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The application of an extract of human placenta in the treatment of rheumatic affections.

Iontophoretic application of Placenta-Lucchini in combination with catalase and mucopolysaccharidase for the treatment of acute rheumatic inflammation results in significant clinical improvement. Such improvement is not seen if Placenta is used alone. Intra-articular application of Placenta in the arthritis joints of patients with rheumatoid arthritis results in a significant improvement as compared with placebo. Its efficacy is, however, inferior to that of Triamcinolone. Intra-articular application of Placenta prolongs local arthritic remissions induced by steroids.

Arthritis, Rheumatoid↗

Caprine placental lactogen: levels of prolactin-like and growth hormone-like activities in the circulation of pregnant goats determined by radioreceptor assays.

Radioreceptor assays for prolactin-like (lactogenic) activity and growth hormone (GH)-like activity have been used to study concentrations of caprine placental lactogen (PL) in the circulation during pregnancy. Both lactogenic and GH-like activities from less than 100 ng/ml (ovine prolactin- and human GH-equivalents) about 60 days after mating to reach peak levels (400-1600 ng/ml) between days 110 and 130 of pregnancy. The levels of both activities increased in essentially the same fashion but during the last 15 days of pregnancy, lactogenic activity declined less than GH-like activity. This divergence was most pronounced at parturition when levels of lactogenic activity increased (approximately 700 ng/ml) despite very low (less than 200 ng/ml) levels of GH-like activity being measured and this probably reflected increased secretion of pituitary prolactin near parturition. When serum from a pregnant goat or a simple alkaline extract of placental cotyledons was fractioned on a column packed with Sephadex G-100, lactogenic and GH-like activities eluted together with distribution coefficients of approximately 0-5-0-6. The possibility that caprine PL serves physiologically as a luteotrophin and/or mammotrophin during pregnancy in goats is discussed.

Animals↗

Placentation in the African elephant, Loxodonta africana. I. Endocrinological aspects.

Placental and fetal tissues were recovered from the uteri of 59 pregnant elephant that ranged in estimated age from day 18 to month 21 of gestation. Incubation of placenta and fetal gonad, alone or in combination, with tritium-labelled cholesterol, pregnenolone and androstenedione failed to yield any labelled progestagens or oestrogens from placenta, but did produce small amounts of labelled progesterone and 5alpha-dihydroprogesterone from fetal gonad. Immunochemical staining of tissues with four antisera specific for enzymes involved in the steroidogenic pathway revealed no staining in sections of placenta but positive labelling for P450 side chain cleavage enzyme (SCC450) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) of the interstitial cells that comprise the bulk of the enlarged fetal gonads during the second half of gestation. Saline extracts of placental tissue showed no activity in three different gonadotrophin assays. In view of this endocrinological inactivity in the zonary elephant placenta and the probable reliance on maternal luteal sources of progestagens for maintenance of the pregnant state, the argument is advanced that uncomplicated abortion would probably follow a single administration of a PGF analogue given at any stage of pregnancy. If so, the treatment might constitute an efficacious method for controlling population increases in elephants maintained in enclosed game parks in Africa.

20-alpha-Dihydroprogesterone↗

Effects of antisera raised against native and denatured human alpha-glucosidase and beta-hexosaminidases on native enzyme activity.

Antisera were raised in rabbits against native and sodium dodecylsulfate denatured forms of human acid alpha-glucosidase and beta-hexosaminidases A and B. Anti-native enzyme antisera were able to precipitate all or nearly all enzyme activity from cell extracts, and to eliminate all stainable activity on electrophoresis. Antisera prepared against denatured enzymes precipitated only a minor part of enzyme activity. Electrophoretic analysis showed that these antisera were able to bind to the enzyme molecule. The result was a slowing down of the anodic migration but not immobilization. The use of variants with hexosaminidase deficiencies helped to clarify the action of the antisera on the various hexosaminidase isozymes.

Animals↗

Stimulation of DNA synthesis in trophoblasts and human umbilical vein endothelial cells by hepatocyte growth factor bound to extracellular matrix.

Hepatocyte growth factor (HGF) promotes the growth not only of hepatocytes but also of several other types of cells such as cytotrophoblasts and endothelial cells. Recent studies have revealed that HGF is trapped in the extracellular (ECM) matrix through heparan sulphate in vivo, thereby acting as a mitogen for hepatocytes in cooperation with heparan sulphate. In this study, we detected HGF protein in chorionic tissue and placental tissue extracts, and found that HGF and heparan sulphate were co-distributed in the endothelial basement membrane and trophoblast basement membrane on immunohistochemical examination. The rates of DNA synthesis in primary cultured cytotrophoblasts and human umbilical vein endothelial cells (HUVEC) cultured on HGF-bound Matrigeltrade mark were 6-8 times those of control cytotrophoblasts and HUVEC. When Matrigeltrade mark dishes were pretreated with heparinase and heparitinase prior to binding of HGF, stimulation of DNA synthesis was markedly decreased. A considerable decrease in stimulation of DNA synthesis was observed following washing of HGF-bound Matrigeltrade mark with 1 m acetic acid, 1 m NaCl and 0.1 per cent trypsin, but not following treatment with chondroitinase ABC. These observations suggest that HGF can be trapped in ECM in vivo, thereby acting as a mitogen for cytotrophoblasts and placental vein endothelial cells in cooperation with heparan sulphate.

Analysis of Variance↗

High-molecular-weight immunoreactive beta-endorphin in extracts of human placenta is a fragment of immunoglobulin G.

A high-molecular-weight protein with beta-endorphin- and adrenocorticotropin-immunoreactivities was isolated from extracts of human placenta after several purification steps, including immunoadsorption with a well-characterized antiserum raised to beta-endorphin. This protein was identified as the heavy chain of the human immunoglobulin class IgG1. These results have led to the recognition of homologies in the amino acid sequences of these physiologically unrelated molecules. They also suggest caution in accepting immunological competence as the sole criterion of the chemical identity of a ligand.

Amino Acid Sequence↗

[Effect of fetoplacental factors on in vitro proliferation of lymphocytes and non-lymphoid cells].

The effect of extracts of placenta, amniotic fluid, serum of pregnant women and newborn infants on PHA-stimulated proliferative activity of lymphocytes and proliferation of non-lymphoid cells (L-cells) was studied. The inhibiting effect decreased in the series: placenta -- amniotic fluid -- serum of pregnant women. The effect on L-cells was similar to that on lymphocytes. It is suggested that the biological samples under study contained factors having non-specific immunosuppressive effect.

Amniotic Fluid↗

Immature human chorionic gonadotropin (hCG) in first trimester placental cells is bound to an ATP-binding protein forming high-molecular-weight hCG.

Human chorionic gonadotropin (hCG) in first trimester placental cells is made up of immature alpha- and beta-subunits containing only N-linked high-mannose sugar chains, which are of 21 kDa for the alpha-subunit and 23 and 19 kDa for the beta-subunit. However, the apparent molecular weight of immature hCG from placental cell extracts has been estimated from gel filtration to be much higher (100-200 kDa; high molecular weight-hCG, HMW-hCG) based on gel filtration than the theoretical value (approximately 44 kDa) of the alpha beta dimer (alpha beta-hCG). We prepared a gel-filtered fraction containing HMW-hCG and investigated treatments for converting it to alpha beta-hCG. We found that the molecular weight of HMW-hCG was decreased to close to that of alpha beta-hCG by treatment with acetone, proteases, or chelating agents. These treatments also shifted the isoelectric point of HMW-hCG from the acidic region (pI = 4-6) to the alkaline (pI = 9-11), approximating to that of alpha beta-hCG. We also found that HMW-hCG, but not acetone-treated HMW-hCG, bound to ATP-agarose resin. These results suggested that the immature alpha beta-hCG molecule in placental cells may be bound to an acidic ATP-binding protein to form HMW-hCG.

Acetone↗

Characterization of glycoprotein hormone free alpha-subunit from human pituitary and placenta extracts.

Standard alpha-subunit dissociated from glycoprotein hormones differs from individual (free) alpha-subunit found in sera or in cell culture media; secreted free alpha-subunit is larger, more acidic and lacks the ability of recombining in vitro with standard hCG-beta. It is unclear whether the large free alpha-subunit is only a secretory product or whether it is also present in tissue. Herein were studied the molecular size, the isoelectric pH, and the recombining activity of free alpha-subunit obtained from pituitary and placenta extracts. Sephadex exclusion chromatography showed the presence of both a large and a small form, and a changing large/small free alpha-subunit ratio in the various extracts. Most of the large form obtained from placenta extracts electrofocused into two peaks of pI 4.8 and 5.1. The large form showed no recombining activity with standard hCG-beta while the small free alpha-subunit recombined as well as did standard hCG-alpha. The observation of three common characteristics (a larger size, a pI 4.8, and a lack of recombining activity) suggests a similarity between the large secreted form and a fraction of the free alpha-subunit in tissue.

Chromatography, Gel↗

Different responses of lymphocyte to human placenta conditioned medium in vitro.

Several cytokines appear to be implicated in peri-implantation events and in maternal-fetal interaction. The majority of these molecules appear in supernatants or in extracts of placental origin. The purpose of this study was to investigate the action of human placenta conditioned medium (HPCM) on peripheral lymphocytes in vitro. Peripheral lymphocytes of women with different numbers of deliveries, of nulliparous and pregnant women, as well as of men, were cultivated in their own plasma with and without HPCM at 5%, 10% and 20%. The index of lymphocyte blast transformation (ILBT) was determined (values of > or = 0.5 indicated stimulation). In women who had had children, the highest lymphocyte blastogenesis was observed with HPCM at 10%, this finding being correlated to the numbers of deliveries (r = 0.828). In pregnant women, even though the highest answer was also obtained with HPCM at 10%, the addition of suboptimal concentration of HPCM (5%) produced blastic transformation (IBLT: x = 0.54 +/- 0.05), which was not observed in women who had had children. Lymphocytes of men and nulliparous women did not respond to any of the HPCM concentrations tested. In conclusion, HPCM at 10% was still the best concentration to produce lymphocyte blastogenesis. In addition, in the plasma of pregnant women, there may be some substance that enhanced the action in vitro of the extract, thus permitting threshold reduction necessary for lymphocyte stimulation.

Adult↗

L-tryptophan as an antioxidant in human placenta extract.

An antioxidant was purified from human placenta extract (HPE) by using gel filtration, liquid-liquid extraction, silicagel column chromatography, and HPLC. The purified antioxidant was identified to be L-tryptophan (L-Trp). L-Trp showed higher inhibitory activity than mannitol and DMSO on the Fenton reaction-induced degradation of 2-deoxy-D-ribose. L-Trp also had much higher inhibitory activity on the cytochrome P-450-dependent lipid peroxidation than the previously identified antioxidants of HPE, L-phenylalanine, L-tyrosine and uracil. On the other hand, the inhibitory effect of L-Trp on the Fenton reaction-induced protein oxidation was smaller than that of uracil. These results suggest that L-Trp is a main antioxidant of HPE of which the effect is based on the suppression of lipid peroxidation in the oxidative stress status.

Analysis of Variance↗

Public determinants of HLA indicated by pregnancy antibodies.

A tentative table of public specificities in the HLA-A and -B loci is provided. It is based on an all-inclusive survey of placenta extracts from 50,000 pregnancies. We postulate that most of the specificities found are directed against public epitopes. In support of this postulate are the facts that certain combinations occur very frequently, monoclonal antibodies have been made to some of the epitopes, and some have already been established by absorption experiments as being a single specificity. The immunogenicity score for each private and public specificity was computed by taking into account the chance of immunization. It was shown that immunogenicity can vary by factors of more than ten between different specificities. Significantly, immunogenicity of the public epitopes was just as high as against the private ones. This indicates that the public epitopes should be considered as independent, separate antigens in transplantation. Establishment of a table of public specificities and the recognition of each by international nomenclature would be the first step in evaluating public epitopes for transplantation matching.

Antilymphocyte Serum↗

The pharmacological basis for the use of dried sheep placenta in traditional obstetric practice in Nigeria.

Dried sheep placenta is sometimes used in traditional medicine to facilitate labour. The effects of an extract of powdered dried sheep placenta with normal saline on guinea-pig uterus, ileum, spontaneously beating atrium and Langendorff heart, rat uterus and hindquarters, and cat blood pressure were therefore examined. It was found that 1 g of dried sheep placenta had, on the guinea-pig uterus, an oxytocic activity equipotent with 0.075-0.32 i.u. of oxytocin. The oxytocic activity was unaffected at pHs between 4 and 10 or by boiling for 30 min or autoclaving for 15 min. Neither atropine nor promethazine inhibited the oxytocic action, but promethazine inhibited, to the same degree, contractions induced in the ileum by equipotent doses of the infusion and histamine. Atropine, however, had no effect on infusion-induced contractions in the ileum. The vasoconstriction induced in the rat hindquarters was antagonized by promethazine and phentolamine. Cat blood pressure was reduced, but it had positive inotropic and chronotropic effects on the spontaneously beating guinea-pig atrium and on the guinea-pig Langendorff heart. It was concluded that the dried placenta contains a chorionic oxytocic substance the action of which is independent of stimulation of H1 receptors or of muscarinic receptors.

Animals↗

Growth-promoting activity of Hominis Placenta extract on regenerating sciatic nerve.

AIM: Extract of Hominis Placenta (HP) has been used in oriental medicine as an agent for improving physiological function. The present study was conducted to investigate whether HP treatment in an experimental sciatic nerve injury animal model produces growth-promoting effects on regenerating peripheral nerve fibers after injury. METHODS: After HP was injected into a sciatic nerve injury site, changes in protein levels were analyzed in the regenerating nerve area by Western blotting and immunofluorescence staining analyses. For quantitative assessment of axonal regeneration, a retrograde tracing technique was used to identify the neuronal cell bodies corresponding to regenerating axons, and the extent of neurite outgrowth in cultured dorsal root ganglia (DRG) sensory neurons prepared from animals that had experienced a sciatic nerve crush injury 7 d before neuron collection was analyzed. RESULTS: Induction levels of axonal growth-associated protein (GAP-43) in the injured sciatic nerves were elevated by HP treatment. HP treatment also upregulated cell division cycle 2 (Cdc2) protein levels in the distal stump of the injured sciatic nerve. Induced Cdc2 protein was detected in Schwann cells, suggesting that Cdc2 kinase activity may be involved in the growth-promoting activity of regenerating axons via Schwann cell proliferation. Cell body measurement by retrograde tracing indicated that HP treatment produced significant increases in regenerating motor axons. Finally, HP treatment of cultured DRG sensory neurons significantly increased neurite arborization and elongation. CONCLUSION: HP promotes the regeneration of injured sciatic axons by upregulating the synthesis of regeneration-related protein factors such as GAP-43 and Cdc2.

Animals↗

Mechanisms of monozygotic (MZ) twinning: a possible role for the cell adhesion molecule, E-cadherin.

Monozygotic (MZ) twins account for about one-third of all twins. Most MZ twins arise sporadically but there are familial occurrences. The causes of MZ twinning are largely speculative but involve both genetic and environmental factors. Genetic factors could include cell adhesion molecules and factors involved in growth and differentiation of the placenta and fetus. In this study, we examine the possible role of E-cadherin, a cell adhesion molecule, in the causation of twinning. A common polymorphism in the E-cadherin gene reduces expression of E-cadherin by up to 68%. We examined the E-cadherin polymorphism in DNA extracted from placental tissue from 121 MZ twins, 99 DZ twins, and 93 singletons. There was no significant difference in the distribution of the E-cadherin polymorphism in MZ twins although there was a slight over-representation of homozygotes for the polymorphism in MZ twins. We conclude that there is insufficient evidence to suggest that decreased E-cadherin expression alone is a causative factor in MZ twinning but it could be a contributory factor along with other genetic and environmental influences.

Cadherins↗