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PubMed1997

Pre-labour rupture of membrane: the histological study of membrane and bacteriological profile.

One hundred two (102) cases of pre-labour rupture of membrane (PROM) were studied and special attention was given to the histological study of the amniotic membrane as well as to the bacteriological study of high vaginal flora, cervical flora and flora of amniotic fluid, in search of probable causes or factors leading to PROM. The incidence was found to be 3.16% in the age group of 20-25 years without any relation to parity; and the duration of gestation was 38 to 40 weeks in most of the cases. The histological study revealed: (a) Focally denuded amniotic epithelium, focally separated amniotic epithelium from chorion layer, lesser density of focal squamoid change of the epithelium and thicker chorion layer probably indicating focal immaturity of the chorio-amnion, (b) lesser thickness of collagen layer, focal hydropic degeneration and mild cellular infiltrate, (c) presence of focal hyaline degeneration and focal calcification of chorio-amnion. Microbial culture revealed: (a) Higher rate of positive culture in high vaginal swab, cervical swab and amniotic fluid showing presence predominantly of Esch coli, Strept haemolyticus, klebseilla species, Staph aureus, Strept non-haemolyticus, proteus species and pseudomonas species against that of positive cultures in the control cases, (b) no anaerobic bacteria from high vaginal swab, cervical swab or from amniotic fluid. It was presumed that focal immaturity of chorio-amnion or focal irregularity in the chorio-amnion at the microscopical level, focal degeneration of collagen superadded with bacterial infection, however mild, could be the factors leading to weakness in the tensile strength of chorio-amnion, again leading to PROM, in the face of stress factors of foetal origin.

Adult↗
PubMed1998

Transfer of steroidal and nonsteroidal compounds across guinea pig fetal membranes.

Transfer of steroidal and nonsteroidal compounds across guinea pig amnion and chorion laeve was investigated as a function of stage of gestation, tissue orientation, steroid specificity, and molecular size. Each fetal membrane was examined at early and late stages of gestation, before and after pubic symphysis relaxation. Early amnion was impermeable to macromolecules and small charged molecules while [3H]estrone and [3H]pregnenolone were transferred, the latter depending on tissue orientation and involving conjugation at the basolateral interface. After symphysis dilation, amnion transferred all substrates tested with the exception of BSA; the molecular weight cutoff was approximately 5,000. Unlike amnion, early chorion transferred both free and conjugated steroids as well as inorganic sulfate. Transfer of estrone involved conjugation and depended on tissue orientation. Transfer of [3H]estrone-sulfate, [3H]estrone-glucuronide, and [3H]pregnenolone-sulfate was similar despite selective deconjugating activity toward estrone-sulfate. Near term, chorion was impermeable to inorganic sulfate and transfer of estrone-glucuronide depended on tissue orientation, involving deconjugation in the maternal to fetal direction. At no stage of gestation did chorion transfer macromolecules. These results suggest that the transfer of free and conjugated steroids across fetal membranes is differentially regulated by tissue, its stage of development, and direction of transfer.

Amnion↗
PubMed1985

Distribution and characterization of anionic sites in the basal lamina of developing human amniotic epithelium.

We have studied the distribution of anionic sites in the basal lamina of developing human amniotic epithelium by using the cationic stain ruthenium red. Amnions at 7-12 weeks of gestation and at term contained ruthenium red-positive granules in a quasi-regular array on both the cellular and interstitial sides of the lamina densa. In order to characterize the anionic sites, small pieces of amnion were incubated in the presence or absence of either chondroitinase ABC, neuraminidase, Streptomyces hyaluronidase, or heparitinase in appropriate buffer systems. Incubation in the presence of heparitinase resulted in the complete disappearance of the basal lamina-associated granules, but other enzymes tested had no demonstrable effect on these granules. We conclude that the anionic sites associated with amnion basal lamina, and demonstrable with ruthenium red, consist of glycosaminoglycans rich in heparan sulfate, probably present as heparan sulfate proteoglycan. Because amniotic fluid has a low protein content and amniotic epithelium (at least at term) lacks tight junctions, we postulate that the heparan sulfate proteoglycan associated with the amnion basal lamina may have an important function as a permeability barrier to anionic macromolecules.

Amnion↗
PubMed1982

Hydromineral balance in the chick embryo: effects of hypophysectomy.

The developing chick embryo exhibits a regular pattern of accumulation and reabsorption of fluid in its extraembryonic compartments. Fluid volume (measured by dye dilution) peaks at Day 14 in both the allantois (12.4 ml) and the amnion (5.1 ml). The standard experimental procedure of removing 2 ml of albumen at 33-36 hr of incubation does not alter this pattern, but leads to a maximum decrease in fluid volume of ca. 1 ml in both amniotic and allantoic compartments. Although [Na] and [Cl] remain essentially constant in the serum, they steadily decrease in the allantois (remaining substantially below serum levels). In the amnion, [Cl] is maintained above that of serum, whereas [Na] falls below serum [Na] by Day 14. In the Day 12 (stage 38) embryo, hypophysectomy (HX) causes a substantial increase in the allantoic fluid volume and [Na] and [Cl], thus resulting in a 166% increase in total Na and Cl over that of sham-operated embryos. By contrast, HX did not alter amniotic fluid volume, although it did decrease [Na] and to a lesser degree, [Cl]. Serum [Na], but no [Cl], was reduced by HX. HX abolished the normally steep ionic gradients maintained between the allantois and both serum and amnion. Thus, the most dramatic effects of HX at Day 12 are on the allantois, which accumulates both Na and Cl at the expense of the amnion and serum. This suggests that pituitary hormones may normally act by Day 12 to reduce urinary volume and electrolyte excretion and/or to increase fluid and electrolyte reabsorption from the allantois.

Allantoin↗
PubMed1976

The "stright-jacket" syndrome in chicks. II. Mechanism of development.

In a longitudinal study including measurement of the pressure in the amniotic cavity, amniotomy, and planimetric evaluation of the size of the amniotic sac, we investigated the development of the "strait jacket" syndrome in White Leghorn chicken embryos injected intraamniotically (ia) or paraamniotically on the fourth day of incubation with histone or embryotoxic serum, with the following results. Hyperlordosis and eventration developed as an outcome of tonic contraction of the amnion, which was observed only three hours after ia administration. Contraction of the amnion caused elevation of the intraamniotic pressure, which, 12 hours after ia injection, attained a mean value of 22.4 Pa (2.3 mm H2O). This value was not only significantly higher than the mean for control embryos (3.9 Pa), but it was critically close to the mean fluid pressure in the brain vesicles. Loss of the latter overpressure caused the vesicles to collapse, and the walls shriveled and exencephaly developed. Paraamniotic injection was not followed by either contraction of the amnion, or significant increase in intraamniotic pressure. This did not prevent heart malformations and cranioschisis of various extent. The majority of cardiovascular malformations were probably the hemodynamic consequence of overfilling of the intraembryonic vascular bed, which was one of the early signs of the effect question. Cranial-vault defects can be causally associated with the formation of amnionic adhesion and fusion with the epidermal ectoderm. This observation stresses the significance of the embryonic membranes and the fluid pressures within them for the development of certain congenital deformities and concentrates attention on teratological study of substances that induce protracted contraction of smooth muscle.

Abnormalities, Drug-Induced↗
PubMed1994

Immunohistochemistry of carbonic anhydrase in human placenta and fetal membranes.

The localization of human carbonic anhydrase (CA) isoenzymes HCA I, HCA II, and rat CA II have been studied in human umbilical cord, chorion laeve including amnion and placenta from first and second trimester and also from term pregnancies. Detection techniques of immunofluorescence and immunoperoxidase were used in cryostat and paraffin sections. Both isoenzymes were found in the villous syncytiotrophoblast throughout pregnancy. HCA I staining patterns in the villous endothelium were highly variable whereas increasing immunoreactivity levels of endothelial HCA II were detected as pregnancy advances. The extravillous cytotrophoblast showed generally weaker levels of immunoreactivity. In amnionic epithelium of membranes, chorionic plate and umbilical cord, higher activities for HCA I, HCA II and rat CA II were found than in all other localizations. Our findings emphasize the importance of enzyme mediated bicarbonate/CO2 removal from the feto-placental unit as opposed to simple bicarbonate diffusion or carrier mediated transport. As effective transfer routes should be considered not only umbilical cord--placental villi--intervillous space, but also fetal kidney--amnionic fluid--amnion--uterine vessels.

Amnion↗
PubMed1993

Maternal decidua and fetal membranes contain immunoreactive neuropeptide Y.

The aim of the present study was to evaluate whether the various intrauterine tissues contain immunoreactive neuropeptide Y (NPY). Previous observations showed that human placenta produces NPY and that it may play a local role. Using a polyclonal NPY antiserum and an immunofluorescent technique, sections of maternal decidua, amnion and chorion collected from a pregnant women at parturition were studied. An intense positive staining for NPY was observed in epithelial amnion cells and in chorionic cytotrophoblast. Some of the maternal decidual cells showed a weaker signal of immunoreactive NPY. In evaluating whether NPY may coexist with other hormones in these tissues, adjacent slices of decidua, amnion and chorion were stained with activin beta B subunit antiserum. In the various tissues a relevant number of cells showed positive signals for both NPY and activin. The present findings showed that the various intrauterine tissues contain NPY and that in a large number of cells of amnion, chorion and decidua the neuropeptide is colocalized with immunoreactive activin. In view of the physiological implications of NPY in the regulation of uterine contractility and of placental hormonogenesis, the present findings indicate a large distribution of NPY in the various intrauterine tissues.

Activins↗
PubMed2000

Tribolium embryogenesis: a SEM study of cell shapes and movements from blastoderm to serosal closure.

Embryogenesis in the beetle Tribolium is of increasing interest to both molecular and evolutionary biology because it differs from the Drosophila paradigm by its type of segment specification (short- vs. long-germ) and by the extensive epithelial envelopes - amnion and serosa - that are typical of most insects but not of higher dipterans. Using scanning electron microscopy of DAPI staged embryos we document development in Tribolium castaneum from blastoderm to completion of the envelopes, recording many details not otherwise accessible; we also provide a time table of the respective stages at 30 degrees C. The nascent blastoderm cells remain basally confluent with the yolksac until after the 13th (=last synchronous) mitotic cycle. The cells in the prospective serosa - the first domain to segregate visibly from the uniform blastoderm - carry surface protrusions likely to contact the overlying vitelline envelope. The embryonic rudiment, the other (and larger) blastodermal domain, gives rise to amnion and germ anlage. In the latter, visible differentiation begins with a "primitive pit" reminiscent of the posterior midgut rudiment of Drosophila. The subsequent invagination of the mesoderm resembles Drosophila gastrulation, except in the head region where the median groove extends through the entire preoral region. The prospective amnion starts differing visibly from the germ anlage during early gastrulation. It then folds underneath the spreading serosa and, advancing with the latter, closes the amniotic cavity at the ventral face of the germband. The largest (=posterior) amniotic fold covers a crestlike protrusion of the yolksac. Together with marked changes in the shape and arrangement of the amnion cells, this protrusion may contribute to the fold's elevation and early progress.

Amnion↗
PubMed1983

Phagocytosis and onset of human labor.

An in vitro experimental model has been developed which allows the study of amnion lysosomal enzyme release under controlled conditions. Briefly, a layer of human amnion membrane mounted on a specially designed reaction vessel serves as the reaction surface. We have noted that the addition of particulate material to these membranes incubating in pseudoamniotic fluid results in an increased release of the lysosomal marker enzyme N-acetylglucosaminidase when compared to the release in the absence of particles. This release is completely inhibited by iodoacetate and slightly by azide. A similar increased release is also noted with the use of term amniotic fluid as incubation medium when compared to centrifuged (30,000 g/20 min) amniotic fluid. Lecithin and lysolecithin were effective in releasing increasing amounts of enzyme. This increased release was noted only from membranes of placentas collected from subjects who had undergone cesarean section prior to labor. Membranes collected from vaginal deliveries after labor showed a baseline increased release but no further stimulation upon the addition of any of the substances. These results suggest that the release of lysosomal enzymes from amnion membranes is brought about by substance(s) present in amniotic fluid. Very probably, these are surfactants. The interaction of these substances with amnion cells would eventually result in an exponential burst of prostaglandin synthesis, which would result in labor.

Acetylglucosaminidase↗
PubMed1987

Amniotic basement membrane: a barrier to neutrophil invasion.

Polymorphonuclear neutrophils activated by N-formylmethionyl-leucylphenylalanine are able to cross the human amnion as shown by Russo et al. (1981). With the same technique, we added polymorphonuclear neutrophils to either the epithelial or stromal surface of the amnion. The lower compartment of the incubation chamber contained 10(-8) mol/L of N-formylmethionyl-leucylphenylalanine. Of 24 amnion tested, only one showed polymorphonuclear neutrophil permeation. Ultrastructural observations indicated that the basal lamina and the zona reticularis of the basement membrane acted as barriers to polymorphonuclear neutrophil invasion. Recent studies indicate that the basal lamina and the zona reticularis of the basement membrane contain collagen type V. No data support the existence of collagenase of polymorphonuclear neutrophil origin that cleaves collagen type V. If enzymatic degradation of extracellular matrix components play a role in polymorphonuclear neutrophil invasion, it is possible that type V collagen acts as a barrier to polymorphonuclear neutrophil invasion in the human amnion.

Amnion↗
PubMed1995

Levels of hepatocyte growth factor in maternal serum and amniotic fluid.

OBJECTIVE: The purpose of our study was to investigate hepatocyte growth factor levels in maternal serum and amniotic fluid during pregnancy. We also demonstrated production and secretion of hepatocyte growth factor by placenta and amnion at different stages of gestation. STUDY DESIGN: Hepatocyte growth factor levels in maternal serum (n = 219), cord blood (n = 20), and amniotic fluid samples (n = 90) were measured by an enzyme-linked immunosorbent assay. The secretion of hepatocyte growth factor by placenta and amnion was evaluated by measuring the amount released into the culture supernatant. RESULTS: Most hepatocyte growth factor levels in maternal serum were below the detection limit before 10 weeks of pregnancy. Levels increased significantly thereafter and continued to increase until term. On the other hand, levels in amniotic fluid were significantly higher between 20 and 29 weeks of gestation than after 30 weeks. Hepatocyte growth factor secretion from the placental tissue per weight seemed unchanged throughout pregnancy. Its secretion from amnion was, however, approximately 300 to 400-fold higher in the second trimester compared with that at term. CONCLUSION: Both placenta and amnion produce and secrete hepatocyte growth factor, suggesting its role in fetal growth and the growth and differentiation of placenta.

Adult↗
PubMed1990

Oxytocin receptor in human fetal membranes at term and during labor.

Human fetal membranes, taken from 30 patients submitted to caesarean section during the final stages of gestation and labor, were examined in order to evaluate the presence and characteristics of the oxytocin receptor. The presence of oxytocin receptors in human fetal membranes, both in the amnion and in the chorion-decidua, was demonstrated in this study. The receptor binding to oxytocin showed a significant increase during early and advanced labor compared with before the onset of labor. When the pre-labor level was taken as the normalized form (control = 100) the increase with respect to the control (10 cases) for the amnion in early labor (2.27 times +/- 0.11, mean +/- SEM, P less than 0.001, 10 cases) and in advanced labor (2.53 times +/- 0.15, 10 cases, P less than 0.001) was highly significant. In the chorion-decidua the increase was 1.61 times +/- 0.09, P less than 0.001 in early labor and 1.66 times +/- 0.19, P less than 0.001 in advanced labor. Scatchard analysis showed a single receptor site for oxytocin in amnion and chorion decidua. The dissociation constant (Kd) did not change during the various stages of labor; the mean values found were 0.228 +/- 0.02 (mean +/- SEM) nM in the amnion and 0.193 +/- 0.03 nM in the chorion-decidua respectively. These findings suggest that human fetal membranes are target organs for oxytocin and that they might play a role in the onset of labor through an increase of receptor binding.

Amnion↗
PubMed1984

Metallothionein-like proteins in human placenta and fetal membranes.

Metallothionein (MT) levels were measured in the term human placenta, chorion, and amnion by the Cd-radioassay method. MT concentrations in the amnion and chorion were 8 to 9 micrograms MT/g tissue while placental levels were approximately half this value. For individual subjects a significant correlation (r2 = 0.96) was found for placental and chorionic MT, while amnionic MT levels did not correlate with either placental or chorionic MT. Gel filtration and uv spectral analysis confirmed the presence of a MT-like protein in cytosol obtained from amnion, chorion, and placenta. MT levels in trophoblasts cultured from chorion were increased upon exposure in vitro to either Cd or Zn.

Amnion↗
PubMed1983

Estradiol-17 beta and 2-hydroxyestradiol-17 beta-induced differential production of prostaglandins by cells dispersed from human intrauterine tissues at parturition.

Prostaglandin (PGE, 6-keto PGF1 alpha) output by cells dispersed from human amnion and decidua in the presence of increasing levels (0-5000 ng/ml) of estradiol-17 beta (E2) or 2-hydroxyestradiol-17 beta (2-OH E2) was studied in relation to parturition. Tissues were obtained from women at term either before (CS) or after (SL) spontaneous labor and vaginal delivery. In the absence of estrogens, the output of both PGs from amnion increased significantly with labor. No significant increase in decidua PG output occurred with labor. Neither estrogen influenced CS amnion PG output. However, both E2 and 2-OH E2 stimulated SL amnion PGE output (2-OH E2 greater than E2) while having no affect on 6-keto PGF1 alpha output. Only the highest dose of 2-OH E2 stimulated PGE output in CS decidua, but both estrogens significantly inhibited 6-keto PGF1 alpha output in this tissue. In SL decidua only 2-OH E2 significantly stimulated PGE, and neither estrogen affected 6-keto PGF1 alpha output. These results might suggest that estrogens modulate PG biosynthesis at the level of endoperoxide to primary PG conversion.

6-Ketoprostaglandin F1 alpha↗
PubMed1994

Progesterone metabolism by guinea pig intrauterine tissues.

Progesterone metabolism by guinea pig amnion, chorion, myometrium, and endometrium was studied at the following gestational stages. Day 45 represents mid-gestation, about 5 days before strong chorion interaction between the entire surface of the chorion and the uterus; days 57-58, 1-2 days after chorion attachment, and 2-3 days before the onset of pubic symphysis relaxation; days +1-+6, 1-6 days after the onset of pubic symphysis relaxation, i.e. within 1 week of parturition. The high metabolic activity of chorion exceeded that by amnion at all stages. Metabolism by endometrium and myometrium was always low. Conversion of progesterone by amnion significantly decreased (P < 0.05) between days 57-58 and days +1-+6. Progesterone metabolites produced by chorion and amnion were identified by TLC, HPLC, and capillary GC/MS. Both tissues converted progesterone to three major products during 60-min incubations. These were 5 alpha-pregnane-3,20-dione, 3 alpha-hydroxy-5 alpha-pregnan-20-one, and 3 beta-hydroxy-5 alpha-pregnan-20-one. The metabolite pattern differed between the two tissues. Three-minute incubations with chorion resulted in a significantly higher proportion of 3 alpha-hydroxy-4-pregnen-20-one (P < 0.01) and 5 alpha-pregnane-3,20-dione (P < 0.025) than at 60 min. The production of 3 beta-hydroxy-5 alpha-pregnen-20-one by chorion decreased (P < 0.05) between days 50-51 and 57-58. The ratio of 3 alpha-hydroxy-5 alpha-pregnan-20-one to 3 beta-hydroxy-5 alpha-pregnan-20-one increased (P < 0.05) between days 45 post-relaxation. The marked conversion of progesterone by chorion, or the formation of one or more of its metabolites, may serve to influence uterine function prior to delivery.

Amnion↗
PubMed2005

Cloning and cellular expression of aquaporin 9 in ovine fetal membranes.

OBJECTIVE: Amniotic fluid (AF) absorption across fetal membranes is essential for AF volume homeostasis, balancing fetal swallowing, urine flow, and lung liquid production. In sheep, AF is absorbed primarily across the amniotic membrane into fetal vasculature situated between the amnion and chorion. Aquaporins (AQPs) are cell membrane proteins that serve as water channels. Recent studies have demonstrated the expression of AQP 1, 3, 8, and 9 in human chorioamniotic membranes and placenta. As AF dynamics continued to be explored primarily in the ovine model, we sought to clone and characterize the expression of ovine AQP9 in fetal membranes. METHODS: Ovine AQP9 gene was cloned with the use of homology reverse transcriptase-polymerase chain reaction (RT-PCR). RT-PCR and Northern analysis were used to determine AQP9 gene expression, and immunohistochemistry (IHC) used to localize AQP9 protein expression in ovine fetal membranes. RESULTS: A 2085-base pair (bp) full-length complementary DNA (cDNA) sequence of ovine AQP9 was cloned. The ovine AQP9 cDNA is 86%, 82%, and 82%, and the predicted amino acid sequence (295 amino acids) is 77%, 71%, and 69% identical to human, rat, and mouse AQP9, respectively. RT-PCR and Northern analysis detected AQP9 messenger RNA expression in ovine amnion and allantois, but not in placenta, chorion, or umbilical cord. Immunohistochemistry localized AQP9 protein in epithelia of amnion and allantois. CONCLUSION: The presence of significant AQP9 messenger RNA and protein expression in ovine fetal membranes suggests that AQP9 may be a major water channel for intramembranous AF resorption in sheep. The cloning of ovine AQP9 and the demonstration of AQP9 expression in amnion and allantois significantly enhances our understanding of ovine AF regulation and offers the potential for therapeutic approaches for the treatment of oligohydramnios and polyhydramnios.

Allantois↗
PubMed1996

Stretching of fetal membranes increases the concentration of interleukin-8 and collagenase activity.

OBJECTIVE: The aim of this study was to determine whether stretching of fetal membranes can increase interleukin-8 concentration and collagenase activity. STUDY DESIGN: Strips of whole fetal membranes, amnion, or muscles of the lower uterine segment were stretched for 2 or 4 hours. Interleukin-8 and collagenase activity were measured in homogenized control and stretched samples. Immunohistochemical staining for interleukin-8 was carried out. RESULTS: The interleukin-8 concentration increased significantly after the whole fetal membranes were stretched for 2 and 4 hours (p <0.0007 and 0.001, respectively). Also, stretching of amnion and muscles of the lower uterine segment for 2 and 4 hours increased the concentration of interleukin-8 significantly (p <0.0007 after 2 and 4 hours, respectively). Collagenase activity was significantly increased after stretching of amnion, amniochorion, and muscles of the lower uterine segment for 4 hours (p <0.0007, 0.006, and 0.0007, respectively). After stretching, samples were darkly stained for interleukin-8 compared with control nonstretched samples. CONCLUSION: Stretching of amnion, amniochorion, and muscles of the lower uterine segment increased interleukin-8 production and collagenase activity.

Amnion↗
PubMed1998

Inhibition of amniotic prostaglandin E release by ampicillin.

OBJECTIVE: The effect of antibiotics in the prevention of preterm labor needs to be further investigated. The aim of this study was to determine the effect of ampicillin on prostaglandin E release from amnion as a possible explanation for its ability to retard preterm labor. STUDY DESIGN: The effect of the beta-lactam antibiotic ampicillin on prostaglandin E release from human amnion was tested under basal and stimulated conditions. RESULTS: Ampicillin dose dependently inhibits basal prostaglandin E release from amnion in both static and dynamic conditions. In our experiments, 10(-7) mol/L ampicillin (a concentration able to significantly inhibit prostaglandin E output) leaves the microbiologic features of the medium substantially unmodified up to 5 hours of incubation. Moreover, the drug reversibly counteracts the prostaglandin E elevation induced by arachidonic acid or oxytocin. CONCLUSION: This finding (i.e., that ampicillin inhibits prostaglandin E release from amnion) may offer an explanation for a beneficial response to ampicillin therapy in the case of preterm labor even in the absence of bacterial infection.

Amnion↗