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PubMed1980

Secretion of lysosomal enzymes by human placental villi in vitro.

Chorionic villi from first trimester and term human placentas have been incubated in vitro and shown to release the lysosomal enzymes, beta-hexosaminidase, alpha-glucosidase and beta-glucuronidase. There was negligible release of the cytoplasmic enzyme, lactate dehydrogenase, under the same conditions. The first trimester villi released proportionally more of their lysosomal enzyme content than did the term villi. Extracellular levels of beta-hexosaminidase were raised and those of alpha-glucosidase and beta-glucuronidase were lowered when tissue was incubated with 1 microM colchicine, suggesting that microtubules are involved in the control of lysosomal enzyme release from placental villi.

Chorionic Villi↗

PubMed1977

Morphology of the placenta in fetal I-cell disease.

Placentas were studied from three interrupted pregnancies of a mother whose first liveborn child had I-cell disease (mucolipidosis II). I-cell disease of the fetus was shown by investigation of the amniotic fluid, fetal cells and the aborted fetus in two pregnancies, but in the third case placenta was the only available product of conception. In every placenta extensive vacuolization of the syncytiotrophoblastic layer of the chorionic villi and chorionic mesenchymal cells was found. In electron microscopy the inclusions were identical to those of other tissues in I-cell disease. The importance of histological study of placenta in unexplained spontaneous abortions needs to be emphasized, since this may be the only way of detecing new cases of lysosomal storage diseases.

Abortion, Habitual↗

PubMed1995

Influence of organic cations on basic amino-acid uptake by human placental villi.

Human placental chorionic villi were incubated for 30 min with [3H]lysine or [3H]arginine and the distribution ratios (intracellular:extracellular concentrations) were determined. The ratios remained unchanged when Na+ in Earle's buffered salt solution was replaced with Li+. When Na+ was replaced with choline there was a significant increase is distribution ratios (lysine 1.34 +/- 0.33 v. 3.99 +/- 0.15, arginine 1.95 +/- 0.37 v. 5.05 +/- 1.16). Leucine, a neutral amino acid with a Na(+)-independent transport system, was unaffected by choline substitution. The distribution ratio for alanine, which is Na(+)-dependent, was reduced (2.50 +/- 0.41 v. 1.45 +/- 0.20). Two other quarternary amines, acetyl-beta-methylcholine and tetraethylammonium chloride (TEA) caused similar increases in the distribution ratios of the basic amino acids. Hordenine, a tertiary amine, was less effective and there was little or no effect with ephedrine, a secondary amine. The choline effect was first observable at concentrations of 105 mM. With TEA, there was a progressive increase in distribution ratios beginning at 29 mM. Lysine efflux was measured after incubation of villi with lysine in Earle's buffer or choline buffer. Lysine was rapidly released to the fresh medium with 25% more retained in choline-exposed villi. The amines may cause alterations in the kinetics of basic amino-acid transporters or may modify other aspects of placental physiology permitting an increase retention of the basic amino acids.

Amino Acids↗

PubMed1987

[Transcervical chorion biopsy--hitherto experiences and results of more than 200 cases].

This is a report on more than 228 chorionic biopsies performed at the Department of Gynaecology of the University of Heidelberg. After having completed the pilot study (about 100 cases before planned termination of pregnancy) with a success rate of 87% in obtaining useful chorionic villi we initiated chorionic biopsy for diagnostic purposes. A cytogenetic result was obtained in 95% of all cases after the villi had been sampled, using the method of transcervical aspiration. In 1% of the cases the obtained tissue could not be used; in another 1% a chromosomal mosaic-like pattern was seen, whereas in 3% of the cases no cytogenetic result was obtained despite the fact that partly the available tissue quantities were quite sufficient. No false sex diagnosis was made in any of the examined cases. In 98% of all instances of sampling of chorionic villi, a sufficient amount of useful chorionic villi tissue was obtained. Vaginal bleeding after chorionic biopsy occurred only in about one-third of the cases within 1-7 days after sampling. In another third of the patients questioned accordingly, no vaginal bleeding was reported following chorionic villi sampling. The remaining patients stated that there had been only short-term haemorrhages after biopsy. 122 of 226 patients have since delivered, 39 are at present in the 16th to 28th week of gestation, 41 beyond the 28th week and the remaining 13 were before the 16th week at the time they were questioned. Abortion or foetal death after chorionic biopsy was seen in four cases only (1.8%). No malformations were seen so far in the delivered infants.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1999

Vasculogenesis in early human placental villi: an ultrastructural study.

In this study we examined the chorionic villi of 5 normal human placentas at 12-14 weeks of gestation ultrastructurally with regard to differentiation of the vascular components. The aim of the present report is to discuss the factors influencing vasculogenesis (in situ formation of blood vessels) at the ultrastructural level. Our observations have led us to think that the cytotrophoblast influences vasculogenesis in human chorionic villi. Mesenchymal-preendothelial cell groups were always found in very close association with the cytotrophoblast at the periphery of the villi, forming blood vessels. The cytotrophoblast probably attracts mesenchymal cells towards the margin of the villi by secreting vascular endothelial growth factor (VEGF). Once cells attach to the trophoblastic basement membrane they begin to differentiate into endothelial cells. This close structural relation between two cell types (cytotrophoblast and mesenchymal cells) may not be the only mechanism controlling vasculogenesis, but it seems to be one of the factors influencing the differentiation of mesenchymal cells into the endothelial cells of blood vessels in early human chorionic villi.

Blood Vessels↗

PubMed1998

Uniparental disomy with and without confined placental mosaicism: a model for trisomic zygote rescue.

In the population of children born after prenatal cytogenetic investigation in chorionic villi at our department from 1992 to 1995 (N = 3940), three are known to us with uniparental disomy. One case of maternal heterodisomy 16 was prenatally discovered because of trisomy 16 in direct chorionic villi with subsequently normal amniotic fluid cells. The other two had normal karyotypes in chorionic villi. Maternal heterodisomy 15 was postnatally detected in one of them because of Prader-Willi syndrome. Maternal hetero/isodisomy 16 was accidentally encountered in the other case in the course of prenatal DNA analysis of the tuberous sclerosis complex 2 region at 16p13.3. A model is presented for the understanding of the various combinations of karyotypes in direct chorionic villi, cultured chorionic villi and the fetus in the case of successful and unsuccessful trisomic zygote rescue.

Adult↗

PubMed2003

Placental villotrophoblastic pulmonary emboli after elective abortion: immunohistochemical diagnosis and comparison with ten control cases.

Although pulmonary trophoblastic embolism is now considered a physiologic phenomenon of normal pregnancy, this phenomenon has not been demonstrated in a living asymptomatic patient. Recently we encountered a 26-year-old woman suspected of pulmonary embolism of villotrophoblastic tissues after therapeutic abortion. Although her serum beta-hCG was low, a computed tomography scan showed multiple nodules in both lungs. Histological examination of a nodule in a lung-biopsy specimen showed granulation tissue surrounding a hemorrhagic mass within which were structures resembling degenerating chorionic villi. Immunohistochemical study on the patient's lung nodule, and a second endometrial-curettage specimen, six control endometrial and tubal specimens containing degenerating chorionic villi, and four endometrial specimens containing viable chorionic villi were performed. The patterns of immunostaining for cytokeratin, human chorionic gonadotropin, human placental lactogen, placental alkaline phosphatase, and inhibin-alpha of the chorionic villus-like structures in the lung nodule were almost identical to those in the degenerating chorionic villi, but different from those of viable villi. This is a unique case of embolism of chorionic villi and trophoblast to the lung in a living patient after therapeutic abortion.

Abortion, Induced↗

PubMed1996

Expression of heparin-binding epidermal growth factor messenger RNA in the human endometrium.

Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the epidermal growth factor family. In rodents HB-EGF has been found to mediate the mitogenic effects of oestrogen on the endometrial glandular cells and those of progesterone on the stromal cells. HB-EGF has also been found to be expressed at the site of implantation before any other discernible sign of blastocyst attachment in rodents. Our aim was to determine whether HB-EGF mRNA was expressed in the human pregnant and non-pregnant endometrial tissues. Normal endometrial tissues at different stages of the menstrual cycle, first trimester chorionic villi and decidua, second trimester chorionic villi and term placental tissues were snap frozen and stored at -80 degrees C. Reverse transcription-polymerase chain reaction (RT-PCR) was used to analyse the expression of HB-EGF mRNA in these tissues. HB-EGF mRNA was found to be expressed in human endometrium throughout the menstrual cycle. It was also present in first trimester chorionic villi and decidua, second trimester villi and term placenta. These findings suggest that HB-EGF may have a role in the growth or function of the human uterus, and that it may be important at the feto-maternal interface throughout pregnancy.

DNA, Complementary↗

PubMed1996

Prenatal diagnosis of succinyl-coenzyme A:3-ketoacid coenzyme A transferase deficiency.

Succinyl-CoA:3-ketoacid CoA transferase (SCOT) deficiency is a rare disorder of ketone body catabolism. In the present study, we prenatally diagnosed SCOT deficiency in a fetus in a family of which the proband was the first patient with SCOT deficiency identified in Japan, by analysis of enzyme activity levels in samples of chorionic villi and cultured amniocytes. In the fetus of the family, SCOT activity was not detected in either chorionic villi or cultured amniocytes. Since the levels of SCOT activity in control chorionic villi were close to our minimal detectable level and were much lower than those in control cultured amniocytes, enzyme assay in cultured amniocytes was more feasible than that in chorionic villi for prenatal diagnosis of SCOT deficiency. No elevated accumulation of 3-hydroxybutyrate or acetoacetate was detected in the amniotic fluid of the fetus. To our knowledge, this report is the first of prenatal diagnosis of SCOT deficiency.

3-Hydroxybutyric Acid↗

PubMed1994

Tubal ectopic pregnancy: a histologic, morphometric, and statistical study.

Pathologists are occasionally asked for medicolegal reasons to estimate the gestational age of an ectopic pregnancy based on morphologic findings. We evaluated the correlation between the gestational age and the maximal diameter of the resected fallopian tube, the average diameter of chorionic villi, the number of chorionic villi per square millimeter of the microscopic section, the number of degenerated villi per square millimeter, the number of syncytial knots per square millimeter, and the average number of layers of cytotrophoblastic cells covering the villi. There was a significant correlation between the diameter of the fallopian tube and the gestational age, with a correlation coefficient of 0.41 and a P value of .05. There was no significant correlation of the gestational age with any of the microscopic parameters measured. The findings indicate that an approximate estimation of the gestational age of tubal ectopic pregnancies may be made using the diameter of the resected fallopian tube.

Chorionic Villi↗

PubMed1999

The predictive value of endometrial stripe thickness in patients with suspected ectopic pregnancy who have an empty uterus at ultrasonography.

UNLABELLED: Prior research suggests that, in patients with empty uteri at ultrasonography, endometrial stripe thickness may be predictive of ectopic pregnancy or the likelihood of obtaining chorionic villi after a dilatation and evacuation procedure (D+E). However, it is unclear whether the predictive value of endometrial stripe thickness is confined to patients with low beta-human chorionic gonadotropin (beta-hCG) values. OBJECTIVE: To determine whether endometrial stripe thickness is predictive of the risk of ectopic pregnancy or the likelihood of obtaining chorionic villi after D+E in patients with beta-hCG values >1,000 mIU/mL or < or =1,000 mIU/mL. METHODS: In an urban academic ED, the authors conducted a retrospective chart review of consecutive ED patients from August 1991 to August 1997 with abdominal pain or vaginal bleeding, a positive beta-hCG value, and an empty uterus by transvaginal ultrasound examination. Patients were divided into four groups-group 1: endometrium thin, beta-hCG value < or =1,000 mIU/mL; group 2: endometrium thick, beta-hCG value < or =1,000 mIU/mL; group 3: endometrium thin, beta-hCG value >1,000 mIU/ mL; and group 4: endometrium thick, beta-hCG value >1,000 mIU/mL. The secondary analysis was limited to patients who had a D+E performed within 48 hours of the ED visit. The risks of ectopic pregnancy and the likelihoods of obtaining chorionic villi after D+E were compared using chi-square or Fishers' exact test where appropriate, with a p-value of 0.05 being significant. RESULTS: 224 patients were enrolled in the initial analysis. Intergroup differences in the frequency of ectopic pregnancy were of borderline significance (p = 0.08). However, when the comparison was limited to the groups with beta-hCG values < or =1,000 mIU/mL, the predictive value of endometrial stripe thickness reached statistical significance (group 1: 27/99 [27%], group 2: 2/28 [7%], p = 0.05). 79 patients had a D+E performed. Intergroup differences in the rate of obtaining chorionic villi were significant (p = 0.002). Group 1 had the lowest frequency of having chorionic villi identified (4/26 [15%]) and was the only group in which villi were obtained in fewer than 50% of cases. CONCLUSION: Endometrial stripe thickness may be predictive of the risk of ectopic pregnancy and the likelihood of obtaining chorionic villi at D+E. However, its predictive value appears to be confined to patients with beta-hCG values < or =1,000 mIU/mL.

Academic Medical Centers↗

PubMed1996

Prenatal paternity testing with deoxyribonucleic acid techniques.

OBJECTIVES: Our purpose was to determine the feasibility and optimal techniques for prenatal paternity testing. STUDY DESIGN: Since January 1989 we have offered prenatal paternity testing by deoxyribonucleic acid testing. We analyzed the ability to complete the testing and the time required to complete the testing and developed polymerase chain reaction-based tests to speed test results. RESULTS: Before April 1990 only five of nine cases could be completed. Since that time 28 consecutive cases were successfully completed before delivery. Introduction of polymerase chain reaction-based testing has allowed us to perform testing on uncultured chorionic villi and to derive results within 3 weeks. CONCLUSION: Analysis of uncultured chorionic villi allows prenatal paternity testing to be completed within the first trimester of pregnancy. Prenatal paternity testing can also be performed on cultured amniocytes and chorionic villi.

Abortion, Therapeutic↗

PubMed2000

Human cytomegalovirus infection of placental cytotrophoblasts in vitro and in utero: implications for transmission and pathogenesis.

Human cytomegalovirus (CMV) is the leading cause of prenatal viral infection. Affected infants may suffer intrauterine growth retardation and serious neurologic impairment. Analysis of spontaneously aborted conceptuses shows that CMV infects the placenta before the embryo or fetus. In the human hemochorial placenta, maternal blood directly contacts syncytiotrophoblasts that cover chorionic villi and cytotrophoblasts that invade uterine vessels, suggesting possible routes for CMV transmission. To test this hypothesis, we exposed first-trimester chorionic villi and isolated cytotrophoblasts to CMV in vitro. In chorionic villi, syncytiotrophoblasts did not become infected, although clusters of underlying cytotrophoblasts expressed viral proteins. In chorionic villi that were infected with CMV in utero, syncytiotrophoblasts were often spared, whereas cytotrophoblasts and other cells of the villous core expressed viral proteins. Isolated cytotrophoblasts were also permissive for CMV replication in vitro; significantly, infection subsequently impaired the cytotrophoblasts' ability to differentiate and invade. These results suggest two possible routes of CMV transmission to the fetus: (i) across syncytiotrophoblasts with subsequent infection of the underlying cytotrophoblasts and (ii) via invasive cytotrophoblasts within the uterine wall. Furthermore, the observation that CMV infection impairs critical aspects of cytotrophoblast function offers testable hypotheses for explaining the deleterious effects of this virus on pregnancy outcome.

Cells, Cultured↗

PubMed2001

Fluorescence in situ hybridization of chorionic interphase cells for prenatal screening of Down syndrome.

OBJECTIVE: Our purpose was to determine the usefulness and reliability of fluorescence in situ hybridization on interphase chorionic villi cells in the prenatal diagnosis of Down syndrome. METHODS: A total of 336 samples of chorionic villi were analysed by direct chromosome preparation and FISH with a DNA probe specific to chromosome 21. The samples were obtained as part of the routine obstetric investigation and management. RESULTS: The sampling and direct karyotyping was successful in all cases. At least 50 cells were valuable by FISH in 331 of 336 samples. Both methods showed Down syndrome in 12 cases. The follow-up investigations showed that there was no false-negative or false-positive result following these procedures. CONCLUSION: Based on these results and the fact that it is possible to analyse by interphase FISH at least ten times more cells than by conventional cytogenetic methods, and these cells originate from different tissues of chorionic villi, it is concluded that FISH increases the reliability of the diagnosis. Nevertheless, more data are needed for correct statistical analysis. Since this method is cheaper and gives diagnosis earlier than cell culture, the combination of direct chromosome preparation and FISH on chorionic villi is offered for prenatal Down syndrome screening.

Chorionic Villi↗

PubMed1985

Immunohistochemical demonstration of placental hormones in the diagnosis of uterine versus ectopic pregnancy.

The authors find that immunohistochemical demonstration of placental hormones in endometrium is useful in the identification of trophoblast independent of the presence of chorionic villi. Human chorionic gonadotropin (HCG) and human placental lactogen (HPL) are markers for trophoblastic cells. The markers were studied in 21 cases of gestational endometrium without villi in which the clinical differential diagnosis was ectopic pregnancy versus missed or incomplete uterine abortion. Trophoblastic cells were identified in four cases by routine microscopy and in an additional seven cases using the markers. In none of these cases was there a subsequent demonstration of ectopic pregnancy. In six of the ten negative cases, ectopic pregnancies subsequently were removed. Thus, the use of these hormone markers in endometrial specimens increases precision in the diagnosis of uterine versus ectopic pregnancy.

Abortion, Incomplete↗

PubMed1987

First trimester prenatal diagnosis of lysosomal storage disease. Study of alpha-L-fucosidase isoenzyme patterns in fetal and maternal tissue.

The normal range of activities of 6 lysosomal enzymes was determined in extracts of chorionic villi samples obtained by a rigid forceps in the first trimester of pregnancy. These activities were compared to those in villi obtained after abortion and in cultured amniotic fluid cells and fibroblasts. For five of the six enzymes tested, the data suggest that first trimester prenatal diagnosis should be possible and reliable. For the sixth enzyme, alpha-L-fucosidase, where on occasion very low activities were found, the results obtained on fresh chorionic villi have to be interpreted with extreme caution. Considerable lysosomal enzyme activities were also found in maternal decidua. Therefore, extreme care must be taken in the preparation of chorionic villi for prenatal diagnosis of lysosomal disorders since even small amounts of maternal tissue could lead to misdiagnosis. This study has allowed us to monitor 2 pregnancies at risk for lysosomal storage disease. Differences in the isoenzyme pattern of alpha-L-fucosidase were found in chorionic villi and maternal decidua. Although further studies are required, this observation could lead to the development of immuno-biochemical methods to evaluate the purity of chorionic villi used for prenatal diagnosis.

Amniotic Fluid↗

PubMed2001

Cytogenetic investigation of fetuses and infants conceived through intracytoplasmic sperm injection.

OBJECTIVE: To determine the incidence of aneuploidy among fetuses and infants conceived through intracytoplasmic sperm injection (ICSI) in our clinic using umbilical cord blood samples. DESIGN: Follow-up study of the cytogenetic outcome of ICSI pregnancies. SETTING: University-based IVF clinic. PATIENT(S): Forty-six couples who underwent ICSI and conceived. INTERVENTION(S): Umbilical cord blood was taken after delivery of the infant for analysis. Samples of chorionic villi and chorion were taken for studies on the spontaneous abortuses. Amniocentesis was performed for couples that chose prenatal diagnosis. MAIN OUTCOME MEASURE(S): The cytogenetic chromosomal status of the pregnancy outcome. RESULT(S): Fifty pregnancies and 55 live births were recorded, with nine spontaneous abortions. Of 43 separate umbilical cord blood samples analyzed, 1 abnormality (2%) was found, 45, XX,+21. Nine births went through prenatal diagnosis alone, with four accepting both forms of analysis-no abnormalities were found. Origin of abnormality was established in two spontaneous abortion cases (45, XO and 45, XY,-21), and the maternal chromosome was lost in both cases. CONCLUSION(S): Using umbilical cord blood obtained after birth, we obtained karyotype results from 78% of the ICSI population in our clinic. Combined with results from five additional cases that underwent prenatal diagnosis but not umbilical cord blood sampling, a chromosomal result was obtained in 87% of our ICSI population. The use of umbilical cord blood for cytogenetic analysis substantially improves the ability to determine rates of chromosomal abnormalities in newborns produced via ICSI clinics.

Adult↗

PubMed1993

Ultrastructure of the tubal chorion.

The chorion of five cases of ampullar pregnancies aged 9-11 weeks was examined in electron microscope. The appearance of chorion is affected by massive haemorrhage into the tubal lumen which occurs in this type of tubal pregnancy and causes its segregation from the tubal wall. The chorionic villi scattered in haematoma show different structure. The electron microscope investigation reveals that many chorionic villi possess the intact microvillous surface and are similar to normal villi of the intrauterine chorion. The development of syncytiotrophoblastic organelles especially as far rough endoplasmic reticulum, Golgi apparatus and mitochondria suggests that the syncytiotrophoblast may for some period be functional under these unfavourable conditions. The chorionic villi imprisoned in blood clot are often covered by fibrin deposits, their syncytium shows considerable folded nuclei with coarse chromatin aggregations, extremely dilated and degranulated endoplasmic reticulum. When syncytium degenerates the fibrinoid sheets invest the isolated portions of the villi. The necrotic villi as well as the cells of cytotrophoblastic islands are embedded in the fibrinoid masses. The cytotrophoblastic cells resembling the cells of basal plate in placenta are found in the tubal wall adjacent to the haematoma.

Chorion↗