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PubMedDate not supplied

Human embryo modulates placental function in the first trimester; effects of neural tissues upon chorionic gonadotropin and progesterone secretion.

We investigated the effect of embryonal neural and adrenal tissues (7-14 weeks gestational age) upon beta hCG secretion by homologous placental explants in static and dynamic cultures. In static co-culture significant inhibition by SC and brain was noted at 7-9 weeks. Similarly, in superfusion, using a novel co-chambering technique there was a significant reduction in the area under the curve but not peak frequency of spontaneous pulsatile beta hCG secretion. Incubations with neural tissues 11 weeks and above caused a stimulatory effect upon beta hCG secretion in both models. The effect of adrenal tissue in static cultures was different, namely slightly inhibitory at 7-9 weeks and inhibitory at 11 weeks and above. In superfusion, the effect of adrenal tissue was not significant. Extracted neural tissue 7-9 weeks incubated with placental explants exhibited inhibitory effects upon beta hCG secretion as well. Buffer-based extracts of neural tissues effect was more pronounced than alcohol-based extracts regarding beta hCG secretion. The effect of extracts was dose-dependent and effects were noted up until a 2000-fold dilution. In contrast, the buffer SC extract had no effect on progesterone (P) secretion while the alcohol extract effect was inhibitory at 7-9 weeks and stimulatory at greater than 11 weeks. Superfused explants pattern of beta hCG secretion was inhibited by one minute pulse of the SC buffer extract. In conclusion, the human neural tissue of embryonal origin may modulate placental hCG and P secretion during early pregnancy.

Adrenal Glands↗

PubMed1982

Demonstration using monoclonal antibodies of inter-locus heteromeric isozymes of human alkaline phosphatase.

Monoclonal antibodies were used to demonstrate hybrid forms of human alkaline phosphatase (ALP) composed of subunits from both placental and intestinal loci. Four main isozymes with alkaline phosphatase activity appear on polyacrylamide gels of the HeLa cell line, Hep 2/5 after electrophoresis. The mobilities of all 4 isozymes are retarded after incubating cellular extracts with a monoclonal antibody specific for placental ALP, while the mobilities of 2 isozymes are also affected by a monoclonal antibody which reacts specifically with intestinal ALP. These 2 isozymes, therefore represent interlocus heteromers (placental/intestinal ALP).

Adult↗

PubMed1999

Cytokine abundance in placental tissues: evidence of inflammatory activation in gestational membranes with term and preterm parturition.

OBJECTIVES: This study of the changes in cytokine concentrations in gestational tissues from women with term and preterm labor was undertaken to assess the extent of inflammatory activation associated with spontaneous labor and delivery. STUDY DESIGN: Extracts of amniotic, chorionic-decidual, and placental tissues from women delivered at term before labor (n = 15), at term after labor (n = 15), and preterm (n = 31) were assayed for interleukin 1beta, interleukin 6, and interleukin 8. RESULTS: In amniotic tissues of women delivered by spontaneous labor at term the median interleukin-6, interleukin-8, and interleukin-1beta concentrations were 3.8 to 5.4 times those of tissues from women delivered at term without labor (P <.05, Mann-Whitney U test). Interleukin-6 and interleukin-8 concentrations were also significantly increased (3. 3-4 times) in chorionic-decidual tissues. Marked increases (approximately 3-6 times) in the concentrations of all 3 cytokines were observed in both amniotic and chorionic-decidual tissues from women with preterm deliveries with respect to those from women with term deliveries after labor. Cytokine concentrations were significantly correlated within amniotic tissues from both women with term delivery after labor and women with preterm delivery and also in preterm chorionic-decidual tissues but not preterm placental tissues. Concentrations of cytokines in the tissues of women delivered preterm were not significantly affected by mode of delivery, treatment with antibiotics, or twin birth. In preterm tissues with evidence of intrauterine infection only amniotic interleukin-1beta concentrations were significantly elevated (P <. 05). Little or no labor-related change in cytokine concentrations was seen within placental tissues. CONCLUSIONS: Increased cytokine abundance in gestational membranes associated with labor supports the view that an inflammatory process is involved in both term and preterm labor. This process does not, however, appear to be evident in the villous placenta.

Cytokines↗

PubMed1984

Lack of correlation between amniotic fluid and maternal plasma contents of beta-endorphin, beta-lipotropin, and adrenocorticotropic hormone in normal and pathologic pregnancies.

Reported are the concentrations of beta-endorphin, beta-lipotropin, and adrenocorticotropic hormone (ACTH) in the amniotic fluid and plasma of 40 healthy pregnant women at different stages of gestation. Moreover, the amniotic fluid levels of the three peptides were evaluated in 20 other pregnant women affected by different pathologic conditions (Cooley's disease, gestosis, diabetes, placental insufficiency, etc.). A silicic acid extraction procedure was performed on the samples. Each extract was subjected to Sephadex G-75 column chromatography, and the two fractions corresponding to beta-lipotropin and beta-endorphin were collected, freeze-dried, and assayed by two specific radioimmunoassays. Levels of ACTH were measured by radioimmunoassay directly on the extracts. Levels of beta-endorphin in amniotic fluid showed the highest values in the first trimester (173 +/- 30 fmol/ml, mean +/- SEM) but were significantly decreased in the second (75.2 +/- 14) and third trimesters (14.3 +/- 1.8). An inverse trend characterized plasma levels of beta-endorphin, which showed a progressive increase from the first trimester to term (10.4 +/- 11.1). Amniotic fluid levels of beta-lipotropin remained stable during the first (48.6 +/- 6.3) and second (54.6 +/- 11.1) trimesters, but decreased significantly in the third trimester (17.9 +/- 2.3). The plasma concentrations of beta-lipotropin showed the highest levels in the first trimester (10.9 +/- 0.9), and decreased significantly at term (8.9 +/- 1.3). Last, amniotic fluid levels of ACTH decreased from 55.3 +/- 4.75 fmol/ml in the first trimester to 12.5 +/- 1.16 in the second trimester, and rose again in the third trimester to 34.4 +/- 6.6 fmol/ml. Plasma levels of ACTH were characterized in the first two trimesters by values twice those recorded for nonpregnant women, and decreased at term to 8.9 +/- 1.4 fmol/ml. In the pregnant patients with fetuses affected by Cooley's disease (second trimester) and in those with edema-proteinuria-hypertension (EPH) gestosis (third trimester), amniotic fluid levels of beta-endorphin, beta-lipotropin, and ACTH were in the same range as those in healthy pregnant women.

Adrenocorticotropic Hormone↗

PubMed1979

Human chorionic gonadotropin in the plasma of normal, nonpregnant subjects.

To determine whether ectopic secretion of a protein hormone can occur normally, we studied plasma from normal, nonpregnant subjects for the presence of a placental hormone, human chorionic gonadotropin. We extracted and purified this hormone from other plasma proteins. We identified the hormone in the final residue on the basis of its dose-response curves in a specific radioimmunoassay and calculated the plasma concentration after correction for losses. Because this assay is sensitive to concentrations as low as 2 pg per milliliter, human chorionic gonadotropin could be detected in the plasma of 12 of 16 blood donors; the median concentration was 19 pg per milliliter (range, less than 2 to 361). This immunologic human chorionic gonadotropin was further characterized from a pool of normal plasma by gel filtration on Sephadex G-100 and was found to be identical to the standard form of the hormone. The concentration in this pool from 13 normal men was 18 pg per milliliter. The source of this ectopic hormone production is unknown, but may be normal, rapidly proliferating nonmalignant cells.

Adult↗

PubMed1982

Coated vesicles from human placenta carry ferritin, transferrin, and immunoglobulin G.

Coated vesicles prepared by using isotonic gradients retain their contents. Ferritin, transferrin, and immunoglobulin G have been detected in human placental coated vesicles. Triton X-100 extraction of crude coated vesicles removes contaminating uncoated membranes, leaving coated particles and their associated proteins apparently intact; this provides an efficient means of preparing large quantities of coated vesicle proteins. The clathrin cages surround "cores" of structural protein containing polypeptides of 100,000 and 50,000 molecular weight.

Clathrin↗

PubMed1987

Differentiation of axon-related Schwann cells in vitro. I. Ascorbic acid regulates basal lamina assembly and myelin formation.

Rat Schwann cells cultured with dorsal root ganglion neurons in a serum-free defined medium fail to ensheathe or myelinate axons or assemble basal laminae. Replacement of defined medium with medium that contains human placental serum (HPS) and chick embryo extract (EE) results in both basal lamina and myelin formation. In the present study, the individual effects of HPS and EE on basal lamina assembly and on myelin formation by Schwann cells cultured with neurons have been examined. Some batches of HPS were unable to promote myelin formation in the absence of EE, as assessed by quantitative evaluation of cultures stained with Sudan black; such HPS also failed to promote basal lamina assembly, as assessed by immunofluorescence using antibodies against laminin, type IV collagen, and heparan sulfate proteoglycan. The addition of EE or L-ascorbic acid with such HPS led to the formation of large quantities of myelin and to the assembly of basal laminae. Pretreatment of EE with ascorbic acid oxidase abolished the EE activity, whereas trypsin did not. Other batches of HPS were found to promote both basal lamina and myelin formation in the absence of either EE or ascorbic acid. Ascorbic acid oxidase treatment or dialysis of these batches of HPS abolished their ability to promote Schwann cell differentiation, whereas the subsequent addition of ascorbic acid restored that ability. Ascorbic acid in the absence of serum was relatively ineffective in promoting either basal lamina or myelin formation. Fetal bovine serum was as effective as HPS in allowing ascorbic acid (and several analogs but not other reducing agents) to manifest its ability to promote Schwann cell differentiation. We suggest that ascorbic acid promotes Schwann cell myelin formation by enabling the Schwann cell to assemble a basal lamina, which is required for complete differentiation.

Animals↗

PubMedDate not supplied

Overexpression of placental tissue protein 17b/TIP47 in cervical dysplasias and cervical carcinoma.

In our previous Western- and Northern-blot investigations, high over-expression of placental protein 17b/TIP47 was detected in extracts of human cervical carcinoma tissues compared to normal conditions of the cervical tissue. PP17b serum levels were also elevated in untreated cervical carcinoma patients compared to healthy controls. In the present study, the expression pattern of PP17 proteins was investigated in various cervical dysplasias and in cervical carcinoma tissue specimens by the streptavidin-biotin immunoperoxidase technique using PP17-specific antiserum. In normal third-trimester human placentas, which served as positive controls, mainly cytoplasmic PP17 immunostaining of syncytiotrophoblasts and chorionic trophoblasts was observed. Normal human uterine cervical squamous and glandular epithelia were negative or weakly positive, while in low grade dysplasias (CIN I-II) only the cytoplasms of dysplastic cells were weakly positive or positive; in high grade dysplasias (CIN III/ISC) cytoplasms of the dysplastic cells were strongly positive. Normal and superficial cells in the differentiated zones were negative in all tissue specimens. In cases of invasive epithelial cervical carcinomas, small basal-type tumour cells were mostly negative whilst cells with squamous differentiation were strongly positive for PP17. Our hypotheses for this newly detected phenomenon are briefly discussed.

Adult↗

PubMed2003

Effects of dioxin (2,3,7,8-TCDD) and PCDDs/PCDFs congeners mixture on steroidogenesis in human placenta tissue culture.

OBJECTIVE: The aim was to compare the direct effect of most toxic 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) as well as of naturally occurring congener mixture of polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) extracted from fly ash on the placental steroidogenesis. The concentration of all 17 toxic congeners was reported and the toxic equivalent (TEQ) was calculated as a 27.7 micrograms-TEQ/kg of fly ash. METHODS: Placental cotyledons were harvested immediately after expulsion of placenta. The cells were prepared according to KLIMAN et al. (1986). To examine TCDD and PCDDs/PCDFs mixture action on cytochrome P450 side change cleavage enzyme (P450 scc) and 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity the placental cells were cultured either in basal conditions or with the addition of 25-hydroxycholesterol (25-OH) or pregnenolone (P5). RESULTS: TCDD in all doses used decreased basal P4 secretion, while did not show any effect on 25-hydroxycholesterol (25-OH) and pregnenolone (P5) supplemented cultures. In all variants of culture PCDDs/PCDFs mixture was without effect on basal and substrate supplemented progesterone (P4) secretion suggesting a reduction in the activity of cytochrome P450scc or 3 beta-HSD. To examine TCDD and PCDDs/PCDFs mixture action on aromatase cytochrom P450 (P450 arom) activity the placental cells were cultured in basal condition or with the addition of dehydroepi-androsterone (DHEA) or testosterone (T). Significant increase of estradiol secretion under the influence of TCDD in DHEA and T supplemented cultures suggests its action on the activity of P450 arom. CONCLUSION: The discrepancy found between the action of pure TCDD and dioxin mixture on placental steroids secretion is possibly due to an additional effect of pentachlorodibenzo-p-dioxin (PeCDD) and pentachlorodibenzo-furan (PeCDF) which covered > 50% of the total toxic equivalents (TEQ) present in this mixture.

Cells, Cultured↗

PubMed1992

[Morphological evaluation of fetal development after administration of Poterium sanguisorba for correction of fetoplacental circulation].

Polyphenol preparation of Poterium sanguisorba was tried for the treatment of experimental pregnancy abnormalities. The teratogenic and embryotoxic effects of the preparation administered in a dose of 0.75 LD50 were studied. The therapeutic doses of the drug are characterized by an immunostimulating effect on the mother and help correct the fetoplacental circulation and improve the fetal morphofunctional status.

Animals↗

PubMed2001

Vascular endothelial growth factor and placenta growth factor concentrations in Down's syndrome and control pregnancies.

Vascular endothelial growth factor (VEGF) and placenta growth factor (PLGF) are considered to play important roles in angiogenesis and vascular permeability during placental development. Since trisomy 21 placentae show trophoblastic hypoplasia and hypovascularity, we investigated PLGF and VEGF synthesis in Down's syndrome pregnancies. Maternal serum was collected from 102 euploid and 24 trisomy 21 pregnancies between 15 and 20 weeks gestation and tested for these two factors by enzyme-linked immunosorbent assays. Protein extracts from 15 normal and six trisomy 21 placentae were also tested. VEGF was not detected in maternal serum, while PLGF increased significantly with gestational age. Serum PLGF, transformed as a multiple of the gestational age median (MoM), in Down's syndrome pregnancies was significantly lower than in euploid controls (mean 0.67 +/- 0.043 MoM versus 1.00 +/- 0.047 MoM, analysis of variance F = 11.605, P < 0.001 ). Both VEGF and PLGF were detected in placental protein extracts without variation according to gestational age. Down's syndrome placentae had significantly less PLGF compared to normal placentae (Mann-Whitney, P < 0.05 ) but no difference was observed in placental VEGF content (Mann-Whitney, P = 0.94 ). Considering the biological properties of PLGF, this decrease may provide new insights into the mechanism(s) leading to the structural and functional anomalies described in trisomy 21 placentae.

Adult↗

PubMed1985

Immunoactive products of placenta. IV. Impairment by placental cells and their products of CTL function at effector stage.

Trophoblast-enriched cell suspensions prepared by collagenase digestion from midterm murine placentae were found resistant to CTL-mediated lysis. Treatment of such cells by trypsin or neuraminidase rendered these cells susceptible to such lytic effectors. Collagenase-prepared cell suspensions could impair CTL action, whereas neuraminidase- or trypsin-treated cells did not retain this property. This effect was also observed with extracts. These results indicate that soluble factors (which we will characterize in another paper) released by trophoblast cells (in fact, spongiotrophoblast) can interfere in a dose-dependent fashion with the action of lytic effectors. We suggest that such active mechanisms are physiologic components of the placental barrier and might be defective in some cases of immunologic abortions.

Animals↗

PubMed1985

Rabbit placental relaxin: purification and immunohistochemical localization.

Rabbit placentas were extracted with 0.7 N HCl-acetone (3:5, vol/vol) containing protease inhibitors. Gel filtration (Sephadex G-50) followed by ion exchange chromatography (carboxymethyl cellulose) separated a bioactive relaxin-like fraction with a specific activity of 8.5 U/mg protein as determined by the in vitro mouse uterus bioassay. Column chromatography using Sepharose CL-4B in 6 M guanidine HCl was employed to purify the bioactive sample. The yield of the purified relaxin-like protein was 12 micrograms/g placenta and the specific activity was 23 U/mg protein. The bioactive sample was also immunoreactive after being electrophoretically transferred to nitrocellulose paper and stained using rabbit antiporcine relaxin serum and peroxidase-antiperoxidase immunochemistry. Isoelectrofocusing of the purified relaxin-like protein revealed one band with an isoelectric point of approximately 6.5. The apparent molecular weight of the rabbit relaxin was approximately 7200 as determined by sodium dodecyl sulfate-urea slab gel electrophoresis. Upon reduction with 5.0% mercaptoethanol and electrophoresis on sodium dodecyl sulfate-urea polyacrylamide gels, both the 7200 dalton rabbit immunoreactive relaxin-like polypeptide and porcine relaxin migrated as a lower molecular weight protein. These results suggest that rabbit relaxin, like pig, rat, shark, and human relaxin, consists of two chains linked by disulfide bonds. At the light microscopy level, immunohistochemical staining with guinea pig antiporcine relaxin serum indicated that relaxin was located in the syncytiotrophoblast cells of the placental labyrinth of day-23 and day-30 pregnant rabbits. The syncytiotrophoblast cells from day 16 of pregnancy did not stain for relaxin.

Animals↗

PubMed1979

Isolation and characterization of human placental chorionic villar extracellular matrix.

The cell-free extracellular matrix of human placental chorionic villi has been prepared by a procedure employing extraction of the terminal villar fragments with the detergents Triton X-100 and sodium deoxycholate. The isolated human placental extracellular matrix retains an intact, but collapsed, histoarchitecture, as observed by scanning and transmission electron microscopy. It remains intact, in large part because of the presence of continuous sheets of villar basement membranes and associated interstitial collagen fibers and scattered patches of fibrin. The staining charcteristics and chemical composition of the isolated human placental extracellular matrlix are similar to those reported for basement membranes in several tissues and indicate the presence of collagen-like and glycoprotein components in this preparation. Gel electrophoresis of urea-SDS-mercaptoethanol extracts of the matrix showed that it consists of several polypeptide components of various molecuar weights, some of which are associated into high molecular weight complexes by disulfide bonds.

Amino Acids↗

PubMed1978

Presence of fibronectin in basement membranes and acidic structural glycoproteins from human placenta and lung.

Cold-insoluble globulin was detected in both trophoblast and alveolar basement membrane preparations, whereas it was not detected in GBM preparations. Acidic structural glycoproteins from both placental villi and lung parenchyma, which were extracted with 0.3 M acetic acid and recovered by adjusting the pH to 4.7, also contained CIg. Fractions of TBM, solubilized by either dilute alkali (0.01 N NaOH), by reduction and alkylation of the disulfide bonds, or by 0.3 M acetic acid extraction, all contained the antigen and possessed properties similar to those of ASG. The ASG fractions also reacted with antifibrinogen, but proof that the two types of determinants occur on a given molecular species is lacking at present. Purified collagenase solubilized CIg from ABM, from lung parenchyma, and from the stroma of placental villi, and this finding is strong evidence for an association of CIg with collagen in these connective tissues.

Basement Membrane↗

PubMed1979

Role of mammalian RNase inhibitor in cell-free protein synthesis.

Addition of the human placental RNase inhibitor at 10 mu/ml to a mixture of wheat germ extract and translation components, prior to the addition of mRNA from dog pancreas or influenza virus-infected cells, resulted in a significant increase in the yield of proteins synthesized. Analysis of the translation products by sodium dodecyl sulfate/polyacrylamide gel electrophoresis indicated that the inhibitor preferentially increased the yield of the larger proteins. In the presence of the inhibitor, yields of the preprocarboxypeptidases were increased 4.5-fold and yields of preamylase were increased 15-fold. Incubation of the wheat germ extract or individual translation components with dog pancreas mRNA, with or without the placental inhibitor, indicated significant RNase contamination among the fractions. Two other in vitro protein synthesis systems-the reticulocyte lysate system and the Krebs ascites system-were found to contain latent RNase activity (RNase in complex with the inhibitor) and an excess of RNase inhibitor. The addition of placental RNase inhibitor did not increase the yield in these systems, except in those cases in which the RNase contamination approached the amount of endogenous inhibitor. When used during the isolation of rat liver cell fractions, the placental inhibitor increased the yield (as measured by A(260)) of rough microsomes and detached polysomes by 24% and 4.6-fold, respectively. Analysis of translation products indicated that detached polysomes isolated in the presence of the inhibitor were intact; those isolated in the absence of inhibitor were degraded.

Animals↗

PubMed1989

Insulin-like growth factor 2 and short-range stimulatory loops in control of human placental growth.

Substructures of the first-trimester human placenta (within 3 months post-conception) display 'pseudo-malignant' properties. We show here, by in situ hybridization, that the insulin-like growth factor 2 (IGF-2) gene expression is particularly active in the cytotrophoblasts, which dominate these structures. Because the majority of placental IGF-2 mRNA is polysomal in extracts of first-trimester placenta, the spatial pattern of IGF-2 transcripts generally also defines the pattern of IGF-2 production. In primary trophoblast cultures, rendered quiescent by serum starvation. IGF-2 performs as a human embryonic growth factor by activating cell cycle entry/progression. Although both type 1 and 2 IGF receptor mRNAs can be found co-distributed with IGF-2 mRNA during placental development (supporting an autocrine role for IGF-2), these occasional patterns are confined to cytotrophoblasts with low proliferative potential. The reciprocity in ligand and receptor expression patterns are discussed in terms of rate-limiting steps in the involvement of IGF-2 in the proliferative phenotype of the early human placenta.

Cell Division↗

PubMed1983

[Placental tissue concentration of pregnancy-associated proteins in early and term pregnancies].

We examined the localization of five pregnancy-associated proteins such as pregnancy specific beta 1-glycoprotein (SP1), human chorionic gonadotropin (HCG), human placental lactogen (HPL), alpha 2-PA-glycoprotein (SP3) and pregnancy-associated plasma protein-A (PAPP-A) in the placentae of early and term pregnancies by means of the PAP method in immunohistochemical technology. It was found that the degree of staining of these proteins did not reflect their concentration in serum at each gestational age. Therefore, the concentrations of these proteins in the solution extracted from the placenta were measured. 1) Placental tissues were fixed with a 10% neutral formaldehyde solution and these were embedded into paraffin blocks. These specimens were used for the PAP method. The localizations of five pregnancy-associated proteins from 10 placentae at the gestational age of 7 to 10 weeks were compared with those at the gestation age of 38 to 41 weeks from 7 patients. The staining degree of SP1 in free villi was pale in both early and term pregnancies. HCG was stained deeply in early pregnancy but pale at term pregnancy. HPL was stained deeply at both gestational ages. Horseradish peroxidase reaction products from SP1, HCG and HPL were located chiefly in the syncytiotrophoblast. SP3 was not stained on the placental tissue. PAPP-A was stained to a greater extent in the cytotrophoblast in early pregnancy but pale in the syncytiotrophoblast at term pregnancy. 2) The concentrations of SP1, HCG, HPL, SP3 and PAPP-A were measured in the placental tissues in both early and term pregnancies. Placental tissues were obtained from 24 normal pregnancy patients aborted artificially at 8 to 11 weeks gestation and from 28 patients terminated by normal deliveries at 38 to 40 weeks gestation. The tissue was homogenated with 3 volumes of 1/2 PBS (0.005M phosphate-buffered 0.07M sodium chloride, pH 7.4). The supernatant was removed after centrifugation and stored at -20 degrees C until assayed. The concentration of SP1 was measured by single radial immunodiffusion. That of HCG was assayed by a directed Latex agglutination test (Gestate Slide Eiken). The concentrations of HPL, SP3 and PAPP-A were quantified by rocket immunoelectrophoresis. No significant difference in SP1 levels was shown between early and term pregnancies, and the SP1 level was 4.3 +/- 1.3 (mean +/- 1 S.D.) mg/dl at term pregnancy. The HCG level was 1,100 +/- 300 IU/ml in early pregnancy and at least 20-fold higher than that at term pregnancy.(ABSTRACT TRUNCATED AT 400 WORDS)

Chorionic Gonadotropin↗