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Chromosomal abnormalities in ectopic pregnancy chorionic villi.

OBJECTIVE: To evaluate the incidence of chromosomal abnormalities in ectopic pregnancy chorionic villi. METHODS: A prospective study of patients with the diagnosis of ectopic pregnancy was conducted, with chorionic villi obtained at the time of surgical therapy cultured and analyzed for karyotype. Review of the patient's medical record and ultrasound evaluation was then completed and findings correlated with karyotype results. RESULTS: Twenty-two patients undergoing surgery for the diagnosis of ectopic pregnancy yielded chorionic villi for culture. Successful culture was performed in 21 patients, with 3 (14%) revealing abnormal karyotypes. Review of the medical record showed ultrasound results consistent with fetal development or a gestational sac in 15 of 18 patients with normal chromosomal analysis. Three of 6 patients without fetal development yielded abnormal chromosomal findings. CONCLUSION: Our results confirm that a high degree of success can be achieved in the karyotype analysis of ectopic pregnancy chorionic villi and that these conceptuses have a rate of abnormality similar to that reported for intrauterine gestations. Our data further suggest that when a gestational sac or fetal pole is identified by ultrasound, there is usually a normal karyotype.

Chorionic Gonadotropin, beta Subunit, Human↗

[Expression of cathepsins B and L in early gestational decidua and chorionic villi].

OBJECTIVE: To study the expression of cathepsins B and L in first-trimester gestational decidua and chorionic villi. METHODS: The decidua and chorionic villi in the first trimester of gestation were obtained from 30 women undergoing induced abortion, 25 with spontaneous abortion, 10 with normal endometrium in secretory phase, and 15 with bydatidiform mole in whom the expression of cathepsins B and L was determined by immunohistochemistry. RESULTS: Positive staining for cathepsins B and L were mainly detected in the trophoblasts and decidual cells. The positive expression rate of cathepsin B in the normal endometrium of secretory phase, decidua in the first trimester of gestation and spontaneous abortion were 10% (1/10), 83.3% (25/30), and 32.0% (8/25), respectively, and the rate of cathepsin L expression in the endometrium in secretory phase, decidua of induced abortion and spontaneous abortion were 0.0%, 63.3% (19/30), and 32.0% (8/25) respectively, showing significant difference in the expression rates of cathepsins B and L between the 3 groups (P<0.05). Strong positive expression of cathepsin B in chorionic villi of induced abortion, spontaneous abortion and hydatidiform mole were detected at the rates of 10.0% (3/30), 0.0%, and 66.7% (10/15), respectively, and cathepsin L at the rates of 26.7% (8/30), 4.0% (1/25), and 80.0% (12/15), respectively, with significant difference in strong cathepsins B and L expressions between the 3 groups (P<0.05). CONCLUSION: The expression of cathepsins B and L differs in normal and abnormal deciduas and chorionic villi of early pregnancy, suggesting that the cathepsins may play important roles in the process of implantation.

Cathepsin B↗

Expression of Ras GTPase-activating protein (GAP) in human normal chorionic villi and hydatidiform mole.

Ras GTPase-activating protein (GAP), an important downregulator of Ras activity, has previously been shown to be abundant in human placenta. The expression of p120 and p100 isoforms of GAP in human normal chorionic villi (n=5) and hydatidiform mole (n=5) was investigated to clarify the involvement of Ras GAP in the growth of chorionic villi in the first trimester of pregnancy. Immunoblot analysis revealed that both p120- and p100-GAP isoforms were remarkably less expressed in mole villi than in normal chorionic villi. The expression of p100-GAP significantly reduced in comparison with that of pl20-GAP in mole villi. Northern blot analysis showed that the amount of GAP mRNA reduced in hydatidiform mole less than one-third of that in normal chorionic villi. The GAP activity, measured by the effect of tissue extract on the hydrolysis of Ras-bound GTP, was significantly lower in hydatidiform mole than in normal chorionic villi. These results suggest that Ras GAP may play an important role in the normal growth and differentiation of human chorionic villi in the first trimester.

Animals↗

[Prenatal diagnosis of hereditary diseases using chorionic villi].

The paper is based on the first results of a preliminary study of the cytogenetic analysis of chorionic villi samples. Chorionic villi were obtained from 31 women undergoing artificial abortion in the first trimester of pregnancy. The tissue was used for direct chromosome preparations and sex-chromatin examinations (X and Y body). The results suggest the possibility of a simple and quick prenatal determination of fetal sex in the first trimester and the provision of a sufficient number of mitosis with well-spread chromosomes. This new method may become the method of choice in prenatal diagnosis after being thoroughly examined and verified.

Chorionic Villi↗

Morphologic alterations and immunohistochemical analysis of alpha-fetoprotein and CD34 in chorionic villi of anembryonic pregnancy.

OBJECTIVE: To investigate the morphology of chorionic villi using light and electron microscopy, especially the expression of alpha-fetoprotein (AFP) in trophoblastic cells and the process of maturation and margination vasculogenesis proper using CD34 immunohistochemistry in tissues from the first trimester of pregnancy loss due to anembryonic pregnancy in comparison with embryonic pregnancy. METHODS: The study consisted of 2 groups: 9 patients with anembryonic pregnancies and 9 patients with embryonic pregnancies between 6 and 10 weeks of gestational age registered at the Department of Gynecology and Obstetrics, University Hospital of Karadeniz Technical University, Turkey, from March 2003 to December 2004. We examined the chorionic villi using light and electron microscopy. For immunohistochemical staining, we used AFP and CD34. RESULTS: Microscopically, pathologic changes were shown in syncytiotrophoblast cells of anembryonic pregnancies and AFP was strongly expressed by villous trophoblastic cells compared to embryonic pregnancies. We determined the CD34 positivity in both groups. In anembryonic pregnancies, vascular elements were much fewer in number compared with embryonic pregnancies (p<0.001) and were located in the formed of hemangioblastic cords. CONCLUSION: Placental vasculogenesis is a basic feature in all types of pregnancy and a relationship exists between trophoblast cells and vessels in the chorionic villi with the potential to influence each other's functions. Defective chorionic villus vascularization is associated with embryonic death. This study may support the hypothesis, as suggested by previous studies, that anembryonic pregnancy results from early embryonic death and subsequent reabsorption rather than from the nondevelopment of an embryo.

Antigens, CD34↗

Ultrastructure of first trimester chorionic villi with regard to the prenatal diagnosis of genodermatoses.

Hopes are held out for chorion villus sampling, a technique which is gaining more and more importance for the first trimester prenatal diagnosis of chromosomal aberrations and metabolic abnormalities. A variety of inherited skin diseases can be diagnosed postnatally and prenatally (in the second trimester) by ultrastructural diagnostic markers. For evaluation of prenatal diagnosis in the first trimester, we investigated chorionic villi derived from the trophoblast layer of the early pregnancy by light microscopy and conventional electron microscopy. The ultrastructure of the cellular layers covering the villi, i.e., the inner cytotrophoblast and the outer syncytiotrophoblast, as well as that of the connective tissue of the inner extraembryonic mesoderm, are thoroughly described in relation to the ultrastructural changes in certain genodermatoses including epidermolyses and keratinization disorders. We found that chorionic villi have only a few of the characteristics differentiated in skin, and none of the structures which are relevant to the diagnosis of genodermatoses. In our view, the ultrastructural approach is not suitable for first trimester prenatal diagnosis of genodermatoses in chorionic villi.

Chorionic Villi↗

Detection of congenital cytomegalovirus infection by using chorionic villi of the early pregnancy and polymerase chain reaction.

OBJECTIVE: To detect congenital cytomegalovirus (CMV) infection of chorionic villi in early pregnancy. METHODS: Extraction of DNA of chorionic villi and amplification of the gene of major immediate-early (MIE) antigen of CMV using a polymerase chain reaction (PCR). RESULTS: Sixty-eight specimens of chorionic villi and 16 specimens were positive for CMV infection by PCR. The incidence of congenital CMV infection in the first trimester of pregnancy was 23.5%. CONCLUSIONS: The risk of transmission of CMV from mother to fetus in early pregnancy is very high and potential CMV carriers may transmit CMV to their fetus in early pregnancy.

Chorionic Villi↗

Interpretation of chromosome mosaicism and discrepancies in chorionic villi studies.

In 3,000 chorionic villi studies (CVS) 33 cases of mosaicism and 7 false-positive cell lines in all cells were seen. The mosaic cell lines were caused by aneuploidy of autosomes (13x), sex chromosomes (9x), and structural anomalies (11x). Mosaics of fetal origin were only 4 cases of trisomy 21 and one 47,XXY mosaic. In 7 cases abnormal karyotype of non-fetal origin was seen in all cells in direct studies, including trisomy 16 (3x) and trisomy 18 (2x). The combined use of direct CVS and cell cultures always uncovered the non-fetal origin of chromosome abnormalities and the study of cultured cells in all cases could have prevented 5 terminations. Complete follow-up studies demonstrated no false-negative results. Therefore, CVS can be nearly 100% accurate when both direct studies and cultures are examined in cases of mosaicism and other cell lines of possible non-fetal origin, such as trisomy 16, trisomy 18, translocation (21;21), and 45,X cells.

Aneuploidy↗

The changes in CAP activity in first trimester placental chorionic villi and maternal serum, and their clinical significance.

We investigated the CAP activity of placental chorionic villi and maternal serum in the first trimester using a new ultramicro fluorometric method and obtained the following results: The CAP activity of the lysosome fraction (C3) was the greatest of the first trimester villi fractions (6.8 +/- 1.5n mol AMC/min/mg protein). No changes in CAP activity were seen in each fraction for the same wet weights of chorionic villi obtained during the different trimesters. The peak volumes (found with Sephadex G200) of the first trimester chorionic villi C3 fraction and the first trimester maternal serum almost coincided. CAP activity in normal healthy pregnant women in the 5th to 13th weeks of gestation rose gradually as pregnancy advanced. Maternal serum CAP activity was found to be lower than normal when abortion was imminent, but in cases in which the prognosis was favorable, the clinical signs, such as genital bleeding and lower abdominal pain, vanished and the values returned to within the normal range. However, when the prognosis was poor, the clinical signs did not improve and the values remained below normal. These findings suggest that first trimester CAP is produced by the lysosome fraction (C3) of the chorionic villi and that its activity in the maternal serum is directly related to placental function.

Abortion, Threatened↗

Confirmation of mosaicism and uniparental disomy in amniocytes, after detection of mosaic chromosome abnormalities in chorionic villi.

Chromosome mosaicism is detected in about 1-2% of chorionic villi samples (CVS), and may be due to a postzygotic nondisjunction event generating a trisomic cell line in an initially normal conceptus (mitotic origin) or the postzygotic loss of one chromosome in an initially trisomic conceptus (meiotic origin and trisomy rescue). Depending on the distribution of the abnormal cell line, the mosaic can be confined to the placenta (CPM) or generalised to the fetus (TFM, true fetal mosaicism). Trisomy rescue could theoretically be associated with a 33.3% probability of uniparental disomy (UPD) in the fetus. The aim of this study was to determine the risk of fetal involvement in a cohort of numerical and structural chromosome mosaics revealed in chorionic villi by means of combined direct and long-term culture analyses; we also determined the incidence of UPD associated with mosaic aneuploidies and supernumerary markers involving imprinted chromosomes. A total of 273 of a consecutive series of 15,109 CVS evaluated during a period of 5 years showed a mosaic condition in direct preparations and/or long-term cultures; confirmatory amniocentesis was performed in 203 cases. The abnormal cell line was extended to the fetus in 12.8% cases in terms of structural and numerical abnormalities involving autosomes and sex chromosomes; the risk of TFM varied and depended on the placental tissue distribution of the abnormal cell line. One of the 51 cases in which the mosaic involved an imprinted chromosome showed UPD, thus indicating a risk of 1.96%.

Amniotic Fluid↗

A comparison in concentration of heat shock proteins (HSP) 70 and 90 on chorionic villi of human placenta in normal pregnancies and in missed miscarriages.

PURPOSE: To investigate the role of heat shock protein (HSP) on the chorionic villi of human placental cells and to compare the concentration of placental HSP70 & 90 in term deliveries and in missed miscarriages. MATERIALS AND METHODS: Fifty products of conception from women who experienced first trimester missed miscarriage and 50 placentas from women who gave birth at term were studied. An immunohistochemical investigation was carried out with which we marked the localization of heat shock proteins 70 and 90 on the syncytiotrophoblastic, cytotrophoblastic, stromal and blood vessel cells, using specific antibodies which can detect the presence of those proteins on light microscopy. We compared their expression with the normal placental tissue of term pregnancies and with material acquired from first trimester missed miscarriages. An indirect immunoperoxidase method was applied using polyclonal antibodies against HSP70 and HSP90 on formalin-fixed paraffin-embedded tissues. RESULTS: Expression of HSP90B was increased in chorionic villi of first trimester missed miscarriages concerning syncytiotrophoblasts, cytotrophoblasts, vessel and stroma cells compared to full-term placentas. There was a statistically significant increase of HSP90A expression in chorionic villi of first trimester missed miscarriages, concerning only the cytotrophoblast cells, compared to full-term placentas. Expression of HSP70 cognate protein was significantly increased in chorionic villi of first trimester missed miscarriages, concerning syncytiotrophoblastic cells only, compared to full-term placentas. Finally, HSP70 inducible protein was significantly increased in chorionic villi of first trimester missed miscarriages concerning syncytiotrophoblasts, cytotrophoblasts, vessel and stroma cells compared to full-term placentas. CONCLUSIONS: The results of the present study have sufficiently shown that there is an increase of HSP70 & 90 expression in chorionic villi of first trimester missed miscarriages compared to full-term placentas and this increase may have an important implication on the miscarriage process.

Aborted Fetus↗

Chorionic villi sampling: clinical experience, immediate complications, and patient attitudes.

To develop chorionic villi sampling as a procedure for prenatal diagnosis, a pilot study was undertaken to perfect the obstetric and laboratory techniques, to evaluate our success with the procedure in continuing pregnancies, and to assess the attitudes of potential users of the procedure. Women about to have elective first-trimester abortions for nongenetic reasons were enrolled in the first phase of the study. Of the patients with a positive pregnancy test, 12.4% were found to have a nonviable pregnancy on ultrasound examination. Samples adequate for cytogenetic analysis were obtained in 130 of the 155 remaining cases, and the success rate was 93% in the 100 most recent cases. Direct cytogenetic analysis was undertaken in those cases successfully sampled, and karyotypes could be prepared in 97%. Immediate complications occurred in 5% of the pregnancies. Eight women at risk of bearing a child with a genetic defect had diagnostic chorionic villi sampling. Cytogenetic analysis was performed successfully on all of them. One had an induced abortion following the procedure because of the fetal diagnosis (a male with a 50% risk of Duchenne's muscular dystrophy). The other pregnancies are continuing uneventfully at 22 to 35 weeks' gestation. Finally, from preliminary analysis of our survey of potential users it appears that women 35 years old or over would prefer chorionic villi sampling to amniocentesis if the risks of the sampling were known to be low.

Adult↗

Efficient direct chromosome analyses and enzyme determinations from chorionic villi samples in the first trimester of pregnancy.

Chorionic villi were obtained by an aspiration technique which proved to be the best of four alternative procedures. We report in detail the series of experiments which led to (1) successful, rapidly growing cell cultures practically free of maternal cell contamination (the use of hormone-supplemented Chang medium greatly increased the growth rate); (2) an efficient direct method to obtain high quality metaphases from the Langhans cells of the cytotrophoblast tissue and with which the fetal karyotype is defined within a few hours of chorionic villi sampling; and (3) successful testing for the activity of eight enzymes directly from the villi samples, thus showing that this material is suitable for a rapid, direct diagnosis of the related metabolic diseases.

Catheterization↗

Prenatal diagnosis of a true fetal tetraploidy in direct and cultured chorionic villi.

Prenatal diagnosis of a true fetal tetraploidy in direct and cultured chorionic villi: Tetraploidy is characterized by four complete sets of chromosomes (4n= 92). Although it has been frequently reported in spontaneous abortions, tetraploidy is extremely rare in term pregnancy. Most of late surviving patients are diploid/tetraploid mosaics and present severe mental and physical impairment. Up to date, only five tetraploidies were ascertained in the prenatal stage in amniocytes and/or fetal blood lymphocytes. No one has been reported in chorionic villi probably because tetraploidy is generally considered in this tissue as a false positive result due to confined placental mosaicism (CPM) or placental culture artefacts. We report here on a case of tetraploidy detected in chorionic villi because of fetal cystic hygroma. We discuss the reliability of this diagnosis and propose guidelines in the follow-up of tetraploidies detected after chorionic villus sampling (CVS). Thus a misdiagnosis of this poor condition will be avoided at best and an appropriate genetic counseling will be given to the parents.

Amniocentesis↗

Endogenous carbon monoxide formation by chorionic villi of term human placenta.

Carbon monoxide (CO) is a novel messenger that is proposed to play a complementary role with nitric oxide in the regulation of placental haemodynamics. In a previous study, CO formation from exogenous haem has been measured in the microsomal fraction of chorionic villi as an index of haem oxygenase activity. The objective of the present study was to determine whether endogenous CO is formed by dissected chorionic villi of term human placenta, to which no exogenous substrate or co-factor had been added. Each sample of freshly isolated chorionic villi (approximately 0.4 g) of term human placenta from caesarean delivery was incubated in a sealed vial containing 1 ml of Krebs' solution (pH 7.4) at 37 degrees C. CO formation was determined by quantitating, using a gas-chromatographic method, the amount of CO released into the headspace gas of the incubation vial. There was time-dependent formation of endogenous CO in chorionic villi incubated at 37 degrees C during a 60-min time course. CO formation was found to be minimal in chorionic villi samples incubated at 4 degrees C and was increased relative to tissue weight. The data demonstrate that there is endogenous CO formation by chorionic villi of term human placenta.

Carbon Monoxide↗

Kinetic analysis, isolation, and characterization of hydrophilic folate-binding proteins released from chorionic villi cultured under serum-free conditions.

Full term placental chorionic villi cultured for 7 days in serum-free medium released hydrophilic folate-binding proteins (S-FBP(PCM) into the conditioned medium; in contrast, hydrophobic FBPs were the only form recovered from chorionic villi. Kinetic studies revealed that (i) S-FBP(PCM) was maximally released by the 3rd day, and this was associated with a proportionate decrease in hydrophobic FBPs; (ii) although cycloheximide inhibited de novo synthesis of [35S]methionine-labeled hydrophobic FBPs and S-FBP(PCM) by greater than 90%, unlabeled net S-FBP(PCM) release was only inhibited by 50%; (iii) EDTA markedly inhibited release of S-FBP(PCM) which was accompanied by a proportionate increase in hydrophobic FBPs; (iv) EDTA effects were completely reversed by 5-fold molar excesses of Mg2+ which led to a 50-fold greater release of S-FBP(PCM) compared to EDTA alone; (v) whereas Mg2+ alone stimulated S-FBP(PCM) release 4-fold greater than basal conditions, addition of cycloheximide to Mg2+ suppressed (by 4-fold) the expected increase observed with Mg2+ alone. Biochemical analyses of isolated S-FBP(PCM) revealed similarities to hydrophobic FBPs with respect to the ligand-binding domain and epitopes but differed in detergent-binding characteristics; furthermore, amino acid and carbohydrate analysis revealed a lower Mr = 25,500 with 12% carbohydrate. Based on kinetic analysis of S-FBP(PCM) release from chorionic villi-associated hydrophobic FBPs as well as structural analysis of S-FBP(PCM), these data continue to support the hypothesis that (a) a significant amount of maternal and probably fetal serum hydrophilic FBPs originate from placental hydrophobic FBPs, and (b) the endogenous hydrophobic FBP-directed Mg(2+)-dependent placental protease plays a major role in their release.

Carrier Proteins↗

Emotional responses of pregnant women to chorionic villi sampling or amniocentesis.

Seventy-four "high-risk" pregnant women interested in participating in a clinical trial comparing chorionic villi sampling and amniocentesis were initially assessed on five background measures and for anxiety, depression, hostility, and concern about abortion. The 61 women who then agreed to be randomized into the chorionic villi sampling and amniocentesis groups were assessed three additional times between 9 and 22 weeks' gestational age. Physical discomfort experienced during the diagnostic procedure was also assessed. Women in the chorionic villi sampling group underwent a reduction in anxiety up to 10 weeks earlier and reported less procedure-related discomfort than women in the amniocentesis group. The group X time interaction for depression was also significant, with women undergoing chorionic villi sampling reporting an earlier decrease in depression than women undergoing amniocentesis. No differences were found between the two groups in hostility or concern about abortion. Findings were discussed in the context of evidence linking prenatal maternal emotionality to an increased risk of obstetric complications.

Amniocentesis↗

Detection of Chlamydia trachomatis intrauterine infection using polymerase chain reaction on chorionic villi.

OBJECTIVE: To detect Chlamydia trachomatis intrauterine infection in the early pregnancy by using chorionic villi. METHOD: The C. trachomatis infection in pregnant women was investigated by cervical specimens and Clearview kits. DNA of chorionic villi was extracted and the gene of a major outer membrane protein of C. trachomatis was amplified by polymerase chain reaction (PCR). RESULTS: 120 cervical specimens of the pregnant women were analyzed and 10 cervical specimens were positive for C. trachomatis infection. In this study, the prevalence of C. trachomatis infection was approx. 8.3%. Fifty-nine specimens of chorionic villi and three positive specimens of C. trachomatis infection were analyzed by PCR. The incidence of C. trachomatis intrauterine infection in the early pregnancy was 5.1%. CONCLUSION: The vertical transmission of C. trachomatis infection in the early pregnancy may be a pathway of intrauterine infection. Chorionic villus sampling in early pregnancy and the PCR method could be developed as a technique for prenatal diagnosis of C. trachomatis intrauterine infection.

Cervix Uteri↗