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Anaphase lag as the most likely mechanism for monosomy X in direct cytotrophoblasts but not in mesenchymal core cells from the same villi.

A 36 year old white female was referred for chorionic villus sampling for advanced maternal age. Direct (cytotrophoblast) preparations of chorionic villi were 45,X, but cultured mesenchymal core cells from the same villi were 46,XX. Study of embryonic and extraembryonic tissues showed the aneuploidy to be limited to cytotrophoblasts from specific placental sites. In aggregate, the cytogenetic findings can best be explained by anaphase lag during development of the cytotrophoblast, suggesting that this cytological mechanism and not non-disjunction is responsible for the common occurrence of monosomy X in villi.

Adult↗

Non-invasive early prenatal molecular diagnosis using retrieved transcervical trophoblast cells.

Fetal DNA was recovered from 17 of 39 (44%) transcervical cell (TCC) samples obtained between 7 and 9 weeks of gestation by endocervical canal flushing. Trophoblast retrieval was adequate for polymerase chain reaction (PCR) amplification of Y chromosome-specific DNA sequences and detection of paternal-specific microsatellite alleles. The fetal sex predicted by PCR in TCCs was confirmed in all cases by karyotype analysis of chorionic villi at 10 weeks of gestation. The absence of the disease-associated paternal alleles in TCC samples from two pregnancies at risk for spinal muscular atrophy and myotonic dystrophy predicted unaffected fetuses in agreement with subsequent results on chorionic villi and newborns' leukocytes. A trisomy 21 fetus was diagnosed in TCCs using fluorescent in situ hybridization (FISH) and semi-quantitative PCR analysis of superoxide dismutase-1 (SOD 1). Present experience indicates that TCC sampling is a promising technique for early prenatal monitoring of Mendelian disorders and chromosome aneuploidy.

Adult↗

The bovine placenta; a source and target of steroid hormones: observations during the second half of gestation.

Apart from estrone-3-sulfate (E1S) the bovine placenta produces progesterone (P4), though the corpus luteum is the major source of P4 responsible for maintaining pregnancy. So far the biological function of placental steroids in cattle is largely unknown. However, since the local availability of free estrone (E1) in the placenta seems to be controlled by sulfatase and sulfotranferase, the hypothesis was developed that placental estrogens and P4 might act as local regulatory factors. To test for such a function placentomes from 150, 220, 240, 270 days (D) pregnant and parturient cows were screened immunohistochemically for progesterone and estrogen receptors (PR, ER). PR were found at all stages in the caruncle in stromal cells and capillary pericytes but only at parturition in arterial walls. Percentage of PR-positive caruncular stromal cells (CSC) increased (P<0.05) from 51.8+/-2.6% at D150 to 58.9+/-1.8% at parturition. ER were detected in CSC, caruncular epithelial (CE) cells and in caruncular capillary pericytes. Mean percentage of ER-positive CSC decreased from 39.0+/-5.9% in pregnant cows to 17.5+/-8.3% at parturition (P<0.05). In CE all cells exhibited positive signals with the exception of those immediately surrounding large primary chorionic villi. Proliferation was assessed immunohistochemically by determining the percentage of Ki67-antigen positive cells. Highest values (P<0.001) were obtained for CE (58.0-68.3%), followed by the trophoblast (23.3-25.4%), CSC (10.6-45.3%) and the stroma of the chorionic villi (2.9-10.5%). A transient depression of proliferation in CSC between D150-270 (P<0.05) paralleled local estrogen tissue concentrations. The results suggest that placental estrogens and P4 are important factors controlling caruncular growth, differentiation and function.

Animals↗

Spatiotemporal expression of three gap junction gene products involved in fetomaternal communication during rat pregnancy.

The expression of three different members of the gap junction multigene family, alpha 1 (Cx43), beta 1 (Cx32), and beta 2 (Cx26), was analysed in the rat implantation chamber (a structural unit containing fetal, extraembryonic and maternal components within the pregnant uterus) during mid- and late stages of gestation as well as in the delivering, post-partum and non-pregnant uterus. A differential, spatiotemporal and cell-type-specific regulation of gap junctional coexpression was observed for beta 1 and beta 2 in all epithelia examined (visceral, luminal and glandular), as well as for alpha 1 and beta 2 in decidual cells and keratinocytes of the fetal epidermis. alpha 1 antigen was detected in the mesometrial stroma, mesometrial myometrium, connective tissue, mesothelia of the amnion and visceral yolk sac and in the allantoic mesodermal layer throughout gestation. In addition, expression of alpha 1 in the placental basal zone and trophoblast giant cells coincided with the differentiation of these cells. beta 2 expression was observed prominently in the chorionic villi of the placental labyrinth. The presence of beta 1 and beta 2 in the visceral epithelium (visceral yolk sac = the primary route for embryonic nourishment prior to the formation of the chorioallantoic placenta) and beta 2 in the chorionic villi (placental barrier = the major fetomaternal exchange route) suggests that gap junctions have an important role in fetomaternal communication.

Animals↗

Right ectopic gestation following in vitro fertilisation: case report.

The management of ectopic gestation has in recent years transformed from the normally accepted laparotomy to the laparoscopic approach. The objective of this case report is to describe a rare occurrence of an ectopic gestation following in vitro fertilisation procedure. A 35-year-old para 0 + 0, presented with lower abdominal pain and per vaginal bleeding six weeks after an in vitro fertilisation was done in South Africa. The patient was admitted with severe lower abdominal pain and per vaginal bleeding at six weeks gestation following an in vitro fertilisation procedure. She had undergone an "evacuation" one-week prior to this episode due to an initial diagnosis of an incomplete abortion. No chorionic villi were reported on histology. The repeat serum BhCG was 777 mimicro/l and at laparoscopy a right unruptured ampullary ectopic gestation (4 cms in size) was evident. A right linear salpingostomy was subsequently performed laparoscopically. Histology confirmed the presence of tubal chorionic villi. The laparoscopic management of ectopic pregnancies is now regarded as the gold standard in many centres in the world. In this patient the ectopic pregnancy resulted following an intra-uterine zygote transfer, and was managed successfully.

Abdominal Pain↗

Pathological studies on the placenta from pre-term and term maternal toxicosis.

Fifty one placentae with maternal pre-term and term toxicosis were studied pathologically. In the severe pre-term maternal toxicosis, the placenta reveals hyperplasia of syncytial cytotrophoblasts(S-cells) with knots, fibrin deposit at their surface, fibrinoid degeneration of the stroma, in addition to marked interstitial fibrosis of chorionic villi. The basement membrane is thick and cytotrophoblasts(C-cells) are scattered beneath it. Decidual arteries are hyalinized, but a few decidual arteries are thick and almost obstructed in their lumina. On the other hand, the placenta of maternal term toxicosis show chorionic villi with few C-cells under the thin S-cell lining. Their basement membrane is thin with well developed vasculosyncytial membrane. Decidual arteries are more hyalinized and widened. These pathological changes seen in term maternal toxicosis are the severer than those of placental aging.

Eclampsia↗

Diffuse cystic change of a term placenta with a normal newborn.

We recently encountered a case of term placenta with a diffuse cystic lesion of the villi. A 19-year-old primipara at 36 weeks of gestation underwent cesarean section due to breech presentation with premature rupture of the membranes; she delivered a mature male baby of 2,502 g with an Apgar score of 9/9. The placenta was 940 g in weight and 29 x 20 x 3 cm in size, and macroscopically had multiple cystic lesions (3-8 mm in diameter) that resembled hydatidiform moles. However, histopathological examination revealed that the severe hydropic change was localized in the stem villi but not remarkable in the terminal chorionic villi. Moreover, abnormal proliferation of the trophoblast was not observed. However, the hypertrophic change was observed in the vascular wall of stem villi, in which hyperplasia of smooth muscle-like cells was present. The urinary hCG levels at 1 month and 2 months after delivery were less than 50 IU/l. These findings indicate that the multiple cystic lesions of the placenta in this case are essentially different from those of a trophoblastic disease, and that the diffuse cystic lesion of the villi might have been secondary to changes in the local circulation.

Actins↗

[Clinical value of antenatal diagnosis of intrauterine active human cytomegalovirus infection with reverse transcription polymerase chain reaction].

OBJECTIVE: To study the clinical value of antenatal diagnosis of intrauterine active human cytomegalovirus (HCMV) infection with reverse transcription polymerase chain reaction (RT-PCR). METHODS: The active HCMV infection of pregnant women was diagnosed with enzyme-linked immunosorbent assay (ELISA), serum Nest-PCR (N-PCR) and RT-PCR. The methods of antenatal diagnosis include villus sampling, amniocentesis and cordocentesis under ultrasound guidance. RT-PCR and N-PCR was used to screen fetal active infection. RESULTS: The infection rate of chorionic villi and cord blood whose mother was in active infection is 53.3% (16/30) and 47.8% (33/69) respectively, whereas that whose mother just had infectious history is 8.3% (4/48), 9.7% (6/62) respectively, the differences are statistically significant respectively (P < 0.001). The active infection rate of chorionic villi and cord blood in maternal active infection group is 30.0% (9/30) and 29.0% (20/69) respectively, whereas that with infectious history is 4.2% (2/48) and 4.8% (3/62), the differences are statistically significant respectively (P < 0.001). In the active intrauterine infection, the incidence of abortion and premature delivery was 41.2% (14/34), significantly higher than that of the latent infection 8.0% (2/25, P < 0.01). CONCLUSIONS: Antenatal diagnosis of intrauterine active HCMV infection with RT-PCR and the means of sampling described are of safety and reliability. It has a great clinical value in management of the fetus whose mother had been in active HCMV infection.

Cytomegalovirus Infections↗

Maternal serum levels of human chorionic somatotropin correlates with transferrin and erythropoietin in pregnancy.

During pregnancy, plasma levels of erythropoietin (EPO) and transferrin (TSF) show a gradual increase till term, when a slight decrease occurs. Human chorionic somatotropin (hCS) is a peptide hormone with a direct effect on the function of the syncytiotrophoblast and on the mass of functional placental tissue. In order to understand the relationships between the placental functions, maternal erythropoiesis and iron transfer to the fetus, the relationships between EPO and hCS and between TSF and hCS were investigated. A group of healthy pregnant women (n = 190) attending the Obstetric Clinic of the University of Siena were studied. They were between 18 and 43 weeks pregnant. A single blood sample was taken from each patient by radial vein puncture on admission into the clinic before the beginning of any therapy, for the determination of haemoglobin, EPO, ferritin, serum iron, TSF and hCS. The highest correlation was that between hCS and TSF (r = 0.336, p < 0.0001). This correlation was significant in anaemic patients and lost its significance in non-anaemic patients. In our study, we also found a significant relationship between hCS and logEPO (r = 0.195; p < 0.01) which reached an elevated significance over the entire survey, but did not reach statistical significance in anaemic women. We hypothesise that in pregnancy chorionic villi develop a great capacity to respond to pathological maternal events with the result of regulating the transport of iron, the oxygen tension and the maternal-fetal transfer of oxygen through haemoglobin. Evidently, the response of the villi serves to balance the lack of iron in favour of the fetus, probably through a mechanism that links the variations of the iron transport on the maternal side and the increased use on the fetal side through a dynamic and consensual response by the chorionic villi.

Adult↗

Localization of mRNA for the beta-subunit of placental hCG by in situ hybridization.

The cellular origin of placental hCG is not yet completely established. Depending on the method used, the syncytium, cytotrophoblast or both types of tissue have been claimed to synthesize hCG. In the present study in situ hybridization was used on sections of chorionic villi in order to detect expression of the gene for the beta-subunit of hCG (beta-hCG). Placental tissue was obtained from the 8th to the 11th weeks of pregnancy, when the concentration of hCG is high. A cDNA clone, encoding the entire amino acid sequence of beta-hCG, was used as a probe. Hybrids of beta-hCG cDNA/mRNA were found only over the syncytiotrophoblast. Background noise was extremely low and no signals above that were detected in the cytotrophoblast cells. It is concluded that at this stage of pregnancy the gene for beta-hCG is activated in the syncytial parts of chorionic villi and not in the cytotrophoblast.

Autoradiography↗

Reliability of frozen section of uterine curettings in evaluation of possible ectopic pregnancy.

STUDY OBJECTIVE: To assess the accuracy of frozen section evaluation of uterine curettings to look for chorionic villi and rule out ectopic pregnancy before laparoscopy. DESIGN: Retrospective review (Canadian Task Force classification III). SETTING: Northeastern medical school-affiliated teaching hospital. PATIENTS: Thirty-six women. Intervention. Dilatation and curettage, laparoscopy, and laparotomy. MEASUREMENTS AND MAIN RESULTS: Of 36 cases, 13 showed evidence of intrauterine pregnancy, either chorionic villi or implantation site, on final permanent sections. Five false negatives were identified in which no villi were identified on frozen section, although villi or evidence of an implantation site was noted on permanent sections (sensitivity 62%, 95% CI 0.32, 0.86). CONCLUSION: Frozen section evaluation of uterine curettings can produce false negative diagnoses, and this should be considered in the operative planning of women with suspected ectopic pregnancy.

Culture Techniques↗

The yolk sac in early pregnancy failure.

An attempt was made to visualize the yolk sac in 845 patients scheduled for chorionic villi sampling. The distribution of yolk sac diameters and the interpolating growth curve up to 11 weeks of development were analyzed in 239 pregnant women who were delivered of normal infants. The highest visualizing rate of the yolk sac in normal pregnancies was 97 at 7 weeks of gestation. A total of 130 miscarriages occurred before chorionic villi sampling. In these cases, the diameter of the yolk sac versus crown-rump length tended to be larger than found in normal pregnancies. The visualizing rate of the yolk sac in miscarriages after the embryo had been formed was significantly higher in those women who demonstrated fetal heart activity (82.1%) than in those who did not (54.5%). On the other hand, the yolk sac was observed in 44% of miscarriages without a visible embryo. These findings suggest different types of missed abortion. An abnormal karyotype was observed in 23 of 29 chromosomal analyses performed on aborted specimens. An abnormal karyotype was observed in all eight cases with only a yolk sac-like structure within the gestational sac.

Abortion, Spontaneous↗

Does plasminogen activator inhibitor-1 (PAI-1) control trophoblast invasion? A study of fetal and maternal tissue in intrauterine, tubal and molar pregnancies.

Urokinase plasminogen activator, its receptor and the inhibitor PAI-1 are believed to control proteolysis and remodelling of maternal tissue during trophoblast invasion. This system appears to be strictly regulated in normal intrauterine pregnancies whereas tubal and molar pregnancies seem to be characterized by an uncontrolled excessive placental invasion. This study evaluates subcellular PAI-1 by immunohistochemistry in the villous placenta, in the basal plate and placental bed, and in the decidual compartments of normal, tubal and molar pregnancies. PAI-1 was present in villous syncytiotrophoblasts and co-localized focally with fibrin-type fibrinoid on the surface of the chorionic villi. Basal plate and placental bed extravillous interstitial trophoblasts, as well as vascular trophoblasts, were also PAI-1 positive. In the decidua parietalis, PAI-1 was observed in the cytoplasm of the non-invaded decidual cells. In the decidua basalis comprising the basal plate, PAI-1 was seen to be membrane-associated or confined to the extracellular matrix (ECM) facing the invasive front of anchoring villi. The ECM of decidua capsularis and chorion laeve displayed the most pronounced PAI-1 expression towards the maternal interface. In contrast, the majority of placental bed decidual cells adjacent to the interstitial and vascular trophoblasts were PAI-1 negative. Only a few stromal cells distant from the implantation site were PAI-1 positive in the tubal pregnancies and decidualization was not present. Likewise, excessive decidual necrosis and fibrinoid deposition devoid of PAI-1 was a common finding in complete molar pregnancies. These results suggest that PAI-1 defines specific extravillous invasive trophoblasts within the maternal decidua. Moreover, maternal cellular lack of PAI-1 in tubal pregnancies and excessive decidual necrosis in molar pregnancies indicate an uncontrolled placental invasion. The present data indicate that trophoblast invasion is primarily regulated by signals from decidual cells.

Chorionic Villi↗

Expression of CDKI p27Kip1 in trophoblastic neoplasm.

OBJECTIVE: To evaluate the role of p27Kip1 in tumorigenesis and the development of trophoblastic cell disease. METHODS: Using immunohistochemistry, the expression of p27-protein was investigated in 10 normal chorionic villi in the first trimester of pregnancy, 15 complete hydatidiform moles (HM), 7 invasive moles (IM) and 7 choriocarcinomas (CC). RESULTS: In all cases, immunohistochemical staining localized p27-protein in the plasma. Decreased expression of p27Kip1 was observed in malignant trophoblastic neoplasms with a positive rate of 21.43%, which is significantly less than that in normal chorionic villi (80%) and in complete HM (73.33%) (P < 0.05). The positive rate of p27Kip1 in those complete HM with large uterine size for gestational age was lower than that in those with normal or small uterus (42.86% vs 100%, P < 0.05). CONCLUSION: p27Kip1 may be involved in the tumorigenesis of gestational trophoblastic neoplasm as a negative regulator of the cell cycle. The expression level of p27Kip1 in trophoblastic cells may be a prognostic factor for complete HM.

Cell Cycle Proteins↗

Enzymohistochemical study on the gestational choriocarcinoma: histochemistry of choriocarcinoma.

Histochemical investigation of Al-p, HSAP, Ac-p, and LDH was performed with tissues of normal chorionic villi, H. mole, invasive mole, and choriocarcinoma with the following conclusions: 1. In the normal villous tissues, Al-p, HSAP, and Ac-p are localized in syncytiotrophoblast, the former two in the rim of the cell, and the latter inside the cytoplasm. LDH displayed higher activity in cytotrophoblast than syncytiotrophoblast, especially in the former of cell column of initial chorionic villi. 2. In H. mole, Al-p, HSAP, and Ac-p are localized in syncytiotrophoblast of cystic epithelium, and in this layer and transitional cells of the chrioblastic part, the former two are localized in the rim of the cell, the latter within the cytoplasm. LDH is localized in cytotrophoblast and the transitional cell of the cystic epithelium and the chorioblastosis. 3. Localization and activities of Al-p, HSAP, Ac-p, and LDH in invasive mole are closely similar to those in H. mole. 4. Localization and activities of Al-p, HSAP, and Ac-p in choriocarcinoma presented various aspects, but more frequently they are localized in syncytiotrophoblasts in the periphery of focus and also partly in cytotrophoblasts and transitional cells. Also localization and activity of LDH showed diversity. However, the activity is stronger in cytotrophoblast. 5. From those findings, choriocarcinoma is hardly comparable with H. mole or invasive mole from an enzymohistochemical viewpoint and is assumed to have specific enzymic deviation invisible in the others and therefore to possess particular characteristics different from those of any other villous tissues.

Acid Phosphatase↗

The localization of dopamine D2 receptor mRNA in the human placenta and the anti-angiogenic effect of apomorphine in the chorioallantoic membrane.

While there are some reports indicating that dopamine (DA) and D2-like receptors (Rc) are detected in human placenta, there is little evidence available regarding the function of DA or the precise localization of its receptors in this organ. In the present study, we confirmed the placental expression of DA D2 Rc transcripts by Northern blot analysis. Using in situ hybridization, we also first revealed that DA D2 Rc mRNA was expressed in cytotrophoblasts, syncytial trophoblasts, vascular endothelial cells, Hafbauer cells, and fibroblasts in the chorionic villi of the human placenta. The expression sites of DA D2 Rc mRNA led us to suspect other functions of DA in the placenta besides the regulation of human placental lactogen. Since the cells expressing DA D2 Rc mRNA are related to proliferation and remodeling of placental tissue, we tried to evaluate a possible involvement of DA in the regulation of placental angiogenesis. To this end, we used the chicken chorioallantoic membrane (CAM) assay. In CAM assay, apomorphine, a potent nonselective agonist of DA, has an anti-angiogenic effect. These results suggest that DA may regulate the vascularization of human placenta through its receptors present in the chorionic villi.

Allantois↗

Telomerase and proliferative activity in placenta from women with and without fetal growth restriction.

OBJECTIVE: To analyze telomerase and proliferative activity in placenta from women with and without fetal growth restriction (FGR). METHODS: Telomerase activity was analyzed in 30 first-trimester chorionic villi specimens (group A) and in 28 second- and third-trimester placenta specimens (group B) from women without FGR. Telomerase activity also was analyzed in 11 placenta specimens from women with asymmetric FGR (group C). The proliferative activity of these 69 specimens was assessed by immunohistochemical staining, using the MIB-1 monoclonal antibody. RESULTS: Telomerase activity was detected in 28 (93.3%) of 30 chorionic villi specimens and in 18 (64.3%) of 28 placenta specimens without FGR. In contrast, no telomerase activity was exhibited in the placenta specimens from any of the 11 women with asymmetric FGR by telomeric repeat amplification protocol assay. Telomerase activity also was detected by in situ telomeric repeat amplification protocol assay in trophoblastic cells from women without FGR but not in trophoblastic cells from women with asymmetric FGR. Thus, telomerase activity was detected significantly more often in groups A and B than in group C (P < .01). The rate of proliferative activity, evident as positive MIB-1 staining in trophoblastic cells, in groups A and B (28.1+/-1.7% and 7.0 +/-2.9%, respectively) was significantly higher than that in group C (1.9+/-0.6%; P < .01). CONCLUSION: Telomerase and proliferative activity were minimal in placenta from women with asymmetrical FGR, suggesting placental senescence with asymmetrical FGR.

Cell Division↗

Secretion and biosynthesis of N-acetyl-beta-D-hexosaminidases by human placental tissues in vitro.

Chorionic villi selectively secrete the B isoenzyme and, to a lesser extent, the A isoenzyme of lysosomal beta-hexosaminidase when incubated in vitro, suggesting that this tissue could contribute to maternal serum beta-hexosaminidase B and A activities in vivo. The results make it unlikely, however, that the placenta is the source of the I2 isoenzyme found in maternal serum. Samples of amnion and term chorion laeve secreted predominantly the B isoenzyme whereas first-trimester chorion laeve secreted the A, B and I2 isoenzymes. Amnion and chorion laeve may, therefore, be a source of the A, B and I2 activities found in amniotic fluid. Chorionic villi supported the cycloheximide-sensitive incorporation of [3H]leucine into immunoreactive beta-hexosaminidase thereby providing direct evidence for enzyme biosynthesis. Newly synthesized beta-hexosaminidase was detectable in the medium only after extended incubation and the secretion of beta-hexosaminidase activity continued at normal rates for several hours in the absence of protein synthesis. These results indicate that enzyme synthesis is not a primary factor in the regulation of enzyme secretion.

Chromatography, DEAE-Cellulose↗