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PubMed2001

Gene analysis of the N-terminal region of the estrogen receptor alpha in preeclampsia.

Alterations in the NH(2)-terminal region of the estrogen receptor alpha (ERalpha) gene expressed in placental bed tissue may be implicated in the development of preeclampsia, the pathogenesis of which involves the spiral arteries. Therefore, mutations and polymorphisms on exons 1 and 2 of the gene encoding ERalpha were studied. Placental bed biopsies were taken from 20 healthy, normotensive pregnant women and 16 preeclamptic patients. DNA was extracted from the tissue and exon 1 and exon 2 were amplified by PCR prior to denaturing gradient gel electrophoresis analysis or to single stranded conformational polymorphism analysis. In exon 1, a codon 10 polymorphism, either homozygous for the wild type gene, homozygous for the mutant type gene, or heterozygous, was revealed in both patients and healthy individuals. A codon 87 polymorphism, homozygous for the wild type gene, was detected in both groups. No mutations or polymorphisms were found in exon 2. The allele distribution for either codon 10 or 87 between patients and healthy individuals showed no significant differences. In conclusion, genetic alterations in the NH(2)-terminal region of the ERalpha molecule are not correlated with preeclampsia.

Codon↗

PubMed2002

Abdominal delivery of the second twin.

In a series of 220 consecutive twin deliveries, 10 second twins were delivered by caesarean section after vaginal birth of the first, but only six survived. The mean time-interval between the birth of twins was 68 minutes. Because a protracted interdelivery interval carries risks of placental separation and cervical contraction, prompt trial of version of the second twin followed by vaginal breech extraction under anaesthesia might improve perinatal outcome.

Apgar Score↗

PubMed1992

Mammary gland differentiation in hypophysectomized, pregnant mice treated with corticosterone and thyroxine.

Swiss Webster mice were hypophysectomized or sham-operated on Day 11 of pregnancy. The animals were fitted s.c. with osmotic minipumps containing either corticosterone (B) dissolved in Molecusol (Pharmatec, Alachua, FL) or the vehicle alone immediately after they were hypophysectomized. Animals in some of the experimental groups also received thyroxine (T4) in their drinking water. The mice were killed on Day 18 of gestation, and mammary tissue was homogenized and extracted for assessment of DNA, RNA, alpha-lactalbumin, and alpha-casein. Serum was assayed for placental lactogen-I (PL-I), and placental lactogen-II (PL-II), B, and T4. The concentration of PL-II in serum was elevated in the hypophysectomized mice, whereas the PL-I concentration did not differ among experimental groups. Hypophysectomy decreased both T4 and B concentrations in serum, and administration of these hormones restored their serum concentrations to normal or, in some cases, somewhat higher than normal levels. Hypophysectomy reduced the total RNA content and RNA/DNA ratio of the mammary gland, but treatment with B alone or with B and T4 restored RNA levels to those of sham-operated animals. T4 alone was ineffective in restoring RNA levels. Sham-operated animals that received hormonal treatment (B and T4) had the highest levels of RNA in the mammary tissue. Hypophysectomized animals had reduced content and concentration of alpha-lactalbumin in the mammary gland as compared to all other experimental groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1985

Familial hyperinsulinemia complicated by extreme insulin resistance during pregnancy: a probable postreceptor defect.

Detailed studies of a family with hyperinsulinemia are reported. The index patient, a 30-yr-old woman with polycystic ovary syndrome, presented with gestational diabetes which was completely resistant to insulin in the presence of severe endogenous hyperinsulinemia. Sensitivity to insulin was regained after delivery. Therapy with cyproterone acetate and ethinyl estradiol for hirsutism exacerbated the hyperinsulinemia toward the levels occurring in pregnancy, with a concomitant deterioration of glucose tolerance. Five other members of her family also were found to have hyperinsulinemia together with high concentrations of circulating C-peptide. Antibodies to insulin and to insulin receptors were not detected, insulin antagonists were not increased, and insulin degradation in the circulation was normal. Insulin extracted from the patient's serum was identical to normal insulin by the criteria of Sephadex chromatography, placental membrane insulin receptor binding, and stimulation of 2-deoxyglucose uptake in isolated rat adipocytes. Although [125I]insulin binding to erythrocytes of all family members and to the patient's placental membranes was markedly reduced, binding to fibroblast cultures from the patient was normal. Insulin-stimulated glucose transport in these fibroblasts also was normal, but there was a mild (20%) reduction in the concentration of cytochalasin B-binding sites in erythrocyte ghosts. Insulin resistance in this family may be due to a partial defect distal to the insulin receptor. This is asymptomatic unless metabolic stresses (pregnancy or steroid administration) are superimposed.

Adult↗

PubMed1978

Placenta-like alkaline phosphatases from human osteosarcoma cells.

Hormone-induced alkaline phosphatases in human osteosarcoma cells (LM) were extracted and purified. Characterization of the purified enzyme showed two distinct isoenzymes. One isoenzyme was heat labile, was homoarginine inhibited, and had the electrophoretic migration of alkaline phosphatase of human osseous origin. Immunodiffusion showed that this isoenzyme reacted positively only against anti-bone alkaline phosphatase antibodies. The second isoenzyme was heat stable, was inhibited by phenylalanie, and had the same electrophoretic migration as did alkaline phosphatase extracted from mature normal human placenta. This second isoenzyme had the same antigenicity as did the normal placental enzyme. Like the D-variant placental phenotype, this second isoenzyme was inhibited by L-leucine and ethylenediaminetetraacetic acid.

Alkaline Phosphatase↗

PubMed1978

Improved liquid chromatographic determination of placental estriol in urine.

Placental estriol in urine can be determined, after enzymatic hydrolysis, by liquid chromatography. A single-step extraction technique results in rapid processing with minimal sample manipulation. Analytical recovery of estriol added to urine was 96 to 106%. Day-to-day precision (CV), established by 10 replicate analyses at a mean concentration of 15.9 mg of estriol per liter, was 3.7%.

Chromatography, High Pressure Liquid↗

PubMed1993

Complement regulatory proteins in early human fetal life: CD59, membrane co-factor protein (MCP) and decay-accelerating factor (DAF) are differentially expressed in the developing liver.

The human fetus appears to be capable of protecting itself from maternal complement (C) from an early stage in development by expressing the C regulatory proteins decay-accelerating factor (DAF), membrane co-factor protein (MCP) and CD59 on fetally derived trophoblast at the feto-maternal interface. In this study we have examined the ontogeny of these proteins within the fetus itself and have focused on the liver which represents a major site of haemopoiesis during development. Immunostaining revealed that DAF, MCP and CD59 are all expressed from at least 6 weeks of gestation in the liver but that these proteins display distinct distribution patterns. CD59 was broadly distributed both within the epithelial and haemopoietic compartments, but expression of C3 convertase regulators was more restricted. DAF expression was limited to isolated cells within haemopoietic nests and the epithelium was DAF-negative. Although MCP expression on haemopoietic cells was also limited, by contrast with DAF the developing hepatic epithelium was strongly MCP-positive. Typical CD59 and MCP components were observed in fetal liver extracts by immunoblotting, although liver MCP components consistently migrated 4000-5000 MW ahead of those observed on placental trophoblast. Differences in the distribution of these proteins were also observed between the fetal and adult liver. In particular, by comparison with fetal hepatic epithelium, there was an apparent loss of MCP expression from adult hepatocytes. Thus, MCP appears to be developmentally regulated in the human liver and is expressed in the absence of DAF on the early hepatic epithelium. Overall, this study suggests that C regulatory proteins, and in particular CD59 and MCP, are required from the very early stages of gestation within the fetus itself.

Adult↗

PubMed1987

Human endometrium and menstrual fluid contain placental protein 10 (PP10).

Placental protein 10 (PP10), a newly isolated protein from the human placenta, was detected by radioimmunoassay in 10 out of 23 extracts of proliferative and secretory endometrium and in all of the 20 menstrual fluids examined. The concentrations were higher in the secretory phase, when the tissue PP10 content ranged from less than 0.2 to 1.3 microgram/g of endometrial cytosol protein. In menstrual fluid, PP10 was found at concentrations of 16 to 270 micrograms/l. In RIA, the dose-response curves of endometrial, menstrual fluid and purified PP10 were parallel. In gel filtration, the major part of PP10 from the endometrium and menstrual fluid eluted in the same fraction as purified placental PP10. Our results add another placental protein to the list of placental proteins which also appear in the endometrium.

Adult↗

PubMed2001

Determination of 3'-azido-2',3'-dideoxyuridine in maternal plasma, amniotic fluid, fetal and placental tissues by high-performance liquid chromatography.

3'-Azido-2',3'-dideoxyuridine (AZDU, Azddu, CS-87) is a nucleoside analog of 3'-azido-3'-deoxythymidine (zidovudine, AZT) that has been shown to inhibit human immunodeficiency virus (HIV-1). AZDU is a potential candidate for treatment of pregnant mothers to prevent prenatal transmission of HIV/AIDS to their unborn children. A rapid and efficient high-performance liquid chromatography (HPLC) method for the determination of AZDU concentrations in rat maternal plasma, amniotic fluid, placental and fetal tissue samples has been developed and validated. Tissue samples were homogenized in distilled water, protein precipitated and extracted using a C-18 solid-phase extraction (SPE) method prior to analysis. Plasma and amniotic fluid samples were protein precipitated with 2 M perchloric acid prior to analysis. Baseline resolution was achieved using a 4.5% acetonitrile in 40 mM sodium acetate (pH 7) buffer mobile phase for amniotic fluid, placenta and fetus samples and with a 5.5% acetonitrile in buffer solution for plasma at flow-rates of 2.0 ml/min. The HPLC system consists of a Hypersil ODS column (150x4.6 mm) with a Nova-Pak C-18 guard column with detection at 263 nm. The method yields retention times of 6.2 and 12.2 min for AZDU and AZT in plasma and 8.3 and 17.6 min for AZDU and AZT in amniotic fluid, fetal and placental tissues. Limits of detection ranged from 0.01 to 0.075 microg/ml. Recoveries ranged from 81 to 96% for AZDU and from 82 to 96% for AZT in the different matrices. Intra-day (n=6) and inter-day (n=9) precision (% RSD) and accuracy (% Error) ranged from 1.48 to 6.25% and from 0.50 to 10.07%, respectively.

Amniotic Fluid↗

PubMed1996

Occurrence of a growth hormone-releasing hormone-like messenger ribonucleic acid and immunoreactive peptide in the sheep placenta.

Growth hormone releasing factor (GHRH) has been described in the rat, mouse and human placentae. This study reports the presence of an immunoreactive GHRH activity (IR-GHRH) in the ovine placenta. This activity was detected by radioimmunoassay from day 50 (D50) until the end of pregnancy. Higher IR-GHRH concentration in placental tissue was observed on days 100 (543 +/- 123 pg/g) and 140 (550 +/- 62 pg/g) and, when compared with the GHRH content of the ovine hypothalamus (1.2 ng/hypothalamus), represents a considerable amount of GHRH per placenta (a mean of 200 ng). Perifused placenta explants released IR-GHRH in vitro at a mean rate of 200 pg/g/h. Depolarization by 55 mM KCl increased the IR-GHRH concentration of the perifusion media 1.7 times over basal values. The elution position of GHRH immunoreactivity in the gel filtration chromatography profiles was the same for placenta and hypothalamus extracts and lay very near to the molecular weight of bovine GHRH. Northern blot hybridization analysis revealed the existence of a placental transcript whose size (0.75 kb) was comparable to the size of the ovine hypothalamus and rat placenta GHRH transcripts. Hybridization signal was observed at each stage studied from D50 until D120 of pregnancy. This study demonstrated the existence of a IR-GHRH peptide in the ovine placenta.

Animals↗

PubMed1980

Increased glycosylation of serum human chorionic gonadotropin and subunits from eutopic and ectopic sources: comparison with placental and urinary forms.

The role of carbohydrate in the heterogeneity of hCG and its subunits is unclear. To study this question, we chromatographed over Sephadex G-100 an extract of term placenta as well as sera from a woman in the first and third trimesters of pregnancy and sera from two patients with nontrophoblastic malignancies. Samples were cochromatographed with radiolabeled urinary standards. hCG from first trimester pregnancy serum showed multiple peaks on G-100. The dominant peak eluted with an apparent molecular weight (72,000) higher than that of hCG from third trimester serum (63,000), urine (6),000), and placenta (59,000). hCG from both malignancy sera eluted with an apparent molecular weight (62,000) similar to that of hCG from third trimester and urinary hCG. Free hCG alpha from all sera eluted with a similar apparent molecular weight (29,000), which was higher than that of placental and urinary free alpha-subunit (22,000) and the alpha-subunit dissociated from intact hCG from all sources (22,000--23,000). The subunits were dissociated in the denaturing medium of 6 M guanidine-HCl, pH 3.0, and chromatographed in this medium over Sepharose CL-6B. This eliminated all of the differences in apparent molecular weight among corresponding forms that were found on G-100. All forms of hCG alpha coeluted with a chemically identified 80% deglycosylated hCG alpha. hCG and free alpha-subunits were incubated with mixed exoglycosidases which lacked detectable protease activity and were then rechromatographed on G-100. After deglycosylation, hCG from different sources eluted with a considerable heterogeneity (mol wt range, 40,000--50,000) not present in the native forms. Despite the heterogeneity of native free alpha-subunit from various sources, deglycosylation produced a common species with apparent molecular weights of 11,000--12,000, close to the chemically determined molecular weight of the polypeptide chain (10,400). These studies suggest that1) ectopic serum hCG and free alpha-subunit are similar to corresponding eutopic forms; 2) serum hCG and free alpha-subunit from all sources are more glycosylated than placental or urinary forms; 3) first trimester hCG is more glycosylated than other forms of hCG; and 4) serum free alpha-subunit is more glycosylated than the alpha-subunit which combines with hCG beta to form intact hCG.

Carcinoid Tumor↗

PubMedDate not supplied

D-glucose transfer rates derived from single-injection and steady state tracer experiments in the isolated guinea-pig placenta.

In isolated guinea-pig placentae the transfer of D-[3H]glucose and L-[14C]glucose was investigated in single injection (SIE) and steady state (SSE, n = 11, placental weight 4.5 +/- 1.14 g) experiments at constant perfusion flow rates (3 ml/min). In SIE, a mixture of D- and L-glucose was injected as a bolus into either the fetal or maternal side of the placenta, uptake curves were obtained and the maximal extraction values Umax were derived. From these the membrane rate constants Kmc and Kfc of either the maternal or fetal side of the trophoblast membrane were calculated. The specific placental transfer rate constants were computed for the maternal-fetal (Kspec,mf) and fetal-maternal (Kspec,fm) direction from the amount of label transferred to the acceptor side in either SIE or in SSE (where labelled D- and L-glucose had been added to the stock solution). The chemical concentration of D-glucose was changed (5, 50 and 100 mmol/l, n = 11) and it was found that all rate constants decreased with increasing D-glucose concentration. At a D-glucose concentration of 5 mmol/l, Kmc was 3.11 +/- 1.59 ml/min (n = 6) and Kfc 2.69 +/- 0.5 ml/min (n = 5), the combined membrane rate constant (which determines the total placental transfer) was estimated to be 1.44 ml/min. This value was not significantly smaller than the mean (both directions) transfer rate constant Kspec in SIE (1.51 +/- 0.89 ml/min) or SSE (1.52 +/- 0.56 ml/min). Thus the results from uptake and transfer experiments are consistent. From the rate constants D-glucose fluxes at different chemical D-glucose concentrations were estimated. Whereas the mean Km values for all fluxes based on the various rate constants were about 30 mmol/l, the maximal flux Vmax was highest at the maternal trophoblast membrane (159.1 +/- 70.2 mumol/min), it was 107.9 +/- 11.6 mumol/min at the fetal side and 67.3 +/- 7.9 mumol/min in SIE or SSE for placental transfer in both directions. It is concluded that D-glucose carriers predominate at the maternal side of the trophoblast, and that maternal-fetal glucose transfer is proportionate to the transplacental glucose concentration difference within the physiological range of glucose concentrations.

Animals↗

PubMed2003

Interventions for treating placental abruption.

BACKGROUND: Placental abruption is an important cause of maternal and fetal mortality and morbidity. OBJECTIVES: To assess the effectiveness and safety of any intervention for the care of women and/or their babies following a diagnosis of placental abruption. SEARCH STRATEGY: Comprehensive electronic search of the Cochrane Pregnancy and Childbirth trials register. Date of last search: October 2002. SELECTION CRITERIA: Randomised and 'quasi-randomised' trials that report clinically meaningful outcomes and present results on an intention to treat basis. DATA COLLECTION AND ANALYSIS: If eligible trials were to be identified, data will be extracted, unblinded, by the reviewer from all studies. MAIN RESULTS: No studies that met the inclusion criteria were identified. REVIEWER'S CONCLUSIONS: The clinical management of placental abruption has to rely on knowledge other than that obtained through randomised clinical trials.

Abruptio Placentae↗

PubMed2001

PPAR gamma/RXR alpha heterodimers are involved in human CG beta synthesis and human trophoblast differentiation.

Recent studies performed with null mice suggested a role of either RXR alpha or PPAR gamma in murine placental development. We report here that both PPAR gamma and RXR alpha are strongly expressed in human villous cytotrophoblasts and syncytiotrophoblasts. Moreover, specific ligands for RXRs or PPAR gamma (but not for PPAR alpha or PPAR delta) increase both human CG beta transcript levels and the secretion of human CG and its free beta-subunit. When combined, these ligands have an additive effect on human CG secretion. Pan-RXR and PPAR gamma ligands also have an additive effect on the synthesis of other syncytiotrophoblast hormones such as human placental lactogen, human placental GH, and leptin. Therefore, in human placenta, PPAR gamma/RXR alpha heterodimers are functional units during cytotrophoblast differentiation into the syncytiotrophoblast in vitro. Elements located in the regulatory region of the human CG beta gene (beta 5) were found to bind RXR alpha and PPAR gamma from human cytotrophoblast nuclear extracts, suggesting that PPAR gamma/RXR alpha heterodimers directly regulate human CG beta transcription. Altogether, these data show that PPAR gamma/RXR alpha heterodimers play an important role in human placental development.

Cell Differentiation↗

PubMed1984

Synthesis of a fluorescent derivative of glucosyl ceramide for the sensitive determination of glucocerebrosidase activity.

A fluorescent derivative of glucosyl ceramide was synthesized by covalently linking a fluorescent fatty acid, 12-[N-methyl-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)] aminododecanoic acid to the amino group of sphingosyl-1-O-beta-D-glucoside, glucosyl sphingosine. For hydrolysis by glucocerebrosidase, this substrate was dispersed in mixed micelles with Triton X-100 and sodium taurocholate or in unilamellar liposomes with phosphatidylcholine and the negatively charged lipid, dicetylphosphate. In either micellar or liposomal dispersions of the fluorescent substrate, reaction rates were linear with time and protein concentration, and saturation kinetics were observed. The rate of hydrolysis of this fluorescent substrate was equal to that obtained with radiolabeled glucosyl ceramide. The fluorescent glucosyl ceramide was used to determine glucocerebrosidase activity in extracts of human leukocytes, cultured skin fibroblasts, and various tissues as well as in partially purified splenic and placental glucocerebrosidase preparations. This fluorescent derivative of the natural substrate was not hydrolyzed by aryl beta-glucosidase(s), thereby facilitating the specific and reliable diagnosis of heterozygotes and homozygotes with Gaucher disease.

4-Chloro-7-nitrobenzofurazan↗

PubMed1983

Constituents of human meconium--I. Identification of 3-hydroxy-etianic acids.

The monohydroxylated fraction of bile acids of human meconium was analyzed by capillary GC-MS. In the sulfate-glucuronide fraction three saturated, and one unsaturated C20 steroidal acids were found. These acids were identified as 3 alpha-hydroxy-5 alpha-, 3 alpha-hydroxy-5 beta-,3 beta-hydroxy-5 alpha-androstane-17 beta-carboxylic, and 3 beta-hydroxyandrost-5-ene-17 beta-carboxylic based on the unequivocal GC-MS comparison with standards of all possible epimers at C-3, 5 and 17. The amount of the major C20 acid, 3 alpha-hydroxy-5 alpha-androstane-17 beta-carboxylic, in meconium was 0.2 nmol/g, i.e. 5 to 10 times the amount of lithocholic acid. To prevent the oxidation of 21-hydroxy-20-oxopregnanes to C20 acids meconium was extracted in the presence of sodium borohydride. In the absence of this reducing agent the amount of 3 beta-hydroxyandrost-5-ene-17 beta-carboxylic acid was increased and its 17 alpha-epimer could be detected. This indicates partial artifactual formation of this C20 acid from 21-hydroxypregnenolone, which is known to be present in human meconium. The amount of the saturated C20 acids was unaffected by the presence of sodium borohydride in the extraction medium, and their native occurence in human meconium was further confirmed by the absence of their 17 alpha-epimers in extracts obtained both with and without borohydride. The probable metabolic origin of C20 acids in the fetal-placental-maternal unit is discussed.

Androstanols↗

PubMedDate not supplied

Transport of radioactive deoxyglucose by the isolated perfused human placental cotyledon as measured by the paired-tracer dilution method. Inconsistency of the transfer rates across the two trophoblast borders with the simultaneously measured rate of transplacental transfer.

The uptake of [3H]2-deoxy-D-glucose ([3H]2DG) by the maternal and fetal borders of the trophoblast was measured in the dually perfused human placental cotyledon by use of the paired-tracer dilution method. The uptake was estimated from the maximum extraction of the tracer. The transcellular component of transplacental transport of [3H]2DG was measured simultaneously. From the rates of uptake by the two trophoblast borders the rate of transcellular transport was predicted. The rate predicted was significantly lower than the rate observed. This was taken to indicate that the paired-tracer dilution method based on the maximum extraction underestimated the rates of uptake.

Biological Transport↗

PubMed1986

[Cesarean section for placenta praevia. Perinatal mortality and morbidity].

The severity of placenta praevia is well known and responsible of a non negligible perinatal mortality. This study relates to 65 children born after a cesarean section for placenta praevia and analyzes the fetal complications, their frequency, their prognosis as well as the obstetrical factors connected to this problem. Mortality is 3%, prematurity remains high and so is the anemia, but hypotrophy is rare. The main complications are infection essentially due to an overlapping placenta praevia, its hemorrhagic nature and the increasing number of hemorrhagic episodes and respiratory complications (respiratory distress, inhalation of amniotic fluid and hyaline membrane disease). From these fetal data, the authors propose a method of management of low placental insertions consisting essentially of bed rest as soon as the diagnosis in made on the sonogram, and extraction at the 35th week of amenorrhea. In case of hemorrhage, extraction must be performed earlier if obstetrical conditions, maternal as well as fetal, do not permit to undertake a conservative approach aimed essentially to decrease prematurity and its consequences.

Cesarean Section↗