabolism
- Abstract
The chorion laeve controls the levels of active prostaglandins within the uterus by NAD-dependent 15-hydroxy prostaglandin dehydrogenase (PGDH). The expression of PGDH in chorion is modulated by glucocorticoids and progesterone. In this study, we investigated glucocorticoid receptor (GR) and progesterone receptor A and B (PRA and PRB) in the regulation of PGDH expression in chorion, and we determined whether reduced PGDH expression in chorion during labor is associated with the changes in GR and PR expression by real-time RT-PCR and Western blot analysis. Dexamethasone (DEX) inhibited PGDH expression whereas progesterone stimulated PGDH expression in chorionic trophoblasts. DEX suppressed PGDH expression in GR overexpression and PR knockdown cells. The inhibitory effect of DEX did not occur in GR knockdown cells. Progesterone inhibited PGDH in GR overexpression and PR knockdown cells and it stimulated PGDH in PRB overexpression cells whereas it suppressed PGDH in PRA overexpression cells. Knockdown of c-Jun resulted in a loss of progesterone- and DEX-induced effects. PGDH was down-regulated in chorion tissues during labor. PRB was decreased whereas PRA and GR were increased in chorion during labor. Glucocorticoids inhibit PGDH expression via GR in chorionic trophoblasts. Progesterone enhances PGDH expression through PRB, whereas it inhibits PGDH expression via GR and PRA. Decreased PGDH expression is associated with increased GR and PRA, although decreased PRB, in chorion during labor., (Copyright © 2013 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.)
- Published
- 2013
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6. Chronic glucocorticoid exposure potentiates placental chorionic plate artery constriction: implications for aberrant fetoplacental vascular resistance in fetal growth restriction.
- Author
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Nugent JL, Wareing M, Palin V, Sibley CP, Baker PN, Ray DW, Farrow SN, and Jones RL
- Subjects
- 11-beta-Hydroxysteroid Dehydrogenases metabolism, 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid pharmacology, Arteries drug effects, Carbenoxolone pharmacology, Chorion blood supply, Chorion enzymology, Dexamethasone pharmacology, Female, Humans, Hydrocortisone pharmacology, Mifepristone pharmacology, Placental Circulation drug effects, Pregnancy, Vasodilation drug effects, Fetal Growth Retardation physiopathology, Glucocorticoids pharmacology, Placenta blood supply, Vasoconstriction drug effects
- Abstract
Fetal growth restriction (FGR) is a serious pregnancy complication, resulting in significant perinatal morbidity and mortality. Increased vascular resistance in the fetoplacental circulation is a hallmark of FGR and is associated with enhanced vasoconstriction of the resistance arteries in the placenta, the chorionic plate arteries (CPAs). Although the cause is unknown, FGR is associated with excess exposure to glucocorticoids (GCs), key mediators of vascular resistance in the systemic circulation. We hypothesized that GCs alter CPA reactivity, thereby contributing to the altered blood flow dynamics seen in FGR. We aimed to examine the acute and chronic effects of GCs on CPA reactivity and the operational mechanisms. Glucocorticoid receptors were highly expressed by CPA. 11β-Hydroxysteroid isoenzyme type 2 was detected within the endothelium, whereas 11β-hydroxysteroid isoenzyme type 1 was absent. Acute GC treatment significantly attenuated U46619-induced constriction. This effect was reversed by cotreatment with mifepristone or an endothelial NOS inhibitor. In contrast, chronic GC treatment potentiated U46619 constriction in a dose-dependent manner, which was partially abolished by mifepristone cotreatment. Similar effects were observed using a novel nonsteroidal glucocorticoid receptor-specific agonist. Chronic treatment with GCs altered the expression of several vasoactive factors, including thromboxane and bradykinin receptors, prokineticin-1, cyclooxygenase-2, and endothelial NOS. In summary, acute and chronic GC treatment exerts contrasting effects on CPA vasoreactivity. These opposing effects are consistent with temporal actions in other vascular beds and reflect activation of distinct nongenomic and genomic pathways. Chronic exposure to elevated GCs may contribute to the raised vascular resistance observed in the fetoplacental circulation in FGR.
- Published
- 2013
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7. 15-Hydroxyprostaglandin dehydrogenase expression and localization in guinea pig gestational tissues during late pregnancy and parturition.
- Author
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Welsh T, Paul J, Palliser HK, Tabatabaee H, Hirst J, Mesiano S, and Zakar T
- Subjects
- Animals, Chorion cytology, Chorion enzymology, Female, Guinea Pigs, Humans, Labor Onset metabolism, Mesoderm cytology, Mesoderm enzymology, Models, Animal, Placenta cytology, Placenta enzymology, Pregnancy, Uterus cytology, Yolk Sac cytology, Yolk Sac enzymology, Hydroxyprostaglandin Dehydrogenases biosynthesis, Parturition metabolism, Uterus metabolism
- Abstract
Prostaglandins are key components of the parturition cascade; however, the mechanisms that regulate prostaglandin concentrations in the uterus during pregnancy are largely unknown. The purpose of this study was to determine the intrauterine expression of the chief prostaglandin-inactivating enzyme, 15-hydroxyprostaglandin dehydrogenase (PGDH), during gestation and labor in the guinea pig, an animal model in which the endocrine control of pregnancy and parturition is analogous to that of women. PGDH messenger RNA (mRNA) abundance decreased significantly in the visceral yolk sac membrane (VYS, the anatomical equivalent of the human chorion laeve) and the amnion throughout the last third of pregnancy. PGDH protein was robustly expressed in the VYS epithelium and mesoderm, correlated strongly with PGDH mRNA levels and exhibited a nadir at term prior to labor onset. PGDH protein was not detected in the amnion. PGDH mRNA and protein levels in the placenta and myoendometrium were variable throughout late gestation. In the placenta, PGDH protein was concentrated in the parietal yolk sac membrane (PYS) lining the placental surface and in placental blood vessels. We observed strong expression of PGDH protein in the endometrial epithelium with comparably little expression in the myometrium. These data indicate that metabolic inactivation of prostaglandins in the pregnant guinea pig uterus takes place in the VYS, PYS, and endometrium. Decreased PGDH expression in the fetal membranes may contribute to the increase in intrauterine prostaglandin concentrations at term, stimulating the onset of labor.
- Published
- 2012
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8. Molecular evidence of a (pro)renin/ (pro)renin receptor system in human intrauterine tissues in pregnancy and its association with PGHS-2.
- Author
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Pringle KG, Zakar T, Yates D, Mitchell CM, Hirst JJ, and Lumbers ER
- Subjects
- Amnion drug effects, Amnion enzymology, Chorion drug effects, Chorion enzymology, Cyclooxygenase 2 genetics, Female, Humans, Labor, Obstetric metabolism, Placenta cytology, Placenta enzymology, Pregnancy, Protein Transport, RNA, Messenger genetics, RNA, Messenger metabolism, Receptors, Cell Surface genetics, Renin genetics, Renin pharmacology, Uterus cytology, Uterus enzymology, Prorenin Receptor, Cyclooxygenase 2 metabolism, Receptors, Cell Surface metabolism, Renin metabolism, Uterus metabolism
- Abstract
Prorenin stimulates decidual prostaglandin (PG) production in vitro, the (pro)renin receptor ((P)RR) may mediate this action. The role of prorenin in amnion PG synthesis has not been examined, despite this being the key site of PG synthesis. To determine if (P)RR, prorenin and PGHS-2 are co-localized in gestational tissues and if expression is altered by labour, term amnion, chorion, decidua and placenta were collected during elective caesarean section or after spontaneous labour. Prorenin, (P)RR and PGHS-2 mRNA abundance was determined by real-time RT-PCR. (P)RR protein was examined by immunohistochemistry. The effect of recombinant human (rh) prorenin on PGHS-2 mRNA abundance in amnion explants was determined. Prorenin and (P)RR mRNA were highest in decidua and placenta, respectively. Decidual prorenin, (P)RR and placental (P)RR mRNA abundance decreased with labour. (P)RR protein was present in all gestational tissues. After labour, decidual prorenin was positively correlated with amnion PGHS-2 mRNA and rh-prorenin significantly increased PGHS-2 mRNA abundance in amnion explants. We conclude that the decidua is the principal source of prorenin and is downregulated with labour. All gestational tissues are targets for prorenin. Decidual prorenin may be involved in the labour-associated increase in amnion PGHS-2 abundance via the (P)RR.
- Published
- 2011
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9. Vascular endothelial expression of indoleamine 2,3-dioxygenase 1 forms a positive gradient towards the feto-maternal interface.
- Author
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Blaschitz A, Gauster M, Fuchs D, Lang I, Maschke P, Ulrich D, Karpf E, Takikawa O, Schimek MG, Dohr G, and Sedlmayr P
- Subjects
- Cell Separation, Chorion cytology, Chorion enzymology, Decidua cytology, Decidua enzymology, Endothelial Cells cytology, Endothelial Cells enzymology, Endothelium, Vascular cytology, Epitopes immunology, Female, Gene Expression Regulation, Enzymologic, HLA-DR Antigens, Humans, Immunohistochemistry, Indoleamine-Pyrrole 2,3,-Dioxygenase genetics, Paraffin Embedding, Pregnancy, Pregnancy Trimester, First metabolism, Protein Transport, RNA, Messenger genetics, RNA, Messenger metabolism, Tryptophan metabolism, Endothelium, Vascular enzymology, Indoleamine-Pyrrole 2,3,-Dioxygenase metabolism, Maternal-Fetal Exchange
- Abstract
We describe the distribution of indoleamine 2,3-dioxygenase 1 (IDO1) in vascular endothelium of human first-trimester and term placenta. Expression of IDO1 protein on the fetal side of the interface extended from almost exclusively sub-trophoblastic capillaries in first-trimester placenta to a nearly general presence on villous vascular endothelia at term, including also most bigger vessels such as villous arteries and veins of stem villi and vessels of the chorionic plate. Umbilical cord vessels were generally negative for IDO1 protein. In the fetal part of the placenta positivity for IDO1 was restricted to vascular endothelium, which did not co-express HLA-DR. This finding paralleled detectability of IDO1 mRNA in first trimester and term tissue and a high increase in the kynurenine to tryptophan ratio in chorionic villous tissue from first trimester to term placenta. Endothelial cells isolated from the chorionic plate of term placenta expressed IDO1 mRNA in contrast to endothelial cells originating from human umbilical vein, iliac vein or aorta. In first trimester decidua we found endothelium of arteries rather than veins expressing IDO1, which was complementory to expression of HLA-DR. An estimation of IDO activity on the basis of the ratio of kynurenine and tryptophan in blood taken from vessels of the chorionic plate of term placenta indicated far higher values than those found in the peripheral blood of adults. Thus, a gradient of vascular endothelial IDO1 expression is present at both sides of the feto-maternal interface.
- Published
- 2011
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10. Differences in expression and activity of 11beta-hydroxysteroid dehydrogenase type 1 and 2 in human placentas of term pregnancies according to birth weight and gender.
- Author
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Mericq V, Medina P, Kakarieka E, Márquez L, Johnson MC, and Iñiguez G
- Subjects
- 11-beta-Hydroxysteroid Dehydrogenase Type 1 metabolism, 11-beta-Hydroxysteroid Dehydrogenase Type 2 metabolism, Chorion enzymology, Chorion metabolism, Female, Fetal Growth Retardation enzymology, Fetal Growth Retardation genetics, Fetal Growth Retardation metabolism, Gene Expression Regulation, Enzymologic, Gestational Age, Humans, Infant, Newborn, Infant, Small for Gestational Age metabolism, Male, Pregnancy, Pregnancy Trimester, Third metabolism, Sex Factors, Term Birth metabolism, 11-beta-Hydroxysteroid Dehydrogenase Type 1 genetics, 11-beta-Hydroxysteroid Dehydrogenase Type 2 genetics, Birth Weight genetics, Placenta metabolism, Pregnancy Trimester, Third genetics, Term Birth genetics
- Abstract
Background: Fetal exposure to maternal glucocorticoids may determine fetal growth and the programing of later disorders. Availability of the glucocorticoids in the placenta is regulated by the 11beta-hydroxysteroid dehydrogenase (11beta-HSDs) enzymes. To date, there are discrepancies with regard to cortisol (F) cord blood levels in fetuses with intrauterine growth retardation in different species. Objective To study the expression and activity of 11beta-HSDs in placentas from full term small for gestational age (SGA), appropriate for gestational age (AGA) and large for gestational age (LGA) newborns, and cortisol cord blood concentration., Methods: Twenty-five placentas from AGA, 24 SGA and 25 LGA were collected., Results: SGA newborns had significantly lower and LGA newborns had significantly higher birth weight, birth length, head circumference, and placental weight than AGA counterparts. We observed a direct correlation between placental weight and birth weight, birth length and head circumference, and higher cord F levels in SGA newborns. The 11beta-HSD1 expression was similar among the SGA, AGA, and LGA placentas. However, within the placentas of SGA newborns, the 11beta-HSD1 mRNA levels were significantly reduced in the chorionic plate compared with basal plate. An inverse correlation between cord F levels and activity of 11beta-HSD1 in the chorionic plate of the SGA placentas was detected. The 11beta-HSD2 activity was seven- to eightfold higher compared with 11beta-HSD1 in the placentas, and there was a lower 11beta-HSD2 activity in females' SGA placentas compared with the male SGA placentas., Conclusion: We observed a lower expression and activity of 11beta-HSD1 in the chorionic plate of the SGA placentas, suggesting a possible compensatory mechanism to diminish the higher cortisol fetal concentrations observed in fetuses with intrauterine growth restriction.
- Published
- 2009
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11. Direct analysis reveals an absence of gamma-carboxyglutamic acid in cancer procoagulant from human tissues.
- Author
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Kaplinska K and Mielicki WP
- Subjects
- 1-Carboxyglutamic Acid immunology, Antibodies, Monoclonal immunology, Anticoagulants pharmacology, Cell Line, Tumor enzymology, Cysteine Endopeptidases pharmacology, Enzyme Activation drug effects, Factor X drug effects, Female, Humans, Melanoma pathology, Neoplasm Proteins pharmacology, Pregnancy, Warfarin pharmacology, 1-Carboxyglutamic Acid analysis, Amnion enzymology, Chorion enzymology, Cysteine Endopeptidases chemistry, Melanoma enzymology, Neoplasm Proteins chemistry
- Abstract
Additional carboxylation of glutamic acid by vitamin K-dependent gamma-carboxylase is a common posttranslational modification of many proteins, including some of blood clotting factors. Vitamin K-antagonists, such as warfarin, are often included in the therapy of malignant disease, decreasing the blood coagulation potential. Cancer procoagulant, a direct blood coagulation factor X activator from malignant tissue, is considered as a vitamin K-dependent protein, so it could serve as one of possible targets for the therapy with warfarin. However, there is still no experimental data demonstrating directly the presence of gamma-carboxyglutamic acid (Gla) in a cancer procoagulant molecule. The presence of Gla in cancer procoagulant isolated from human amnion-chorion membranes and from human malignant melanoma WM 115 cell line was analyzed directly, using specific anti-Gla monoclonal antibodies. There was no detectable amount of Gla in cancer procoagulant isolated from fetal or malignant tissue. Cancer procoagulant from human tissues does not contain Gla-rich domain. The finding indicates that cancer procoagulant is rather a poor target for warfarin therapy of malignant disease.
- Published
- 2009
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12. Direct contribution of inducible nitric oxide synthase expression to apoptosis induction in primary smooth chorion trophoblast cells of human fetal membrane tissues.
- Author
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Yuan B, Ohyama K, Takeichi M, and Toyoda H
- Subjects
- Amnion cytology, Amnion enzymology, Cells, Cultured, Down-Regulation, Enzyme Activation, Female, Heme Oxygenase-1 biosynthesis, Heme Oxygenase-1 genetics, Humans, Immunohistochemistry, MAP Kinase Signaling System, Nitric Oxide Synthase Type II genetics, Nitric Oxide Synthase Type II metabolism, Oxidative Stress, Transfection, p38 Mitogen-Activated Protein Kinases metabolism, Apoptosis physiology, Chorion cytology, Chorion enzymology, Nitric Oxide Synthase Type II biosynthesis, Trophoblasts cytology, Trophoblasts enzymology
- Abstract
We have previously demonstrated that apoptosis induction is observed only in smooth chorion laeve trophoblast cells, and not in amnion epithelial cells of human fetal membrane tissues prepared at the term. Apoptosis induction was suppressed by the presence of an inhibitor specific for inducible nitric oxide synthase (iNOS), suggesting that intracellular oxidative stress plays a critical role in this process. In this study, we transfected the iNOS gene into primary cultured chorion and amnion cells to examine the direct contribution of iNOS gene expression to the apoptosis induction in these cells. We identified a significant increase in the levels of iNOS protein expression and nitrite accumulation in both chorion and amnion cells after the iNOS gene transfection. However, the induction of apoptosis was observed in an approximately 70% of chorion cells transfected with iNOS gene. Transfection of the iNOS gene into chorion cells resulted in the activation of p38 mitogen-activated protein kinase (MAPK) and downregulation of hemeoxygenase-1 protein expression, whereas no such events were observed in the transfected amnion cells. These results suggest that apoptosis induced in the chorion trophoblast cells by the iNOS gene expression is closely linked to a physiological consequence, such as the rupture of fetal membranes.
- Published
- 2009
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13. [Fetal membranes: embryological development, structure and the physiopathology of the preterm premature rupture of membranes].
- Author
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Pasquier JC and Doret M
- Subjects
- Amnion enzymology, Chorion enzymology, Decidua enzymology, Female, Fetal Membranes, Premature Rupture enzymology, Humans, Matrix Metalloproteinases metabolism, Pregnancy, Extraembryonic Membranes embryology, Extraembryonic Membranes enzymology, Fetal Membranes, Premature Rupture physiopathology
- Abstract
Fetal membranes development is a complex process. The amniotic and exo-celomic cavities are appearing first. The rapid growth of the amniotic cavity is leading to the disappearance of the exo-celomic cavity and the chorion is merging with the decidua. Fetal membranes consist of three layers: the amnion and the chorion, issued from fetal tissues and the decidua issued from maternal tissue. A balance between the synthesis and the degradation of membranes components is physiologic throughout the gestation. Two main mechanisms are involved in the degradation process: apoptosis in the cellular compartment and matrix metalloproteinase (MMP) in the extracellular matrix. Regulation of MMP is depending on factors increasing their expression (cytokines) and factors decreasing their activity tissue inhibitor of metalloproteinases (TIMPS). Particular conditions can induce an unbalance between synthesis and degradation leading to the weakening of the membranes. Different factors can be associated to induce this unbalance: infection, hormonal factors, default in membranes fusion, oxidative stress and mechanic factors. In fine, the spontaneous rupture of the membranes is always occurring in regard of the uterine cervix after a process started several weeks before.
- Published
- 2008
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14. Design of stable and uniform single nanoparticle photonics for in vivo dynamics imaging of nanoenvironments of zebrafish embryonic fluids.
- Author
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Nallathamby PD, Lee KJ, and Xu XH
- Subjects
- Animals, Contrast Media, Zebrafish metabolism, Body Fluids cytology, Chorion cytology, Chorion enzymology, Image Enhancement methods, Nanoparticles chemistry, Zebrafish anatomy & histology, Zebrafish embryology
- Abstract
We report here the use of a simple washing approach to reduce the ionic strength of the solution, which increased the thickness of the electric double layer on the surface of silver (Ag) nanoparticles and thereby enhanced their surface zeta-potential. This approach allowed us to prepare optically uniform (75-99%) and purified Ag nanoparticles (11.3 +/- 2.3 nm) that are stable (nonaggregation) in solution for months, permitting them to become robust and widely used single nanoprobes for in vivo optical imaging. These Ag nanoparticles show remarkable photostability and serve as single nanoparticle photonic probes for continuous imaging nanoenvironments of segmentation-stage zebrafish embryos for hours. Unlike other particle tracking experiments, we utilized size-dependent localized surface plasmon resonance spectra (LSPRS) (colors) of single Ag nanoparticles to determine given colored (sized) nanoparticles in situ and used the monodisperse color (size) of nanoparticles to simultaneously measure viscosities and flow patterns of multiple proximal nanoenvironments in segmentation-stage zebrafish embryos in real time. We found new interesting counterclockwise flow patterns with rates ranging from 0.06 to 1.8 microm/s and stunningly high viscosity gradients spanning two orders of magnitude in chorion space of the embryos, with the highest viscosity observed around the center of chorion space and the lower viscosity at the interfacial areas near the surface of both chorion layers and inner mass of the embryos. This study demonstrates the possibility of using individual monodisperse nanophotonics to probe the roles of embryonic fluid dynamics in embryonic development.
- Published
- 2008
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15. Cellular localisation of the pregnancy-associated glycoprotein family (PAGs) in the synepitheliochorial placenta of the European bison.
- Author
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Majewska M, Panasiewicz G, Szafranska B, Gizejewski Z, Majewski M, and Borkowski K
- Subjects
- Animals, Aspartic Acid Endopeptidases metabolism, Cattle, Chorion cytology, Epithelial Cells enzymology, Female, Glycoproteins metabolism, Immunohistochemistry, Maternal-Fetal Exchange physiology, Pregnancy, Tissue Distribution, Bison metabolism, Chorion enzymology, Placenta enzymology, Pregnancy Proteins metabolism
- Abstract
This paper describes the cellular immuno-localisation of the PAG family in synepitheliochorial (cotyledonary) placenta of the European bison (Eb). Uteri were harvested from pregnant wild Eb (n=4; 45-150 days post coitum-dpc); and additionally from cattle (30, 45 dpc) and pigs (42 dpc)--both domestic species were used as positive controls for cellular PAG immunodetection. Placentas were sectioned, fixed, dehydrated and subjected to double fluorescent immunohistochemistry (dF-IHC) with the use of Alexa 488 fluorochrom (A488) and propidium iodide (PI). Native positive EbPAG signals were detected by heterologous (ht; cross-species) dF-IHC with primary rabbit anti-PAG polyclonals against native or recombinant porcine PAG antigens (anti-pPAG); then visualised with secondary anti-rabbit goat immunoglobulins--conjugated to A488. Our htdF-IHC indicated an unequivocal localisation to the mono- and bi-nuclear trophectoderm (chorionic epithelium) cells expressing the PAGs (A488-green) among all placental cells, in which PI (red) stained nuclei. This is the first paper reporting the EbPAG family expression examined by htdF-IHC at the feto-maternal interface in wild Pecoran species. The cross-reactivity of anti-pPAG polyclonals with the EbPAGs suggests that shared epitopes are present in these molecules. It seems that the EbPAG family, which is robustly expressed in mono- and bi-nucleated trophectoderm cells, is associated with events taking place during placenta development. Our study also provided a proficient ht-system to identify various PAGs that could be useful as prenatal protein markers for pregnancy diagnoses, which is essential for effective reproductive management of endangered mammals.
- Published
- 2008
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16. The effect of progesterone on gelatinase expression in the decidua and fetal membranes before and after contractions.
- Author
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Goldman S, Weiss A, and Shalev E
- Subjects
- Amnion enzymology, Cells, Cultured, Chorion enzymology, Dose-Response Relationship, Drug, Female, Humans, Luciferases genetics, Matrix Metalloproteinase 2 metabolism, Matrix Metalloproteinase 9 metabolism, Matrix Metalloproteinases metabolism, Reverse Transcriptase Polymerase Chain Reaction, Transfection, Decidua enzymology, Extraembryonic Membranes enzymology, Gelatinases metabolism, Progesterone pharmacology, Progestins pharmacology, Uterine Contraction drug effects, Uterine Contraction physiology
- Abstract
Objective: This study was aimed to explore the effect of progesterone on gelatinase expression in the decidua and fetal membranes before and after contractions., Study Design: Zymography was conducted for matrix metalloproteinase (MMP) secretion. Semiquantitative reverse transcriptase-polymerase chain reaction was performed to examine MMP2 transcripts, and the effect of progesterone on MMP2 promoter activity was determined with the use of luciferase activity., Results: Progesterone decreased pro-MMP2 secretion, expression, and promoter activity in decidua before contractions began. The effect of progesterone was reversed completely by mifepristone (RU486). Progesterone failed to inhibit MMP2 expression in the amnion and chorion before contractions began. After contractions, progesterone failed to inhibit MMP2 expression in both the decidua and fetal membranes., Conclusion: MMP2 expression is inhibited by progesterone only in the decidua and only before contractions begin.
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- 2007
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17. Heparanase expression correlates with metastatic capability in human choriocarcinoma.
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Jingting C, Yangde Z, Yi Z, Huining L, Rong Y, and Yu Z
- Subjects
- Cell Line, Tumor, Cell Proliferation drug effects, Chorion enzymology, Female, Glucuronidase genetics, Humans, Oligodeoxyribonucleotides, Antisense pharmacology, Pregnancy, RNA, Messenger metabolism, Reverse Transcriptase Polymerase Chain Reaction, Choriocarcinoma enzymology, Choriocarcinoma pathology, Glucuronidase metabolism, Neoplasm Metastasis, Uterine Neoplasms enzymology, Uterine Neoplasms pathology
- Abstract
Objective: In this report, we studied the role of Hpa in metastatic capability of human choriocarcinoma. At the same time, we investigated the effect of Hpa antisense oligodeoxynucleotide (ASODN) on inhibition of invasiveness of human choriocarcinoma., Methods: The different invasion ability between JEG-3 and JAR cell lines was proved by Matrigel invasion assay in vitro. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot analyses were carried out respectively to determine Hpa gene and protein expression; the localization of this molecule was demonstrated by immunohistochemistry. Finally, Hpa antisense oligodeoxynucleotide (ASODN) was transfected into JEG-3 cells and Hpa mRNA and protein were quantified by RT-PCR and Western blot. The effect of ASODN on the metastatic capability of JEG-3 was evaluated by Matrigel invasion assay., Results: (1) We proved that the invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P<0.05). (2) We found that the Hpa gene and protein in JEG-3 and JAR cell lines were significantly higher than those in normal chorion (P<0.05). On the other hand, we detected that JEG-3 expressed much more Hpa than JAR (P<0.05). (3) Both in JEG-3 cell and in JAR cell, we found that Hpa protein express in cytoplasm. (4) After transfection of Hpa ASODN, Hpa mRNA and protein expression in JEG-3 cell decreased 4- and 5-fold. At the same time, we also observed that the invasion ability of JEG-3 cell was weakened than before (P<0.05)., Conclusion: The current study demonstrated that the expression of Hpa plays an important role in metastatic capability of human choriocarcinoma and reducing the expression of Hpa can help weaken the invasion ability of human choriocarcinoma.
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- 2007
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18. [Correlation between expression of heparanase and invasion of choriocarcinoma].
- Author
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Yu R, Zhang Y, and Cai JT
- Subjects
- Blotting, Western, Cell Line, Tumor, Choriocarcinoma enzymology, Chorion enzymology, Chorion pathology, Cytoplasm enzymology, Female, Humans, Immunohistochemistry, Neoplasm Invasiveness, Neoplasm Metastasis, Trophoblasts enzymology, Trophoblasts pathology, Uterine Neoplasms enzymology, Choriocarcinoma pathology, Glucuronidase metabolism, Uterine Neoplasms pathology
- Abstract
Objective: To investigate the association between the expression of heparanase (Hpa) and the invasion of choriocarcinoma by studying the expression of Hpa in human choriocarcinoma cell lines JEG-3 and JAR and human chorionic villous tissues., Methods: (1) Matrigel invasion assays were used to detect in vitro invasive ability of JEG-3 cells and JAR cells. (2) Expression of Hpa protein in the human chorionic villous tissues and choriocarcinoma cell lines (JEG-3 cells and JAR cells) were detected by immunocytochemistry and western blot., Results: (1) The invasive cell number was significantly larger in JEG-3 cells than in JAR cells (191 +/- 17 vs 106 +/- 13, P < 0.05). (2) Hpa protein mainly located in cytoplasm by immunocytochemistry. (3) Hpa protein expression was stronger in JEG-3 cells than in the JAR cells (1.560 +/- 0.180 vs 0.610 +/- 0.170, P < 0.05); the Hpa protein expression was significantly stronger in choriocarcinoma cell lines than in human chorionic villous tissues (0.190 +/- 0.008) by western blot (P < 0.05). (4) The data suggested that there were significantly positive correlations between Hpa and the invasiveness of choriocarcinoma cells (r = 0.89, P < 0.05)., Conclusions: (1) Hpa protein expression is significantly stronger in choriocarcinoma cell lines than in the human chorionic villous tissues. (2) Activation of Hpa enhances the invasion capability of choriocarcinoma. (3) Overexpression of Hpa may be related to the oncogenesis of choriocarcinoma and Hpa may play an important role in invasion of choriocarcinoma.
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- 2007
19. 15-Hydroxyprostaglandin dehydrogenase protein expression in human fetal membranes with and without subclinical inflammation.
- Author
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Rizek RM, Watson CS, Keating S, Tai HH, Challis JR, and Bocking AD
- Subjects
- Acute Disease, Adult, Amnion pathology, Analysis of Variance, Antibodies, Blotting, Western, Chorioamnionitis enzymology, Chorioamnionitis pathology, Chorion pathology, Female, Humans, Immunohistochemistry, Inflammation pathology, Isoenzymes metabolism, Mesoderm enzymology, Mesoderm pathology, Obstetric Labor, Premature enzymology, Pregnancy, Trophoblasts enzymology, Trophoblasts pathology, Amnion enzymology, Chorion enzymology, Fetal Membranes, Premature Rupture enzymology, Hydroxyprostaglandin Dehydrogenases metabolism, Inflammation enzymology, Premature Birth enzymology
- Abstract
Prostaglandins play a central role in the stimulation and maintenance of both term and preterm labor. 15-Hydroxyprostaglandin dehydrogenase (PGDH), localized primarily to chorion trophoblasts, is the key enzyme responsible for the metabolism of prostaglandins. In preterm chorion, levels of PGDH protein and activity were lower when compared to term and were further reduced with the presence of infection, but effects of subclinical inflammation and membrane rupture on PGDH expression are not known. Our objectives were (1) to determine the relative expression of PGDH in amnion and chorion and (2) to determine the effect of preterm premature rupture of membranes (PPROM) and (3) subclinical inflammation on PGDH protein expression in preterm fetal membranes. Fetal membranes were collected from women with idiopathic preterm labor. Patients were divided into preterm birth (1) <32 weeks with PPROM (n = 6), (2) <32 weeks with intact membranes (n = 11), (3) >or=32 and <37 weeks with PPROM (n = 10), and (4) >or=32 and <37 weeks with intact membranes (n = 10). Different antibodies were used to detect protein expression and localization of PGDH in amnion and chorion from these patients using both Western blotting and immunohistochemistry. Antibody T (AbT) localized PGDH to chorion trophoblasts, whereas antibody C (AbC) detected immunoreactive (ir) PGDH predominantly in the amnion mesenchyme. By Western blot, AbT showed a stronger 29-kDa ir-PGDH band whereas with AbC, a stronger 55-kDa ir-PGDH signal was detected. 55-kDa ir-PGDH was significantly higher in PPROM amnion, specifically in the <32 weeks group (P < .05) and with PPROM >24 hours (P < .05). No change was detected in the 29-kDa ir-PGDH in either amnion or chorion with gestational age or the presence and absence of PPROM. In addition, neither form of ir-PGDH was altered significantly with or without subclinical inflammation. ir-PGDH is detectable in both chorion trophoblasts and amnion, especially in the mesenchyme; however, the predominant form of the enzyme differs in the 2 tissues. PPROM and subclinical inflammation do not appear to affect the levels of 29-kDa ir-PGDH protein in the fetal membranes. The differential expression of 55-kDa ir-PGDH in preterm amnion with and without PPROM supports the need for a better understanding of the different forms of PGDH.
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- 2007
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20. [Expression of cyclooxygenase-2 and 15-prostaglandin dehydrogenase of placenta and fetal membranes in patients of preterm labor].
- Author
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Ding YL and Li YJ
- Subjects
- Adult, Chorion enzymology, Decidua enzymology, Female, Gestational Age, Humans, Immunohistochemistry, Labor, Obstetric metabolism, Pregnancy, Cyclooxygenase 2 metabolism, Extraembryonic Membranes enzymology, Hydroxyprostaglandin Dehydrogenases metabolism, Obstetric Labor, Premature enzymology, Placenta enzymology
- Abstract
Objective: To investigate the effect of cyclooxygenase 2 (COX-2) and 15-prostaglandin dehydrogenase (15-PGDH) on preterm labor., Methods: The expressions and localizations of COX-2 and 15-PGDH in placentas and fetal membranes were examined by immunohistochemical (IH) two-step assay (IH scores were expressed as the sum of the percentage of immunoreactivity and the stained degree of cells), respectively. The samples were obtained from 14 preterm delivery (PL), 18 term in labor (TL) and 17 term not in labor (control group)., Results: (1) The immunoreactivity of COX-2 was found in the amniotic epithelial cells, chorion cells and decidual cells. IH scores of COX-2 in amniotic epithelium of placenta or fetal membranes in PL were 4.6 +/- 1.2, 4.7 +/- 0.9, respectively; in TL were 3.2 +/- 1.0, 3.6 +/- 1.0, respectively, and in control group were 2.2 +/- 0.6, 2.5 +/- 0.9, respectively. IH scores in PL and TL were obviously higher than those in control group (P < 0.01). It was higher in PL than in TL (P < 0.01). The expression of COX-2 was not significantly different between amnion of placenta and amnion of fetal membranes among the three groups (P > 0.05). (2) IH scores of COX-2 in chorion of placenta in PL were 4.9 +/- 1.0, in TL were 3.9 +/- 1.2 and in control group were 2.3 +/- 0.7. IH scores in PL and TL were obviously higher than those in control group (P < 0.01), and it was higher in PL than in TL (P < 0.01). The expression of COX-2 was not significantly different among the three groups in chorion of fetal membranes (P > 0.05) and it was not significantly different among the three groups in decidual cell of placenta (P > 0.05), either. (3) The immunoreactivity of 15-PGDH was found in the amniotic epithelial cells, chorion cells and decidual cells. The expression of 15-PGDH was not significantly different among the three groups in amnion of placenta or fetal membranes (P > 0.05). IH scores of 15-PGDH in chorion of placenta and fetal membranes in PL were 1.5 +/- 0.6, 2.3 +/- 0.8, respectively, in TL were 2.6 +/- 0.8, 3.0 +/- 0.7, respectively, and in control group were 4.4 +/- 1.1, 4.1 +/- 1.2, respectively. IH scores in PL and TL were obviously lower than those in control group (P < 0.01), and it was lower in PL than in TL (P < 0.05). IH scores of 15-PGDH in decidua of placenta in PL were 2.1 +/- 0.7, in TL were 2.8 +/- 0.8 and in control group were 4.5 +/- 1.0. IH scores in PL and TL were obviously lower than those in control group (P < 0.01); it was lower in PL than in TL (P < 0.05)., Conclusions: The higher expression of COX-2 in amniotic epithelium and the lower expression of 15-PGDH in chorion and decidual cell are relevant with the onset of premature labor. They play a role in the pathogenesis of premature labor.
- Published
- 2006
21. Biodiversity of multiple Pregnancy-Associated Glycoprotein (PAG) family: gene cloning and chorionic protein purification in domestic and wild eutherians (Placentalia)--a review.
- Author
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Szafranska B, Panasiewicz G, and Majewska M
- Subjects
- Animals, Animals, Domestic, Animals, Wild, Aspartic Acid Endopeptidases metabolism, Cloning, Molecular, DNA, Complementary metabolism, Female, Glycoproteins metabolism, Molecular Sequence Data, Pregnancy, Pregnancy Proteins metabolism, Artiodactyla genetics, Aspartic Acid Endopeptidases genetics, Chorion enzymology, Glycoproteins genetics, Placenta enzymology, Pregnancy Proteins genetics
- Abstract
This review presents a broad overview of chorionic glycoproteins encoded by the Pregnancy-Associated Glycoprotein (PAG) gene family and also serves to illustrate how the recent discovery of the PAG family has contributed to our general knowledge of genome evolution, placental transcription and placental protein expression. The complex and large PAG family is restricted to the Artiodactyla order, although single PAG-like genes have also been identified in species outside the Artiodactyla. The PAGs are members of the aspartic proteinase (AP) superfamily. Unexpectedly, however, some members of the PAG family possess amino acid substitutions within and around the active site that likely render them unable to act as proteinases. This paper summarises the available information regarding biodiversity of PAG gene expression based on cDNA cloning, mRNA localisation studies and the structural organisation of the PAG genes with a particular emphasis on PAG promoters. It also compares available data regarding PAG protein purifications, sequencing and their N-glycodiversity. Finally, it discusses the scientific relevance, possible functional roles of the PAGs and describes possible profitable applications related to the detection of PAG proteins in the blood of pregnant domestic and wild species.
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- 2006
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22. Changes in matrix metalloproteinase (MMP)-2 and MMP-9 in the fetal amnion and chorion during gestation and at term and preterm labor.
- Author
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Yonemoto H, Young CB, Ross JT, Guilbert LL, Fairclough RJ, and Olson DM
- Subjects
- Adult, Blotting, Western, Female, Humans, Pregnancy, Premature Birth, Amnion enzymology, Chorion enzymology, Labor, Obstetric metabolism, Matrix Metalloproteinase 2 metabolism, Matrix Metalloproteinase 9 metabolism, Obstetric Labor, Premature metabolism
- Abstract
Increased matrix metalloproteinase (MMP)-9 proteolytic activity is associated with term birth, preterm birth and premature rupture of membranes. However, most studies show no changes with MMP-2, which binds tightly to cell and matrix proteins. We hypothesized better protein extraction would reveal new MMP patterns. Human amnion and chorion were collected from 25 patients at preterm or term, extracted with 2% SDS (a high concentration), and the MMP protein levels and pro-enzyme activities were determined by Western immunoblotting and zymography. MMP-2 protein and MMP-2 and -9 pro-enzyme activities in the amnion increased significantly (p<0.05) with labor at term, and were higher than at preterm labor (p<0.05), when extracted with high SDS concentration. There were no changes in chorion MMPs under any condition. These associations suggest MMP-2 may be another regulator of membrane rupture and other labor-associated mechanisms at term parturition, and its role(s) should be examined further.
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- 2006
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23. Contribution of inducible nitric oxide synthase and cyclooxygenase-2 to apoptosis induction in smooth chorion trophoblast cells of human fetal membrane tissues.
- Author
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Yuan B, Ohyama K, Bessho T, and Toyoda H
- Subjects
- Cells, Cultured, Cyclooxygenase 2 genetics, Gene Expression Regulation, Humans, Nitric Oxide Synthase Type II genetics, Oxidation-Reduction, Oxidative Stress, RNA, Messenger genetics, Apoptosis, Chorion enzymology, Cyclooxygenase 2 metabolism, Nitric Oxide Synthase Type II metabolism, Trophoblasts cytology, Trophoblasts enzymology
- Abstract
We examined the contribution of apoptosis- and oxidative stress-associated genes to apoptosis induction in trophoblast cells of human fetal membrane tissues undergoing apoptosis during in vitro incubation. RT-PCR analyses demonstrated an increased level of HO-1, Mn-SOD, Cox-2, iNOS, TNFalpha, TNFR1, IL-1beta, IL-6, Bax, Bak, and Bad gene expression, while Bcl-2 mRNA expression level decreased. Western blot analyses demonstrated an increase in iNOS, Cox-2, and HO-1 protein levels; a decrease in pro-caspase-3 and 9, proform-PARP, and Apaf-1 protein levels; a leakage of cytochrome c from the mitochondria. An antioxidative reagent, general and selective Cox-2 inhibitors, and an iNOS inhibitor suppressed in vitro progression of the apoptosis. Furthermore, an NO donor reagent induced apoptosis in primary cultured trophoblast cells. Therefore, we concluded that the induction of apoptosis in the smooth chorion trophoblasts is mediated through oxidative stress induction followed by mitochondria damage, suggesting that iNOS and Cox-2 play an important role in the apoptosis induction in trophoblasts of human fetal membrane tissues.
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- 2006
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24. Mechanisms regulating prostaglandin H2 synthase-2 mRNA level in the amnion and chorion during pregnancy.
- Author
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Johnson RF, Mitchell CM, Giles WB, Bisits A, and Zakar T
- Subjects
- Cyclooxygenase 2 metabolism, Female, Gene Expression, Humans, Labor, Obstetric metabolism, Pregnancy, Pregnancy Trimester, First, Pregnancy Trimester, Third, RNA, Messenger analysis, Reverse Transcriptase Polymerase Chain Reaction, Amnion enzymology, Chorion enzymology, Cyclooxygenase 2 genetics, RNA, Messenger metabolism
- Abstract
Increasing prostaglandin H(2) synthase (PGHS)-2 expression in the fetal membranes is implicated in the production of prostaglandins (PGs) that stimulate labour. We have determined the activity of the PGHS-2 gene in the amnion and chorion throughout gestation and defined the contribution of transcriptional and post-transcriptional mechanisms to the increase of PGHS-2 mRNA levels. We also measured PGHS-1 mRNA abundance to assess the participation of the two isoenzymes in fetal membrane PG-production during pregnancy. Amnion and chorion were collected from non-labouring women at 10-19 weeks (early), at 28-36 weeks (preterm) and at term (37-41 weeks). We determined PGHS-1 and -2 mRNA abundance and assessed PGHS-2 gene activity by measuring PGHS-2 heterogeneous nuclear RNA levels using real-time RT-PCR. PGHS-2 gene activity and mRNA levels were up-regulated in both tissues with advancing gestation. Path analysis demonstrated that the PGHS-2 mRNA up-regulation involved both transcriptional and post-transcriptional components. PGHS-2 mRNA abundance increased 9-11 fold between the early (10-19 weeks) and preterm (28-36 weeks) groups and remained high at term. The underlying mechanism was predominantly transcriptional in the amnion and post-transcriptional in the chorion. PGHS-1 mRNA expression precipitously decreased between early gestation and term. Thus, PGHS-2 mRNA abundance is up-regulated well in advance of term and is not a trigger for labour. There is a switch in PGHS mRNA expression during pregnancy with PGHS-1 dominating in the early period and PGHS-2 dominating at term.
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- 2006
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25. [The glutathione-dependent system of placenta antioxidant defense in miscarriage].
- Author
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Prokopenko VM, Partsalis GK, and Burmistrov SO
- Subjects
- Antioxidants metabolism, Chorion enzymology, Female, Humans, Pregnancy, Abortion, Spontaneous enzymology, Glutathione Peroxidase analysis, Glutathione Reductase analysis, Glutathione Transferase analysis, Placenta enzymology
- Published
- 2006
26. The enzymatic component of Drosophila melanogaster chorion is the Pxd peroxidase.
- Author
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Konstandi OA, Papassideri IS, Stravopodis DJ, Kenoutis CA, Hasan Z, Katsorchis T, Wever R, and Margaritis LH
- Subjects
- Animals, Chorion enzymology, Gene Expression Regulation, Insect Proteins metabolism, Peroxidases metabolism, Drosophila melanogaster enzymology, Insect Proteins chemistry, Peroxidases chemistry
- Abstract
In the present study, we demonstrate the isolation and characterization of the Pxd cDNA clone, which codes for the Drosophila melanogaster chorion peroxidase. This specific peroxidase is involved in the chorion hardening process, through protein crosslinking mediated by the formation of di- and tri-tyrosine bonds. The Pxd gene product has been identified in crude protein extracts from adult flies as three immunoreacting, with the anti-rAePO polyclonal antibody, bands of 77, 67 and 55 kDa, while in larvae and purified chorions as a unique 55 kDa band. Moreover, the mature form of the Pxd recombinant protein was specifically recognized by the anti-rAePO antibody as a 77 kDa band, while in the presence of H2O2 was able to convert tyrosine residues to di-tyrosine moieties. Northern blotting analysis of total RNA preparations revealed distinct molecular weight patterns of the Pxd RNA transcripts among adult flies, ovaries and larvae. The in situ hybridization clearly shows that the Pxd mRNA is specifically expressed in follicle cells during the late stages of oogenesis 11-14, while the reverse transcription reactions dictate the stage-specific developmental regulation of the Pxd gene. The immunolocalization approach, using the anti-rAePO polyclonal antibody, has revealed that the Pxd peroxidase is selectively localized in the chorion structures and particularly in the endochorion and innermost chorionic layer (ICL).
- Published
- 2005
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27. Purification and gene cloning of Fundulus heteroclitus hatching enzyme. A hatching enzyme system composed of high choriolytic enzyme and low choriolytic enzyme is conserved between two different teleosts, Fundulus heteroclitus and medaka Oryzias latipes.
- Author
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Kawaguchi M, Yasumasu S, Shimizu A, Hiroi J, Yoshizaki N, Nagata K, Tanokura M, and Iuchi I
- Subjects
- Amino Acid Sequence, Animals, Chorion enzymology, Chorion ultrastructure, Cloning, Molecular, Conserved Sequence, DNA, Complementary genetics, Exons, Fundulidae embryology, Gene Expression Regulation, Developmental, Gene Expression Regulation, Enzymologic, In Situ Hybridization, Introns, Isoenzymes genetics, Isoenzymes isolation & purification, Microscopy, Electron, Molecular Sequence Data, Oryzias embryology, Phylogeny, Sequence Homology, Amino Acid, Species Specificity, Fundulidae genetics, Fundulidae metabolism, Metalloendopeptidases genetics, Metalloendopeptidases isolation & purification, Oryzias genetics, Oryzias metabolism
- Abstract
Two cDNA homologues of medaka hatching enzyme -- high choriolytic enzyme (HCE) and low choriolytic enzyme (LCE) -- were cloned from Fundulus heteroclitus embryos. Amino acid sequences of the mature forms of Fundulus HCE (FHCE) and LCE (FLCE) were 77.9% and 63.3% identical to those of medaka HCE and LCE, respectively. In addition, phylogenetic analysis clearly showed that FHCE and FLCE belonged to the clades of HCE and LCE, respectively. Exon-intron structures of FHCE and FLCE genes were similar to those of medaka HCE (intronless) and LCE (8-exon-7-intron) genes, respectively. Northern blotting and whole-mount in situ hybridization showed that both genes were concurrently expressed in hatching gland cells. Their spatio-temporal expression pattern was basically similar to that of medaka hatching enzyme genes. We separately purified two isoforms of FHCE, FHCE1 and FHCE2, from hatching liquid through gel filtration and cation exchange column chromatography in the HPLC system. The two isoforms, slightly different in molecular weight and in MCA-peptide-cleaving activity, swelled the inner layer of chorion by their limited proteolysis, like the medaka HCE isoforms. In addition, we identified FLCE by TOF-MS. Similar to the medaka LCE, FLCE hardly digested intact chorion. FHCE and FLCE together, when incubated with chorion, rapidly and completely digested the chorion, suggesting their synergistic effect in chorion digestion. Such a cooperative digestion was confirmed by electron microscopic observation. The results suggest that a hatching enzyme system composed of HCE and LCE is conserved between two different teleosts Fundulus and medaka.
- Published
- 2005
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28. Characterization of N-linked oligosaccharides in chorion peroxidase of Aedes aegypti mosquito.
- Author
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Li JS and Li J
- Subjects
- Amino Acid Sequence, Animals, Carbohydrate Conformation, Carbohydrate Sequence, Glycosylation, Mass Spectrometry methods, Molecular Sequence Data, Monosaccharides analysis, Peroxidase isolation & purification, Polysaccharides chemistry, Aedes enzymology, Chorion enzymology, Oligosaccharides chemistry, Peroxidase chemistry, Peroxidase metabolism
- Abstract
A peroxidase is present in the chorion of Aedes aegypti eggs and catalyzes chorion protein cross-linking during chorion hardening, which is critical for egg survival in the environment. The unique chorion peroxidase (CPO) is a glycoprotein. This study deals with the N-glycosylation site, structures, and profile of CPO-associated oligosaccharides using mass spectrometric techniques and enzymatic digestion. CPO was isolated from chorion by solubilization and several chromatographic methods. Mono-saccharide composition was analyzed by HPLC with fluorescent detection. Our data revealed that carbohydrate (D-mannose, N-acetyl D-glucosamine, D-arabinose, N-acetyl D-galactosamine, and L-fucose) accounted for 2.24% of the CPO molecular weight. A single N-glycosylation site (Asn328-Cys- Thr) was identified by tryptic peptide mapping and de novo sequencing of native and PNGase A-deglycosylated CPO using matrix-assisted laser/desorption/ionization time-of-flight mass spectrometry (MALDI/TOF/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS). The Asn328 was proven to be a major fully glycosylated site. Potential tryptic glycopeptides and profile were first assessed by MALDI/TOF/MS and then by precursor ion scanning during LC/MS/MS. The structures of N-linked oligosaccharides were elucidated from the MS/MS spectra of glycopeptides and exoglycosidase sequencing of PNGase A-released oligosaccharides. These CPO-associated oligosaccharides had dominant Man3GlcNAc2 and Man3 (Fuc) GlcNAc2 and high mannose-type structures (Man(4-8)GlcNAc2). The truncated structures, Man2GlcNAc2 and Man2 (Fuc) GlcNAc2, were also identified. Comparison of CPO activity and Stokes radius between native and deglycosylated CPO suggests that the N-linked oligosaccharides influence the enzyme activity by stabilizing its folded state.
- Published
- 2005
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29. 5 Beta-dihydroprogesterone and steroid 5 beta-reductase decrease in association with human parturition at term.
- Author
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Sheehan PM, Rice GE, Moses EK, and Brennecke SP
- Subjects
- 5-alpha-Dihydroprogesterone blood, Amnion enzymology, Chorion enzymology, Female, Humans, Myometrium enzymology, Oxidoreductases genetics, Placenta enzymology, Pregnancy, RNA, Messenger metabolism, 5-alpha-Dihydroprogesterone metabolism, Labor, Obstetric metabolism, Oxidoreductases metabolism, Parturition metabolism
- Abstract
The role of progesterone withdrawal in human parturition continues to provoke controversy. One possible mechanism by which functional progesterone withdrawal may be achieved is by a decrease in the circulating concentration of its bioactive metabolites. The progesterone metabolite 5beta-dihydroprogesterone (5betaDHP) has been shown to be a potent tocolytic in vitro. We quantified plasma concentrations of 5betaDHP in association with the onset of spontaneous labour in women at term and steroid 5beta-reductase mRNA expression in placenta, myometrium, chorion and amnion in relation to parturition, using real time RT-PCR. Serial blood samples were obtained from patients late in pregnancy, before term labour, during term labour and within the first 24 h postpartum. Following organic solvent extraction, steroids including 5betaDHP were separated by high-performance liquid chromatography (HPLC) and then quantified by radioimmunoassay (RIA). 5betaDHP concentration decreased two-fold (P = 0.00001, n = 25) from 0.317 +/- 0.039 nmol/ml to 0.178 +/- 0.017 nmol/ml in association with active labour. Tissue 5beta-reductase mRNA-relative abundance was determined in placenta, myometrium, chorion and amnion obtained from labouring and non-labouring women. In placenta and myometrium, relative expression decreased significantly in association with labour, by about two-fold and 10-fold, respectively. These data are consistent with a possible role for 5betaDHP in the onset of spontaneous human labour. Further studies exploring this hitherto unrecognized endocrinological pathway are indicated.
- Published
- 2005
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30. Evidence for a role of phosphodiesterase 4 in lipopolysaccharide-stimulated prostaglandin E2 production and matrix metalloproteinase-9 activity in human amniochorionic membranes.
- Author
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Oger S, Méhats C, Dallot E, Cabrol D, and Leroy MJ
- Subjects
- 3',5'-Cyclic-AMP Phosphodiesterases antagonists & inhibitors, Adjuvants, Immunologic pharmacology, Adjuvants, Immunologic physiology, Amnion drug effects, Amnion metabolism, Chorion drug effects, Chorion metabolism, Cyclic AMP antagonists & inhibitors, Cyclic AMP physiology, Cyclic Nucleotide Phosphodiesterases, Type 4, Cyclooxygenase 2, Enzyme Activation drug effects, Enzyme Activation physiology, Enzyme Precursors metabolism, Female, Humans, Immune Sera pharmacology, Interleukin-10 immunology, Interleukin-10 metabolism, Membrane Proteins, Phosphodiesterase Inhibitors pharmacology, Pregnancy, Prostaglandin-Endoperoxide Synthases biosynthesis, Rolipram pharmacology, Tissue Culture Techniques, Tumor Necrosis Factor-alpha antagonists & inhibitors, Tumor Necrosis Factor-alpha metabolism, Up-Regulation drug effects, Up-Regulation immunology, 3',5'-Cyclic-AMP Phosphodiesterases physiology, Amnion enzymology, Amnion immunology, Chorion enzymology, Chorion immunology, Dinoprostone biosynthesis, Lipopolysaccharides immunology, Matrix Metalloproteinase 9 metabolism
- Abstract
Chorioamniotic infection is a leading cause of preterm premature rupture of fetal membranes (amnion and chorion). Bacterial infection induces an inflammatory response characterized by elevated production of proinflammatory cytokines; the latter activate the production of both PGs that stimulate uterine contractions, and matrix metalloproteinases (MMPs) that degrade the extracellular matrix of the chorioamniotic membranes. The inflammatory response is under the control of cAMP content, which is partly regulated by phosphodiesterases (PDE). In this study, we investigated the role of the PDE4 family in the inflammatory process triggered by LPS in a model of amniochorionic explants. We found that PDE4 family is the major cAMP-PDE expressed in human fetal membranes and that PDE4 activity is increased by LPS treatment. Selective inhibition of PDE4 activity affected LPS signaling, because PDE4 inhibitors (rolipram and/or cilomilast) reduced the release of the proinflammatory cytokine TNF-alpha and increased the release of the anti-inflammatory cytokine IL-10. PDE4 inhibition reduced cyclooxygenase-2 protein expression and PGE(2) production and also modulated MMP-9, a key mediator of the membrane rupture process, by inhibiting pro-MMP-9 mRNA expression and pro-MMP-9 activity. These results demonstrate that the PDE4 family participates in the regulation of the inflammatory response associated with fetal membrane rupture during infection. The PDE4 family may be an appropriate pharmacological target for the management of infection-induced preterm delivery.
- Published
- 2005
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31. Initial characterization of the microenvironment that regulates connective tissue degradation in amniochorion during normal human labor.
- Author
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Estrada-Gutierrez G, Zaga V, Gonzalez-Jimenez MA, Beltran-Montoya J, Maida-Claros R, Giono-Cerezo S, and Vadillo-Ortega F
- Subjects
- Amnion cytology, Amnion enzymology, Chorion cytology, Chorion enzymology, Connective Tissue enzymology, Female, Humans, Leukocyte Count, Matrix Metalloproteinase 2 genetics, Matrix Metalloproteinase 2 metabolism, Matrix Metalloproteinase 9 genetics, Matrix Metalloproteinase 9 metabolism, Pregnancy, Tissue Culture Techniques, Amnion metabolism, Chorion metabolism, Connective Tissue metabolism, Labor, Obstetric metabolism
- Abstract
Extracellular matrix degradation in fetal membranes leading to its rupture is coupled to myometrial activity and cervical ripening during human normal labor. Mechanisms which modulate collagen degradation in amniochorion during labor have not been elucidated. Initial characterization of the effect of different blood compartments on connective tissue degradation in amniochorion during human labor was explored. Amniochorion explants were stimulated with plasma of maternal venous blood, umbilical cord blood or placental blood, obtained from women with pregnancies at term, with or without labor. MMP-2 and MMP-9 activities were quantified in conditioned media by gelatin-zymography as an index of connective tissue degradation. Collagen content was measured in tissue explants and collagen fibrils distribution was examined by electron microscopy. Placental plasma from term pregnancies, with or without labor, is enriched with soluble signals that enhance the in vitro MMP-9 production by amniochorion. Accompanying ultrastructural distortion of collagen fibers and demonstration of collagen degradation fragments confirmed induction of extracellular matrix degradation. Control experiments in which MMP-9 activity was blocked with TIMP-1 resulted in inhibition of all the above mentioned changes. These results suggest that placental intervillous space is a functional compartment in which mediators capable to induce collagen degradation in amniochorion are selectively expressed during human labor.
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- 2005
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32. Expression of CYP4A isoforms in developing rat placental tissue and rat trophoblastic cell models.
- Author
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Xu Y, Knipp GT, and Cook TJ
- Subjects
- Allantois growth & development, Animals, Cell Line, Chorion growth & development, Cytochrome P-450 CYP4A genetics, Female, Gene Expression, Immunoenzyme Techniques, Isoenzymes, Pregnancy, RNA, Messenger metabolism, Rats, Reverse Transcriptase Polymerase Chain Reaction, Allantois enzymology, Chorion enzymology, Cytochrome P-450 CYP4A metabolism, Trophoblasts enzymology
- Abstract
Maintaining fatty acid homeostasis during pregnancy is critical for normal fetal development. As an organ that controls nutrient supply from the mother to the fetus, the placenta plays a significant role in guiding fatty acid transfer to the developing fetus. The cytochrome P450 4A (CYP4A) subfamily of metabolizing enzymes is a group of structurally and functionally conserved proteins that are specialized in the omega/omega-1 hydroxylation of saturated and unsaturated fatty acids and their derivatives. To understand the function of the CYP4A system in the placenta and its significance in maintaining fetal fatty acid homeostasis, information about the placental expression of individual CYP4A isoforms is required. In the present study, we have elucidated the temporal and spatial patterns of expression of the four known rat CYP4A isoforms (CYP4A1, CYP4A2, CYP4A3, and CYP4A8) in the junctional and labyrinthine zones of the developing rat chorioallantoic placenta as well as two rat trophoblastic cell lines, HRP-1 and Rcho-1, using semi-quantitative RT-PCR and immunohistochemical analyses. The mRNA from the four rat CYP4A isoforms was detected in the developing rat placenta with CYP4A1 exhibiting the strongest expression (4A1 > 4A2 >> 4A3 approximately equal to 4A8). CYP4A1 was also detected by immunohistochemical staining in the developing rat placenta. We also observed CYP4A1 in both HRP-1 and Rcho-1 cells by RT-PCR, suggesting the utility of these cells as in vitro tools to study the effects of xenobiotics on placental fatty acid metabolism. Establishing the expression of CYP4A isoforms in these tissues and cell models provides a framework for further investigation of their functional and physiological significance in guiding proper fetal development., ((c) Elsevier Ltd. All rights reserved.)
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- 2005
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33. Regulation of 15-hydroxyprostaglandin dehydrogenase (PGDH) gene activity, messenger ribonucleic acid processing, and protein abundance in the human chorion in late gestation and labor.
- Author
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Johnson RF, Mitchell CM, Clifton V, and Zakar T
- Subjects
- Female, Humans, Hydroxyprostaglandin Dehydrogenases analysis, RNA, Messenger analysis, Chorion enzymology, Hydroxyprostaglandin Dehydrogenases genetics, Labor, Obstetric metabolism, Pregnancy metabolism, RNA, Messenger metabolism
- Abstract
The prostaglandin (PG)-inactivating enzyme 15-hydroxyprostaglandin dehydrogenase (PGDH) is highly expressed in the chorion leave. To assess the involvement of PGDH in the regulation of intrauterine PG levels, we have determined the mechanisms that control chorionic PGDH expression in women at term and preterm labor. PGDH gene activity decreased at term and during normal labor. PGDH mRNA abundance also decreased at term due to changing splice variant distribution. Gene activity predicted PGDH mRNA abundance preterm and after normal labor, but not at term before labor. PGDH mRNA decayed rapidly in cultured tissues and was stabilized by transcriptional arrest. PGDH protein levels varied without being significantly different between the patient groups. PGDH mRNA levels predicted PGDH protein levels at term, but not preterm and after labor. PGDH gene activity, mRNA variant, and immunoreactive protein levels were not different between the preterm labor and preterm not in labor groups. Thus, PGDH mRNA is transiently down-regulated before term labor by a posttranscriptional mechanism(s). Protein turnover controls PGDH protein abundance at preterm and after normal labor. At term, PGDH protein levels become dependent on the rapidly turning over PGDH mRNA. This may allow rapid changes in PGDH protein abundance and uterotonic PG concentrations promoting labor.
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- 2004
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34. Carbonic anhydrases in chick extra-embryonic structures: a role for CA in bicarbonate reabsorption through the chorioallantoic membrane.
- Author
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Gabrielli MG
- Subjects
- Absorption, Animals, Chick Embryo, Chorion cytology, Chorion enzymology, Immunohistochemistry, Bicarbonates metabolism, Carbonic Anhydrase IV metabolism, Chorion metabolism
- Abstract
The villus cavity cells, a specific cell type of the chick chorioallantoic membrane, express both cytosolic carbonic anhydrase in their cytoplasm and HCO3(-)/Cl(-) anion exchangers at their basolateral membranes. By immunohistochemical analysis, we show here that villus cavity cells specifically react with antibodies directed against the membrane-associated form of carbonic anhydrase, CAIV. Staining is restricted to the apical cell membranes, characteristically invaginated toward the shell membrane, as well as to endothelia of blood vessels present in the mesodermal layer. The occurrence of a membrane-associated CA form at the apical pole of villus cavity cells, when definitively confirmed, would be fairly consistent with the role proposed for these cells in bicarbonate reabsorption from the eggshell so to prevent metabolic acidosis in the embryo during development.
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- 2004
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35. Intracrine control of estrogen action in human gestational tissues at parturition.
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Madsen G, Zakar T, Manuelpillai U, Wallace E, Kwek K, Yeo GS, Smith R, and Mesiano S
- Subjects
- 17-Hydroxysteroid Dehydrogenases genetics, Chorion enzymology, Decidua enzymology, Estradiol metabolism, Estrone metabolism, Female, Gene Expression, Humans, Isoenzymes metabolism, Labor, Obstetric, Myometrium enzymology, Pregnancy, RNA, Messenger analysis, Reverse Transcriptase Polymerase Chain Reaction, 17-Hydroxysteroid Dehydrogenases metabolism, Estrogens pharmacology, Parturition
- Abstract
Objective: We examined whether estrogen action in human parturition is regulated by an intracrine mechanism mediated by target tissue expression of specific 17beta-hydroxysteroid dehydrogenase (17betaHSD) isozymes that interconvert estrone (E1) and estradiol (E2), such that the onset of labor is associated with an increase in local E2 bioavailability., Methods: The extent of 17betaHSD-1, -2, -3, -4, -5, and -7 expression (measured by quantitative reverse transcriptase polymerase chain reaction) and the capacity to interconvert E1 and E2 were compared in amnion, chorion, placenta, decidua, and myometrium obtained from women at term before (n = 6) and after (n = 6) the onset of labor., Results: In chorion, abundance of 17betaHSD-1 (converts E1 to E2) mRNA decreased 2.7-fold (P <.05) in association with labor onset. In myometrium, 17betaHSD-1 and 17betaHSD-4 (converts E2 to E1) mRNAs increased two-fold and five-fold, respectively, with the onset of labor (P <.05 for each). No other statistically significant labor-associated change in 17betaHSD expression was observed. In chorion, 17betaHSD oxidative (E2 to E1) and reductive (E1 to E2) activities and the net E2 synthetic capacity increased with labor. In decidua, both activities decreased with the onset of labor, but there was no change in net E2 synthetic capacity. The capacity to interconvert E1 and E2 did not change in the other tissues., Conclusion: The increase in E2 synthetic capacity in the chorion might contribute to an increase in local estrogen bioactivity in association with the onset of labor. However, it cannot be explained by changes in 17betaHSD isozyme expression and is unlikely to account for the increased estrogen action at parturition. These data show that intracrine mechanisms based on 17betaHSD isozyme expression play a minor role, if any, in controlling estrogen action in gestational tissues during human parturition.
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- 2004
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36. Antioxidants inhibit angiogenesis in vivo through down-regulation of nitric oxide synthase expression and activity.
- Author
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Polytarchou C and Papadimitriou E
- Subjects
- Animals, Anti-Inflammatory Agents pharmacology, Cell Membrane enzymology, Chick Embryo, Cyclic N-Oxides metabolism, Dexamethasone pharmacology, Down-Regulation, Enzyme Inhibitors pharmacology, Hydrogen Peroxide pharmacology, NADPH Oxidases antagonists & inhibitors, NG-Nitroarginine Methyl Ester pharmacology, Nitric Oxide Synthase antagonists & inhibitors, Nitric Oxide Synthase Type II, Spin Labels, Sulfones pharmacology, Superoxide Dismutase pharmacology, Allantois enzymology, Antioxidants pharmacology, Chorion enzymology, Neovascularization, Physiologic physiology, Nitric Oxide metabolism, Nitric Oxide Synthase metabolism, Reactive Oxygen Species metabolism
- Abstract
Although reactive oxygen species (ROS) participate in many cellular mechanisms, only few data exist concerning their involvement in physiological angiogenesis. The aim of the present work was to elucidate possible mechanisms through which ROS affect angiogenesis in vivo, using the model of the chicken embryo chorioallantoic membrane (CAM). Superoxide dismutase (SOD) and its membrane permeable mimetic tempol, dose dependently decreased angiogenesis and down-regulated inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production. The NADPH oxidase inhibitors, 4-(2-aminoethyl)-benzenesulfonyl fluoride (AEBSF) and apocynin, but not allopurinol, also had a dose dependent inhibitory effect on angiogenesis and NO production in vivo. Catalase and the intracellular hydrogen peroxide (H2O2) scavenger sodium pyruvate decreased, while H2O2 increased in a dose-dependent manner the number of CAM blood vessels, as well as the expression and activity of iNOS. Dexamethasone, which down-regulated NO production by iNOS and L-NAME, but not D-NAME, dose dependently decreased angiogenesis in vivo. These data suggest that antioxidants affect physiological angiogenesis in vivo, through regulation of NOS expression and activity.
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- 2004
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37. Regulation of phospholipase isozymes by nuclear factor-kappaB in human gestational tissues in vitro.
- Author
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Lappas M, Permezel M, Georgiou HM, and Rice GE
- Subjects
- Amnion enzymology, Amnion metabolism, Chorion enzymology, Chorion metabolism, Cyclooxygenase 1, Cyclooxygenase 2, Cytosol enzymology, DNA metabolism, Decidua enzymology, Decidua metabolism, Dinoprost metabolism, Electrophoresis, Enzyme-Linked Immunosorbent Assay, Extraembryonic Membranes enzymology, Female, Humans, In Vitro Techniques, Isoenzymes metabolism, Membrane Proteins, NF-kappa B genetics, Phospholipases A metabolism, Placenta enzymology, Pregnancy, Prostaglandin-Endoperoxide Synthases metabolism, Sulfasalazine pharmacology, Transcription Factor RelA, Dinoprost analogs & derivatives, Extraembryonic Membranes metabolism, NF-kappa B physiology, Phospholipases metabolism, Placenta metabolism
- Abstract
Phospholipid-derived mediators are implicated in the initiation and progression of human labor and delivery, particularly in relation to infection-induced preterm labor. We previously demonstrated that, in human intrauterine tissues, lipopolysaccharide (LPS)-stimulated nuclear factor-kappaB (NF-kappaB) DNA binding activity, and subsequent cytokine release can be suppressed by sulfasalazine (SASP) concentrations greater than 5 mM. The aim of this study was to elucidate the effect the SASP on secretory type II phospholipase A(2) (PLA(2)), cytosolic PLA(2) (cPLA(2)), cyclooxygenase (COX) isozymes, and subsequent prostaglandin F(2alpha) (PGF(2alpha)) production in human gestational tissues. Human placenta, amnion, and choriodecidua (n = 4-9 separate placentas) were incubated in the presence of SASP (0.1, 1, 5, and/or 10 mM) under either basal or LPS (10 microg/ml) conditions. After 6 h incubation, the tissues were collected and assayed for type II PLA(2) by ELISA and cPLA(2), COX-1, and COX-2 content by Western blotting. The incubation medium was collected and assayed for type II PLA(2) and 13,14-dihydro-15-keto PGF(2alpha) release by ELISA and PGF(2alpha) by RIA. Treatment of placenta, amnion, and choriodecidua with SASP concentrations greater than 5 mM significantly inhibited basal and/or LPS-stimulated type II PLA(2) content and release, 13,14-dihydro-15-keto PGF(2alpha) release, and cPLA(2) protein content (ANOVA, P < 0.05); however, no effect of SASP was observed on basal or LPS-stimulated COX-1 or COX-2 protein. Although no effect of SASP was observed on basal and LPS-stimulated PGF(2alpha) release from placenta and amnion, it significantly increased both basal and LPS-stimulated PGF(2alpha) release from choriodecidua. In addition, SASP concentrations of 5 mM or greater significantly suppressed NF-kappaB DNA binding activity. These data are consistent with the hypothesis that NF-kappaB regulates the expression and release of phospholipase isozymes.
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- 2004
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38. Matrix metalloproteinase-8 is expressed in human chorion during labor.
- Author
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Arechavaleta-Velasco F, Marciano D, Díaz-Cueto L, and Parry S
- Subjects
- Blotting, Western, Case-Control Studies, Enzyme-Linked Immunosorbent Assay, Female, Humans, Immunohistochemistry, In Situ Hybridization, Matrix Metalloproteinase 8 genetics, Pregnancy, RNA, Messenger metabolism, Reverse Transcriptase Polymerase Chain Reaction, Chorion enzymology, Labor, Obstetric metabolism, Matrix Metalloproteinase 8 metabolism
- Abstract
Objective: The purpose of this study was to investigate the expression of matrix metalloproteinase-8 by human fetal membranes during labor., Study Design: Fetal membranes were obtained from women who underwent normal labor or elective cesarean delivery at term. Matrix metalloproteinase-8 levels in fetal membranes were determined by enzyme-linked immunosorbent assay and Western blot; the expression of the matrix metalloproteinase-8 gene was detected by reverse transcription-polymerase chain reaction. Immunohistochemistry and in situ hybridization was performed to localize matrix metalloproteinase-8 protein and messenger RNA in intact membranes., Results: Matrix metalloproteinase-8 protein levels were increased 5-fold in fetal membranes from labor compared with membranes that were obtained from cesarean delivery. Western blots confirmed the presence of matrix metalloproteinase-8 in protein extracts. Reverse transcription-polymerase chain reaction, in situ hybridization, and immunohistochemistry demonstrated that matrix metalloproteinase-8 messenger RNA and protein were expressed almost exclusively in the chorion after labor., Conclusion: We conclude that matrix metalloproteinase-8 is produced primarily by chorionic cells in human fetal membranes and that the level of matrix metalloproteinase-8 protein and messenger RNA expression in fetal membranes increases during labor.
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- 2004
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39. [Properties of the chorioamnios zone inducing premature membranes rupture].
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Meraz Cruz MC, Beltrán Montoya J, Bustos López H, Flores Pliego A, Espejel A, Buendía Díaz G, and Vadillo-Ortega F
- Subjects
- Adult, Culture Techniques, Female, Humans, Matrix Metalloproteinase 9 metabolism, Pregnancy, Amnion enzymology, Amnion pathology, Chorion enzymology, Chorion pathology, Fetal Membranes, Premature Rupture etiology
- Abstract
Premature membrane rupture (PMR) is one of the most serious public health problems in the world, ocurring in 10% of all pregnancies. PMR has important adverse effects on maternofetal morbidity-mortality, as it has been estimated that it accounts on the whole for 70% and 40% of neonatal morbidity and mortality, respectively. PMR treatment is empirical, as its aetiology is unknown and its physiopathogenic description has just been initiated. This work analyzes the possibility of documenting functional differences in human chorio-amnios, comparing the zone where rupture most frequently occurs in PMR with some other distant chorio-amnionic zones and with equivalent zones of fetal membranes obtained from nine month pregnancies which have not undergone labor. The membrane zone which was nearest to the cervical os was identified and marked to be analyzed later for extracellular matrix metalloprotease (MMP) activity, histology and topographical MMP distribution. The MMP expression was quantitatively determined in explant culture media from membrane fragments using specific immuno-enzymatic essays (ELISA) and zymography. In addition, immuno-histochemistry methods were used to reveal MMP expression in the different tissues. This methods allowed us to show the existence of a decreasing MMP activity gradient, with the greatest value corresponding to the zone nearest to the cervical os in the membranes obtained from PMR cases. In membranes obtained from cesarean operations no characteristic pattern was documented and values were always lower than those obtained for PMR tissues. We conclude that there is a chorio-amnionic zone in which connective tissue degradation is specifically induced and which coincides with the membrane zone in contact with the cervical os.
- Published
- 2003
40. Chorionic expression of heterogeneous products of the PAG (Pregnancy-Associated Glycoprotein) gene family secreted in vitro throughout embryonic and foetal development in the pig.
- Author
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Szafranska B, Panasiewicz G, Majewska M, and Beckers JF
- Subjects
- Animals, Aspartic Acid Endopeptidases immunology, Aspartic Acid Endopeptidases metabolism, Blotting, Western veterinary, Chorion enzymology, Cross Reactions, Culture Techniques, Electrophoresis, Polyacrylamide Gel veterinary, Female, Immune Sera immunology, Immune Sera isolation & purification, Multigene Family genetics, Pregnancy, Pregnancy Proteins immunology, Pregnancy Proteins metabolism, Trophoblasts metabolism, Aspartic Acid Endopeptidases genetics, Chorion metabolism, Gene Expression Regulation, Developmental genetics, Pregnancy Proteins genetics, Pregnancy, Animal physiology, Swine embryology
- Abstract
Porcine PAG (pPAG) are placental products of a multigene family that is strongly expressed in the chorionic epithelium (trophoblast and trophectoderm). The objective of this study was to define a pattern of the pPAG proteins, secreted in vitro by chorionic explants harvested on 16-77 days of pregnancy. Trophoblastic and trophectodermal explants were collected from pregnant (PR) gilts (n = 27) and used for protein in vitro production (8-261 h). Endometrial explants of luteal-phase gilts (E10, n = 4) and pseudopregnant gilts (PsE, n = 2) were used as negative controls for protein immunoblotting. Proteins (PR, E10, PsE) were isolated mainly from incubation media, fractionated, dialysed and separated by SDS-PAGE. Heterogeneous Western blotting with various polyclonal anti-PAG sera raised against bovine or ovine antigens (anti-bPAG, or anti-oPAG) initially identified the pPAG proteins. Such blotting of fractionated chorionic proteins allowed for the isolation of porcine antigens that were employed as immunogens to raise several homologous antisera (anti-pPAG). Crude antisera were adsorbed on endometrial extracts or proteins of non-PR pigs, to remove non-relevant antibodies. The patterns of pPAG proteins secreted in vitro varied throughout pregnancy (35-72 kDa). During implantation, approximately 43 kDa (Day 16) or approximately 68.1 kDa (Days 17-25) pPAG proteins were detected. During placentation and as pregnancy advanced (Days 31-77), approximately 72.3 kDa pPAG proteins were observed. The secretions of parallel multiple smaller proteins (35.4-47.2 kDa), presumably, as forms of processed pPAG precursors, increased with the progress of gestation. In conclusion, the pPAG protein family plays a very important role during implantation, placenta formation and embryonic/foetal development in the pig.
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- 2003
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41. Late gestation increase in 11beta-hydroxysteroid dehydrogenase 1 expression in human fetal membranes: a novel intrauterine source of cortisol.
- Author
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Alfaidy N, Li W, MacIntosh T, Yang K, and Challis J
- Subjects
- 11-beta-Hydroxysteroid Dehydrogenase Type 1, Female, Fetus enzymology, Humans, Labor, Obstetric physiology, Pregnancy, Pregnancy Trimester, Third, Amnion enzymology, Chorion enzymology, Hydrocortisone metabolism, Hydroxysteroid Dehydrogenases metabolism, Placenta enzymology
- Abstract
Late human gestation is associated with an increase in the concentration of cortisol (F) in the fetal circulation and amniotic fluid. It had been assumed that most of the F measured in the amniotic fluid came from the fetal adrenal gland. However, local production of F can also occur in human intrauterine tissues from inactive cortisone under the influence of the enzyme 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1. Recent studies have shown that 11beta-HSD 1 activity is up-regulated by prostaglandins (PG) E2 and F2alpha, hormones that are produced in the fetal membranes (FM) at term. In the present study, we hypothesized that 11beta-HSD 1 expression would increase in FM during pregnancy and at labor, creating the potential for local increase in F production at term. We examined 11beta-HSD 1 expression in placenta and FM obtained during normal pregnancy from nonlaboring women [26-28 wk (n = 3); 29-30 wk (n = 3); 32-33 wk (n = 3); 35-36 wk (n = 3)] and from uncomplicated term pregnancies after elective cesarean section (n = 6). 11beta-HSD 1 expression was also examined in amnion and chorionic tissues in relation to term labor (n = 12). Immunohistochemistry and Western blot analysis were used to examine 11beta-HSD 1 localization and expression. 11beta-HSD 1 activity was also measured in microsomal fractions prepared from whole fetal membranes. At term, immunoreactive 11beta-HSD 1 expression was localized predominantly to the chorion trophoblast cells, attached decidua, and amnion epithelial cells. 11beta-HSD 1 expression in FM increased with gestational age and reflected increased enzyme reductase activity. No change in 11beta-HSD 1 expression was found in placental tissue from the same patients. There was a significant increase in 11beta-HSD 1 expression in amnion but not in chorion with the onset of labor. We suggest that increases in 11beta-HSD 1 expression/activity by intrauterine membranes during late gestation may result in increased potential for a local increase in F production and that FM should be considered as an extraadrenal source of F during late gestation. This local F production may be involved in different pathways contributing to the regulation of parturition.
- Published
- 2003
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42. Vascular endothelial growth factor increases heme oxygenase-1 protein expression in the chick embryo chorioallantoic membrane.
- Author
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Fernandez M and Bonkovsky HL
- Subjects
- Allantois physiology, Animals, Chick Embryo, Chorion physiology, Heme Oxygenase (Decyclizing) genetics, Heme Oxygenase-1, Humans, Membrane Proteins, Up-Regulation physiology, Allantois enzymology, Chorion enzymology, Gene Expression Regulation, Enzymologic physiology, Heme Oxygenase (Decyclizing) biosynthesis, Vascular Endothelial Growth Factor A physiology
- Abstract
(1) Vascular endothelial growth factor (VEGF) is a potent angiogenic factor. It has been recently suggested that the inducible heme oxygenase (HO-1) isoform may play a role in angiogenesis. (2) The aims of this study were to determine, in chicken embryo chorioallantoic membranes (CAM), whether VEGF increases HO-1 protein expression, and, if so, by which molecular mechanism, and whether HO-1 activity is required for VEGF-induced angiogenesis. (3) Treatment of CAMs with VEGF for 48 h caused a significant increase in HO-1 protein expression, simultaneously with angiogenesis. (4) VEGF-stimulated angiogenesis in CAMs was markedly attenuated by the HO inhibitor zinc mesoporphyrin (ZnMP). This inhibitory effect of ZnMP was not observed with copper mesoporphyrin (CuMP), a metalloporphyrin that has a similar structure to ZnMP but does not inhibit HO enzymatic activity. (5) Overexpression of HO-1 protein elicited by VEGF in CAMs was significantly attenuated by the intracellular calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester (BAPTA-AM). The effects of BAPTA-AM were, in turn, compensated by the calcium ionophore A-23187. (6) In addition, the protein kinase C inhibitor staurosporine significantly attenuated, in a dose-dependent manner, the VEGF-stimulated HO-1 induction observed in CAMs. (7) These results demonstrate, for the first time, that VEGF upregulates HO-1 protein expression in vivo in CAMs by a mechanism dependent on an increase in cytosolic calcium levels and activation of protein kinase C. Our findings also suggest that HO-1 activity is necessary for VEGF-induced angiogenesis in CAMs.
- Published
- 2003
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43. The control of prostaglandin endoperoxide H-Synthase-2 expression in the human chorion laeve at term.
- Author
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Johnson RF, Mitchell CM, Giles WB, Walters WA, and Zakar T
- Subjects
- Cyclooxygenase 2, Female, Humans, Isoenzymes genetics, Membrane Proteins, Pregnancy, Prostaglandin-Endoperoxide Synthases genetics, RNA Stability physiology, RNA, Heterogeneous Nuclear chemistry, RNA, Heterogeneous Nuclear genetics, RNA, Messenger biosynthesis, RNA, Messenger genetics, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic physiology, Chorion enzymology, Gene Expression Regulation, Enzymologic physiology, Isoenzymes biosynthesis, Labor, Obstetric metabolism, Prostaglandin-Endoperoxide Synthases biosynthesis
- Abstract
Objective: Prostaglandin endoperoxide H synthase-2 (PGHS-2), the key enzyme of prostaglandin biosynthesis in gestational tissues, is expressed in the chorion laeve at term. We have determined the mechanisms that control the level of PGHS-2 mRNA in the chorion membrane in order to assess the significance of chorion-derived prostaglandins in term labor., Methods: Chorion membranes were collected after elective cesarean delivery (CD, n = 21) and after spontaneous labor (SL, n = 20) at term. The PGHS-2 gene transcription rate was measured by transcriptional run-on, and PGHS-2 mRNA and heterogeneous RNA (hnRNA) abundance was determined by quantitative real-time reverse transcriptase polymerase chain reaction. PGHS-2 mRNA stability, PGHS-2 hnRNA processing rate, and the short-term dynamics of the two RNA species were characterized in 0-24-hour-long tissue incubations., Results: The transcriptional activity of the PGHS-2 gene predicted (P <.02) the abundance of PGHS-2 mRNA and hnRNA in individual tissues. PGHS-2 gene activity and hnRNA processing rate were not different in the CD and SL groups. PGHS-2 mRNA was constitutively stable before and after labor, and its abundance spontaneously increased sixfold in tissues incubated for 24 hours. At the same time, PGHS-2 gene activity decreased by 80% within 2 hours and rebounded to 60% of its initial level by 24 hours., Conclusions: PGHS-2 mRNA is highly stable, and its abundance is transcriptionally controlled in the chorion laeve at term. Labor is not associated with changing PGHS-2 gene activity. Endogenous factors drive PGHS-2 gene transcription in the chorion, and the stable PGHS-2 mRNA accumulates in the tissue at term. This accumulation has little or no impact on the timing of labor.
- Published
- 2003
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44. Expression of membrane prostaglandin E synthase in human placenta and fetal membranes and effect of labor.
- Author
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Alfaidy N, Sun M, Challis JR, and Gibb W
- Subjects
- Adult, Amnion enzymology, Autoradiography, Blotting, Western, Chorion enzymology, Decidua enzymology, Female, Gene Expression Regulation, Enzymologic genetics, Humans, Immunohistochemistry, In Situ Hybridization, Pregnancy, Prostaglandin-E Synthases, RNA, Messenger biosynthesis, Extraembryonic Membranes enzymology, Intramolecular Oxidoreductases biosynthesis, Labor, Obstetric physiology, Placenta enzymology
- Abstract
Initiation and maintenance of labor in humans is associated with an increase in prostaglandin synthesis by intrauterine tissues. The objective of the present study was to characterize the distribution of membrane-bound PGES (mPGES) protein and mPGES mRNA in human placenta, fetal membranes, and decidua at term and to determine whether any changes occurred with labor. Immunoreactive mPGES was found to be highly concentrated in amnion epithelial cells and the chorion laeve trophoblasts, with lower levels in the mesenchymal layers. The enzyme was at very low levels or undetectable in the decidual tissue. Much lower levels of mPGES protein and mRNA were found in placenta than in fetal membranes. mPGES was associated with the syncytiotrophoblast and in cells surrounding blood vessels. The expression of mPGES mRNA did not change with labor in full membranes or placenta, but Western analysis showed an increase in mPGES protein in chorion laeve and a decrease in mPGES protein in placenta during labor, with no change in the amnion. The differences in expression found among placenta, chorion, and amnion before and after labor would indicate that this enzyme is differentially regulated in these tissues at this time.
- Published
- 2003
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- View/download PDF
45. Regulation of 15-hydroxy prostaglandin dehydrogenase by corticotrophin-releasing hormone through a calcium-dependent pathway in human chorion trophoblast cells.
- Author
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McKeown KJ and Challis JR
- Subjects
- 8-Bromo Cyclic Adenosine Monophosphate pharmacology, Antibodies pharmacology, Cells, Cultured, Chelating Agents pharmacology, Colforsin pharmacology, Corticotropin-Releasing Hormone immunology, Corticotropin-Releasing Hormone pharmacology, Dinoprost pharmacology, Egtazic Acid pharmacology, Female, Gestational Age, Humans, Peptide Fragments pharmacology, Pregnancy, Calcium pharmacology, Chorion enzymology, Corticotropin-Releasing Hormone physiology, Egtazic Acid analogs & derivatives, Hydroxyprostaglandin Dehydrogenases metabolism, Trophoblasts enzymology
- Abstract
Prostaglandins (PGs) play a crucial role in mediating parturition events, and their synthesis and metabolism are regulated by PG H synthase and 15-hydroxy-PG dehydrogenase (PGDH), respectively. Within the chorion tissue, it is the actions of PGDH that predominate. Throughout gestation, the fetal membranes secrete increasing amounts of CRH. We hypothesized that CRH, produced locally in the chorion, could act to modulate PGDH activity throughout gestation. To investigate this, we obtained Percoll-purified human chorion and placental trophoblast cells from uncomplicated term pregnancies and cultured them for 72 h. Activity of PGDH was assessed by incubation (4 h) with PGF(2alpha) (282 nM) and measurement of conversion to 13,14-dihydro-15-keto PG F(2alpha). Dose-response curves were constructed for the chorion cell cultures with CRH or 8-bromo-cAMP. To investigate the role of CRH and calcium, cells were treated with either astressin, a CRH antibody, BAPTA, or EGTA. CRH (0-1 micro M) had no effect on PGDH activity; however, cells treated with astressin (10 micro M), with or without exogenous CRH (1 micro M), and cells treated with a CRH antibody showed a significant decrease in PGDH activity. 8-Bromo-cAMP (0-1 mM) had no effect on 13,14-dihydro-15-keto PG F(2alpha) output in chorion trophoblast cells but significantly decreased output from placental trophoblast cells. Cells treated with either BAPTA-AM or EGTA had significantly reduced PGDH activity; and, at intermediate concentrations of chelator, exogenous CRH restored PGDH activity. We suggest that, in chorion trophoblast cells, endogenously produced CRH exerts a tonic stimulatory effect on PGDH activity and may help maintain a metabolic barrier, preventing the transfer of bioactive PGs from the chorioamnion to the myometrium.
- Published
- 2003
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46. Changes in the isoforms of the sodium pump in the placenta and myometrium of women in labor.
- Author
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Esplin MS, Fausett MB, Faux DS, and Graves SW
- Subjects
- Adult, Amnion enzymology, Chorion enzymology, Female, Gestational Age, Humans, Pregnancy, Tissue Distribution, Isoenzymes metabolism, Labor, Obstetric metabolism, Myometrium enzymology, Placenta enzymology, Sodium-Potassium-Exchanging ATPase metabolism
- Abstract
Objective: We determined whether changes in sodium pump isoform abundance accompanied active human labor., Study Design: Specimens of placenta, amniochorion, and myometrium were collected from women in active spontaneous labor and from those not in labor. The abundance of the three sodium pump alpha-isoforms was determined by Western blot analysis., Results: Levels of the alpha1 and alpha2 isoforms were comparable in the three tissues for women in labor and not in labor. However, alpha3 isoform abundance in placenta and myometrium (but not amniochorion) was significantly decreased in women in active labor compared with women not in labor (sodium pump alpha3 in placenta: no labor 91.2 +/- 27.6 vs labor 46.9 +/- 3.6 density units, P =.002. Sodium pump alpha3 in myometrium: no labor 52.3 +/- 7.7 vs labor 19.8 +/- 1.6 density units, P =.0002)., Conclusion: Because reductions in sodium pump number can result in hormone release from secretory tissues and in contraction of muscle, this suggests that the sodium pump may play a significant role in the initiation or maintenance of human labor.
- Published
- 2003
- Full Text
- View/download PDF
47. [Sequential changes of extracellular matrix metalloproteins in pregnancy and labor in the rat chorio-allantois].
- Author
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Meraz-Cruz N, Molina Delgado G, and Vadillo-Ortega F
- Subjects
- Animals, Female, Pregnancy, Rats, Rats, Wistar, Allantois enzymology, Chorion enzymology, Labor, Obstetric metabolism, Matrix Metalloproteinases metabolism, Pregnancy, Animal metabolism
- Abstract
Participation of matrix metalloproteinases (MMP) and their tissue inhibitors (TIMP) in the rupture of fetal membranes (FM) during labor has been proposed. We describe and characterize sequentially the activity of the enzymes that degrade connective tissue in the FM of the rat during pregnancy and labor. Pregnant Wistar rats were sacrificed on different days of gestation and samples of amniotic fluid (AF) and MCA were collected to be analyzed for proteolytic activity, zymography, western blot and northern blot. Results showed that during labor, there is a significant increase in the proteolytic activity in the MCA and in the AF. MMP-2 was identified from day 15 of pregnancy and it increased near labor. MMP-9 was only identified two days before labor. MMP-2 and MMP-9 mRNA expression were coincident with enzymatic activity and western blot data. During labor, TIMP-1 decreased and TIMP-2 did not change. These results allow us to conclude that there exists a quantitative and qualitative differential expression of MMPs during gestation, especially during labor.
- Published
- 2003
48. Expression and localization of prostaglandin E synthase isoforms in human fetal membranes in term and preterm labor.
- Author
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Meadows JW, Eis AL, Brockman DE, and Myatt L
- Subjects
- Amnion cytology, Chorion cytology, Cytosol enzymology, Female, Humans, Isoenzymes metabolism, Microsomes enzymology, Pregnancy, Prostaglandin-E Synthases, Tissue Distribution, Amnion enzymology, Chorion enzymology, Intramolecular Oxidoreductases metabolism, Labor, Obstetric metabolism, Obstetric Labor, Premature enzymology
- Abstract
Increased prostaglandin (PG) synthesis by fetal membranes occurs at parturition. PGE(2) synthesis from arachidonic acid involves multiple enzymes and two isoforms of the terminal enzyme of this biosynthetic pathway, PGE synthase (PGES), were recently identified. Cytosolic PGES (cPGES) is identical to the heat shock protein 90 chaperone, p23, and is reportedly functionally coupled to constitutive PG endoperoxide H synthase-1. Microsomal PGES (mPGES) is inducible by proinflammatory cytokines such as IL-1 beta. We have studied expression and localization of both enzyme isoforms in human fetal membranes either at term or preterm, with or without labor. The cPGES was immunolocalized in the amnion epithelium, and associated with fibroblasts and macrophages in the choriodecidual layer, whereas mPGES was localized in the amnion epithelium as well as the chorion trophoblast. Both enzymes were found to be associated with lipid particles present in the amnion epithelium, which are more prevalent in term tissues. Western blot analysis of the amnion and choriodecidua showed no differences in amounts of either cPGES or mPGES at term or preterm, with or without labor, in either tissue with advancing gestation. It does not appear that expression of PGES is the rate-limiting step in PGE2 synthesis in fetal membranes at labor.
- Published
- 2003
- Full Text
- View/download PDF
49. Production of matrix metalloproteinase-9 in lipopolysaccharide-stimulated human amnion occurs through an autocrine and paracrine proinflammatory cytokine-dependent system.
- Author
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Arechavaleta-Velasco F, Ogando D, Parry S, and Vadillo-Ortega F
- Subjects
- Antibodies pharmacology, Cesarean Section, Chorion enzymology, Chorion metabolism, Culture Techniques, Female, Humans, Interleukin-1 biosynthesis, Interleukin-1 immunology, Interleukin-1 pharmacology, Pregnancy, Tumor Necrosis Factor-alpha biosynthesis, Tumor Necrosis Factor-alpha immunology, Tumor Necrosis Factor-alpha pharmacology, Amnion enzymology, Cytokines pharmacology, Lipopolysaccharides pharmacology, Matrix Metalloproteinase 9 biosynthesis
- Abstract
The objective of this study was to determine the presence of autocrine/paracrine regulation of matrix metalloproteinase-9 (MMP-9) expression mediated by proinflammatory cytokines in human fetal membranes. Fetal membranes obtained from women who underwent cesarean delivery before labor were manually separated into amnion and chorion layers and maintained in culture. These explants were stimulated with tumor necrosis factor alpha (TNFalpha), interleukin-1beta (IL-1beta), and either lipopolysaccharide (LPS) alone or LPS with anti-TNFalpha or anti-IL-1beta-neutralizing antibodies. Levels of proMMP-9 in culture media were evaluated by zymography. Enzyme-linked immunosorbant assay was performed to measure the quantity of IL-1beta, TNFalpha, and tissue inhibitor of matrix metalloproteinases-1 (TIMP-1) after LPS stimulation. ProMMP-9 activity was upregulated after stimulation of the amnion by LPS, TNFalpha, and IL-1beta. The increased activity of proMMP-9 resulting from LPS stimulation in the amnion was blocked by the addition of TNFalpha neutralizing antibody but not with anti-IL-1beta. No significant effect of LPS, TNFalpha, or IL-1beta on proMMP-9 expression was observed in the chorion; however, the chorion produced both cytokines when stimulated with LPS. In contrast, TIMP-1 levels remained unchanged in all cultures incubated in the presence of LPS. Therefore, these data indicate that proMMP-9 is produced by the amnion but not the chorion in response to LPS. Because anti-TNFalpha-neutralizing antibody inhibits proMMP-9 activity in the amnion, TNFalpha appears to upregulate proMMP-9 production by the amnion in an autocrine fashion. Meanwhile, TNFalpha and IL-1beta produced by the chorion may upregulate amnionic proMMP-9 production in a paracrine manner.
- Published
- 2002
- Full Text
- View/download PDF
50. Amifostine protects blood vessels from the effects of ionizing radiation.
- Author
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Giannopoulou E, Katsoris P, Parthymou A, Kardamakis D, and Papadimitriou E
- Subjects
- Actins metabolism, Alkaline Phosphatase metabolism, Allantois blood supply, Allantois cytology, Allantois enzymology, Animals, Apoptosis drug effects, Apoptosis radiation effects, Cell Communication drug effects, Cell Communication radiation effects, Chick Embryo, Chorion blood supply, Chorion cytology, Chorion enzymology, Glioma blood supply, Glioma pathology, Rats, Sulfhydryl Compounds metabolism, Tubulin metabolism, Tyrosine metabolism, X-Rays adverse effects, Amifostine pharmacology, Neovascularization, Physiologic drug effects, Neovascularization, Physiologic radiation effects, Radiation-Protective Agents pharmacology
- Abstract
Amifostine (WR-2721) is a well-known radioprotective drug, selective for normal cells. The purpose of the present study was to define whether amifostine protects the vascular network from the effects of X-rays. We used the in vivo system of chicken embryo chorioallantoic membrane (CAM) as a model of angiogenesis. Amifostine reversed the early X-rays- induced decrease in the number of CAM blood vessels and reversed the early radiation-induced apoptosis of CAM cells. It also inhibited the increase in tyrosine nitration of actin and a-tubulin, which was observed 6 hours after CAM irradiation, when there was a significant decrease in non-protein SH groups. Furthermore, C6 rat glioma cells were inoculated on CAM and tumor growth, as well as tumor-induced angiogenesis, was estimated on haematoxylin-eosin-stained paraffin sections. Amifostine inhibited the post irradiation increase of C6 tumor-induced angiogenesis. These data suggest that amifostine protects CAM cells and blood vessels from the effects of X-rays, through mechanisms that do not depend solely on its free radical scavenging properties.
- Published
- 2002
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