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PubMed1990

[Assessment of the mutagenic effect of maternal factors on human chorionic villi by micronucleus test].

Rapid determination of DNA damage by micronucleus test is well accepted. Animal bone marrow cells or human peripheral lymphocytes used in most studies could not directly reflect the influence of the mutagenic effect on the offsprings by environmental factors. Human chorionic villi micronucleus test to detect directly the mutagenic effect of environmental factors has not been reported in the literature. Direct determination of human chorionic villi micronuclei (CVMN) was established in our laboratory, to study the mutagenic effect of mother's age, gravidity, gestation age, abortion history, contraception (condom, diaphragm, rhythm, oral contraceptives, spermicide or IUD), smoking and drinking on the offsprings. Cross investigation and micronucleus test were used in 507 couples undergoing artificial abortion. Micronuclei were scored according to Countryman's standard 2,000 interphases were observed in each subject for CVMN frequency (%). Arcsine transformation (arcsine [Sqr (P)]) was used in transforming CVMN frequency and the analysis of variance were used for statistics. No correlation between CVMN frequency and mother's age, gravidity, gestation age, abortion history, and contraception was found. Neither smoking nor drinking habit was found among the women of this study. The CVMN frequency of husband smoking was 0.7645 +/- 0.0561%, of husband non-smoking-drinking was 0.5522 +/- 0.0616%, of husband drinking was 0.5667 +/- 0.2004%, of husband smoking and drinking was 0.7944 +/- 0.0754%. There was a statistical difference in CVMN frequency between husband smoking and non-smoking (F = 2.78 DF = 408 P less than 0.05). No significant difference was found between husband drinking and non-drinking.

Alcohol Drinking↗
PubMed2006

Expression and localization of tumor necrosis factor receptor 1 protein in the chorionic villi in early normal and spontaneous abortion.

BACKGROUND: Increasing tumor necrosis factor alpha (TNFalpha) expression is found to be positively associated with spontaneous abortion. TNFalpha acts through the binding of two types of transmembrane receptors called TNFR1 and TNFR2. Here we compare membrane TNFR1 (mTNFR1) protein expression on chorionic villi in women with spontaneous abortion (SA) and viable pregnancy. METHODS: In a prospective study, 31 women with SA and 30 with a viable pregnancy were studied. Chorionic villous membrane TNFR1 was detected by confocal laser scanning microscopy and immunohistochemistry. RESULTS: The mTNFR1 protein is expressed in villous stromal, vessel endothelial, syncytiotrophoblast, and cytotrophoblast cells in early pregnancy. The intensity of mTNFR1 fluorescence (mean+/-S.D.) in villous stromal cells was higher in the abortion groups than in the control group (35.99+/-6.35 versus 32.64+/-5.01; p<0.05). In the abortion groups, villous cytotrophoblast cells were intensely positive for mTNFR1, whereas mTNFR1 staining of vessel endothelial and syncytiotrophoblast cells was significantly lower (p<0.001). CONCLUSION: Abundant mTNFR1 expression in the cytotrophoblast cells in women with SA may mediate TNFalpha to induce programmed cell death in the mTNFR1-expressing cytotrophoblasts to limit their growth. The over-expressing mTNFR1 in villous stromal cells, mediating TNFalpha effects, may cause pathological changes or tissue damage in chorionic villi locally in nonviable pregnancies.

Abortion, Spontaneous↗
PubMed2006

Incestuous paternity detected by STR-typing of chorionic villi isolated from archival formalin-fixed paraffin-embedded abortion material using laser microdissection.

Microscopic examination of a blood clot expelled by a physically and mentally disabled woman taken to the emergency room because of genital bleeding revealed the presence of chorionic villi encircled by decidua, hemorrhage, and necrosis. In order to identify the father of the product of conception, sections of formalin-fixed, paraffin-embedded abortion material were subjected to laser microdissection: DNA extraction from chorionic villi selectively isolated from the surrounding tissues allowed successful STR-typing of fetal cells, which was otherwise prevented by excess maternal DNA. The large number of homozygous genotypes in the fetal profile suggested incestuous paternity. Analysis of reference DNA samples from male relatives excluded the woman's father, paternal grandfather, and maternal grandfather, whereas the obligate paternal alleles of the fetus were constantly present in the genotypes of the woman's brother, clearly demonstrating brother-sister incest (probability of paternity > 99.99999%).

Aborted Fetus↗
PubMed1992

First-trimester biochemical and molecular diagnoses using chorionic villi: high accuracy in the U.S. collaborative study.

The accuracy of biochemical and molecular prenatal diagnoses using chorionic villi as the fetal source was assessed by seven centres participating in the NICHD collaborative study on the safety and accuracy of chorionic villus sampling (CVS) and amniocentesis. Of 601 pregnancies studied, biochemical methods were used to determine the diagnosis in 283 fetuses at risk for 35 different metabolic disorders. Fifteen different lysosomal storage diseases accounted for 81 per cent of the biochemical prenatal diagnoses performed, with 57 per cent of these pregnancies at risk for Tay-Sachs disease. No errors were made in the biochemical diagnoses that predicted affected or unaffected fetuses. However, the diagnoses of certain disorders (e.g., mucopolysacchariodosis type IH, metachromatic leukodystrophy, and Krabbe disease) occasionally required confirmatory studies in cultured amniocytes because the enzyme results were inconclusive in direct and/or cultured villi or due to the presence of a pseudodeficiency allele. Of these, only the diagnosis of a fetus at risk for Krabbe disease remained inconclusive after special studies to discriminate between mutant and pseudo-deficiency alleles. Recombinant DNA techniques were used to predict the diagnosis of 318 fetuses at risk for 16 different disorders in which the defective disease gene could be detected either directly or by linkage analysis to a nearby polymorphic marker. Of these, 32 per cent were for haemoglobinopathies, 25 per cent for cystic fibrosis, 24 per cent for Duchenne or Becker muscular dystrophy, and 7 per cent for haemophilias. Pregnancies at risk for known disorders with specific molecular lesions (e.g., sickle cell disease) were accurately diagnosed in direct and/or cultured villi. Diagnoses requiring analyses with closely linked polymorphic markers were occasionally uniformative or inconclusive. Maternal contamination was not reported in any biochemical or molecular-based diagnosis. These studies document the high accuracy and rapidity of both biochemical and mutation-specific prenatal diagnoses with direct and cultured chorionic villi.

Adult↗
PubMed1995

Cytogenetic study employing chorionic villi in ectopic pregnancy.

Maternal factors such as salpingitis and peritubal adhesion are known to be associated with ectopic pregnancy; however, a few studies have considered the chromosomal complements of ectopic conceptuses. We studied 16 ectopic conceptuses obtained by surgical resection. The karyotyping of chorionic villi was performed using direct and culture technique. Among 16 studied cases, 14 cases showed normal karyotype (nine with 46, XY; five with 46, XX). One case showed trisomy 16(47, XY, + 16) and another showed variation from normal chromosomal complement (46, XY, 14s+), resulting in 6.3% incidence of the structural abnormalities of the chromosome. On the basis of our study, we determined the possibility of chorionic villi karyotyping in ectopic pregnancy. This ectopic conceptuses are no more likely to show chromosomal abnormalities than in utero conceptuses of comparable gestational age. Therefore, maternal factors such as salpingitis and peritubal adhesion are the most likely explanations for ectopic pregnancy.

Adult↗
PubMed1989

[Study of the choice between sampling of the chorionic villi and the amniotic fluid in prenatal diagnosis].

We are reporting the results of a 21-month study during which 653 couples were seen in consultation at the prenatal diagnostic center of the University Hospital in Amiens, referred by their physician. 171 patients presented a theoretical term under 11 weeks of amenorrhea, for whom the choice between chorionic villi biopsy or amniotic fluid tap was possible. The different situations and results are compared for each method. The rate of fetal death was 5.4 per cent for chorionic villi biopsy and 1.5 per cent for amniotic fluid tap.

Amniocentesis↗
PubMed1989

Direct preparations from chorionic villi--relationship between villous morphology and mitotic index.

It has been postulated that chorionic villi with abundant sprouts have a higher mitotic index and are therefore preferable for obtaining direct chromosome preparations from chorionic villus samples. This theory was tested by correlating villous morphology with mitotic index. Surprisingly, no statistically significant relationship was found. Choice of culture medium, however, was found to be important, with serum-free RPMI yielding a higher mitotic index than 40 per cent FCS in MEM. We conclude that villous morphology, as assessed in this study, is not a major factor in determining the success of direct chromosome preparations.

Cells, Cultured↗
PubMed1990

Physiological values of glucose-6-phosphate-dehydrogenase (G-6-PD) in chorionic villi.

The physiological activity values of glucose-6-phosphate dehydrogenase (G-6-PD) from chorionic villi homogenized in Triton X was evaluated. The frozen, non-cultivated chorionic samples were obtained from artificial abortion in an early period of gestation (6-11 weeks). The mean G-6-PD activity was 0.43 U/mg protein. The G-6-PD enzyme activity showed no correlation with the week of gestation.

Chorionic Villi↗
PubMed1984

First-trimester diagnosis on chorionic villi obtained by direct vision technique.

An improved technique for direct vision chorionic biopsy that gives a clear view of the amniotic sac was developed. With this technique, used in 48 women prior to vacuum aspiration and in six cases for diagnosis (karyotyping or enzyme analysis), it was possible to obtain chorionic villi free from contamination by maternal tissue. It was also possible to pick out villi (rich in blood vessels and with abundant buds on their surface) found to be most capable of growing in vitro. In the diagnostic cases, the pregnancies have continued uneventfully since the sampling; one pregnancy is now in the 32nd week.

Adult↗
PubMed1989

[Experiences with transcervical chorionic villi biopsy in 274 studies].

In the years 1985-1987, 274 transcervical chorionic villi biopsy samples were taken at the Klinikum Steglitz Medical Center of the Free University of Berlin. We have information on the outcome of 264 pregnancies (96.3%). 229 cases (83.6%) yielded diagnostically usable results. Multiple biopsy samples were taken in 59 (21.5%) cases. Abnormal results were found in 14 patients. Pregnancy was terminated in 6 cases (2.2%) for proven chromosomal aberrations. In two others, spontaneous abortion occurred before the intended induced abortion. In 4 cases of mosaics, amniocentesis was performed subsequently in the 16th week of pregnancy; three of those pregnancies were carried to full term, and the children are healthy. In one case, we saw a spontaneous abortion in the 26th week of pregnancy. In two cases, we found balanced translocations. Miscarriage was observed in 16 CVS patients (5.8%). Abortion was febrile in two cases, septic in two others.

Chorionic Villi Sampling↗
PubMed1994

Experimental in vivo and in vitro formation of type VI collagen periodic fibrils in chorionic villi of human placenta.

The distribution of type VI collagen in the chorionic villi of normal human term placenta was studied. Since 100 nm periodic fibrils (type VI collagen periodic fibrils) are known to form in the tendon, cornea and cultured fibroblasts by incubation with acidic ATP solution, we incubated the placental tissue with 20 mM ATP, fixed it with Karnovsky's fixative and examined under an electron microscope. Type VI collagen periodic fibrils widely occurred in the interstitial connective tissue, especially close to the basal lamina. By cryo-immunoelectron microscopy, labeling for collagen type VI was found in the area between the trophoblastic basal lamina and the endothelial basal lamina. Furthermore, we extracted type VI collagen from the human placenta and brought both extract and residues into ATP treatment. Periodic structure formation was rare in extracted preparation compared with residues. The results served to clarify the mechanism of fibril formation in the tissue.

Adenosine Triphosphate↗
PubMed2005

The source of endogenous carbon monoxide formation in human placental chorionic villi.

Carbon monoxide (CO) is proposed to play a role in placental vascular control, as the placenta produces and responds to CO. The mechanism by which CO is formed by the placenta is unclear but could be through heme oxygenase (HO) degradation of heme, lipid peroxidation, or both. Human placental cotyledons were perfused with Kreb s solution to remove blood. Chorionic villi segments were prepared for measurements of CO production in the absence/presence of an exogenous supply of heme substrate (methemalbumin), inhibitors of HO, or inhibitors of lipid peroxidation. HO inhibitors used were chromium mesoporphyrin (CrMP) (0.1 mM, 0.3 mM), and azalanstat (0.1 mM, 0.3 mM). The lipid peroxidation inhibitors used were EDTA (0.1 mM, 0.3 mM) and deferoxamine (0.1 mM). Incubation of villi segments with methemalbumin (0.15 mM, 0.3 mM, 0.45 mM) resulted in a concentration-dependent increase in CO formation above the basal, endogenous rate. CrMP and azalanstat inhibited basal endogenous CO production, whereas EDTA and deferoxamine enhanced CO formation above basal level. These results demonstrate that endogenous CO was formed by human chorionic villi from heme, primarily through the action of HO, and are consistent with the hypothesis that HO plays a role in the regulation of placental vasculature by the formation of heme-derived CO.

Carbon Monoxide↗
PubMed2006

(Potential) false-negative diagnoses in chorionic villi and a review of the literature.

OBJECTIVES: To assess the incidence of (potential) false-negative findings of cytogenetic diagnosis in STC-villi and/or LTC-villi and to determine the best strategy for karyotyping chorionic villi in order to avoid false-negative results. METHODS: 2476 chorionic villus samples were received for prenatal cytogenetic investigations. Karyotyping was routinely performed on STC- and LTC-villi preparations by G-banding. Fluorescence in situ hybridization (FISH) analyses were performed in addition to standard chromosome analysis when necessary. Sometimes follow-up investigations like amniocentesis were performed before a definite prenatal cytogenetic result could be reported. RESULTS: In 2389/2476 (96.5%) of the cases, both STC- and LTC-villi were investigated. Normal STC- with abnormal LTC-villi results and finally an abnormal fetal karyotype were detected in ten cases (10/2389; 0.42%); in 9/10 of the cases the indication was fetal ultrasound abnormalities. Normal STC- and LTC-villi and finally an abnormal fetal karyotype were detected in two cases (2/2389; 0.08%). CONCLUSION: The most reliable technique for prenatal diagnosis after chorionic villus sampling (CVS) is the combination of the analysis of both STC- and LTC-villi to reduce the incidence of false-negative findings to a minimum. In the case of fetal ultrasound abnormalities with a small amount of villi available, the investigation of LTC-villi is recommended over that of STC-villi.

Chorionic Villi Sampling↗
PubMed1997

Gene expressions of keratinocyte growth factor and its receptor in the human endometrium/decidua and chorionic villi.

Keratinocyte growth factor (KGF) is secreted from mesenchymal fibroblasts and has a mitogenic specificity for epithelial cells in a paracrine fashion. In order to clarify the biological significance of KGF in the human endometrium which undergoes dynamic changes under the influence of sex steroid hormone, we investigated the gene expressions of KGF and its receptor (KGF-R) in the human endometrium in various sex steroid hormone milieus and chorionic villi, by RT-PCR and Northern blot hybridization. The secretory phase endometrium had a KGF mRNA level 10-fold greater than that of the proliferative phase endometrium. Similarly abundant KGF mRNA was found in decidua and pseudopregnant endometrium compared with proliferative phase endometrium. The KGF-R mRNA was detected by RT-PCR in chorionic villi from early pregnancy. These results indicate that the gene for KGF expressed in the human endometrium is mainly regulated by progesterone and that KGF might have a role in the interaction between decidua and chorion in early pregnancy in man.

Blotting, Northern↗
PubMed1990

Soluble factors produced by isolated first-trimester chorionic villi directly inhibit proliferation of T cells.

Supernatants from human trophoblast cultures have been reported to be both suppressive and stimulatory of mitogen-induced T-lymphocyte transformation. These discordant observations have been attributed to methodologic differences among laboratories, but may instead relate to inadvertant cellular contamination during long-term culture. Retrieval of chorionic villi for genetic analysis has provided an opportunity to evaluate the immunoregulatory activity of the isolated villus preparation, unencumbered by extended culture techniques. Primary cytotrophoblast cultures were established from karyotypically and clinically normal pregnancies and the supernatants were recovered. Allogenic mononuclear cells, stimulated by phytohemagglutinin, were uniformly inhibited by trophoblast supernatant (mean inhibition, 34.3%). This inhibition was verified in one-way, mixed lymphocyte cultures and suppression of T cells occurred in a dose-dependent fashion over a range of trophoblast supernatant concentrations. No evidence of suppressor T-cell induction by trophoblast supernatant was noted with the use of both phenotypic studies of lymphocytes incubated with trophoblast supernatant and a functional suppressor T-cell inducer assay. We believe that the immunoregulatory properties of trophoblast supernatants identified with the use of these methods more closely approximate the paracrine function of the in situ villus.

Analysis of Variance↗
PubMed1991

First-trimester diagnosis of Hunter syndrome: very low iduronate sulphatase activity in chorionic villi from a heterozygous female fetus.

Iduronate sulphatase activity was determined in uncultured chorionic villi from four fetuses at risk for Hunter syndrome. All fetuses were shown to be female by chromosome analysis. Biopsy material from three fetuses showed iduronate sulphatase activity within normal limits whilst the fourth fetus showed activity reduced to 7 per cent of our control mean. The importance of fetal sexing in prenatal diagnosis of this condition is emphasized as female carrier fetuses may show iduronate sulphatase activity reduced to levels observed in affected males.

Chorionic Villi Sampling↗
PubMed1990

Identification of an intermediate form of beta-N-acetylhexosaminidase in chorionic villi.

A minor form of beta-N-acetylhexosaminidase has been found in chorionic villi, in addition to the major forms A and B. This form does not hydrolyze the 4-methylumbelliferyl-2-acetamido-2-deoxy-beta-D-glucopyranoside-6-sulpha te substrate, is thermostable, has a higher mol mass (120,000) than A and B (100,000) and on analytical isoelectric focusing, it shows a microheterogeneity with values ranging between 6.3 and 7.0. For these characteristics, it resembles beta-N-Acetylhexosaminidase P from pregnancy serum, from which is chromatographically indistinguishable.

Anions↗
PubMed1986

Immunoelectron microscope observations on secretion of human placental lactogen (hPL) in the human chorionic villi.

Immunoelectron microscope localization of human placental lactogen (hPL) was investigated in the chorionic villi from week 7 to full-term gestation with the protein A-gold technique. With specific antiserum against hPL, immuno-reactive gold particles were found to be preferentially located in Golgi-derived, electron-dense small granules of 80-180 nm in the syncytiotrophoblast. Our electron micrographs indicate that these small granules increase in number in the course of gestation and are released by exocytosis from the apical cell surface. The present study reveals that hPL is segregated from the Golgi apparatus, stored in the syncytiotrophoblast as secretory granules, and released into the maternal blood.

Chorionic Villi↗