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PubMed1983

Assessment of a transcervical aspiration technique for chorionic villus biopsy in the first trimester of pregnancy.

A cannula attached to a syringe was passed through the cervical canal into the uterine cavity in an attempt to aspirate chorionic villi just before termination of a first trimester pregnancy in 82 patients. Chorionic villi were obtained from 40% of these patients. In view of the value of this technique for the first trimester diagnosis of genetic disorder, development of the technique and further trials of transcervical aspiration of chorionic villi would seem warranted.

Biopsy, Needle↗
PubMed2000

Prenatal diagnosis of cystic fibrosis: a case of twin pregnancy diagnosis and a review of 5 years' experience.

We performed prenatal diagnoses for cystic fibrosis in 32 high risk (1:4) couples (including a dizygotic pregnancy). Chorionic villi sampling did not cause abortion or fetal malformation in any case. The preliminary analysis of 9 short tandem repeats always excluded maternal contamination of the DNA extracted from chorionic villi and confirmed paternity. Twenty-two prenatal diagnoses were made by direct analysis of the mutations. In seven cases diagnosis was made by the analysis of intragenic polymorphisms; in three cases, we analyzed two extragenic polymorphisms. The prenatal diagnosis (including genetic counselling) was completed within 24 h from the sampling. Seven prenatal diagnoses revealed an affected fetus; all couples opted for therapeutic abortion. In 17 cases the fetus was heterozygote, and in seven cases it was non carrier of mutated alleles. In the twin pregnancy, mutations were DeltaF508/N1303K. Direct analysis of the DNA extracted from the two independent samples of chorionic villi revealed one fetus non carrier of mutated alleles and the other a carrier of the N1303K mutation. Analysis of the HPRT locus predicted both the fetuses as males. Furthermore, the genotype of each fetus was defined after birth. The prenatal diagnosis with chorionic villi sampling plays a key role in the prevention of cystic fibrosis. The laboratories must be equipped for both the direct analysis of mutations and for the analysis of a large number of polymorphisms. The preliminary analysis of short tandem repeats is recommended both to exclude maternal contamination and to confirm parentage.

Chorionic Villi Sampling↗
PubMed2000

Use of the endometrial pipelle in the diagnosis of early abnormal gestations.

OBJECTIVE: To determine the sensitivity and specificity of the endometrial pipelle (Unimar, Wilton, CT) in detecting intrauterine chorionic villi, as compared to standard suction dilation and curettage (D and C). DESIGN: Prospective clinical study. SETTING: U. S. Navy Hospital near Tokyo, Japan. Small community hospital. PATIENT(S): All patients presenting for the surgical management of an early abnormal gestation were enrolled prospectively. The patient's age, gravidy, parity, estimated gestational age (EGA), diagnosis, preoperative beta human chorionic gonadotropin (betahCG) level, and preoperative transvaginal ultrasound (TVUS) data were recorded. INTERVENTION(S): An endometrial biopsy was performed in the operating room prior to standard suction dilatation and curettage (D and C). MAIN OUTCOME MEASURE(S): The presence of chorionic villi in the endometrial biopsy and curettage specimens were recorded for each patient. RESULT(S): The sensitivity of the endometrial pipelle in detecting intrauterine chorionic villi was 63% and the specificity was 80%. CONCLUSION(S): In the diagnosis of an early abnormal gestation, particularly in excluding an ectopic pregnancy, it would be ideal to replace the more invasive D and C with the outpatient endometrial biopsy in detecting intrauterine chorionic villi. However, the limited sensitivity of the endometrial biopsy limits its application in this clinical scenario. Our results compare with two previous studies and add to the limited patient database on this topic.

Adult↗
PubMedDate not supplied

Analysis of full fragile X mutations in fetal tissues and monozygotic twins indicate that abnormal methylation and somatic heterogeneity are established early in development.

The fragile X syndrome, the most common cause of inherited mental retardation, is characterized by unique genetic mechanisms, which include amplification of a CGG repeat and abnormal DNA methylation. We have proposed that 2 main types of mutations exist. Premutations do not cause mental retardation, and are characterized by an elongation of 70 to 500 bp, with little or no somatic heterogeneity and without abnormal methylation. Full mutations are associated with high risk of mental retardation, and consist of an amplification of 600 bp or more, with often extensive somatic heterogeneity, and with abnormal DNA methylation. To analyze whether the latter pattern is already established during fetal life, we have studied chorionic villi from 10 fetuses with a full mutation. In some cases we have compared them to corresponding fetal tissues. Our results indicate that somatic heterogeneity of the full mutation is established during (and possibly limited to) the very early stages of embryogenesis. This is supported by the extraordinary concordance in mutation patterns found in 2 sets of monozygotic twins (9 and 30 years old). While the methylation pattern specific of the inactive X chromosome appears rarely present on chorionic villi of normal females, the abnormal methylation characteristic of the full mutation was present in 8 of 9 male or female chorionic villi analyzed. This suggests that the methylation mechanisms responsible for establishing the inactive X chromosome pattern and the full mutation pattern are, at least in part, distinct. Our results validate the analysis of chorionic villi for direct prenatal diagnosis of the fragile X syndrome.

Adult↗
PubMed1997

[Early diagnosis of ectopic pregnancy by uterine curettage with serum hCG assay].

OBJECTIVE: To explore the value of uterine curettage combined with serum hCG assay in the diagnosis of early ectopic pregnancy (EP). METHODS: Judgement was done by uterine curettage pathologic examination of uterine content and determination of serum hCG in 61 patients with suspected EP with clinical symptoms, signs and auxillary examinations. RESULTS: Intrauterine pregnancy was confirmed in 17 patients by the presence of chorionic villi in the uterine contents. 26 patients were diagnosed as having EP by the absence of chorionic villi and elevated levels of hCG 24 hours after curettage. 18 patients who had no chorionic villi and the level of hCG fall down spontaneously after curettage were diagnosed having either intrauterine pregnancy or EP, mostly no further intervention. CONCLUSION: Diagnostic curettage with serum hCG determination can be used for differentiating intrauterine pregnancy from EP and it can also provide a reference for further arrangement.

Adult↗
PubMed2002

Investigations into the biosynthetic pathways for classical and ring B-unsaturated oestrogens in equine placental preparations and allantochorionic tissues.

In on-going studies of 'classical' and ring B-unsaturated oestrogens in equine pregnancy, the products of metabolism of [2,2,4,6,6-2H(5)]-testosterone and [16,16,17-2H(3)]-5,7-androstadiene-3 beta,17 beta-diol with equine placental subcellular preparations and allantochorionic villi have been identified. Using mixtures of unlabelled and [2H]-labelled steroid substrates has allowed the unequivocal identification of metabolites by twin-ion monitoring in gas chromatography-mass spectrometry (GC-MS). Two types of incubation were used: (i) static in vitro and (ii) dynamic in vitro. The latter involved the use of the Oxycell cartridge (Integra Bioscience Systems, St Albans, UK) whereby the tissue preparation was continuously supplied with supporting medium plus appropriate cofactors in the presence of uniform oxygenation. [2H(5)]-Testosterone was converted into [2H(4)]-oestradiol-17 beta, [2H(4)]-oestrone and [2H(3)]-6-dehydro-oestradiol-17 alpha in both placental and chorionic villi preparations, but to a greater extent in the latter, confirming the importance of the chorionic villi in oestrogen production in the horse. On the basis of GC-MS characteristics (M(+) m/z 477/482 (as O-methyl oxime-trimethyl silyl ether), evidence for 19-hydroxylation of testosterone was found in static incubations, while the presence of a 6-hydroxy-oestradiol-17 alpha was recorded in dynamic incubations (twin peaks in the mass spectrum at m/z 504/507, the molecular ion M(+)). It was not possible to determine the configuration at C-6. The formation of small, but significant, quantities of [2H(4)]-17 beta-dihydroequilin was also shown, and a biosynthetic pathway is proposed. In static incubations of placental microsomal fractions, the 17 beta-dihydro forms of both equilin and equilenin were shown to be major metabolites of [2H(3)]-5,7-androstadiene-3,17-diol. Using static incubations of chorionic villi, the deuterated substrate was converted into the 17 beta-dihydro forms of both equilin and equilenin, together with an unidentified metabolite (base peak, m/z 504/506). The isomeric 17-dihydroequilins were also obtained using the dynamic in vitro incubation of equine chorionic villi, together with the 17 beta-isomer of dihydroequilenin. Confirmation of the identity of 17 beta-dihydroequilin and 17 beta-dihydroequilenin was obtained by co-injection of the authentic unlabelled steroids with the phenolic fraction obtained from various incubations. Increases in the peak areas for the non-deuterated steroids (ions at m/z 414 (17 beta-dihydroequilin) and 412 (17 beta-dihydroequilenin) (both as bis-trimethyl silyl ether derivatives) were observed. Biosynthetic pathways for formation of the ring B-unsaturated oestrogens from 5,7-androstadiene-3 beta,17 beta-diol are proposed.

Androstenediol↗
PubMed2005

3-D ultrastructural distribution of collagen in human placental villi at term in relation to vascular tree.

In order to understand the 3-D distribution of collagen in relation to vascularization, chorionic villi of human placentae, belonging to normal pregnancies at term, were studied by scanning electron microscopy (SEM) after alkali maceration techniques, and by transmission electron microscopy (TEM). The villous tree appeared made of an uninterrupted structure of collagen fibres. The collagen fibres connected the chorionic villi axis with their basal plates and organised differently according to the various levels of villous branching. The collagen of stem villi showed copious fibres. The external fibres (facing the villous surface) were arranged mainly longitudinally. The central core of the villi (inner fibres) were arranged concentrically around the wall of the fetal vessels. Both external and internal fibres formed stratified lamellae or small parallel bundles. The inner core of stem villi showed small holes housing capillary spaces. Mature intermediate and terminal villi showed a scarce amount of collagen arranged in thin concentric layer within the villous core, surrounding numerous dilated capillary and sinusoid spaces.These observations demonstrated that the extracellular matrix of human chorionic villi is highly compartmentalised and shows a variable structural 3-D distribution depending on the branching level of the villous tree, such a distribution ensures the most favourable microenvironment for feto-maternal exchanges and it is likely able to provide a modulated support to the developing chorionic fetal vessels and trophoblastic layer as well.

Adult↗
PubMed2004

Norepinephrine transporter (NET), serotonin transporter (SERT), vesicular monoamine transporter (VMAT2) and organic cation transporters (OCT1, 2 and EMT) in human placenta from pre-eclamptic and normotensive pregnancies.

Pre-eclampsia is one of the most common causes of perinatal and maternal morbidity and mortality. High blood pressure and proteinuria are important clinical signs of pre-eclampsia. Sympathetic overactivity and elevated level of circulating vaso active substances, such as monoamines has been shown. Extracellular concentrations of monoamines are normally kept low by specific transporter proteins of which many are expressed in the placenta. In this study we used in situ hybridization and real-time PCR to study the gene expression of monoamine transporters, such as NET, SERT, VMAT2, EMT and OCT1/2, in normal as well as in pre-eclamptic placentae. We demonstrated high expression of NET mRNA in the trophoblast cells of the anchoring villi and a lower expression intensity in the chorionic villi. SERT mRNA was mainly detected in chorionic villi. VMAT2 mRNA was not detected in the central part of the placenta but was present in the spiral arteries of placenta bed biopsies, in cytokeratin positive cells. EMT mRNA was mainly detected in the intra lobular septa and together with OCT1 and OCT2 mRNAs also expressed in scattered cells of placental vessel adventitias. Moreover, quantitative analysis showed a significant lower expression of NET and EMT mRNAs in pre-eclamptic placentae as compared to the control group. A defective gene expression or function of these monoamines transporters might explain the elevated concentrations of monoamines in pre-eclamptic patients. Monoamine transporters may serve as a protective mechanism preventing vasoconstriction in the placental vascular bed and thereby securing a stable blood flow to the fetus.

Adult↗
PubMed1999

Expression of telomerase subunits and localization of telomerase activation in hydatidiform mole.

Activation of telomerase compensating for the loss of telomeres has been implicated in human cell immortalization and carcinogenesis. Telomeric repeat amplification protocol (TRAP) assay can be used to detect telomerase activity in a variety of malignant tumours, including those of the female reproductive tract which have been found to have high levels of telomerase activity. However, it is unclear whether all the cells or only a subset of cells within a tumour have telomerase activity. To determine the regulation mechanism of telomerase activity in hydatidiform moles, we studied telomerase activity at the single cell level (using an in situ TRAP assay), and expression of TLP1 (telomerase protein 1), TERC (telomerase RNA component) and TERT (telomerase reverse transcriptase component). Expression of TERC and TLP1 was observed in all normal chorionic villi, as well as in trophoblastic diseases, and various cell lines irrespective of telomerase activity. TERT expression was observed in trophoblastic diseases and normal chorionic villi with telomerase activity but not in normal chorionic villi without telomerase activity, except in some cases in the present series, indicating that TERT expression is closely associated with telomerase activity. Upregulation of TERT expression may thus play an important role in telomerase reactivation.

Carrier Proteins↗
PubMed2002

Prenatal diagnosis for multiple pregnancies.

In multiple pregnancies, first trimester ultrasound is crucial to diagnose chorionicity, to detect major structural defects, and to screen for chromosomal abnormalities based on nuchal translucency measurement. The efficacy of nuchal translucency measurement screening in twins might be improved when combined with first trimester maternal serum screening. In twins as in singletons, the risk of fetal loss attendant to chorionic villi sampling and to amniocentesis are similar. When an invasive procedure is indicated in twins, chorionic villi sampling has, over amniocentesis, the advantage of allowing selective termination to be performed in the first trimester, when the procedure related risk of miscarriage is minimal. It has the disadvantage of leading to ambiguous results in up to 2% of cases. While chorionic villi sampling is the choice technique in pregnancies at very high risk, amniocentesis is still indicated in cases at more moderate risk. In monochorionic pregnancies, selective termination can now be performed using a variety of techniques including bipolar or monopolar cord coagulation, and, in acardiac twins, alcohol ablation. However, selective termination remains more hazardous in monochorionic than in dichorionic pregnancies. The outcome of the twin-to-twin transfusion syndrome has been substantially improved by laser photocoagulation of placental shunts and by amniodrainage, but randomized trials are needed to establish the optimal therapeutic strategy, and further pathophysiologic research might result in new treatments.

Amniocentesis↗
PubMed1996

Prenatal diagnosis of Sanfilippo A syndrome: experience in 35 pregnancies at risk and the use of a new fluorogenic substrate for the heparin sulphamidase assay.

We have investigated the use of a 4-methylumbelliferone (MU)-derived artificial substrate, MU-alpha-D-N-sulphoglucosaminide, for the sulphamidase assay in chorionic villi and amniotic fluid cells. In the new two-step enzyme assay, fluorescent MU is released by the successive action of endogenous sulphamidase and an added yeast enzyme preparation which hydrolyses the MU-alpha-glucosaminide intermediate. Optimal conditions for a sensitive, accurate, and convenient procedure for use in the prenatal diagnosis of Sanfilippo A syndrome are described. Previously, prenatal diagnosis of Sanfilippo A syndrome has been achieved by a radioactive sulphamidase assay in chorionic villi or in cultured amniocytes and by two-dimensional electrophoresis of glycosaminoglycans in amniotic fluid. Our experience using these methods in 35 pregnancies at risk is reported. The feasibility of the new fluorogenic assay was evaluated by retrospective testing of stored homogenates of chorionic villi and amniotic fluid cells from 22 pregnancies at risk. Unequivocal assignment of the fetal status in five affected pregnancies and 17 pregnancies with a normal outcome confirms the reliability of the new sulphamidase assay, which is in every respect more convenient than the conventional method using 35S-radiolabelled heparin.

Amniocentesis↗
PubMed2000

Detection of a human parvovirus intrauterine infection with the polymerase chain reaction.

OBJECTIVE: To detect human parvovirus B19 intrauterine infection in pregnancy with the polymerase chain reaction (PCR). STUDY DESIGN: DNA of chorionic villi and amniotic fluid was extracted and the gene of human parvovirus B19 amplified with PCR. RESULTS: The study analyzed 61 specimens of chorionic villi and 26 specimens of amniotic fluid and found two positive specimens of chorionic villi and 1 positive specimen of amniotic fluid. CONCLUSION: The vertical transmission of human parvovirus B19 infection in early pregnancy may be a pathway of intrauterine infection. Chorionic villus sampling in early pregnancy and PCR could be developed as a method of prenatal diagnosis of human parvovirus B19 intrauterine infection.

Adult↗
PubMedDate not supplied

Cell surface antigen expression of first trimester chorionic villus samples.

First trimester chorionic villi obtained by chorionic villus sampling at approximately 9 weeks of gestation were investigated by indirect immunofluorescence to demonstrate trophoblast cell surface antigen expression. Villous trophoblast expressing the trophoblast specific markers transferrin receptor, human placental lactogen, and cytokeratin was also found to express a monomorphic major histocompatibility complex class I determinant recognized by the monoclonal antibody W6/32. W6/32 positive regions included sparsely scattered regions of villous trophoblast and fanning outgrowths of trophoblast. The class I antigenic determinant expressed by first trimester trophoblast was found to be recognized exclusively by W6/32 when assayed with a panel of anti-class I determinant monoclonal antibodies. Trophoblast W6/32 determinant expression was not increased after 24 hour organ culture in the presence of 200 U of interferon gamma. Exposure to interferon gamma resulted in increased class I antigen expression by mesenchyme and low level de novo mesenchyme class II antigen expression. These data suggest that early gestational stage villous trophoblast express non-classical class I antigens which do not seem to be subject to the regulatory effects of interferon gamma.

Antigens, Surface↗
PubMed1991

Prenatal diagnosis of Duchenne muscular dystrophy by restriction fragment length polymorphism analysis with pERT 87 intragenomic deoxyribonucleic acid probes.

Prenatal diagnosis of DMD was performed with three intragenic genomic probes and chorionic villus sampling. A total of 8 unrelated families with at least one DMD were analysed. DNA was extracted from peripheral white blood cells for carrier testings (50 individuals). For prenatal detection, it was extracted from chorionic villi obtained by chorionic villus sampling at 9 menstrual weeks. DNA was digested with an appropriate restriction enzyme followed by overnight electrophoresis in 1% agarose gels. DNA was transferred from the gel to nylon membrane according to the protocol of an alkaline transfer method. The pERT 87 probes were labeled by nick translation. The membranes were hybridized overnight after prehybridization. After washing, the membranes were exposed to X-ray films to make autoradiograms for restriction fragment length polymorphism analysis. Fetal sex was determined by a rapid screening test with a Y chromosome-specific repeat sequence. Out of 8 fetuses, 4 were males and 4 were females. All of 4 male fetuses were determined to be unaffected. Out of 4 female fetuses, 3 were diagnosed as non-carriers, and the carrier status of the remaining one was not able to be determined because her mother was not informative for all testings.

Chorionic Villi Sampling↗
PubMed1992

[Beta-mannosidase of trophoblast and humana skin fibroblasts].

Conditions for assay of beta-mannosidase activity in human chorionic villi were studied using the fluorogenic substrate 4-methylumbelliferyl-beta-D-mannopyranoside. Comparison of the biochemical properties of the chorionic villi beta-mannosidase with those of the enzyme from human cultured fibroblasts showed their similarity. Like the enzyme from skin fibroblasts, the chorionic villi beta-mannosidase had rather high activity. Both enzymes had virtually the same pH optimum (4.2-4.7) and Km value. The data presented suggest that chorion biopsy specimens can be used for prenatal determination of beta-mannosidase activity at the early stage of development.

Biopsy↗
PubMed1994

Expression and localization of aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV in the human placenta and fetal membranes.

OBJECTIVE: Our purpose was to determine the distribution of membrane-bound cell surface peptidases, namely aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV in the human placenta and fetal membranes. STUDY DESIGN: Frozen tissue sections of the first-trimester chorionic villi, term placentas, and term fetal membranes were stained by indirect immunofluorescence with specific monoclonal antibodies. RESULTS: In the first trimester chorionic villi cytotrophoblasts expressed both neutral endopeptidase and dipeptidyl peptidase IV, but syncytiotrophoblasts expressed only neutral endopeptidase. Stromal cells in the chorionic villi expressed the three peptidases at various intensities. In the term placentas villous syncytiotrophoblasts expressed neutral endopeptidase weakly, and the villous stromal cells expressed large amounts of both aminopeptidase N and dipeptidyl peptidase IV but neutral endopeptidase weakly or faintly. In the term fetal membranes amniotic epithelial cells and chorion laeve expressed both neutral endopeptidase and dipeptidyl peptidase IV. Decidual cells in the decidua parietalis moderately or highly expressed aminopeptidase N. CONCLUSION: Three peptidases, aminopeptidase N, neutral endopeptidase, and dipeptidyl peptidase IV, are expressed by different cell populations in the human placenta and fetal membranes, suggesting their respective and important roles at the maternofetal interface.

Aminopeptidases↗
PubMed1987

A case of first trimester chromosomal mosaicism confined to the cultivation of the gestational products.

The advantages of the emergence and development of chorionic villi sampling (CVS) for early prenatal diagnosis are evident, but there are a host of new diagnostic problems caused by the use of extraembryonic tissues. We report a case in which 45X/46XY mosaicism was diagnosed by cultivation of chorionic villi and fetal cells. Direct chromosomal preparations of chorionic villi failed to diagnose the abnormality.

Adult↗