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At least 19 recordsLinked to original sources

Localization and distribution of tissue type and urokinase type plasminogen activators and their inhibitors Type 1 and 2 in human and rhesus monkey fetal membranes.

Fetal membranes consist of 10 distinct layers including components of amnion, chorion and decidua, the latter being of maternal origin. They form mechanically integrated sheets capable of retaining amniotic fluid and play an essential role in protecting fetal growth and development in the pregnant uterus. The extracellular matrix, substrate for plasminogen activators (PAs), is an important supportive framework of the fetal membranes. Fetal membranes from women with preterm premature rupture of membranes may differ in their protease activity compared with normal membranes. To identify the presence of PAs and their inhibitors (PAI) and their possible role in the process of fetal membrane rupture, this study investigated the distribution and localization of both protein and mRNA for tissue (t) and urokinase (u) PA and their inhibitors type 1 (PAI-1) and type 2 (PAI-2) in amniochorion of human and rhesus monkey using conventional and confocal immunofluorescence microscopy. In situ hybridization analysis showed that the distribution and localization of mRNAs for tPA, uPA, PAI-1 and PAI-2 were similar in the fetal membranes of human and rhesus monkey; no obvious species difference was observed. Evidence of tPA mRNA was detected in amniotic epithelium, trophoblast cells and nearly all cells of the decidual layer. Strong expression of uPA mRNA was noted in the decidual cells which increased in intensity as the abscission point was approached. Weak staining in chorion laeve trophoblast was also detected. In situ hybridization experiments showed PAI-1 mRNA to be concentrated mainly in the decidual cells, some of which were interposed into the maternal-facing edge of the chorion laeve. Maximal labelling of the decidua occurred towards the zone of abscission. Weak expression of PAI-1 mRNA was also noted in some cells of the chorion laeve. The distribution of PAI-2 mRNA in amniochorion was also concentrated in the cells of the decidual layer, maximum expression of the mRNA was in the level of abscission. No detectable amount of mRNAs for tPA, uPA, PAI-1 and PAI-2 was found in the fibroblast, reticular and spongy layers. Distribution of the proteins of tPA, uPA and PAI-1 in the fetal membranes of these two species was consistent with the distribution of their mRNA. Anti-PAI-2 immunofluorescence was found to be strongly concentrated in the amniotic epithelium, but PAI-2 mRNA was negative in this layer, suggesting that the epithelium-associated PAI-2 is not of epithelial origin. These findings suggest that a local fibrinolysis in fetal membranes generated by precisely balanced expression of PAs and their inhibitors via paracrine or autocrine mechanisms may play an essential role in fetal membrane development, maturation and in membrane rupture. Following an analysis of the distribution and synthesis of activators and inhibitors it was found that they may play a role in abscission during the third stage of labour.

Adult↗

The effects of a cytokine suppressive anti-inflammatory drug on the output of prostaglandin E(2) and interleukin-1 beta from human fetal membranes.

Fetal membranes are a primary source of prostaglandins and pro-inflammatory cytokines implicated in human parturition, so the inhibition of inflammatory pathways may be of benefit in pregnancies complicated by premature labour. We have therefore investigated the effects of a cytokine-suppressant anti-inflammatory drug (CSAID) on the output of prostaglandin E(2) (PGE(2)) and interleukin (IL)-1 beta from human fetal membranes in vitro. Bacterial endotoxin increased the expression of mRNA for IL-1 beta and type-2 cyclo-oxygenase (COX-2), and there were corresponding increases in the output of IL-1 beta protein and PGE(2). The CSAID decreased IL-1 beta protein, COX-2 expression and PGE(2) output, but not mRNA for IL-1 beta, indicating a post-translational effect on the production of IL-1 beta and a transcriptional affect on COX-2, with an overall reduction in PGE(2). These findings are consistent with the effects of CSAIDs in other systems, and indicate that they are of possible use in premature labour.

Anti-Inflammatory Agents, Non-Steroidal↗

Plasma concentrations of estradiol 17beta and PGF2alpha metabolite and placental fatty acid composition and antioxidant enzyme activity in cows with and without retained fetal membranes.

Fetal membrane retention is one of the most common problems in Holstein cattle after parturition. To investigate mechanisms involved, the following parameters were studied in the peri-parturition period: plasmatic concentrations of estradiol-17beta (E2) and PGFM (PGF2alpha metabolite), activity of antioxidant enzymes (superoxide dismutase-SOD, catalase-CAT and glutathione peroxidase-GSH-Px), thiobarbituric acid reagent substances (TBAR) concentrations and fatty acid composition of the placentae. E2 at parturition in the NPR group (control cows, n = 10) was higher than in PR cows (placental retention, n = 10) (P < 0.05). Activity of SOD in fetal tissue of NPR animals was higher than that of the PR group. In contrast, there was no difference between the two groups in activity of GSH-Px and CAT and the TBAR content of placental tissues. PR maternal tissues had proportionally more arachidonic and linoleic acid than tissues from NPR cows. Therefore, a complex of sequential events may cause placenta retention, starting with an unbalance of antioxidant capacity of the placenta, followed by a decrease in production of estrogen, which leads to the accumulation of arachidonic and linoleic acid in placental tissues.

Animals↗

[Immunohistochemical studies of the nature of connective tissue in fetal membranes].

Fetal membranes at the end of pregnancy have been examined indirectly with the help of FITC labelled antibodies for the occurrence of different collagen types (I, II, III, IV) and their initial steps as well as fibronectin and laminin. The connective tissue of amnion and chorion is composed of collagen type I and p-III. Collagen type IV occurs only rarely. Despite of the borderline reticulin fibres are not the component of these mechanical resistant fibre-mats. In the PAS positive material, seen between the cells of the trophoblast, laminin and type IV collagen were found. It can be stated that the compound of the intercellular substance is of the same kind as the basement membrane.

Amnion↗

[Amniocentesis following premature rupture of fetal membranes and suspected premature rupture of fetal membranes in late pregnancy].

Amniocentesis during the third pregnancy trimenon in case of suspected premature rupture is performed for two reasons: on the one hand, the maturity parameters can be determined as usual from the amniotic fluid, and on the other hand it serves to determine the presence of germs that may have caused the premature rupture. At the same time, it is possible to confirm the suspicion of premature hydrorrhoea gravidarum by instillation of dyestuff into the amniotic cavity. Premature rupture was confirmed in 21 of 65 cases where clinical findings had not supplied safe evidence; in the remaining 44 cases this method permitted the exclusion of premature hydrorrhoea. An evaluation was performed by means of a retrospective study of parturitions between 1975 and 1985 with regard to late amniocenteses in cases of suspected premature rupture, the investigated parameters being the course of pregnancy and of birth, postpartal state of well-being, bacteriological findings in the amniotic fluid, and results of instillation of blue dyestuff.

Amniocentesis↗

Initiation of human parturition. VI. Identification and quantification of progesterone metabolites produced by the components of human fetal membranes.

The fetal membranes play a central role in the initiation of human parturition, a role that may be subserved in part by quantitative changes in progesterone metabolism. In order to identify and to quantify the metabolites of progesterone produced by the components of the human fetal membranes, amnion and chorion laeve tissues and homogenates were incubated with [3H]progesterone in the presence of added NADPH. The radioactive metabolites, 20alpha-hydroxy-4-pregnen-3-one, 5alpha-pregnane-3,20-dione and 3beta-hydroxy-5alpha-pregnan-20-one, were isolated and identified by derivative formation, a combination of chromatographic techniques, and by crystallization to constant specific activity after addition of authentic standards. A simplified technique of metabolite quantification was developed that involves one thin layer chromatographic procedure and liquid scintillation assay of radioactivity. The accuracy of this method was established by comparison with data obtained using classical techniques. This study demonstrates the presence of human fetal membranes 5alpha-reductase, 20alpha-hydroxysteroid oxidoreductase, and 3beta-hydroxy-steroid oxidoreductase, and a qualitative difference between amnion and chorion laeve.

20-alpha-Dihydroprogesterone↗

Fetal membrane distention: determination of the intrauterine surface area and distention of the fetal membranes preterm and at term.

OBJECTIVE: This study was undertaken to obtain an accurate measurement of the intrauterine surface area and the degree of distention of the apposed fetal membranes preterm and at term. STUDY DESIGN: Serial longitudinal images of the uterus in 23 women between 25 and 41 weeks' gestation were obtained by ultrasonography. A Mathematica (Wolfram Research, Inc, Champaign, Ill) program assembled a 3-dimensional image and calculated the intrauterine surface area for each patient. The surface areas of the placental amnion and membranes were measured in vitro after delivery. From these measurements the degree of distention of each fetal membrane in vivo was calculated. RESULTS: The mean calculated intrauterine surface areas were as follows: 1037 +/- 70 cm(2) (25-29 weeks' gestation, n = 4), 1376 +/- 121 cm(2) (30-34 weeks' gestation, n = 4), and 1876 +/- 307 cm(2) (37-41 weeks' gestation, n = 15, P =.0021 by Wilcoxon rank sum test). The surface areas of the expelled membranes at 25 to 29, 30 to 34, and 37 to 41 weeks' gestation were 737 +/- 61 cm(2), 855 +/- 77 cm(2), and 1115 +/- 149 cm(2), respectively. The ratios of intrauterine surface area to the area of the expelled membrane and hence a measure of the degree of distention in vivo were 1.4 +/- 0.05 at 25 to 29 weeks' gestation (n = 4), 1.6 +/- 0.2 at 30 to 34 weeks' gestation (n = 4), and 1.7 +/- 0.3 at term (n = 15). CONCLUSION: The intrauterine surface area in vivo increases during gestation. The surface area of the fetal membranes as measured in vitro increases to a lesser extent. The fetal membranes are therefore distended in vivo.

Amnion↗

Antibiotic inhibition of bacterially induced fetal membrane weakening.

A fetal membrane model was used to evaluate in vitro the efficacy of two antibiotics, erythromycin and clindamycin, in preventing bacterial protease-induced weakening of amniochorion. Standardized inocula of protease-producing bacteria (10(9) colony-forming units [cfu]/mL Staphylococcus aureus, incubated at 37C for 20 hours) reliably reduced fetal membrane structural integrity as reflected by bursting tension and work to rupture. Supraminimal inhibitory concentrations (supra-MICs) (erythromycin 0.23 microgram/mL; clindamycin 0.56 microgram/mL) and subminimal inhibitory concentrations (sub-MICs) (erythromycin 0.13 microgram/mL; clindamycin 0.06 microgram/mL) of both antibiotics prevented fetal membrane impairment due to test bacteria. Supra-MICs of both antibiotics prevented bacterial cell growth and release of protease. Sub-MICs of both antibiotics allowed bacterial cell growth of test microorganisms but inhibited protease release and subsequent fetal membrane damage. These findings suggest that inhibitory and even subinhibitory doses of antibiotics such as erythromycin and clindamycin may be effective in reducing the occurrence of premature rupture of membranes and subsequent preterm birth mediated by susceptible microorganisms.

Clindamycin↗

The endothelin system and renin in human fetal membranes.

Term human fetal membranes express prorenin, a key enzyme within the renin-angiotensin system. High levels of another vasoactive peptide, endothelin-1 (ET-1), are found in human amniotic fluid. To address the question of the relationship between these two vasoactive systems, we analyzed the expression of the components of the ET-1 system in fetal membranes in which cell types had been identified using different markers. Immunohistochemistry was performed with antibodies raised against the human proteins of the ET system. Term fetal membranes displayed ubiquitous labeling of endothelin-converting enzyme-1 (ECE-1) and ET-1. ETA receptors were detected in the chorionic connective tissue and the attached decidua; ETB receptors were localized to chorionic trophoblast cells and decidua. The localization of the ET-1 receptor subtype was confirmed by in-situ receptor binding. Renin immunoreactivity was detected in the chorionic connective tissue and the decidua. These findings suggest that ET-1 is produced ubiquitously in human fetal membranes, and its targets may be, trophoblast cells following ETB receptor activation, vascular structures and fibroblasts in the connective tissue and decidua via ETA and ETB receptors. It appears possible that renin and ET may contribute to the pathophysiological changes associated with premature labor and preeclampsia.

Binding Sites↗

Regional and cellular localization of osteonectin/SPARC expression in connective tissue and cytotrophoblastic layers of human fetal membranes at term.

Fetal membranes overlying the cervix in patients prior to and during labour, and within the rupture tear after spontaneous delivery at term, exhibit altered morphology. In this study we report that in comparison to mid-zone fetal membranes biopsies, these regions are characterized by increased expression of the matricellular protein osteonectin or SPARC (Secreted Protein Acidic and Rich in Cysteine). In the reticular layer, the percentage of vimentin positive mesenchymal cells immunoreactive for osteonectin increased in these regions from 3-4% to 25-33% and represented a fraction of the alpha-smooth muscle actin positive myofibroblasts elevated in the same regions. In the fibroblastic layer, the percentage of osteonectin positive cells increased from 1-5% to 8-13%; however, these did not exhibit the same relationship to the alpha-smooth muscle actin positive myofibroblasts in this layer. In the cytotrophoblastic layer the percentage of cytotrophoblastic cells immunoreactive for osteonectin increased from 1% to 6-12%. Elevation of in-situ detectable mRNA was also observed in the same cellular populations in this region. The incidence of cells positive for osteonectin mRNA or protein in the reticular layer correlated with morphological changes. Osteonectin has been implicated in the regulation of extracellular matrix turnover, and its pattern of expression suggests a role in the regional connective tissue and cytotrophoblastic changes proposed to be involved in the cleavage and rupture of fetal membranes.

Connective Tissue↗

Peripartum changes in the bovine placenta related to fetal membrane retention.

To understand fetal membrane retention in dairy cattle, we examined these tissues in the immediate peripartum period before tissue separation. Placentomes were collected at 270-280 d of gestation from pre-partum Holstein cows (n = 5) and at 1, 3, 6, and 12 h postpartum from a) cows releasing fetal membranes in less than 12 h (n = 7), b) cows retaining fetal membranes for more than 12 h (n = 5), and c) cows induced to calve with dexamethasone (releasing membranes in more than 12 h; n = 5). Subjective evaluations of necrosis, distribution, and condition of binucleate giant and of principal cells were made. Necrotic foci and binucleate giant cells were counted for each interval for which tissue was available. Necrosis, existing prepartum, was identical to that observed at 1 h postpartum in all treatment groups (P > 0.05). Necrosis gradually increased in all treatment groups with time (P < 0.05); thus necrosis is unrelated to initiation of calving and is indirectly related to fetal membrane retention. Many binucleate giant cells were observed prepartum. In cows releasing fetal membranes normally, a significant (P < 0.05) decline in these cell numbers had occurred by 1 h postpartum. When fetal membrane retention occurred, no such decrease in binucleate giant cells was observed before 12 h postpartum. Loss of binucleate giant cells appears necessary for, or must occur with, separation of fetal membranes.

Journal Article↗

Transfer of cytokines through human fetal membranes.

Intact human fetal membranes (amnion, chorion and decidua) were incubated with 125I-labelled cytokines added to the fetal or maternal sides of the membrane. The transfer of 125I-labelled interleukin-6 (IL-6), 125I-labelled tumour necrosis factor alpha (TNF-alpha), 125I-labelled interleukin-1 alpha (IL-1 alpha) and 125I-labelled interleukin-1 beta (IL-1 beta) was determined by measurement of radioactivity in a gamma counter and the integrity of the cytokines was assessed by acid precipitation and by radioimmunoassay. IL-1 alpha and IL-1 beta were transferred through human fetal membranes in both feto-maternal and materno-fetal directions at similar rates. Only 2-4% of the cytokine originally added appeared to be intact on the opposing side of the membrane after 24 h of culture. Transfer of intact TNF-alpha (5-7%) and IL-6 (8-17%) was greater than that of the IL-1 isomers. Low but variable amounts of the four cytokines tested may be transferred through the human fetal membrane. This finding suggested that concentrations of cytokines in amniotic fluid would not reflect those produced by decidua if the fetal membranes are intact.

Biological Transport↗

Evolutionary transformations of fetal membranes and reproductive strategies.

Fetal membranes (such as the chorioallantois and yolk sac) are essential to embryonic development, and have contributed importantly to the evolutionary and ecological diversity of vertebrates. Since the mid-19th century, many scientific careers have been devoted to investigations of their structure, function, and development. However, significant gaps remain in our understanding of the diversity and evolution of fetal membranes. This symposium volume focuses on the use of cladistic principles and phylogenetic relationships to reconstruct the evolutionary morphology of fetal membranes. The main goal of the present paper is to introduce the application of such methods to the evolution of vertebrate fetal membranes, as well as to provide the reader with background information on relevant concepts and terminology. Contributions within this journal issue draw upon studies of metatherian and eutherian mammals, as well as sauropsid reptiles (notably squamates). Particular attention is given to historical transformations of fetal membranes associated with the evolution of such phenomena as placentation and matrotrophy, and reproductive strategies such as viviparity. What emerges from the contributed papers is a broad sampling of contemporary research on fetal membranes, and an overview of how these membranes have evolved to support embryonic life in diverse terrestrial and intra-uterine environments.

Animals↗

Micro-trabeculae, macro-plaques or mini-basement membranes in human term fetal membranes?

Immunocytochemical confocal laser scanning microscopy and ultrastructural analysis, including immunoelectron microscopy, reveals the distribution of structures in human term amniochorion similar in some respects to basement membranes but with unusually restricted dimensions. On the basis of their immunoreactivity, these trabecular structures, found on the fibroblast layer side of the spongy layer of human term amniochorion and adjacent reticular layer, have been shown to contain type IV collagen, laminin, and nidogen. The origin of these components may be from primitive epithelial structures which pumped fluid into the lakes that eventually coalesced to form the extraembryonic coelom separating the extraembryonic somatic mesoderm from the extraembryonic splanchnic mesoderm. Such a theory of their origin might link them with the mysterious 'cellular layer', a single-cell-thick layer of cells which is usually no longer present in fetal membranes at term. The similarity in composition but not in size of these structures to anchoring plaques for type VII collagen is possible support for the view that these structures are integrators of extracellular matrix polymeric proteins. The 'pseudobasement' membrane associated with the trophoblast layer, on investigation, appears to be typical by six criteria. 'Coiled' fibrous structures in the extracellular matrix of the spongy layer may aid adjustments under tension at this shear surface by a detachable 'Velcro' or 'two spring' fastening system. The coils are rich in fibronectin. The suggestion is made that the compact layer is a giant lamina reticularis associated with the amniotic epithelial basement membrane.

Basement Membrane↗

The amnion produces little of the prostaglandin E2 detected on the decidual side of human fetal membranes.

Cultured intact fetal membrane disks initially produced high levels of prostaglandin E2 (PGE2) on the fetal and maternal sides which declined during four days of culture. The transfer of a bolus of PGE2 from the fetal side to the maternal side of the membrane ranged from 1% to 3% after 24 hours of culture, and was a minimum over the period of 48-72 hours from the start of the incubation. To assess the handling of PGE2 synthesized by the amnion, 3H-arachidonic acid was incorporated into cultured amnion and into the amnion side of cultured intact fetal membrane disks. Labelled amnion released 3H-PGE2 on both sides of the tissue, whereas similarly labelled cultured intact fetal membrane only had detectable levels of 3H-PGE2 on the fetal side. It was calculated that no more than 9.7 +/- 1.4% of the PGE2 synthesised by the amnion crossed to the maternal side of the membrane without being metabolised during the transfer through the membrane. These results are consistent with similar indirect methods which suggested that PGE2 from the amnion may have only a limited role in human labor, and indicates the importance of using appropriate culture systems to investigate intra-uterine prostaglandin production.

Amnion↗