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Chorionic villi sampling: cytogenetic and clinical findings in 500 pregnancies.

The cytogenetic findings were analysed in a series of 500 pregnancies in which chorionic villi sampling was performed. In all cases a direct method was used, karyotyping being successful in 481 cases (96.2%). The main indication for sampling was maternal age over 36 (412 cases; 82.4%). Abnormal laboratory findings resulted in 24 terminations of pregnancy (4.8%); in addition five unexpected balanced chromosome rearrangements were detected. Twelve of 15 cytogenetic discrepancies were detected at amniocentesis, two after termination, and one at spontaneous abortion. Complete follow up data were available for the first 250 patients, among whom nine pregnancies (3.6%) ended in spontaneous abortion before the 20th week. There were no false negative findings. Seventy additional chromosome studies were performed because of failure of chorionic villi sampling or equivocal results, or for confirmation. Counselling before chorionic villi sampling should include the possibility that subsequent amniocentesis may be needed should mosaicism or other unexpected abnormalities be found. The success rate and accuracy of karyotyping chorionic villi samples by the direct method are acceptable but distinctly less than those of karyotyping cultured amniotic fluid cells.

Abortion, Spontaneous↗

Localization of transferrin receptor in the chorionic villi of complete molar pregnancy.

Transferrin receptors have been identified on the villous trophoblast of normal chorionic villi by immunohistochemical techniques. The current study investigated the expression of transferrin receptors on molar chorionic villi by using a monoclonal antibody in immunofluorescence assays. The villous trophoblast of molar chorionic villi was brightly positive for transferrin receptor in the immunofluorescence assay. The presence of transferrin receptors on molar villous trophoblast may influence the immunologic relationship between molar and host tissues.

Antibodies, Monoclonal↗

Relationship between tissue damage and heme oxygenase expression in chorionic villi of term human placenta.

Heme oxygenase (HO) catalyzes the oxidation of heme to carbon monoxide (CO), biliverdin, and iron and is thought to play a role in protecting tissues from oxidative damage. There are three isoforms of HO: HO-1 (inducible), HO-2 (constitutive), and HO-3 (unknown function). Preeclampsia is characterized by an inadequately perfused placenta and areas of tissue damage. We hypothesized that damaged areas of placentas from women with PE and uncomplicated pregnancies are associated with an alteration in HO expression. Compared with microsomes isolated from morphologically normal and peri-infarct chorionic villi of pathological placentas, microsomes from infarcted chorionic villi from the same placentas had decreased HO activity measured under optimized assay conditions. There was no correlation between microsomal HO levels and activity and tissue damage in uncomplicated pregnancies. Whereas there was no significant difference in HO-1 protein levels across all regions of uncomplicated and mildly preeclamptic pregnancies, HO-2 protein levels were decreased (P < 0.05) in peri-infarct regions and infarcted chorionic villi of mildly preeclamptic pregnancies. Immunohistochemical analysis revealed an apparent decrease in both HO-1 and HO-2 protein expression in damaged tissues. HO-1 and HO-2 were immunolocalized in the syncytiotrophoblast layer of the chorionic villi, the underlying cytotrophoblast, and in the vascular endothelium. This study suggests that the ability of the chorionic villi to oxidize heme to CO, biliverdin, and iron may be compromised in areas of tissue damage in the placenta of women with preeclampsia.

Blotting, Western↗

Immunocytochemistry of human chorionic gonadotropin in human chorionic villi.

The immunocytochemical localization of human chorionic gonadotropin was investigated in chorionic villi from the seventh to twelfth week of gestation. By the light microscopic peroxidase-antiperoxidase technique, positive reactions of human chorionic gonadotropin were found exclusively in the syncytiotrophoblast. Immunoelectron microscopy by means of the protein A-gold technique reveals localization of the immunoreactive gold particles in two kinds of membrane-bound granular inclusions in this cell; one type is granules of 200 to 300 nm in diameter with moderate electron density and the other is large electron-dense bodies of 500 to 1000 nm. The former seems to be Golgi-derived secretory granules that play a role in the release of human chorionic gonadotropin from the syncytiotrophoblast. Although the origin of the latter is still uncertain, a certain amount of this hormone might be stored or treated by lysosomal digestion in the large bodies during these stages.

Antigen-Antibody Reactions↗

Chorion villi sampling (CVS) for prenatal diagnosis of genetic disorders: first results and future research.

Chorion villi sampling (CVS) in the first trimester of pregnancy has become available recently as an alternative method to second trimester amniocentesis for prenatal diagnosis of genetic defects. Currently there are six different tissue sampling techniques being investigated in different centers around the world, but there are very few trials in ongoing pregnancies. From chorionic villi material cytogenetic and biochemical studies as well as DNA analyses can be performed. Different methods of chromosome analysis are being investigated at the University of California San Francisco and elsewhere to determine the most efficient and reliable techniques. Larger studies will be needed to establish the efficacy and the safety of the chorionic villi sampling procedure for the mother and the developing fetus. Although CVS is preferable to amniocentesis for psychological and medical reasons (earlier elective termination of a fetus with a genetic disorder), many questions remain to be answered in carefully controlled studies.

Adult↗

Direct and short-term culture preparation of chorionic villi. Is any one method best?

Chorionic villi from 20 diagnostic cases were prepared for cytogenetic analysis by a direct and two short-term culture methods. No significant differences were found between the methods in the quality and quantity of metaphases obtained. A further study using villi from 20 pre-termination patients indicated an inherent variation in the quality of villi resulting in inconsistent processing and variation in the response of a sample to the methods. This suggests that it would be advantageous to process the villi by more than one method.

Cells, Cultured↗

[Effects of mifepristone on proliferation and apoptosis in early chorionic villi].

OBJECTIVE: To study the effects of mifepristone on proliferation and apoptosis in early pregnant chorionic villi. METHODS: Proliferation and apoptosis in early pregnant choionic villi from surgical aspiration and mifepristone induced abortion were studied. Marker for proliferation is proliferating cell nuclear antigen (PCNA) immunohistochemical staining, apoptotic cell death was detected using DNA in situ terminal deoxynucleotidyl transferas-mediated dUTP-biotin nick ending labeling (TUNEL). RESULTS: In early pregnant chorionic villi, proliferating index of cytotrophoblasts was 53.74% +/- 1.34%, not only nuclear but also cytoplasma were PCNA positive staining, syncytiotrophoblasts were negative. There existed apoptotic cell death in trophoblasts, the apoptotic index were 2.52% +/- 0.86% in syncytiotrophoblasts and 0.52% +/- 0.26% in cytotrophblasts, respectively. After 2 days mifepristone treatment, the proliferating index of cytotrophoblasts decreased slightly and PCNA positive cytoplasmic expression disappeared; while apoptotic cell increased significantly, the apoptotic indices were 22.16% +/- 2.26% in syncytiotrophoblasts and 20.12% +/- 1.74% in cytotrophoblasts, respectively. CONCLUSION: Mifepristone may inhibit proliferation and promote apoptosis of trophoblasts in early pregnant chorionic villi.

Abortifacient Agents, Steroidal↗

Tubuloreticular inclusions in placental chorionic villi of rhesus monkeys after maternal treatment with interferon.

Tubuloreticular inclusions were observed in placental chorionic villi of rhesus monkeys after pregnant female monkeys were injected intramuscularly with recombinant leukocyte A interferon (25 X 10(6) units/kg). They were identified in endothelial cells, fibroblasts, and Hofbauer cells of the chorionic villi, providing evidence that interferon crossed at least part of the maternal-fetal partition. Induction of tubuloreticular inclusions in these cells by exogenous interferon has not been previously reported. Tightly packed regular tubular arrangements appeared in endothelial and Hofbauer cells and loosely organized tubular arrays in fibroblasts. The maximum dimension of the tubuloreticular inclusions measured 3 micron and the diameter of the tubules was 20 nm. The tubuloreticular inclusions were continuous with, and surrounded by, smooth endoplasmic reticulum that was often connected to rough endoplasmic reticulum. The tubuloreticular inclusions were not detected in placental chorionic villi from monkeys not treated with interferon or from interferon-treated monkeys 30 days after cessation of treatment. These results indicate that the formation of tubuloreticular inclusions in rhesus monkey placentas was a transient response associated with elevated serum levels of interferon.

Animals↗

A comparison of chorionic villi sampling and amniocentesis: acceptability of procedure and maternal attachment to pregnancy.

Sixty-one women with high-risk pregnancies participated in a clinical trial comparing chorionic villi sampling and amniocentesis. They were assessed initially on five background measures and maternal attachment to the fetus, then were randomly assigned to either chorionic villi sampling or an amniocentesis group. Women were further assessed for maternal attachment three times during pregnancy. During the final assessment, those who were planning future pregnancies were asked which of the two procedures they would prefer. No differences between groups were found on any of the five background measures. All 31 women who considered future pregnancies indicated a preference for chorionic villi sampling, regardless of the procedure they underwent during the study. Women receiving chorionic villi sampling reported significantly greater attachment during the second trimester than did women receiving amniocentesis, who viewed themselves as being less attached than other pregnant women during this period. The findings were interpreted as evidence for attachment suppression among women undergoing amniocentesis.

Amniocentesis↗

Iron-containing granules in the syncytiotrophoblast of the human chorionic villi.

The syncytiotrophoblast of the human chorionic villi during earlier stages of gestation contains abundant granules derived from Golgi complexes. The granules often include very electron-dense lamellae in their interior, and X-ray microanalysis revealed the presence of iron in these lamellae. It is, therefore, supposed that iron particles absorbed into the syncytiotrophoblast are transported to Golgi complexes and integrated into these lamellae. No evidences that the granules are released from the cells by exocytosis have been proved. Thus, one possibility to their nature might be considered, that is they play a role in lysosomal storage of iron during earlier stages when the capillaries in the chorionic stroma are undeveloped and so have little ability to transport iron into the fetal circulation.

Chorionic Villi↗

First trimester chorionic villi sampling and direct chromosome preparations.

Chorionic villi sampling was performed on 52 patients prior to elective termination of their pregnancies. Villi were obtained in 42, and direct chromosome preparations were successful in 41 of them. The use of a mixture of 0.075 M potassium chloride and 1% sodium citrate in the ratio of 2:1 for hypotonic treatment and 40% acetic acid for cell dispersal yielded chromosomes with good morphology and G-bands.

Biopsy, Needle↗

Prevalence of tetraploid metaphases in semidirect and cultured chorionic villi.

OBJECTIVE: Investigation of the normal frequency of tetraploid metaphases in semidirect (STC) and cultured (LTC) chorionic villi. METHODS: Fifty metaphases in STC- and in LTC-villi slides of 100 women of advanced maternal age were screened for tetraploidy. RESULTS: Up to three tetraploid metaphases were encountered in 27% of the STC-villi preparations; the scores fitted a Poisson distribution. In all LTC-villi preparations tetraploid cells were seen; the scores fitted a log-Gaussian distribution. CONCLUSIONS: On the basis of these distributions, we propose a protocol for the management of tetraploid metaphases in chorionic villi, strongly reducing the number of prenatal follow-up investigations.

Cells, Cultured↗

Transabdominal fine needle biopsy from chorionic villi in the first trimester.

A technique for sampling first trimester chorionic villi for prenatal diagnosis by transabdominal fine needle biopsy is described. Specimens of chorionic villi were obtained from 49 out of 58 women, a success rate of 84.5 per cent. No fetal or maternal complications were demonstrated in the period before abortion. The procedure is useful for obtaining fetal tissue for culturing, DNA analysis and direct chromosome analysis.

Abdomen↗

Identification of trophoblast in chorionic villi biopsy samples.

We have investigated a test for rapid discrimination between foetal and maternal origin of chorionic villi biopsy samples. A monoclonal antibody named H315 reacting against a specific antigen present on the surface of foetal trophoblastic cells, plus a double-colour staining technique (FITC + PI), have been used for the identification of foetal cells (H315-positive) and for visualization of nucleate (PI-positive) and anucleate (PI-negative) structures of chorionic villi. This test could be useful in differentiating foetal and maternal cells in chorionic villi biopsy samples currently used for prenatal diagnostic purposes.

Antibodies, Monoclonal↗

Telomerase activity in human chorionic villi and placenta determined by TRAP and in situ TRAP assay.

Telomerase activity (TA) was analysed in human chorionic villi and placenta in normal and abnormal pregnancy using the telomeric repeat amplification protocol (TRAP) and in situ TRAP assay. Twenty chorionic villi specimens and 25 placenta specimens from normal pregnancies were examined as well as placenta specimens from 10 cases of intrauterine growth retardation (IUGR; nine asymmetric and one symmetric). TA was detected in 18 of the 20 (90 per cent) chorionic villi specimens and in 18 of the 25 (72 per cent) placenta specimens from normal pregnancy. However, no or only weak TA was exhibited in the placenta specimens of the nine asymmetric IUGR cases. In situ TRAP assay detected TA in trophoblastic cells from normal pregnancy, but not in trophoblastic cells from cases of asymmetric IUGR.

Adult↗

Proteins of human chorionic villi tissue: construction of a protein map based upon fractionation by two-dimensional gel electrophoresis.

Proteins of human chorionic villi from the first trimester of pregnancy were studied by high-resolution two-dimensional gel electrophoresis. One hundred and ninety-eight proteins spots were identifiable and allowed the plotting of a two-dimensional map of the proteins of human chorionic villi from the eighth week of pregnancy. The proteins of human chorionic villi from the twelfth week of gestation and of villi from term placenta were also studied. A comparison of protein compositions at different gestational ages was made. The findings from two-dimensional electrophoretic analysis of villi proteins have been summarized and stored in a computer data base.

Chorionic Villi↗