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PubMed1984

Insulin-like growth factor II receptors. Molecular radius and molecular weight determination using quantitative polyacrylamide gel electrophoresis.

High resolution quantitative polyacrylamide gel electrophoresis was employed under nondenaturing conditions to calculate a molecular weight, Mr, for rat placental membrane receptors that bind insulin-like growth factor II (IGF-II). An n-octylglucoside-soluble extract of receptors that had been enriched 20-fold during Sephacryl S-300 gel chromatography and designated peak 1 (Perdue, J. F., Chan, J. K., Thibault, C., Radaj, P., Mills, B., and Daughaday, W. H. (1983) J. Biol. Chem. 258, 7800-7811) was incubated with 125I-IGF-II in the presence or absence of an excess of unlabeled IGF-II and electrophoresed in glass tubes containing highly cross-linked polyacrylamide gels at concentrations ranging from 5 to 12% (w/v). A specifically labeled peak of IGF-II binding activity was identified by freezing, slicing, and counting the gels. The proteins eluted from the region of the gel when cross-linked to 125I-IGF-II with disuccinimidyl suberate and electrophoresed in the presence of sodium dodecyl sulfate have the same molecular weight, Mr, as the previously described IGF-II receptor. From the slope of a plot of log of the relative mobility, RF, at each of 6 to 7 gel concentrations for the 125I-IGF-II-receptor and for each of seven standard proteins (Ferguson-Hedrick plot), a retardation coefficient, KR, was determined. Using a reference curve constructed from a plot of square root KR versus the molecular radius, R, of the standard proteins, the IGF-II receptor was estimated to have a R of 4.13 nm and a calculated Mr of 250,000. Thus, quantitative procedures that separate native proteins based on their size and charge have provided information which is in good agreement with the results of studies of the IGF-II-receptor by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and supports the premise that it is a monomeric single chained glycoprotein, constrained by intradisulfide bonds and with a mass of 250 kDa.

Animals↗
PubMed1996

The effect of placental malaria infection on perinatal mortality in rural Malawi.

Perinatal deaths (fetal or infant deaths from the 28th week of pregnancy up to the seventh day after birth) occur as a result of adverse conditions during pregnancy, labor, and delivery, or in the first few days of life. Placental malaria infection is known to increase the risk of delivery of a low birth weight infant, thus, potentially increasing the risk of perinatal and infant mortality. To better understand the relationship among the adverse events in pregnancy, including placental malaria infection, adverse conditions in labor, and birth weight to perinatal mortality, we investigated the perinatal mortality among a cohort of infants born to rural Malawian women for whom placental malaria infection status and birth weight were documented. Among the 2,063 mother-singleton infant pairs, there were 111 perinatal deaths (53.8 perinatal deaths per 1,000 births). The risk of perinatal death increased as birth weight decreased. Risk factors identified for perinatal mortality among all infants excluding birth weight included abnormal delivery (cesarean section, breech, or vacuum extraction), a history of a late fetal or neonatal death in the most recent previous birth among multiparous women, reactive maternal syphilis serology, nulliparity, and low socioeconomic status. Placental malaria infection was not associated with increased perinatal mortality, but was associated with lower perinatal mortality among normal birth weight (> or = 2,500 g) infants (odds ratio = 0.35, 95% confidence interval = 0.14, 0.92). Interventions to address these risk factors could have a substantial impact on reducing perinatal mortality in this population.

Adolescent↗
PubMedDate not supplied

Placental cadmium and progesterone concentrations in cigarette smokers.

Cadmium and progesterone concentrations were evaluated in term placentas collected from 56 healthy parturients in the city of Zagreb. Concentrations of lead, iron, zinc, and copper in placentas were analyzed. Data collected by questionnaire identified 29 nonsmoking and 27 smoking women. From each placenta, three samples from different locations were taken. Metals were measured by atomic absorption spectrometry. Progesterone was determined by specific radioimmunoassay in homogenized and lyophilized tissue samples after steroid extraction with ethanol. No effect of sample location was found. In placentas of smoking women an increase in cadmium, reduced progesterone and a decrease in iron concentrations were found. Placental copper and zinc concentrations were not altered. In conclusion, the results present new evidence that maternal smoking reduces placental progesterone content and support the established association of smoking with placental cadmium.

Adult↗
PubMed1992

17 beta-hydroxysteroid dehydrogenase gene expression in human breast cancer cells: regulation of expression by a progestin.

The expression of the 17 beta-hydroxysteroid dehydrogenase (17-HSD) gene in a series of human breast cancer cell lines was studied by Northern blot hybridization with a cDNA probe and by a time-resolved immunofluorometric assay using polyclonal antibodies against the enzyme protein. The 17-HSD enzyme protein concentration was measured in the 800 x g cell extract. A high concentration was measured in the BT-20 cell line, corresponding to one-fourth of the average concentration in placental tissue. Western blot analysis indicated that the antigen corresponded to a single Mr 35,000 band. In 2 other cell lines (MDA-MB-361 and T-47D), the 17-HSD protein concentration was much lower, but still measurable, whereas in the remaining 5 cell lines (HBL-100, MCF-7, MDA-MB-231, MDA-MB-468, and ZR-75-1) it was below the detection limit of the assay. Treatment of the cells for 5 days with the synthetic progestin, ORG2058, resulted in an increase of the 17-HSD protein concentration only in the T-47D cell line. By Northern blot analysis, a low level of 2.3-kilobase mRNA transcripts was detected in all 8 cell lines. In addition, a 1.3-kilobase 17-HSD mRNA was present in the samples from the 3 cell lines containing measurable amounts of 17-HSD protein in the cell extract, and the band intensities were proportional to the amount of protein measured with the immunofluorometric assay. Only in the T-47D cell line did progestin treatment correspond to an increased amount of the 17-HSD 1.3-kilobase mRNA. These results suggest that the 1.3-kilobase mRNA for 17-HSD is the form most closely associated with protein expression and is also the only form responding to the progestin induction of the 17-HSD gene.

17-Hydroxysteroid Dehydrogenases↗
PubMed1993

Placental growth hormone levels in normal pregnancy and in pregnancies with intrauterine growth retardation.

To assess the possible role of placental growth hormone (GH) in fetoplacental growth, we measured placental and pituitary GH (GHN) in maternal plasma by means of two RIA using two MAb (5B4 recognizing both placental GH and GHN, and K24 recognizing only GHN) during pregnancy. IGF-I also was measured by RIA in the same samples after extraction. A transverse study of 186 samples obtained between 8 wk of amenorrhea (WA) and term confirmed the reported rise in GH immunoreactivity with 5B4 after 24 to 25 WA from 12.3 +/- 2.0 mU/L (mean +/- SEM) to a plateau of 27.5 +/- 3.4 mU/L at 34 to 35 WA together with the decrease in GHN to undetectable levels by 24 to 25 WA. IGF-I levels increased from 164.0 +/- 44.6 micrograms/L at 24 to 25 WA to 331.6 +/- 63.6 micrograms/L at term. A longitudinal study of 31 normal pregnant women confirmed this hormonal pattern and the reported placental GH plateau after 35 WA. A drastic decrease in placental GH was observed with the onset of labor (from 26.9 +/- 2.1 to 2.7 +/- 1.1 mU/L), whereas the decrease in IGF-I was not significant (from 212.9 +/- 26.5 to 162.4 +/- 16.9 micrograms/L).(ABSTRACT TRUNCATED AT 250 WORDS)

Birth Weight↗
PubMed1987

Transuterofetoplacental conversion of pregnenolone to progesterone in antiestrogen-treated baboons.

The present study was designed to characterize the dynamics of progestin metabolism peripherally and across the uterus during normal baboon pregnancy and to determine whether the decline in placental progesterone (P4) production which results from administration of the antiestrogen ethamoxytriphetol (MER-25) to baboons reflects a decrease in conversion of pregnenolone (P5) to P4 and thus delta 5-3 beta-hydroxysteroid dehydrogenase activity. To examine this possibility, the conversion of [3H]P5 to [3H]P4 was determined by the constant infusion method on day 100 (midgestation) and day 175 (near term) of gestation in baboons that received MER-25 (25 mg/day X kg BW, po) on days 95-100 or 140-175 (term = 184 days). Baboons were sedated with ketamine HCl, then received a constant iv infusion of [3H]P5 (1.0 mu Ci/0.388 ml X min) and [14C]P4 (0.2 mu Ci/0.388 ml X min for 110 min. Radiolabeled progestins were purified from blood samples withdrawn from saphenous, uterine, and umbilical vessels, and the MCR of P4 and P5, uterine extraction of P5, and transfer constants (rho) for the peripheral, transuterofetoplacental, and transuteroplacental conversion of P5 to P4 were determined. The formation of P4 from P5 by incubates of placental cells obtained on day 175 from untreated and MER-25-treated baboons was also assessed. During normal baboon pregnancy the mean (+/- SE) % P5 extracted (i.e. metabolized) by the uterus was 31.0 +/- 3.3 at midgestation and 45.7 +/- 5.6 late in gestation. Peripheral and transuterofetoplacental rho values of P5 to P4 in untreated baboons were 6.9 +/- 1.8% and 37.3 +/- 7.9%, respectively, at midgestation and 6.1 +/- 0.6% and 46.8 +/- 10.1%, respectively, near term. The transuteroplacental rho of P5 to P4 was only slightly lower than the transuterofetoplacental values, indicating minimal conversion of P5 to P4 by the fetus. The peripheral contribution of P5 production to the total production rate of P4 at term in baboons was 1%. The contribution of uteroplacental conversion of P5 to P4 to the total conversion of P5 to P4 at midgestation was estimated to be 22%. MER-25 caused a 53% decline (P less than 0.01) in serum P4 concentrations from a mean (+/- SE) of 12.5 +/- 2.4 ng/ml during the pretreatment period to 5.4 +/- 0.3 ng/ml between days 140 and 175 of gestation.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMedDate not supplied

Structure and permeability of human placental microvasculature.

Endothelial paracellular junctions are important structures for the regulation of vascular permeability, junctional organisation being systematically related to the functional properties of the endothelium. Electron microscopic studies, immunocytochemistry, and single-passage permeability measurements have established that the placental microvessels resemble the fairly tight continuous microvessels of skeletal muscle both in structure and permeability. The endothelial paracellular clefts of these microvessels contain two distinct junctional entities which may influence permeability: the tight junction and the adherens junction. These clefts impose a substantial restriction to molecules above RMM 1000 Da and large haemproteins cannot cross the clefts. The 18 nm-wide zones of the clefts possess the transmembrane adhesion molecules PECAM-1 and VE-cadherin, which have been implicated in junctional assembly and permeability. Inflammatory mediators such as histamine and tumour necrosis factor cause a redistribution of these adhesion molecules to non-junctional regions, and histamine (100 microM) causes a rapid and sustained rise in extraction of radio-labeled tracers. Electron microscopy has also revealed possible first indications of tight junctional disassembly. Both the endothelia of larger placental vessels and isolated placental microvascular endothelial cells express cadherins and PECAM-1 and contain an extensive F-actin cytoskeleton, which is implicated in changes of cell shape and junctional assembly/disassembly. Thus, the human placental endothelium, using perfusion techniques and in vitro experiments, offers a valuable model for vascular permeability studies.

Female↗
PubMed1998

Proton NMR spectroscopy of the phospholipids in human uterine smooth muscle and placenta.

The parameters regulating the fluidity of myometrial and placental phospholipids include double bonds, fatty acid chain lenght and the cholesterol/phospholipid ratio. The transformation of these parameters was studied during pregnancy and labor. Myometrial and placental tissue samples were collected from 24 patients: 6 were nonpregnant, 6 early-pregnant, 6 late-pregnant not in labor and 6 in labor. After butanol extraction, tissue cholesterol and lipid phosphorus were determined. Proton NMR spectroscopy of the phospholipids was performed at 500 MHz. The myometrial cholesterol/phospholipid ratio was slightly elevated in pregnant patients not in labor. The uterine muscle of the nonpregnant patients contained more CH=CH groups in the phospholipids than that of the late-pregnant patients. There were 29 more double bonds in placental than in uterine tissue per 100 fatty acid molecules. The average fatty acid chain length varied from 14.0 to 18.8. The placenta has longer fatty acid chains than the uterine smooth muscle. The myometrial carbon chain was shortened on the average by 1.4 and the placental by 1.0 carbon atoms, when the patient went into clinical labor. These findings suggest fluidity changes in myometrial and placental phospholipids during human pregnancy and labor.

Adult↗
PubMed1975

In vivo evidence for the functional heterogeneity of transferrin-bound iron. II. Studies in pregnant rats.

Iron atoms from the two iron-binding sites of transferrin in the maternal plasma were selectively transferred to fetal tissues across the placenta of the pregnant rat. Rat serum transferrin was selectively double-labeled with 55Fe bound to the A-site and 59Fe bound to the B-site and injected intravenously into pregnant rats at 11 to 20 days gestation. Ratios of 55Fe to 59Fe were measured in various maternal and fetal tissues sampled from 30 to 120 minutes after injection. In the maternal tissues preponderance of A-site 55Fe was observed in circulating red blood cells and in heme extracted from bone marrow and spleen; B-site 59Fe predominated in the liver parenchymal cells and proximal small intestine. As predicted by the Fletcher-Huehns hypothesis, placental transfer of radioiron resulted in a selective concentration of A-site 55Fe in all the fetal tissues including the fetal placenta, yolk sac, whole fetus, and in the separately analyzed liver heme and ferritin and in fetal blood. When the placenta was bypassed by direct injection of the selectively double-labeled transferrin into the umbilical vein, a reversal of the 55Fe/59Fe ratios of heme extracted from the fetal liver was observed. These studies confirm the concept of functional heterogeneity of iron atoms bound to transferrin and indicate that the placenta of rats between the eleventh and twentieth day of pregnancy selectively removes erythroblast-oriented iron from transferrin and diverts it to fetal tissues.

Animals↗
PubMed1985

Distribution of placental protein 14 in tissues and body fluids during pregnancy.

Placental protein 14 (PP14) levels were measured in serum samples from non-pregnant and pregnant women, amniotic fluid, cord blood, and extracts of placenta, decidua and fetal membranes. The levels were low (15-40 micrograms/l) in serum of non-pregnant women. In four pregnancies following in-vitro fertilization, the serum PP14 levels started to rise 2-12 days after embryo replacement. In normal pregnancy, the highest serum PP14 concentrations (up to 2200 micrograms/l) were detected between 6 and 12 weeks. After 16 weeks the level decreased and plateaued at 24 weeks to around 200 micrograms/l. In amniotic fluid, the highest PP14 levels (232 mg/l) were found between 12 and 20 weeks, being considerably higher than those in maternal serum throughout pregnancy. In cord blood, the levels were low (15-22 micrograms/l) or undetectable. In early pregnancy decidua, the PP14 content was higher (41-160 mg/g total protein) than in late pregnancy decidua (60-2700 micrograms/g total protein). In amnion and chorion laeve, the PP14 concentration varied from 50 to 750 and 50 to 1000 micrograms/g protein, respectively. Early pregnancy placenta contained 0.25-15 mg/g and late pregnancy placenta 3-430 micrograms/g protein of PP14. These results show that the levels of PP14 in pregnancy serum have a similar profile to hCG, but in contrast to other placental proteins, the amniotic fluid PP14 levels are remarkably high. This may be explained by suggesting that decidua is a source of PP14.

Amniotic Fluid↗
PubMed1993

Rat amnion: a novel site of oxytocin production.

Fetal membranes, in addition to serving as passive barriers, possess secretory capacities and have a role in amniotic fluid production. In the present study, we demonstrate that rat amniotic/chorionic membranes have the capacity to synthesize oxytocin (OT). Analysis of RNA extracted from rat amnion/chorion membranes by RNA blotting and polymerase chain reaction (PCR) revealed the presence of a single species of OT mRNA. The transcript was smaller than the hypothalamic OT transcript and equaled the size of uterine and placental OT mRNA. The abundance of amniotic OT mRNA decreased by 37% from Day 14 to Day 21 of pregnancy. Over the same time, amniotic/chorionic OT immunoreactivity, as assessed by RIA, increased from 0.17 +/- 0.03 to 1.36 +/- 0.18 ng/g tissue weight. HPLC analysis of amniotic/chorionic membrane extracts revealed two peaks of immunoreactive OT (ir-OT). The first peak (36% of total ir-OT) co-eluted with synthetic OT, while the second peak (64%) corresponded to a noncovalent complex, most likely between OT and neurophysin-I. Using the selective OT receptor ligand, 125I-d(CH2)5 [Tyr(Me)2,Thr4,Tyr-NH2(9)]OVT, we detected high numbers of OT binding sites in rat uterine tissues, but no OT binding sites in amniotic/chorionic or placental membranes. We conclude that the rat amniotic/chorionic membranes represent an extra-hypothalamic site of OT gene expression in the rat during pregnancy and thus represent a possible source of OT present in amniotic fluid.

Amnion↗
PubMed1993

The pattern of respiration with increasing metabolism in a small dasyurid marsupial.

Previous studies have indicated that the pattern of respiration in marsupial and placental mammals may be different. Some marsupials have larger tidal volumes and slower respiratory rates under basal conditions. This study examined the respiratory responses of a small marsupial, Dasyuroides byrnei, to increasing metabolic demand. The highest metabolic rate elicited by cold exposure in a helium-oxygen atmosphere was 10.4 times the basal metabolic rate. Basal tidal volumes and respiratory rates were 138% and 46% respectively of the values predicted for placental mammals. The increasing oxygen demands of metabolism were met by increases in ventilation rather than by changes in oxygen extraction. Initially, tidal volume increased until it reached a maximum value 2.6 times that of basal tidal volume. Subsequently, ventilation was augmented by an increase in respiratory frequency. Ventilatory accommodation to an increasing oxygen demand indicated that D. byrnei has an excellent respiratory capacity to deal with the thermogenic demands of a cold environment.

Animals↗
PubMed1999

Heparin-binding property of human prolactin: a novel aspect of prolactin biology.

Prolactin (PRL) shares several characteristics with growth factors and cytokines, many of which are known to bind to heparan sulfate proteoglycans. In this study we examined the heparin-binding properties of selected members of the PRL/GH family, using heparin affinity columns followed by gel electrophoresis/Western blotting. Purified human PRL and its cleaved 16K fragment, but not human GH or placental lactogen, were retained on the heparin column and were displaced by 0.5 M NaCl. Native PRL in human pituitary extracts and amniotic fluid showed a similar binding affinity to heparin as the purified hormone. None of the other hormones tested, e.g., rat, ovine and bovine PRL, glycosylated ovine PRL or rat GH, bound to heparin. Two consensus heparin-binding sequences are present in human PRL but not in the other hormones included in this study. We postulate that the heparin-binding capability of PRL affects its biological activity as a growth factor and the angiostatic actions of its 16K fragment.

Amniotic Fluid↗
PubMedDate not supplied

Pumpless respiratory assistance using a membrane oxygenator as an artificial placenta: a preliminary study in newborn and preterm lambs.

Newborns suffering from severe respiratory difficulties and not responding to conventional methods have been successfully treated by extracorporeal circulation with a membrane oxygenator (ECMO). However, the technique needs a highly specialized staff, excellent laboratory support, and continuous surveillance of the procedure to prevent complications. In a series of experiments on newborn and preterm lambs, we have investigated a relatively simpler technique of respiratory support that involves a pumpless arteriovenous bypass by cannulating both umbilical arteries and the umbilical vein. A highly efficient microporous membrane oxygenator (MO) with very low resistance was selected. This type of perfusion that mimics the placental circulation, besides providing an additional amount of oxygen to the blood, has proven to be very effective for CO2 extraction. Before its application in humans, however, improvements in the catheters to be inserted in the umbilical vessels, some modifications in the design of the MO, and improvements in the blood compatibility of all foreign surfaces in contact with blood are needed.

Animals↗
PubMed1999

[Scarred uterus: is routine exploration of the cesarean scar after vaginal birth always necessary?].

OBJECTIVE OF THE STUDY: To determine the risks and benefits of routine transcervical revision of previous cesarean uterine scar in patients with successful vaginal delivery. MATERIAL AND METHODS: Retrospective study, over a 10-year period, of a routine palpation practice in the two units of our Obstetric department. Then, a 30-month prospective study comparing, in each unit, two different attitudes toward uterine revision (routine exploration vs symptomatic patients exploration) was conducted. RESULTS: The retrospective part of our study led us to report 3 uterine ruptures (0.43% of all scarred uterus) and 14 dehiscences (2%) during the ten years. All uterine ruptures were sufficiently symptomatic in order to be suspected prior to scar exploration. No dehiscence needed surgical treatment. Some patients with bloodless dehiscence and no repair had subsequent vaginal deliveries with no scar separation found on uterine exploration. In the prospective part of our study, we found a significative difference in the occurrence of fever (18.9% vs 9.9%; p < 0.05) and antibiotic treatment (22.8% vs 12.7%; p < 0.05) between the two groups based on attitude toward uterine revision. CONCLUSION: These data suggest that transcervical revision of previous cesarean uterine scar should be performed only in symptomatic patients (persistent suprapubic pain, placental retention, excessive bleeding during labor or delivery) or when risk factors are present (prolonged labor, prolonged expulsive efforts, instrumental extraction).

Adult↗
PubMed2003

Labour-associated changes in adrenomedullin content in human placenta and fetal membranes.

The aim of the present study was to determine the effects of labour and mode of delivery on human placental and fetal membrane content of adrenomedullin (AdM). Placentas and fetal membranes were collected either at term or pre-term gestation from women either in labour or not in labour, and AdM was measured in tissue extracts by specific RIA. There were significant increases in AdM concentrations in amnion and choriodecidua for the in-labour group compared with the not-in-labour group for both pre-term and term gestations. There was no difference in AdM concentration in placental tissue between labour groups. This study provides evidence that fetal membrane AdM is increased in amniotic and choriodecidual tissues in response to labour, and suggests that it may play a role during human labour.

Adrenomedullin↗