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Mechanical failure of human fetal membrane tissues.

Mechanical integrity of the chorioamnion membrane, and the component chorion and amnion layers, was assessed with biaxial puncture testing. Fetal membranes were obtained from term placentas following labored natural delivery or scheduled cesarean section. Preterm specimens were obtained from deliveries prior to 37 weeks gestation. Dividing and peripheral membranes were obtained from multiple gestation pregnancies. Specimens were gripped between parallel plates with circular openings and loaded with an instrumented, hand-held blunt probe until rupture occurred. Peak force was recorded and rupture sites were examined. Defects in multi-layered membranes differed in both size and shape in the individual layers. Compared with chorion and whole chorioamnion, amnion was more mechanically sensitive to different obstetrical conditions. Amnion varied in response at different physical locations within the same patient. Membrane and component puncture force data were used to calculate biaxial failure strength. Membrane stresses arising from amniotic fluid pressure were computed as a function of gestational age, and compared to membrane strength to examine the criterion for membrane failure in vivo. Possible mechanical conditions for preterm membrane rupture were examined.

Amnion↗

Study of human epithelial cell detachment and damage: development of a model.

Detachment of epithelial structures from underlying basement membrane (BM) represents an important component of a number of human disease processes e.g. airway and alveolar diseases, gastrointestinal ulceration, and retinal diseases. This study describes a method of evaluating human epithelial cell detachment from BM that is simple, rapid, inexpensive, quantifiable and which, because it utilizes BM rather than tissue culture plastic, more closely mimics the in vivo situation than other methods. In this model human amnionic epithelial cells attached to their underlying BM are isolated from fresh placentae and mounted in a multi-well chemotaxis assembly. These membranes can be studied with the epithelial cell monolayer intact. Protease-induced detachment of the epithelial cells from the underlying BM was readily quantifiable using light microscopy and spectroscopy. Following removal of the native amnionic epithelial cells, immunoperoxidase staining for the BM attachment proteins laminin, fibronectin, and type IV collagen demonstrated that these molecules remain intact. The BM could also be used as an attachment surface to reconstitute other epithelial cell monolayers. Cultured human amnionic cells and human respiratory epithelial cells were both able to attach to the denuded BM in the absence of serum (% attachment = 85 +/- 15% and 92 +/- 8% respectively, P = 0.8). Natural BM was a better substrate for epithelial cell attachment than tissue culture plastic in that, in the absence of serum, cultured epithelial cell attachment to tissue culture plastic was 20 +/- 4% of the value for BM (P less than 0.05). Furthermore, cells attached to plastic adhered less effectively than to BM in that trypsin concentrations required to induce 50% cell detachment were 0.72 +/- 0.4 for plastic and 62 +/- 13 BAEE U/ml for BM (P less than 0.001). In view of the complex protein interactions known to be involved in the anchorage of human epithelial cells to BM, it is likely this model will be a useful tool for evaluating the mechanisms underlying human epithelial cell attachment and detachment in a variety of normal and disease situations.

Amnion↗

Activin A exerts both pro- and anti-inflammatory effects on human term gestational tissues.

There is a growing appreciation of the importance of activin as a modulator of immune function. The aim of the present study was to determine whether activin A exerts any effects on cytokine and prostaglandin (PG) production by the tissues of pregnancy. Explant cultures were established for amnion, choriodecidual and placental tissues derived from pregnancies delivered at term by Caesarean section (n=5 placentae). Explants were treated with activin A (0.5, 5 and 50 ng/ml) in serum-free Ham's F12/DME media for 24 h (n=3-4 replicates). Production rates of interleukin (IL)-1beta, IL-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and PGE(2)were determined using immunoassay. Differences between treatment groups were analysed by ANOVA followed by Dunnett's test;P< 0.05 was considered to be significant. Amnion IL-6 production exhibited biphasic responses to activin A: at 5 ng/ml activin A, IL-6 production was significantly stimulated (to 246+/-74.6 per cent of control (mean+/-sem), while at 50 ng/ml it was significantly inhibited (to 46+/-7.4 per cent of control). IL-8 and PGE(2)production by amnion showed significant responses to activin A that were similar to those of IL-6. No significant effects of activin A were observed on choriodecidual and placental IL-6, IL-8 and PGE(2)production. However, TNF-alpha production was significantly inhibited by 50 ng/ml activin A in both choriodecidual and placental explants (to 43+/-9.7 per cent and 51+/-6.7 per cent of control, respectively). Placental IL-1beta production was not altered by treatment with activin A at any concentration. These findings support the concept of activin as an immune modulator in tissues of pregnancy.

Activins↗

Lysyl oxidases: expression in the fetal membranes and placenta.

The cross-linking of the connective tissues in the fetal membranes and placenta is important for their tensile strength and elasticity. We have studied the expression of lysyl oxidase (LOX) because it is the classical enzyme responsible for the cross-linking of collagen and elastin. We have also studied the two recently described, genetically distinct lysyl oxidase-like genes and proteins, lysyl oxidase-like (LOXL) and lysyl oxidase-like 2 (LOXL2), of unknown functions. Specific antisera have been used for immunolocalization in fetal membranes and placentae from early pregnancy terminations and after caesarean section at both preterm and term, prior to labour. In addition, the steady state mRNA levels of the three genes has been quantitated in separated amnion, chorion, decidua and placentae collected at term before labour. The immunocytochemistry shows that the spatial expression of the three lysyl oxidases is similar in early pregnancy in both the fetal membranes and placentae. However, by preterm this pattern had diverged and becomes greatest at term. The expression of the genes found at term was similar to the results of protein expression obtained by immunocytochemistry, with the exception of LOXL which had high placental gene expression, but low levels of immunolocalized protein. Thus by term, LOX was expressed predominantly in the amniotic epithelium, with little expression in the placenta, while LOXL showed highest gene expression in the placenta and lowest expression in the amnion. LOXL2 expression was again different and was expressed predominantly in the chorionic cytotrophoblast of the membranes with low expression in both the amnion and placentae. These results suggest that these three members of the lysyl oxidase family may have similar roles in early pregnancy during the development of the placenta and fetal membranes, but their divergence as pregnancy advances to term, may reflect changes in substrate specificity and connective tissue composition.

Abortion, Legal↗

Fetal esophageal ligation induces expression of vascular endothelial growth factor messenger ribonucleic acid in fetal membranes.

OBJECTIVE: Obstruction of the fetal esophagus does not always produce the expected polyhydramnios. This is because of increased intramembranous absorption of amniotic fluid into the fetal circulation. A possible mediator for this increased absorption is vascular endothelial growth factor (VEGF). The present objective was to explore whether VEGF gene expression and action would be induced in fetal membranes and placentas of ovine fetuses after esophageal ligation. STUDY DESIGN: Five late-gestation fetal sheep underwent esophageal ligation and 5 served as control animals. On postoperative day 9, amnion, chorion, and placenta were collected for cellular localization and quantitation of VEGF messenger ribonucleic acid by in situ hybridization and Northern blot analysis. Reverse-transcription polymerase chain reaction was used to identify the VEGF molecular forms. Immunostaining with Ki-67 antibody was used to determine the proliferation of vascular endothelium in the fetal membranes and placentas. RESULTS: VEGF messenger ribonucleic acid was localized in amniotic epithelium, chorionic cytotrophoblast, and cytotrophoblast of the placenta. VEGF164 was the major transcript expressed in these tissues. The abundance of VEGF messenger ribonucleic acid in the amnion and chorion, but not in the placenta, was significantly increased in the ligated fetuses in comparison with the control fetuses. The proliferation of the intramembranous blood vessel endothelium was greater in the ligated fetuses than in the control fetuses. CONCLUSION: The levels of VEGF messenger ribonucleic acid and the proliferation of vascular endothelium in the amnion and chorion increased after fetal esophageal ligation. This provides a possible mechanism for the enhanced intramembranous absorption of amniotic fluid through increased vascularity and permeability of the fetal membranes, thus ameliorating the development of polyhydramnios. We speculate that the signal(s) that mediate the increase in VEGF expression is present in either the fetal urine or the fetal lung secretions, or both.

Amnion↗

Proteoglycans and hyaluronan in human fetal membranes.

OBJECTIVE: The aim of this study was to describe the distributions of major extracellular matrix components, such as proteoglycans, collagen and hyaluronan, in the fetal membranes at term. STUDY DESIGN: Fetal membranes were obtained from elective cesarean deliveries at term. Guanidinium extracts were analyzed for proteoglycans with alcian blue precipitation, sodium dodecyl sulfate- polyacrylamide gel electrophoresis, and Western blotting and for hyaluronan with a radioimmunoassay. Collagen was measured by estimating hydroxyproline content. Tissue sections were immunostained for decorin and biglycan and stained for hyaluronan with a biotin-labeled hyaluronan-binding protein. RESULTS: The fetal membranes contained predominantly smaller proteoglycans, such as biglycan and decorin. The amnion consisted of typical fibrous connective tissue with a high concentration of collagen. The amnion was dominated by decorin located in close connection with the collagen fibrils. The chorion was composed of a fibroblastic part containing collagen and decorin and a trophoblastic part mainly containing biglycan. In addition, large amounts of hyaluronan were found, especially in the amnion and in the decidual cell layers. CONCLUSION: The distributions of proteoglycans, collagen, and hyaluronan in human fetal membranes may explain the biomechanical properties of this tissue. We suggest that changes in the relative proportions of these extracellular molecules are crucial for the proposed maturation process in the fetal membranes during the last weeks of pregnancy.

Alcian Blue↗

Expression of aquaporin-8 in human placenta and chorioamniotic membranes: evidence of molecular mechanism for intramembranous amniotic fluid resorption.

OBJECTIVE: The intramembranous pathway of amniotic fluid absorption has recently been recognized as a critical regulatory path for amniotic fluid resorption, which contributes importantly to amniotic fluid volume homeostasis. Yet, the underlying molecular and cellular mechanisms for water absorption across the amniotic membranes remain unknown. We hypothesize that amniotic fluid water is absorbed across aquaporin water channels in the chorioamniotic membranes. Recently the water channel aquaporin-8 complementary DNAs for mice, rats, and humans have been cloned; and aquaporin-8 has been found to be expressed in the placenta and in other organs. The objective of the present study was to determine whether aquaporin-8 is expressed in human chorioamniotic membranes. STUDY DESIGN: Freshly frozen human amnion, chorion, placenta, and umbilical cord of a normal term pregnancy were used in this study. Reverse transcriptase-polymerase chain reaction was used to determine the aquaporin-8 gene expression. In situ hybridization with biotin-labeled aquaporin-8 riboprobe was undertaken to localize the cellular expression of aquaporin-8 gene in these human fetal membranes. RESULTS: Using reverse transcriptase-polymerase chain reaction, we have demonstrated that aquaporin-8 is expressed in human amnion, chorion, and placenta. Further studies of in situ hybridization of aquaporin-8 identified aquaporin-8 gene expression in epithelial cells of chorion and amnion and of the syncytiotrophoblasts and outer layer trophoblasts of placenta. CONCLUSION: This is the first study that demonstrates the expression of aquaporin-8 water channel in human chorioamniotic membranes. These results suggest that aquaporin-8 may be a water channel that mediates amniotic fluid resorption by way of the intramembranous pathway.

Amnion↗

Pre-B-cell colony-enhancing factor, a novel cytokine of human fetal membranes.

OBJECTIVE: Our purpose was to determine whether pre-B-cell colony-enhancing factor (PBEF) is expressed in the human fetal membranes during normal gestation and parturition in the absence of infection and to show its effects on the expression of interleukin (IL)-6 and IL-8. STUDY DESIGN: PBEF was immunolocalized in the fetal membranes from early pregnancy, at preterm, and at term. Its expression was quantitated by Northern analysis in separated uninfected amnion, chorion, decidua, and placenta of patients at term before labor and in full-thickness membranes before and after spontaneous labor at preterm and at term. Amnion-like epithelial (WISH) cells and fetal membrane explants were treated with recombinant PBEF (rhPBEF), and the expression of IL-6 and IL-8 was quantitated. RESULTS: PBEF was immunolocalized throughout gestation in the amniotic epithelium and mesenchymal cells as well as the chorionic cytotrophoblast and parietal decidua. Northern analysis showed significantly more (P <.01) PBEF expressed in the amnion than in either chorion or placenta. Its expression increased after labor at both preterm and term and correlated with that of IL-8 (r = 0.87). rhPBEF treatment of WISH cells significantly increased IL-6 (P <.05) and IL-8 (P <.01) gene expression after 4 hours and of IL-8 protein after 24 hours (P <.01); similar 4-hour treatment of fetal membrane explants significantly increased IL-6 (P <.01) and IL-8 (P <.05) gene expression. CONCLUSION: PBEF is a novel cytokine constitutively expressed by the fetal membranes during pregnancy. It increased the expression of IL-6 and IL-8 and may be important in both normal spontaneous labor and infection-induced preterm labor.

Amnion↗

Regulation of amniotic fluid volume by intramembranous absorption in sheep: role of passive permeability and vascular endothelial growth factor.

OBJECTIVE: During long-term intravascular fluid infusion in the ovine fetus, a large increase in fetal urinary flow rate occurs while amniotic fluid volume increases only slightly because of increased intramembranous absorption. The current study tested the hypotheses that passive intramembranous permeability increases in response to fetal intravascular saline solution infusion and that the increased intramembranous absorption occurs in parallel with an increase in vascular endothelial growth factor gene expression in the amnion, chorion, and placenta. STUDY DESIGN: Chronically catheterized fetal sheep that average 126 +/- 1 (SE) days of gestation either were infused intravascularly with 7 L of normal saline solution over 3 days (n = 8 sheep) or served as time controls (n = 6 sheep). Amniotic fluid volume and fetal urinary flow rate were measured daily. Intramembranous diffusional permeability was estimated daily as being equal to the clearance of intra-amniotically injected technetium 99m. Vascular endothelial growth factor messenger RNA abundance in the amnion, chorion, and placenta was determined by Northern blot analysis. Statistical analyses included analysis of variance. RESULTS: In the infused fetuses, amniotic fluid volume and urinary flow increased (P <.01) by 891 +/- 144 mL and 3488 +/- 487 mL per day, respectively, on infusion day 3 compared with no changes over time in the control fetuses. In the infused fetuses, estimated intramembranous absorption increased by 4276 +/- 499 mL during the 3-day infusion. Intramembranous technetium 99m permeability was similar over time in the two groups. In the infused group, vascular endothelial growth factor messenger RNA levels in the amnion, chorion, and placenta increased 2- to 4-fold compared with the control group (P <.001). CONCLUSION: The up-regulation of vascular endothelial growth gene expression may mediate the increase in the intramembranous absorption that is induced by volume-loading diuresis; however, this does not occur by passive mechanisms. We speculate that vascular endothelial growth mediates the increased intramembranous absorption by increasing vesicular transport.

Absorption↗

Significant decrease in parathyroid hormone-related protein concentrations in amniotic fluid with labour at term but not preterm.

It has been determined whether amniotic fluid concentrations of parathyroid hormone-related protein (PTHrP) change with labour. An evaluation of which cells from intrauterine tissues might produce PTHrP has also been conducted. Amniotic fluid was obtained by amniocentesis from women: (1) at term, not in labour; (2) in normal term labour; (3) in preterm labour, undelivered within one week; (4) in preterm labour, delivered within one week; (5) in preterm labour associated with clinical chorioamnionitis; and (6) who were gestation-matched controls for chorioamnionitis patients-women in this group were similar to those in Group 4 but were different patients. Amnion, chorion, and decidual cells were grown by standard techniques and incubated with interleukin-1 beta (IL-1 beta). PTHrP was assayed in duplicate samples of amniotic fluid or tissue culture media using an immunoradiometric assay. There was a significant reduction in amniotic fluid concentrations of PTHrP during labour at term. Preterm labour was not associated with significant changes in amniotic fluid concentrations of PTHrP although a trend for reduced concentrations was observed. Amnion and chorion produced measurable quantities of PTHrP and rates of production were increased by treatment with IL-1 beta. Decidual cells did not produce detectable amounts of PTHrP. Hence, labour at term is associated with a decrease in amniotic fluid PTHrP concentrations that may reflect reduced amnion production, which in turn may play a permissive or active role in the mechanism(s) of parturition. These data support the view that the mechanisms that control term and preterm labour may be regulated differently.

Amniocentesis↗

Type III collagen can be present on banded collagen fibrils regardless of fibril diameter.

Monoclonal antibodies that recognize an epitope within the triple helix of type III collagen have been used to examine the distribution of that collagen type in human skin, cornea, amnion, aorta, and tendon. Ultrastructural examination of those tissues indicates antibody binding to collagen fibrils in skin, amnion, aorta, and tendon regardless of the diameter of the fibril. The antibody distribution is unchanged with donor age, site of biopsy, or region of tissue examined. In contrast, antibody applied to adult human cornea localizes to isolated fibrils, which appear randomly throughout the matrix. These studies indicate that type III collagen remains associated with collagen fibrils after removal of the amino and carboxyl propeptides, and suggests that fibrils of skin, tendon, and amnion (and presumably many other tissues that contain both types I and III collagens) are copolymers of at least types I and III collagens.

Actin Cytoskeleton↗

Inflammation of choriodecidua induces tumor necrosis factor alpha-mediated apoptosis of human myometrial cells.

The present study investigated the ability of human choriodecidua to induce myometrial cell apoptosis through the secretion of tumor necrosis factor alpha (TNF). The secretion of TNF was evaluated in the culture supernatants of amnion and choriodecidua explants that were exposed to the bacterial endotoxin lipopolysaccharide (LPS) to mimic inflammation. The choriodecidua explants produced more TNF than the amnion explants in response to LPS stimulation, despite the fact that the choriodecidua had lower levels of TLR4 expression. Moreover, conditioned medium obtained from LPS-treated choriodecidua explants, but not that from amnion explants, decreased the number of viable cultured myometrial cells and induced cell apoptosis by inducing the overexpression of the proapoptotic protein BAX and by decreasing the expression of the anti-apoptotic protein BCL2. Neutralization of TNF in the choriodecidua-conditioned medium reversed this effect. Exogenous TNF mimicked LPS-treated choriodecidua-conditioned medium in that it induced myometrial cell apoptosis, reduced BCL2 expression, and increased BAX expression. Using neutralizing antibodies against both subtypes of TNF receptors, we found that only TNFRSF1A participates in TNF-induced myometrial cell apoptosis. Our in vitro model of LPS-induced inflammation of human fetal membrane explants suggests a mechanism by which TNF secreted by choriodecidua governs human myometrial cell apoptosis at the end of pregnancy. These data support the hypothesis that TNF participates in the complex network of signaling processes associated with uterine involution.

Amnion↗

Prostaglandin production during spontaneous labor and after treatment with RU486 in pregnant rhesus macaques.

The production of prostaglandins from amnion, chorion, decidua, and myometrium was studied in a superfusion system to determine the level of prostaglandin production during late pregnancy, during spontaneous labor, and after in vivo treatment with RU486. Tissues were divided into three groups: those from pregnant control animals, those from animals receiving RU486 (20 mg/kg/day for 3 days) in vivo, and those from animals in spontaneous term labor. Tissues were collected at cesarean section and placed in the superfusion system. After a 30-min equilibration period, fractions of media were collected after passing through the tissue chambers and were assayed for prostaglandin F2 alpha (PGF2 alpha) and prostaglandin E2 (PGE2). The results showed a significant increase in PGF2 alpha from decidua of animals treated with RU486 compared to pregnant controls (pregnant: 3.01 +/- 0.45 ng/g/min; RU486: 4.45 +/- 0.28 ng/g/min; p < 0.05). There was a dramatic increase in PGE2 production by amnion from animals in spontaneous labor but not after RU486 treatment (pregnant: 0.21 +/- 0.06 ng/g/min; RU486: 0.56 +/- 0.09 ng/g/min; spontaneous labor: 5.83 +/- 0.43 ng/g/min; p < 0.01). We conclude that while progesterone is important for maintaining uterine quiescence during pregnancy, progesterone receptor blockade by RU486 does not lead to an increase in PGE2 production by amnion as is shown during normal spontaneous labor.

Amnion↗

Expression of a pemphigoid gestationis-related antigen by human placenta.

We studied the antigen binding of sera from 25 patients with pemphigoid gestationis (PG) using skin and amnion as substrates in an indirect immunofluorescence technique. In addition, skin was studied after prior incubation with I M NaCl which allows BMZ cleavage through the lamina lucida. Linear BMZ staining was obtained with sera from patients with PG using human amnion as substrate. This was seen with both full-term and second trimester placentae, but not with first trimester placentae. In NaCl incubated skin all sera bound to the epidermal aspect of the separated tissue. The pattern of staining in skin, chemically split skin and amnion was identical to that previously seen with the sera of patients with bullous pemphigoid suggesting that the antigen against which circulating anti-BMZ antigens are directed may be the same in the two conditions.

Amnion↗

Incubation of human chorioamniotic membranes with Candida albicans induces differential synthesis and secretion of interleukin-1beta, interleukin-6, prostaglandin E, and 92 kDa type IV collagenase.

Ascendant colonization of pathogenic microorganisms from the vagina to the uterus is strongly associated to preterm labour and premature rupture of membranes. This study evaluated the secretion of interleukin (IL)-1beta, tumour necrosis factor (TNF)alpha, IL-6, prostaglandin E(2) (PGE(2)), and metalloproteinases 9 and 2 by the human chorioamnion stimulated with Candida albicans. Chorioamniotic membranes were obtained after delivery by elective Cesarean section from women at 37-40 weeks of gestation without evidence of active labour. The membranes were mounted in Transwell devices that form two independent compartments, which allow testing the individual responses and contributions of the amnion and choriodecidua. One million CFU ml(-1) of C. albicans was added to either the amniotic or choriodecidual surface and secretions of the markers were measured in both compartments using specific enzyme-linked immunosorbent assay and zymography. Fetal membranes followed different secretion patterns of proinflammatory cytokines depending on the side to which the stimulus was applied. IL-1beta was produced in higher amounts in the presence of C. albicans when applied to the choriodecidual side; TNFalpha and IL-6 secretion did not change in either the amnion or choriodecidual region. PGE(2) synthesis depicted a different pattern, the amniotic tissue was more responsive than the choriodecidual tissue, and this response tended to be higher even when only the amniotic side was stimulated. Matrix metalloproteinases (MMP)-9 increased after stimulation, being the choriodecidua its main source. Selective stimulation with C. albicans induced a differential secretion of IL-1beta, PGE(2), and MMP-9, resulting from a cooperative and bidirectional communication between the amnion and the choriodecidua.

Amnion↗

Equine amniotic membrane transplantation for corneal ulceration and keratomalacia in three horses.

PURPOSE: Amniotic membrane has antifibrotic, anti-angiogenic and antiprotease properties. This retrospective study was designed to investigate the use of equine amniotic membrane transplants (AMT) at preserving vision, maintaining the structural integrity of the globe and maximizing cosmesis in equine eyes with corneal ulceration and severe keratomalacia. METHODS: Equine amnion had previously been aseptically harvested from a 12-year-old Thoroughbred mare during an elective Cesarean section. Sections of amnion were stored at -80 degrees C and thawed as needed. Records of equine cases at the University of Florida with keratomalacia that received an AMT without adjunctive conjunctival grafting were examined. Clinical description, details of medical and surgical treatment, globe survival and visual outcome were documented. Etiologies were determined by cytology, culture or histology. RESULTS: Three horses with corneal ulceration and severe keratomalacia received an AMT without conjunctival graft between December 2002 and April 2003. Pseudomonas spp. were cultured from all three eyes, with evidence of a concurrent fungal infection in two eyes. The three ulcers were 50, 72, and 76% of corneal diameter, and each one worsened in the face of aggressive medical therapy. In all three cases, the AMT sloughed over a 4 to 6-week period. At last follow-up, all three eyes receiving AMT were comfortable and receiving no medication, with light perception and an inconsistent location-dependent menace response. All three horses returned to their prior work. CONCLUSIONS: Results of a small number of equine AMT suggest that amnion can be used successfully to preserve both globe structure and limited vision, as well as optimize cosmesis, in horse eyes with corneal ulceration and severe keratomalacia.

Amnion↗

The concentrations of prostaglandin E2 and prostaglandin F2alpha in tissues within the fetoplacental unit after spontaneous or induced labour.

The concentrations of prostaglandin E2 (PGE2) and prostaglandin F2alpha (PGF2alpha) were measured by radioimmunoassay in portions of umbilical cord, placenta, amnion, chorion and decidua. The samples were obtained during 12 confinements immediately after the delivery of a healthy baby. Labour had either occurred spontaneously (6 subjects) or had been induced approximately two weeks after the estimated date of delivery. The results showed that the concentration of PGE2 was significantly higher (Student's 't' test) in samples of placenta (P less than 0-05), decidua (P less than 0-025), umbilical cord (P less than 0-0125) and amnion (P less than 0-0025), from those patients who had started labour spontaneously. In contrast, the concentration of PGF2alpha was only raised to a significant extent (P less than 0-025) in the amnion. The findings are discussed.

Amnion↗

Production of active and inactive renin by cultured explants from the human female genital tract.

Myometrium, decidua, chorio-decidua and amnion were collected from women undergoing caesarean section and from pregnant women at hysterectomy and hysterotomy for comparison with myometrium, cervix and endometrium taken from non-pregnant women at hysterectomy. Homogenates of chorio-decidua and amnion contained the largest concentrations of active and inactive renin. Cultured explants of all tissues except amnion were found to produce both types of renin. The percentage of active renin produced before 20 days of culture was significantly greater in cultures of chorio-decidua than in those of myometrium (P less than 0.0005) or decidua vera (P less than 0.025), which suggests that renin synthesis in vivo is probably confined to the chorio-decidua.

Amnion↗