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PubMed1991

[Bacteriologic findings before and in transcervical chorionic villi biopsy and their clinical relevance].

Between August 1987 and May 1989, bacteriological examinations of smears from the cervical canal was performed in 358 pregnant women, who underwent transcervical sampling of chorionic villi. The outcome of 349 of these pregnancies is documented. 12 of these patients (3.5%) had an artificial abortion for genetic reasons. In 12 other patients, the pregnancy ended before 28 weeks (one child surviving). Microbiological examinations showed that in 187 women (52%), it was possible to culture one or more microorganisms from the cervical canal. The most frequently detected pathogen was Chlamydia trachomatis (111 women, 31%). In pregnancies, where more than one microorganism could be cultured, the risk of following abortion was 8.9 times higher than in those, where no pathogens could be found. In only two of the 12 women with a subsequent spontaneous abortion (16.6%), no microorganism could be found. The bacterial contamination of the cervical canal during TC-CVS seems to be a risk factor for subsequent abortion. Therefore, a bacteriological examination of cervical flora should be performed before as well as during TC-CVS, and antibiotic therapy should be initiated in case of positive results.

Abortion, Septic↗
PubMed1993

Cytokine production in first trimester chorionic villi: detection of mRNAs and protein products in situ.

Growth factor cytokines must play important roles in placental growth and differentiation. We used cDNA-mRNA in situ hybridization to examine whether human first trimester chorionic villi could produce these substances locally. IL-1 beta and IL-4 mRNA was found in both trophoblast layers, whereas CSF-1 and TNF alpha were found mostly in syncytiotrophoblast with much less localization to cytotrophoblast areas. IFN-gamma message was exclusively found in the cytotrophoblast. Immunohistochemical analysis confirmed that cytokines are produced by placental tissue, and the patterns of expression correlated well with the data obtained by in situ hybridization. These data are discussed regarding possible roles of cytokines in regulation of MHC antigen expression by trophoblasts and immunomodulation of decidual lymphocytes.

Chorionic Villi↗
PubMed2005

Biochemical analysis of cultured chorionic villi for the prenatal diagnosis of peroxisomal disorders: biochemical thresholds and molecular sensitivity for maternal cell contamination detection.

OBJECTIVES: The prenatal diagnosis of peroxisomal disorders is most often performed by biochemical analysis of cultured chorionic villus sample (CVS) or amniocytes. We aimed to (a) highlight the risk of maternal cell contamination (MCC) in biochemical prenatal diagnosis, (b) establish the threshold of these biochemical assays to MCC, and (c) document the sensitivity of PCR based genotyping of microsatellites for the detection of MCC in prenatal diagnosis of inborn errors by biochemical analysis. METHODS: The threshold of each biochemical assay was assessed by co-cultivating fibroblasts from known affected and normal individuals. Genotypes for three polymorphic loci were determined by PCR and GeneScan analysis. The sensitivity of the molecular test was determined by DNA mixing experiments and isolation of DNA from co-cultivated fibroblasts. RESULTS: MCC was detected in 2.5% of at risk CVS cultures (n = 79). Co-cultivation of defective and normal fibroblasts demonstrated that the peroxisomal biochemical assays were accurate at 25% contamination. Very low level DNA or cell contamination (1-5%) was detectable by genotyping, but an allele did not yield a definitive peak based on morphology until approximately 10% contamination. Furthermore, we demonstrated that other inborn errors of metabolism might be more susceptible to diagnostic error by low level MCC. CONCLUSION: The sensitivity of the microsatellite analysis (> or =10%) is well within the threshold of peroxisomal biochemical assays. Although peroxisomal biochemical assays would not be predicted to introduce a false positive or negative result if MCC <10% were present but not recognised by molecular analysis, the same may not be true for other inborn errors of metabolism.

Cell Culture Techniques↗
PubMed1999

A case of discordant related abnormal karyotypes from chorionic villi and amniocytes.

A case of three discordant cell lines in prenatal diagnosis is described, of which two were abnormal related structural abnormalities of chromosome 11. One of the abnormal cell lines was seen in all metaphases examined from direct preparations of chorionic villi, the cultured preparations showing an apparently normal male karyotype; the other abnormal cell line was seen in conjunction with a normal cell line in cultured amniocytes. Prenatal diagnosis offered solely on chorionic villus sampling would have yielded a mistakenly normal result on the basis of established criteria for distinguishing true mosaicism from confined placental mosaicism.

Adult↗
PubMed1992

[Phenylalanine hydroxylase-like antibody in human chorionic villi].

An antigen similar by electrophoretic mobility to liver phenylalanine hydroxylase (PH) and cross-reacting with monoclonal antibody PH8 against liver PH was detected in extracts of soluble proteins in 6 from 23 samples of chorionic villi. An antigen with electrophoretic mobility corresponding to 40-41 kDa was detected in extracts of membrane proteins from these 23 samples by immunoblotting with monoclonal antibody PH8. Its molecular weight was similar to that of major chymotryptic peptide of human liver PH. The content of the antigen varied with samples and was less than 20 ng/mg of the extracted protein. Two-dimensional gel electrophoresis revealed only 1 spot of the antigen. The antigen did not react with monoclonal antibodies PH7 and PH9 epitopes of which were located in N-terminal fragment of liver PH. These data suggest that the antigen of membrane fraction could be a PH protein without N-terminal domain.

Antibodies↗
PubMed1984

Some new findings about Hofbauer cells in the chorionic villi of the human placenta.

The cytological structure of the Hofbauer cells was investigated in human placentas of the first and second trimesters of gestation. These cells are found in the stromal channel system of the chorionic villi core. Their walls, which are supported by collagen fiber bundles, are produced by reticulum cells and fibroblasts. The cytoplasmic processes of the Hofbauer cells are in contact with the walls of the channels without being associated with them by desmosomal complexes. Some of these cells have features in common with macrophages, such as cytoplasmic processes, larger vacuoles, many pinocytotic vesicles and intracytoplasmic granules. This system of vacuoles and vesicles enables micropinocytotic activity and phagocytosis. This type of Hofbauer cell resembles the typical macrophages. These cells may play a role in the regulation of stromal water content, transportation of ions and the flow of interstitial fluid. The most original finding of this study are long tubes observed in some Hofbauer cells and extending between the nucleus and the extracellular ground substance through the cytoplasm. One of these tubular formations resembles a cilium in structure with three limiting membranes and is filled with a slightly electron-dense substance. This type of Hofbauer cell may transport information between the nucleus and the extracellular ground substance by means of these tubular structures.

Chorionic Villi↗
PubMed1994

Rapid detection of chromosome aneuploidies in fetal blood and chorionic villi by fluorescence in situ hybridisation.

OBJECTIVE: Evaluation of fluorescence in situ hybridisation in the detection of numerical aberrations involving chromosomes X, Y, 13, 18 and 21. SETTING: Harris Birthright Research Centre for Fetal Medicine. SUBJECTS AND METHODS: Chorionic villi (n = 45) or fetal blood (n = 34) were obtained from 79 pregnancies undergoing fetal karyotyping at 10 to 39 weeks of gestation because of ultrasonographic markers of fetal chromosomal abnormality. Karyotyping was performed by both traditional cytogenetics and fluorescence in situ hybridisation, using commercially available kits which utilise a heterochromatic Y probe and the alpha satellite repeat probes for chromosomes X, 18, and 13/21. The frequency distributions of the number of signals obtained by fluorescence in situ hybridisation in the chromosomally normal and abnormal fetuses were compared. RESULTS: Traditional cytogenetic analysis demonstrated that the fetal karyotype was normal in 47 cases and abnormal in 32 (including 24 with trisomies 21, 18 or 13, three with triploidy, one with Turner's syndrome and four with translocations, deletions or mosaicism). With fluorescence in situ hybridisation it was possible to obtain accurate diagnosis of trisomy 18, Turner's or triploidy within six hours of sampling; signal distributions with these chromosomal abnormalities were very different from those of normals. However, for trisomies 21 and 13 there was an overlap in values with those from normals. CONCLUSIONS: In detection of fetal numerical chromosomal abnormalities the use of the combined 13/21 probe cannot provide sufficiently accurate results to justify abandonment of traditional cytogenetics in favour of fluorescence in situ hybridisation.

Aneuploidy↗
PubMedDate not supplied

Cost-effectiveness ratio of chorionic villi biopsy for the prenatal diagnosis of chromosomal aberrations as compared with amniocentesis.

The observed cost of amniocentesis for the prenatal diagnosis of chromosomal aberrations in pregnancies at risk because of advanced maternal age was compared with the estimated cost of chorionic villi biopsy (CVB). The cost of CVB was estimated to be 22% less than that of amniocentesis when the cost of the sampling procedure, the laboratory charges and the cost of spontaneous and therapeutic abortions were considered.

Abortion, Induced↗
PubMed1989

[Analysis of trophoblast subpopulations on touch smears of chorionic villi during normal pregnancy].

Trophoblasts of normal human pregnancy are classified into syncytiotrophoblasts (STs), cytotrophoblasts (CTs) and intermediate trophoblasts (ITs) which exist in cell columns and so forth. We analyzed the trophoblast subpopulations which appear on touch smears of chorionic villi morphologically and immunohistochemically, using the uterine contents of 37 cases of induced abortion. Troma 1, a rat monoclonal antibody, was utilized as a trophoblast marker and a mouse monoclonal antibody to HLA-A,B,C was employed to discriminate ITs from CTs. The results were as follows: 1. Two sorts of cells were positive for Troma 1 and therefore were considered to be trophoblastic: Multinuclear cells of various sizes and relatively large mononuclear cells. 2. Multinuclear trophoblasts were thought to be STs because of their characteristic cytomorphology and negative reaction for HLA-A,B,C. 3. The lower the gestational age was, the more ITs were observed. 4. The cellular and nuclear size of ITs varied and their chromatin was coarsely granular with aggregates. In addition one or more marked nucleoli were noted. Consequently it would be important to take care not to misdiagnose ITs as trophoblasts of a malignant nature.

Antibodies, Monoclonal↗
PubMed1986

Identification of chorion villi in abortion specimens.

The incidence of ectopic pregnancy is rising. All patients with a diagnosis of spontaneous abortion should be observed until they have been proved not to have an ectopic pregnancy. To help in identifying patients with an ectopic pregnancy, sensitive human chorionic gonadotropin tests, ultrasound, and laparoscopy have been used, yet there has been a tendency to forget the simple method of looking for chorion villi. In material specimens of 272 patients with the initial diagnosis of spontaneous abortion, the gynecologist could identify villi in 50% and the pathologist could identify another 30%. Thus, only 20% had to be observed by other methods and one-third proved to have an ectopic pregnancy.

Abortion, Incomplete↗
PubMed2002

Timing of the absence of FMR1 expression in full mutation chorionic villi.

Fragile X syndrome is caused by the expansion of the CGG repeat in the 5' untranslated region of the FMR1 gene. This expansion leads to methylation of the FMR1 promoter region thereby blocking FMR1 protein (FMRP) expression. Prenatal diagnosis can be performed on chorionic villi samples (CVS) by Southern blot analysis. Alternatively, for males, an immunohistochemical method has been introduced for CVS. In this study, we have used this immunocytochemical method for CVS in full mutation male fetuses at different times of gestational age, varying from 10.0-12.5 weeks, and in two cases of full mutation female fetuses (>13 weeks). FMRP expression studies in CVS from full mutation male fetuses (10.0-12.5 weeks) illustrate the timing of the disappearance of FMRP expression in these CVS. Until approximately 10 weeks of gestation, FMRP is expressed normally in full mutation male CVS, whereas FMRP is completely absent at 12.5 weeks of gestation. FMRP expression in full mutation female CVS (>13 weeks) is completely absent in a number of villi, whereas other villi show normal FMRP expression. Unlabelled villi can only be present in the absence of the expression of the full mutation FMR1 gene on one X-chromosome together with the X-inactivation of the normal X allele. FMRP positive villi can be explained by an active normal X allele. The presence of both positive and negative villi indicates that X-inactivation in human CVS is a random process. No villi are found with a mixture of both FMRP-expressing and non-FMRP-expressing cells. This indicates that X-inactivation occurs very early in development, before the villi start to proliferate, and that X-inactivation in villi is a clonal process. In addition, our results indicate that the timing of both X-inactivation and full mutation FMR1 allele inactivation is different, i.e. X-inactivation occurs earlier in development than inactivation of the full mutation.

Alleles↗
PubMed1994

Onset and distribution of factor XIII-containing cells in the mesenchyme of chorionic villi during early phase of human placentation.

To learn more about the distribution and possible function of factor XIII (FXIII)-containing cells of human placenta, paraffin embedded and frozen sections of placenta samples from the first trimester of pregnancies--terminated by legal abortions--were studied by single and double labelling immunomorphological techniques. It was observed that at the fifth gestational week in the chorionic mesenchyme, FXIII-containing small mononuclear, round shaped cells start to appear. The relative amount of the FXIII-containing cells rapidly increased up to the seventh gestational week, reaching nearly 30 per cent of all mesenchymal cells. Simultaneously these cells differentiated into large stellate cells having numerous vacuoles in their cytoplasm. These cells were characterized in double labelling experiments and proved to be macrophages (CD 14+, KiM7+, labelled with antimacrophage monoclonal antibody). In the fifth-seventh weeks of gestation, these cells were homogenously scattered in the immature mesenchymal connective tissue, but from the eight gestational week they tended to accumulate in the peripheral part of chorionic villi while the central mesenchyme showed intense fibrotic changes. The abundance and characteristic distribution of the FXIII-positive macrophages in the chorionic mesenchyme during the first trimester of pregnancy suggest that these cells may have an active role in the formation of connective tissue in the early phase of placentation.

Chorionic Villi↗
PubMed1989

[Microfilaments and intermediate filaments in the fetal capillary endothelial cells of the human chorionic villi--ultrastructural and immunohistochemical study].

Augmentation of cytoskeletons in fetal capillary endothelial cells is one of the well known pathologic changes in toxemic placentae. But we can find very few reports on these filaments in placentae, especially on their ontogeny. An ultrastructural study on human chorionic villi from 55 normal and 25 toxemic pregnancies, covering all periods of gestation, revealed that endothelial cells already had a small amount of intermediate filaments and microfilaments from 6 weeks' gestation and that they gradually increased as pregnancy progresses. In toxemic cases, intermediate filaments were markedly augmented, occasionally forming bundles, to occupy the cytoplasm of endothelial cells, so that there was often no space for cytoplasmic organelles. Intermediate filaments were predominant in thinner parts of endothelial cells; both intermediate filaments and microfilaments were easily recognized in thicker parts. These filaments seemed to determine the arrangement of organelles such as rough endoplasmic reticulum and mitochondria. Immunohistochemical and immunoelectron microscopic studies with monoclonal antibodies proved that endothelial cells contained vimentin and actin, not cytokeratin and desmin. In toxemic placentae, augmentation of intermediate filaments may disturb normal movements of endothelial cells, and may cause an increase in vascular resistance to placental microcirculation and intrauterine growth retardation in the fetus.

Actin Cytoskeleton↗
PubMed1995

Somatic stability in chorionic villi samples and other Huntington fetal tissues.

We have studied different tissues from two affected fetuses with Huntington's disease (HD). In the first case the analysis was performed at 11 weeks of pregnancy; CAG repeats from seven different tissues were compared with the results obtained in the chorionic villi sample (CVS). We found 42 CAG repeats in all samples. In the second case the study was done at 12 weeks; eight tissues (including brain) were studied and compared with the CVS; in all of them, 44 CAG repeats were obtained. Our results show a somatic stability in the different analyzed tissues and suggest that mitotic instability can be a secondary consequence of neuronal degeneration and gliosis. Likewise, our data show great viability in the prenatal diagnosis (PD) of Huntington's disease using samples from any tissue.

Abortion, Therapeutic↗
PubMed1995

Pallister-Killian syndrome: normal karyotype in prenatal chorionic villi, in postnatal lymphocytes, and in slowly growing epidermal cells, but mosaic tetrasomy 12p in skin fibroblasts.

We report on two patients with Pallister-Killian syndrome: an 18 month old male infant followed since the neonatal period and a 4 year old boy. Prenatal diagnosis by chorionic villi sampling (CVS) in the first case showed a normal karyotype without mosaicism. Chromosome analysis on peripheral lymphocytes of the newborn also showed a normal karyotype. The clinical diagnosis of Pallister-Killian syndrome was made after the first year of life because of the typical facial dysmorphism and other characteristic clinical features, such as frontotemporal alopecia, depigmented area of the skin, sensorineural hearing loss, and severe psychomotor retardation. Chromosome analysis from skin fibroblasts now showed an isochromosome 12p mosaicism. The origin of the extra chromosome was confirmed by in situ hybridisation using a chromosome 12 specific library. In the second case chromosomal analysis from peripheral lymphocytes at the age of 19 months showed a normal karyotype 46,XY. Following the clinical diagnosis of Pallister-Killian syndrome a superficial skin biopsy was performed which showed very poor and slow growth of cells and a normal karyotype. Because of the typical symptoms a larger and deeper skin biopsy was performed from which there was rapid growth of fibroblasts. Now the diagnosis was established on the basis of the presence of an i(12p) extra chromosome in 69% of the metaphases.

Abnormalities, Multiple↗