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PubMed1987

Isolation from human chronic-lymphocytic-leukaemia cells of membrane glycoproteins associated with Fc-receptor functions. Physical parameters and production of polyclonal antibodies.

Two membrane glycoproteins that bound immune complexes and inhibited Fc-receptor- (FcR-)mediated functions in vitro were purified from human FcR+ chronic-lymphocytic-leukaemia cells. A multi-step purification was developed, consisting essentially in: (i) Tween 40 extraction of crude cell membranes; (ii) solubilization of membrane fragments by Renex-30; (iii) isolation of glycoproteins by affinity chromatography on Lens culinaris haemagglutinin-Sepharose; (iv) papain treatment of the eluted glycoproteins followed by gel-filtration chromatography; (v) purification by polyacrylamide-gel electrophoresis of two molecular species from the protein-size fraction enriched for immune-complex-binding activity. The two electrophoretically isolated components displayed apparent molecular masses of 70 and 45 kDa by SDS/polyacrylamide-gel electrophoresis and restricted charge heterogeneity by two-dimensional analysis. Two-dimensional peptide mapping revealed the presence of many peptides in common between the two proteins and the absence of a number of peptides in the 45 kDa component. These two polypeptides were used as immunogens to produce polyclonal antibodies that cross-reacted with both proteins and specifically inhibited FcR-mediated reactions in vitro. Furthermore, FcR-related components from detergent-extracted lysates of the human K562 and U937 cell lines or human placental membranes were revealed by the putative anti-FcR antibodies adsorbed on Protein A-Sepharose.

Antigen-Antibody Complex↗
PubMed2004

Placenta percreta causing rupture of an unscarred uterus at the end of the first trimester of pregnancy: case report.

Reports on placenta percreta in early pregnancy leading to a spontaneous rupture of the uterus are rare. We report a case of this potentially life-threatening complication in the 14th week of pregnancy in an otherwise healthy woman who underwent a manual extraction of the placenta during a previous delivery but who had no history of severe pathology that could have potentially resulted in uterine damage. The occurrence of severe abdominal pain and the presence of a large quantity of free fluid in the abdomen necessitated an emergency laparotomy, revealing a haemoperitoneum due to rupture of the uterus, which was followed by a hysterectomy. This case demonstrates that in patients with a history of placenta accreta and subsequent manual extraction of the placenta, a close investigation of the uterine wall and placentation should be performed in the first trimester in order to anticipate a placenta percreta.

Abdominal Pain↗
PubMed1998

Diagnosis of toxoplasma abortion in ewes by polymerase chain reaction.

Eighteen oestrus-synchronised ewes were infected experimentally with 1500 sporulated oocysts of Toxoplasma gondii between 80 and 90 days of gestation. The infection induced pyrexia and specific antibody in all the ewes. One ewe resorbed its fetus, five ewes aborted and 12 delivered live, congenitally-infected lambs whose pre-colostral serum was antibody-positive. Tissues from the aborted fetuses and placentae from the live lambs were examined for toxoplasma infection by polymerase chain reaction (PCR) amplification of the B1 gene and by mouse inoculation. Using a simple protocol of tissue preparation without DNA extraction and a nested format, PCR was as sensitive as mouse inoculation. Placental cotyledon gave a higher sensitivity of detection than brain, lung or liver, and 16 of 19 placentae were positive by PCR compared with 13 of 18 by mouse inoculation. In mock-infected tissues, as few as 10 tachyzoites could be detected. The PCR could be applied to tissues unfit for mouse inoculation.

Abortion, Veterinary↗
PubMed2002

Clearance and disposition of indometacin in chronically instrumented fetal lambs following a 3-day continuous intravenous infusion.

Indometacin is used in pregnancy for the treatment of premature labour, but there are limited data on the disposition of the drug in the fetus. In order to elucidate fetal indometacin pharmacokinetics at plasma levels and duration comparable with those occurring with use of the drug for tocolysis in humans, indometacin was administered at doses of 1.9 (low dose, LD; n = 5) or 7.5 (high dose, HD; n = 9) microg min(-1) to steady state over a 3-day period in chronically instrumented fetal lambs. Indometacin concentrations in biological fluid samples were analysed by a sensitive capillary gas chromatography-electron capture detection method. The mean steady-state fetal arterial plasma indometacin concentrations were 68.6+/-16.5 ng mL(-1) in the LD infusion and 230.3+/-28.8 ng mL(-1) in the HD infusion. Indometacin concentrations in amniotic fluid were approximately 10% of those in fetal plasma, and below assay detection limits in tracheal fluid. Total body clearance (TBC) in the LD and HD infusions were not different and the overall mean was 11.3+/-1.2 mL min(-1) kg(-1). In the 11 experiments where paired fetal arterial and umbilical venous samples were collected, the extraction of indometacin across the placenta averaged only 5.2+/-1.1%, indicating low placental permeability to the drug in sheep. However, fetal placental clearance (CLpl) of indometacin (10.0+/-2.5 mL min(-1) kg(-1), n = 10) averaged 115.1+/-41.2% of TBC in these animals and the calculated value for fetal non-placental clearance (0.6+/-2.8 mL min(-1) kg(-1)) was not significantly different from zero. Fetal renal clearance of intact indometacin (3.8+/-1.1 microL min(-1) kg(-1); n = 12) was also very low. However, treatment of fetal urine with glucuronidase indicated the presence of glucuronide conjugates and these comprised 69.9+/-8.2% of the total drug concentration (i.e. intact+conjugated) in urine. Thus, the fetal lamb appears to be able to glucuronidate indometacin, but the contribution of this and other non-placental routes to overall fetal elimination of the drug appear minimal. CLpl of the drug is also low owing to the physicochemical properties of indometacin (high polarity) and the permeability characteristics of the sheep placenta.

Animals↗
PubMed1993

Retinoic acid-induced differentiation of the developmentally pluripotent human germ cell tumor-derived cell line, NCCIT.

BACKGROUND: Germ cell tumors are empirically divided into seminomas and nonseminomatous germ cell tumors (NSGCT). Some authorities consider seminomas to be the precursors of NSGCT, whereas others consider them as distinct and unrelated neoplasms. Here, we report that the human NSGCT-derived stem cell line, NCCIT has hybrid features of seminoma and embryonal carcinoma, and suggest that this cell line could be useful for studying the relationship of seminoma to NSGCT. EXPERIMENTAL DESIGN: NCCIT, a developmentally pluripotent permanent cell line derived from a mediastinal NSGCT was karyotyped and characterized morphologically, immunochemically, and biochemically. The cells were grown under standard tissue culture conditions and were also exposed to retinoic acid to induce differentiation. RESULTS: The dividing NCCIT stem cell populations consist of vimentin-positive, keratin-negative cells that do not express desmoplakin or cadherin E (uvomorulin) and are not interconnected with one another. These cells have a high nucleocytoplasmic ratio and contain few cytoplasmic organelles, except for free ribosomes and a small number of mitochondria. Lacto- and globoseries oligosaccharide antigens recognized with antibodies to murine stage specific antigens 1, 3 and 4 (SSEA-1, SSEA-3 and SSEA-4), and human teratocarcinoma mucin-like antigen TRA-1-60 and TRA-1-81 are coexpressed on the cell membranes of a considerable number of stem cells. On most cells alkaline phosphatase can be detected by enzyme histochemistry. The placental isoenzyme of alkaline phosphatase was demonstrated by Western blotting in cell extracts. The liver/bone/kidney isoenzyme of alkaline phosphatase is immunochemically detected on 40% of cells. The culture supernatants also contain chorionic gonadotropin and alpha-fetoprotein, presumably derived from trophoblastic and yolk sac-like cells. The cells are hyperdiploid (chromosome range from 54 to 64) and show prominent structural chromosomal aberrations, mostly deletions and isochromosomes. Retinoic acid treatment inhibited the growth of NCCIT cells and induced stem cell differentiation into keratin, glial fibrillary acid protein, and neurofilament-positive somatic cells. The differentiation was associated with the disappearance of oligosaccharide surface antigens typical of the undifferentiated stem cells; a loss of proteins typical of undifferentiated cells and the appearance of new proteins; and the deposition of extracellular matrix. CONCLUSIONS: NCCIT is a developmentally pluripotent cell line that can differentiate into derivatives of all three embryonic germ layers (i.e., ectoderm, mesoderm, and endoderm) and extraembryonic cell lineages. We suggest that this cell line could be a malignant replica of human cleavage stage embryonic cells with features intermediate between seminoma and embryonal carcinoma.

Adult↗
PubMed1982

Measurement of somatomedin-related peptides in fetal, neonatal, and maternal rat serum by insulin-like growth factor (IGF) I radioimmunoassay, IGF-II radioreceptor assay (RRA), and multiplication-stimulating activity RRA after acid-ethanol extraction.

Previous measurements of somatomedins (Sms) and insulin-like growth factors (IGFs) in maternal and fetal serum have yielded contradictory results. We have, therefore, measured maternal, fetal, and neonatal rat serum with two highly specific assays: 1) IGF-I/Sm-C RiA and 2) a highly specific IGF-II/rat placental membrane radioreceptor assay (RRA). In addition, we have made measurements with a less specific multiplication-stimulating activity (MSA)-rat placental membrane RRA. To avoid possible serious artifacts created by Sm-binding proteins, preliminary acid-ethanol extraction of serum was performed. Results are expressed in terms of a reference human serum with an assigned potency of 1 U/ml. Maternal RIA IGF-I fluctuated between 1.1-1.4 U/ml from the 17th day of pregnancy to the 25th day after delivery (nonpregnant rat serum pool, 1.25 +/- 0.22 U/ml). On day 21 of gestation, fetal serum radioimmunoassayable IGF-I was 1.03 +/- 0.03 U/ml. After birth, radioimmunoassayable IGF-I fell and reached .19 +/- 0.03 U/ml at 18 days of age, but rose to 0.71 +/- 0.04 U/ml at 25 days of age. At term, maternal radioreceptor assayable IGF-II was 2.18 +/- 0.27 U/ml (nonpregnant female pool, 1.4 +/- 0.12). By the 25th postpartum day, radioreceptor assayable IGF-II was 1.39 +/- 0.12 U/ml. Radioreceptor assayable IGF-II in fetal serum on day 19 was 3.26 +/- 0.48 U/ml and rose to 5.37 +/- 0.66 U/ml on the day of delivery. A further rise to 8.92 +/- 1.03 occurred on day 5. A subsequent fall to 2.41 +/- 0.05 U/ml was observed on day 25. The patterns of results of the MSA RRA in fetal and neonatal rat serum were similar to that obtained with the IGF-II RRA. We now conclude that radioimmunoassayable IGF-I is present in higher concentrations than previously reported in term fetal rat serum and that radioreceptor assayable IGF-II is selectively elevated in rat fetal and neonatal life and may have unique metabolic and growth-promoting significance.U

Animals↗
PubMed1987

Relationship between the two immunologically distinguishable forms of glucocerebrosidase in tissue extracts.

Extracts of human spleen contain two immunologically distinguishable forms of glucocerebrosidase: form I is precipitable by polyclonal or monoclonal anti-(placental glucocerebrosidase) antibodies, whereas form II is not [Aerts, J. M. F. G., Donker-Koopman, W. E., Van der Vliet, M. F. K., Jonsson, L. M. V., Ginns, E. I., Murray, G. J., Barranger, J. A., Tager, J. M. & Schram, A. W. (1985) Eur. J. Biochem. 150, 565-574]. The proportion of form II glucocerebrosidase was high in extracts of spleen, liver and kidney and low in extracts of brain, placenta and fibroblasts. Furthermore, the proportion of form II enzyme was higher in a detergent-free aqueous extract of spleen than in a Triton X-100 extract of total spleen or splenic membranes. When form II glucocerebrosidase in a splenic extract was separated from form I enzyme by immunoaffinity chromatography and stored at 4 degrees C, a gradual conversion to form I enzyme occurred. The conversion was almost immediate if 30% (v/v) ethylene glycol was present. In the denatured state both forms of glucocerebrosidase reacted with anti-(placental glucocerebrosidase) antibodies. Form I glucocerebrosidase was stimulated by sodium taurocholate or sphingolipid-activator protein 2 (SAP-2), whereas form II enzyme exhibited maximal activity in the absence of the effectors. The pH activity profile of form II glucocerebrosidase was almost identical to that of form I enzyme in the presence of SAP-2. In the native state, form I glucocerebrosidase had a molecular mass of 60 kDa whereas that of form II glucocerebrosidase was about 200 kDa. After gel-permeation high-performance liquid chromatography of splenic extracts, the fractions with form II glucocerebrosidase contained material cross-reacting with both anti-(placental glucocerebrosidase) and anti-(SAP-2) antibodies. Preincubation of form I glucocerebrosidase with SAP-2 at pH 4.5 led to masking of the epitope on glucocerebrosidase reacting with monoclonal anti-(placental glucocerebrosidase) antibody 2C7. Furthermore, preincubation of form I glucocerebrosidase with monoclonal antibody 2C7 prevented activation of the enzyme by SAP-2. We propose that form I glucocerebrosidase is a monomeric form of the enzyme, whereas form II glucocerebrosidase is a high-Mr complex of the enzyme in association with sphingolipid-activator protein 2.

Antibodies, Monoclonal↗
PubMed1981

Characterization of placental luteinizing hormone-releasing factor-like material.

High pressure liquid chromatography (HPLC) and radioimmunoassay were employed to characterize luteinizing hormone-releasing factor (LRF)-like material in the human placenta. Methanol extracts of the placenta were washed with acetic acid and chloroform, further purified on coarse octadecylsilane columns, fractionated on HPLC, and tested by radioimmunoassay. In HPLC, placental LRF had the same retention time as synthetic LRF, and such fractions gave an inhibition curve which was parallel to that of synthetic LRF in radioimmunoassay. It is concluded that human placental LRF is similar or identical to LRF in the central nervous system.

Chromatography, High Pressure Liquid↗
PubMed1985

Identification of proliferin mRNA and protein in mouse placenta.

Proliferin is a recently described, prolactin-related protein whose mRNA appears in several murine cell lines during active growth. We have surveyed a number of mouse organs or tissues for the presence of mRNAs that hybridize to cloned proliferin cDNA. Of the tissues tested, only the placenta yielded proliferin-related mRNA. This placental RNA is about 1 kilobase in length, increases sharply between days 8 and 10 of pregnancy, and then gradually declines through day 18. It is more abundant in RNA extracted from the fetal, compared to the maternal, part of the placenta. From a cDNA plasmid library prepared from poly(A)+ placental RNA, two types of proliferin-related clones were isolated, differing in intensity of hybridization to proliferin cDNA. By nucleotide sequence analysis, a strongly hybridizing clone was found to be nearly identical to the proliferin cDNA clone isolated from a library prepared from mRNA of a growing mouse fibroblastic cell line. Using an antiserum prepared against a synthetic proliferin fusion protein, we show that proliferin is secreted as a glycoprotein by minced placental tissue and that it differs from mouse placental lactogen. We conclude that proliferin is a placental hormone that is synthesized in certain mouse cell lines during active growth. Its function during pregnancy and during the growth of cultured cells is presently unknown.

Amino Acid Sequence↗
PubMed1996

Anti-phospholipid antibodies in preeclampsia and their binding ability for placental villous lipid fractions.

OBJECTIVE: In order to investigate the significance of the anti-phospholipid antibodies (APAs) in preeclampsia, 3 kinds of APAs-namely, the anti-cardiolipin antibody (ACA), the anti-phosphatidylserine antibody (APSA), and the anti-phosphatidylcholine antibody (APCA)-were measured, and the APA-binding ability for placental villi using villous lipid fractions was studied. METHODS: Serum samples were taken from 70 cases of preeclampsia, including 42 cases of the severe type, and from 45 normal pregnant women. The APAs were measured by an enzyme linked immunoabsorbent assay (ELISA). Placental villous membranes (PVMs) from normal placenta were purified and the lipid fractions were extracted from PVMs. The antibody-binding ability for PVM lipid fractions was evaluated by the ELISA method. RESULTS: In IgG-antibody measurements, the rate of ACA-positive, APSA-positive and APCA-positive cases were 10.0%, 18.1%, and 7.1%, respectively. The presence of APAs was revealed in preeclampsia. Using thin-layer chromatography, negatively charged phospholipids were found in the lipid fractions obtained from PVMs. Antibody binding for the lipid fractions of PVMs was detected in 9 of 12 APA-positive cases in preeclampsia. CONCLUSION: The results suggest that one cause of preeclampsia is the production of antiphospholipid antibodies that bind to placental villous membrane antigens.

Antibodies, Anticardiolipin↗
PubMed2000

Hormonal placental function tests for fetal assessment in high risk pregnancies.

BACKGROUND: Biochemical tests of fetal well-being such as placental hormone levels have not always shown direct benefits for mothers and babies. OBJECTIVES: The objective of this review was to assess the effects of measuring placental hormone levels during high risk pregnancies. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and MEDLINE. We also contacted study authors. SELECTION CRITERIA: Adequately controlled trials comparing availability with no availability of hormone placental tests in high risk pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by two reviewers and this was checked by a colleague. Study authors were contacted for additional information. MAIN RESULTS: Two trials involving 3355 women were included. Methodological quality of the trials was acceptable, but results for only 230 out of 2733 women (abnormal hormone levels) were available from one trial. One trial compared oestriol levels reported promptly with levels measured but not reported. The overall perinatal mortality rate was 30.5 per 1000 live births weighing 500 grams or more. No beneficial effects of oestriol measurement on fetal outcome could be identified (odds ratio of perinatal death 0.87, 95% confidence interval 0.35 to 2.18). For abnormal test results only, the reported oestriol group showed a statistically non-significant trend to less perinatal mortality (odds ratio 0.48, 95% confidence interval 0.15 to 1.60). The other trial compared human placental lactogen measurements reported promptly with measurements not reported until after pregnancy. For abnormal test results, the reported group showed significantly less perinatal mortality with four (3.4%) deaths compared to 17 (15%) in the control group (odds ratio 0.25, 95% confidence interval 0.10 to 0.61). Data from normal test results was not available for this trial. REVIEWER'S CONCLUSIONS: The measurement of human placental lactogen levels may be of some value in high risk pregnancies, but there is not enough evidence to evaluate the use of hormone placental tests generally.

Female↗
PubMed2003

Multiple luteinizing hormone receptor (LHR) protein variants, interspecies reactivity of anti-LHR mAb clone 3B5, subcellular localization of LHR in human placenta, pelvic floor and brain, and possible role for LHR in the development of abnormal pregnancy, pelvic floor disorders and Alzheimer's disease.

Distinct luteinizing hormone receptor (LHR) protein variants exist due to the posttranslational modifications. Besides ovaries, LHR immunoreactivity (LHRI) was also found in other tissues, such as the brain, fallopian tube, endometrium, trophoblast and resident tissue macrophages. The 3B5 mouse monoclonal antibody was raised against purified rat LHR. In rat, porcine and human ovaries, the 3B5 identified six distinct LHR bands migrating at approximately 92, 80, 68, 59, 52 and 48 kDa. Characteristic LHRI was detected in rat, human and porcine corpora lutea. During cellular differentiation, subcellular LHR distribution changed from none to granular cytoplasmic, perinuclear, surface, nuclear and no staining. There were also differences in vascular LHR expression--lack of LHRI in ovarian vessels and strong staining of vessels in other tissues investigated. In normal human term placentae, villous LHRI was associated with blood sinusoids and cytotrophoblast cells, and rarely detected in trophoblastic syncytium. In all abnormal placentae, the LHRI of sinusoids was absent, and syncytium showed either enhanced (immature placental phenotypes) or no LHRI (aged placental phenotype). LHRI in human brain was identified in microglial cells (CD68+ resident macrophages). Protein extracts from human vaginal wall and levator ani muscle and fascia showed strong approximately 92 and 68 kDa species, and LHRI was detected in smooth muscle cells, fibroblasts, resident macrophages and nuclei of skeletal muscle fibers. Our observations indicate that, in contrast to the theory on the role of vascular hormone receptors in preferential pick up of circulating hormones, there is no need to enhance selective pick up rather only prevent LH/CG transport to inappropriate sites. Abnormal placental LHR expression may play a role in the development of abnormal pregnancy. Expression of LHR in the pelvic floor compartments suggests that high LH levels in postmenopausal women may contribute to the pelvic floor relaxation and increased incidence of pelvic floor disorders. Since chorionic gonadotropin increases secretion of a variety of cytokines by monocytes, and induces their inflammatory reaction and phagocytic activity, high LH levels in aging individuals may also activate microglia (mononuclear phagocyte system in the central nervous system) and contribute to the development of Alzheimer's disease and other inflammation-mediated neurodegenerative diseases.

Alzheimer Disease↗
PubMed1989

Human fibroblast tissue factor is inhibited by lipoprotein-associated coagulation inhibitor and placental anticoagulant protein but not by apolipoprotein A-II.

Studies of proteins that inhibit tissue factor activity have generally been conducted using either an extracted tissue homogenate ("thromboplastin") or tissue factor protein reconstituted into phospholipid vesicles rather than with tissue factor expressed in cell membranes (its physiological environment). In the present study, a human fibroblast cell strain was used to evaluate the effects of lipoprotein associated coagulation inhibitor (LACI), placental anticoagulant protein (PAP), and apolipoprotein A-II (apo A-II) on human tissue factor in cell membranes. LACI was tested from 7.8 to 500 pmol/L on fibroblasts cultured at cell densities ranging from 3,500 to 9,925 cells/well, and caused a progressive inhibition of tissue factor activity. PAP was tested from 3.9 nmol/L to 1 mumol/L at cell densities ranging from 4,500 to 15,400 cells/well and caused up to 83% inhibition of tissue factor activity. Inhibition by these proteins appeared to be influenced by cell density as well as whether the cells were intact or disrupted. Apo A-II, up to 1 mumol/L, did not inhibit the tissue factor activity of intact or disrupted fibroblasts at any cell density examined even though it did inhibit the activity of tissue factor in phospholipid vesicles. Of these inhibitors of tissue factor-dependent activation of factor X, LACI was the most effective in suppressing the generation of factor Xa activity. The effects obtained with apo A-II are clearly dependent on the nature of the tissue factor preparation with which it is tested. The disparity between the inhibitory effect of apo A-II on the activity of tissue factor reconstituted into lipid vesicles and the absence of effect on the activity of tissue factor remaining in cell membranes serves to reemphasize the necessity of reexamining results obtained with model systems using as nearly physiological reagents as possible.

Annexins↗
PubMed1991

Immunological and molecular characterization of plasminogen activator inhibitors 1 and 2 in baboon (Papio anubis) placental tissues.

Two major plasminogen activator inhibitors (PAI-1 and PAI-2) increase in the peripheral circulation during pregnancy in humans. PAI-1 is of vascular endothelial origin whereas PAI-2 is produced primarily by human placental tissues. This study was undertaken to determine a) if PAI-1 and PAI-2 are also present in the baboon and b) their association with pregnancy. Citrated plasma was obtained from pregnant baboons sequentially at 15 +/- 3-day intervals between Days 30 and 140 of pregnancy. PAI activity increased significantly (p less than 0.05) at Day 120 (15.3 IU/ml) and 140 (21.8 IU/ml) of gestation and returned to baseline (2.6 IU/ml) 48 h post cesarean section. Placental tissues obtained at cesarean section during the third trimester were either placed in explant culture, fixed for immunocytochemistry, or frozen for RNA extraction. Western blot analysis of tissue culture media (TCM) indicated that the polyclonal antibody to PAI-1 reacted with a major band (Mr 47 000) in TCM from placental tissues while the PAI-2 antibody reacted primarily with a doublet (Mr 67 000 and 69 000) in these same media. PAI-1 was immunocytochemically localized primarily in the chorioamniotic tissue (CAM-D) and PAI-2 was found predominantly in placental villi. Slot blot hybridization with cDNAs to PAI-1 and PAI-2 indicated that the mRNA for PAI-2 was found primarily in placental villi, whereas the mRNA for PAI-1 was present in all three tissue compartments.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2000

Chemopreventive effects of coumaperine from pepper on the initiation stage of chemical hepatocarcinogenesis in the rat.

This study was designed to investigate the chemopreventive action of three natural products, coumaperine, aurapten and an extract from rosemary, against the initiation stage of rat hepato-carcinogenesis. Coumaperine has been isolated from white pepper as a naturally occurring antioxidative agent, but its potential modifying effects on carcinogenesis remain unclear. In experiment 1, a modification of the model developed by Tsuda et al. was applied, with assessment of numbers and areas of induced glutathione S-transferase placental form (GST-P)-positive hepatocellular foci in male F344 rats. Coumaperine, aurapten and the extract from rosemary were administered i.g. at 100 mg / kg / day once daily for 5 days with initiation by diethylnitrosamine (DEN) on day 4 (20 mg / kg, i.p.). Numbers and areas of GST-P-positive foci in each group given test chemicals tended to be decreased as compared to the vehicle control group values, significance being achieved for number with coumaperine. Experiment 2 was planned to investigate the mechanism of the inhibitory effects of coumaperine. Livers at 8 h after initiation by DEN were examined with coumaperine administered at 100 mg / kg / day once daily for 3 days. Proliferating cell nuclear antigen (PCNA)-positive cells tended to be decreased as compared to the vehicle control, but no effects on apoptosis or cytochrome P-450 (CYP) 2E1 expression were apparent. Our results suggest that coumaperine provides protection against initiation of hepatocarcinogenesis, and that this is related to inhibition of cell proliferation.

Animals↗
PubMed1985

Abruptio placentae and disseminated intravascular coagulation.

Several parameters of hemostasis have been studied in 19 patients suffering from abruptio placentae. In 10 of them severe hemostatic alterations were detected and in 5, disseminated intravascular coagulation was observed. The patients were divided into four groups according to the severity of their clinical picture. The degree of placental separation was related to the severity and course of the clinical history and to the alterations in hemostasis detected at the most critical clinical moment. The analytical parameters were evaluated after extraction of the thromboplastic material. A good correlation was observed between the severity of the clinical picture and the degree of placental separation and the greatest analytical alteration, especially with cross-linked soluble fibrin monomer complexes (SFMC). In 9 of the 19 patients who showed analytical and/or clinical alterations, an improvement was detected in these alterations after evacuation of the uterus.

Abruptio Placentae↗
PubMed1977

Human placental lactogen mRNA and its structural genes during pregnancy: quantitation with a complementary DNA.

A complementary DNA (cDNA) strand was transcribed from human placental lactogen (hPL) mRNA. Based on alkaline sucrose gradient centrifugation, the size of the cDNA was about 8 S, which would represent at least 80% of the hPL mRNA. Previously we showed that four to five times more hPL was synthesized in cell-free extracts derived from term as compared to first trimester placentas. Hybridization of the cDNA with RNA derived from placental tissue revealed that there was about four times more hPL mRNA sequences in total RNA from term placenta than in a comparable quantity of total first trimester RNA. Only background hybridization was observed when the cDNA was incubated with RNA prepared from human kidney. To test if this differential accumulation of hPL mRNA was the result of an amplification of hPL genes, we hybridized the labeled cDNA with cellular DNA from first trimester and term placentas and with DNA isolated from human brain. In all cases, the amount of hPL sequences was approximately two copies per haploid genome. Thus, the enhanced synthesis of hPL mRNA appears to result from a transcriptional activation rather than an amplification of the hPL gene. The increase likely reflects placental differentiation in which the proportion of syncytial trophoblast increases at term.

DNA↗
PubMed1987

Calcium transport across the isolated dually perfused human placental lobule.

1. Movements of 45Ca and 3H2O in maternal to fetal (M----F) and fetal to maternal (F----M) directions across the dually perfused isolated human placental lobule were measured under steady-state conditions. 2. M----F values of the clearances (CR) and extractions (ER) of 45Ca relative to 3H2O were 0.371 +/- 0.056 and 0.492 +/- 0.086 (mean +/- S.E. of mean) respectively. The corresponding values for F----M movements were 0.277 +/- 0.017 and 0.251 +/- 0.010 respectively. The F----M perfusion flow ratio (QF/QM) was 0.34 +/- 0.01 throughout. Comparison with previously published data indicated a significant degree of membrane limitation to Ca transfers. 3. There was evidence of a mismatch between tissues receiving a fetal and those receiving a maternal perfusion. 4. The relative extraction ER was markedly and reversibly enhanced when perfusate total Ca was reduced from 2.4 to 0.1 mM. The effect was present in both M----F and F----M transfers and provided evidence for carrier-mediated uptake of Ca on both aspects of the placental barrier. Small and transient decreases in the relative clearance CR were observed on changing from 2.4 to 0.1 mM-Ca in M----F and to a lesser extent F----M transfers while transient increases were seen on changing from 0.1 back to 2.4 mM-Ca. 5. Measurement of net changes in Ca levels in closed-circuit studies indicated a significant release of both ionized (Ca2+) and total (CaT) Ca into the fetal perfusate at total Ringer solution concentrations of 1.4, 1.9 and 2.4 mM-Ca. Release of Ca into the maternal circuit was also observed using 1.4 mM-Ca Ringer solution but when 1.9 and 2.4 mM-Ca Ringer solution was used a net uptake occurred. 6. These findings strongly suggest that mechanisms by which Ca is transferred from M----F circulations in vivo are at least partly preserved in the in vitro human placental preparation. They indicate that this preparation is suitable for the study of these mechanisms and their regulation by hormonal and other factors.

Biological Transport↗