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Origin and differentiation of the yolk sac and extraembryonic mesoderm in presomite human and rhesus monkey embryos.

Reexamination of presomite human and rhesus monkey embryos in the Carnegie Collection provides no evidence to corroborate the hypothesis that the trophoblast is the source of all extraembryonic tissues in these embryos. Instead, the present study indicates that the developmental pattern of the yolk sac and extraembryonic mesoderm is homologous to that in other eutharian mammals. The primary yolk sac of 10- to 11-day human blastocysts is partially filled with a meshwork of extraembryonic endoderm, whereas such a meshwork is absent in the rhesus monkey. It is suggested that this endodermal meshwork develops as the result of interstitial implantation in the human embryo. A small secondary yolk sac develops in 12- to 13-day human and macaque embryos as the result of pinching off of a portion of the larger primary yolk sac. Development of a secondary yolk sac in higher primates appears to be related causally to differential rates of expansion of the blastocyst and primary yolk sac within the simplex uterus. The caudal margin of the primitive streak develops precociously in 12- to 14-day human and macaque embryos, and this appears to be the source of all the extraembryonic mesoderm of the chorion, chorionic villi, and body stalk. It is suggested that the peripheral spread of extraembryonic mesoderm plays in inductive role in the development of chorionic villi, similar to other types of epithelial-mesenchymal inductive interactions. In contrast to previous hypotheses, the human and macaque trophoblasts appear to give rise only to additional trophoblast.

Amnion↗

Prenatal diagnosis and fetopathological findings in a fetus with ring chromosome 18.

We report on a fetus with ring chromosome 18 and monosomy 18 mosaicism [mos 45,XX,-18/46,XX,r(18)] detected in cultured amniotic fluid cells at 15 weeks' gestation. Chromosome analysis of fetal blood, cultured chorionic villi and fetal fibroblasts confirmed the aberrant karyotype. The aberrant chromosome complement could not be detected in a short-term culture of chorionic villi (46,XX in 21 metaphases). These findings suggest that the aberrant karyotype was a postzygotic event. Fetal ultrasound was normal and the parents decided to terminate the pregnancy at 21 weeks' gestation. Autopsy revealed multiple abnormalities including facial dysmorphy, partial agenesis of the corpus callosum and an interatrial septal defect.

Adult↗

[Morphological appearance of cellular immune reaction in the endometrium in spontaneous miscarriage].

Morphometric analysis of lymphocyte, macrophage and polymorphonuclear leukocyte levels in the endometrial stroma was performed in 67 spontaneous and 30 induced abortions during 5-12 weeks of gestation. The results were correlated with the degree of structural maturity of decidual tissue and terminal chorionic villi and showed activation of lymphocyte macrophage reaction mostly pronounced in case of incomplete decidual transformation of endometrial stroma and hypoplastic alterations of chorionic villi. It was concluded that the aforementioned changes were secondary to a decreased function the gravid yellow body and subsequent activation of cell-mediated immune response in the endometrium during spontaneous abortion.

Abortion, Spontaneous↗

Implantation and placental development in somatic cell clone recipient cows.

Successful somatic cloned animal production has been reported in various domesticated species, including cattle; however, it is associated with a high rate of pregnancy failure. The low cloning yield could possibly arise from either an abnormal and/or poorly developed placenta. In comparison to control cows, fewer placentomes were found in somatic cell nuclear recipient (NT) cows at day 60 of gestation, suggesting a retardation of fetal/placental growth in these animals. NT cows not only had fewer numbers of chorionic villi but also had poorly developed caruncles. Macroscopic examination revealed atypical development of the placentome in terms of shape and size. Histological disruption of chorionic villi and caruncular septum was found in NT cows. Of particular interest was that the expression of genes, as well as proteins in the placentome, was disparate between NT and artificially inseminated cows, especially placental lactogen (PL) and pregnancy-associated glycoprotein (PAG). In contrast, prolactin-related protein-1 (PRP-1) signals were comparable across cows, including NT cows carrying immotile fetuses. The expression of extracellular matrix degrading molecule, heparanase (HPA), in NT cows was divergent from that of control cows. Microarray data suggest that gene expression was disorientated in early stages of implantation in NT cows, but this was eliminated with progression of gestation. These findings strongly support a delay in trophoblast development during early stages of placentation in NT cows, and suggest that placental specific proteins, including PLs, PAGs, and HPA, are key indicators for the aberration of gestation and placental function in cows.

Animals↗

Ultrasound guided chorion biopsy in antenatal diagnosis of genetic disorders.

The technique of transcervical chorion biopsy was evaluated in women about to undergo therapeutic termination of pregnancy. The trophoblast biopsy catheter (Portex) under real-time ultrasound guidance was the most reliable in obtaining chorionic villi. The overall success rate for chorion biopsy was 73% in the first 100 patients studied.

Biopsy↗

Cell differentiation and endogenous cyclic adenosine 3',5'-monophosphate regulate osteopontin expression in human trophoblasts.

Integrin receptors and their extracellular matrix ligands have been implicated in the molecular and cellular mechanisms of trophoblast adhesion and migration. In the present series of experiments, the regulation of expression of osteopontin (OPN), a secretory extracellular matrix protein that mediates cell adhesion by binding to members of the alpha V family of integrins was investigated. Human chorionic villi were obtained during the mid and late first trimester, early second trimester, and late third trimester of pregnancy. In addition, cytotrophoblasts were isolated from chorionic villi, and the effects of exposure of cultured cytotrophoblasts to cAMP agonists or antagonists were assessed. OPN messenger RNA (mRNA) was expressed by human trophoblasts in the first trimester and throughout pregnancy. Using immunolocalization in prepared tissue sections, cytotrophoblasts stained intensely for OPN, but syncytial trophoblasts did not. This differentiation-dependent expression was confirmed in vitro by demonstrating that freshly isolated mononuclear cytotrophoblasts exhibited a high level of OPN mRNA, but as the cells aggregated and fused to form multinucleated syncytia in vitro, mRNA levels decreased dramatically. 8-Bromo-cAMP inhibited the expression of OPN mRNA, whereas the cAMP antagonist Rp-cAMP inhibited the OPN mRNA decrease during the in vitro differentiation of the cells. This study demonstrates the regulated expression of OPN by human trophoblasts. In addition, our data suggest that this expression is dependent upon the state of cellular differentiation of the trophoblasts and is regulated by endogenous cAMP. We speculate that binding of OPN to its alpha V beta 3-integrin receptor may be a critical signaling pathway contributing to the integrity of the chorionic villus and may also play a role in maternal-embryonic communication during the process of placentation.

8-Bromo Cyclic Adenosine Monophosphate↗

Expression of the fusogenic HERV-FRD Env glycoprotein (syncytin 2) in human placenta is restricted to villous cytotrophoblastic cells.

Recently, the expression of a human endogenous retrovirus HERV-FRD, able to encode a fusogenic envelope protein (syncytin 2), has been observed in human placenta. The aim of the present study was to localize the expression of syncytin 2 in first trimester placenta. In addition, we investigated the presence of HERV-FRD transcripts during the in vitro differentiation of isolated villous and extravillous trophoblastic cells from first trimester chorionic villi. Using a monoclonal antibody specifically raised against the HERV-FRD Env protein, syncytin 2 was immunolocalized only in the villous trophoblast of the chorionic villi, at the level of cytotrophoblastic cells. Interestingly, immunostaining was not observed in all cells but only in some of them, and was detected, more frequently, at the membrane level at the interface between the cytotrophoblastic cells and syncytiotrophoblast. Labeling was observed neither in the syncytiotrophoblast nor in the mesenchymal core of the villi nor in the extravillous trophoblast. In vitro detection of HERV-FRD transcripts was restricted to villous trophoblastic cells and decreased significantly with time in culture. These results suggest that syncytin 2 might play a role in human trophoblastic cell fusion.

Cells, Cultured↗

Human decidual cells activity in women with spontaneous abortions of probable CMV aetiology during the first trimester of gestation. An immunohistochemical study with CMV-associated antigen.

AIM: To determine the expression of CMV-associated antigen in the human decidual endometrial stromal cells in spontaneous abortions with no evidence of maternal relapse during the first trimester of gestation. EXPERIMENTAL DESIGN: We examined 15 placentas resulting from intrauterine fetal death after spontaneous abortion during the 8th, 10th, and 12th week of gestation respectively, and in which CMV reactivation was ruled out from serological evaluation of the pregnant women at admission, versus equal controls after voluntary abortion following well-documented maternal viral recurrence. In addition, a panel of monoclonal antibodies for the identification of leukocytes (CD45/LCA), B-lymphocytes (CD20/L-26), and T-lymphocytes (CD45RO/UCHL1), was performed. All women received hormonal medication to support gestation, in the cases of spontaneous abortions. RESULTS: Immunohistochemical examination using a specific antibody against cytomegalovirus showed large multinucleated infected cells with intranuclear inclusions, located primarily in the decidual stroma within a lymphoplasmacytic infiltrate in the cases of spontaneous abortions. No evidence of infection was observed in the chorionic villi. In the cases of voluntary abortions same findings were observed in the relevant areas, and a strong evidence of infection was observed in the chorionic villi. CONCLUSION: This study demonstrates 1) that the decidual endometrial stromal cells can express the CMV-associated antigen prior to serological manifestation of the viral replication, 2) the expression of the antigen is higher in cases of hormonal administration to support gestation. In these cases a mild mononuclear infiltrate of UCHL1 (T marker) positive cells, accompanies the CMV-associated antigen positive cells.

Abortion, Spontaneous↗

Secretion of human chorionic gonadotropin in early pregnancy.

Remarkable secretion from the syncytial trophoblast of chorionic villi was observed in early pregnancy. Histology and histochemistry revealed many cytoplasmic protrusions containing numerous secretory granules, which were of basophilic glycoprotein in nature. Also, immunohistochemistry proved that these were human chorionic gonadotropin (hCG). Electron microscopy clarified the overall secretory process, which started with the production of small granules in the Golgi complex; then, the granules migrated toward the cell surface, increasing their size by fusing with each other, and gathered in cytoplasmic protrusions, then liquefied to be liberated by a mode of exocytosis into the maternal bloodstream (intervillous space). This remarkable hCG secretion, morphologically represented by the many cytoplasmic protrusions containing numerous secretory granules, arose synchronously on the surface of almost all chorionic villi. This phenomenon was limited to the eighth and ninth weeks of gestation, concurring with the maximal hCG concentration of maternal blood serum during pregnancy.

Chorionic Gonadotropin↗

[Aspiration of chorionic tissue: diagnostic potential in prenatal (fetal) diagnosis].

The authors have investigated the possibilities of obtaining chorionic villi from legal pregnancy termination patients between the 7th and 11th week and the method of direct chromosome preparation from the villi as well. According to their investigations under continuous "real time" ultrasound guidance and with the immediate microscopic checking of the obtained material it is possible to receive chorionic villi of necessary quantity in nearly 100% of the cases, and after 2 or 3 hours the evaluation in a great deal of mitosis of good quality can be carried out. The method may revolutionize the prenatal (fetal) diagnosis, but its routine application may not be proposed until the rate of fetal risk is exactly known.

Biopsy, Needle↗

In routine diagnostic practice, how many sections should we examine from cases of products of conception?

In order to determine whether a single representative section taken from routine specimens of products of conception would contain sufficient material to trigger a more comprehensive search for the features of a hydatidiform mole, cases of gestational trophoblastic disease submitted over a five year period were reviewed. Partial hydatidiform moles were either suspected or diagnosed on the first histological section in 46 (92%) cases. In the remaining four cases, although abundant placental tissue showing diagnostic features was available in later blocks, most of the tissue in the first block consisted of endometrial tissue with only occasional chorionic villi being identified. The current study shows that a molar gestation can be suspected after examining a single tissue block, providing that it contains a representative number of chorionic villi. If a molar pregnancy is suspected clinically or pathologically, additional blocks should be examined to secure the diagnosis and classify the condition.

Adult↗

The significance of cystic hygroma in fetuses.

Cystic hygroma of the neck was observed in seven spontaneously aborted, severely autolyzed female fetuses. Four of five karyotyped cases had the 45,XO karyotype; one was a normal female, 46,XX. Diagnostic features associated with monosomy X in fetuses, which were observed in six of these cases, include large cystic hygromas, generalized edema, edematous chorionic villi with scalloped borders and trophoblastic inclusions, and, possibly, a small heart. In contrast, the single euploid fetus in this series had a small cystic hygroma, no hydrops, and chorionic villi of normal size. These features may differentiate cystic hygromas associated with the XO karyotype, which are not associated with recurrence risk, from those associated with a familial syndrome.

Abortion, Spontaneous↗

Retained trophoblastic tissue in fallopian tubes: a consequence of unsuspected ectopic pregnancies.

Five patients had retained trophoblastic tissue within the Fallopian tubes, which suggested remote ectopic pregnancies. Four remote ectopic pregnancies were identified in patients after delivery. One patient underwent salpingo-oophorectomy after the identification of a small ovarian mass and 1.0 cm "necrotic area" in the fallopian tube at cesarean section. The other three patients underwent tubal ligation, and one was noted to have a 1.0 cm "calcified" nodule in the distal fallopian tube. The fifth patient underwent laparotomy and was found to have an acute ectopic pregnancy in one fallopian tube and a clinically unsuspected 3.0 cm mass in the other, which proved to be an ectopic pregnancy with ghost outlines of chorionic villi and trophoblast. Histopathologically, all five patients showed foci of viable-appearing intermediate trophoblast and surrounding abundant eosinophilic hyalinized material in the fallopian tubes; four patients also had hyalinized ghost outlines of chorionic villi. None of the mothers had a history of previous ectopic pregnancy. The natural history of clinically unsuspected ectopic tubal pregnancies is not well understood, but these cases illustrate that trophoblast may persist in fallopian tubes and potentially result in clinical confusion as well as tubal pathology.

Adult↗

An updated protocol for abortion surveillance with ultrasound and immediate pathology.

OBJECTIVE: To modify and improve a protocol for surveillance of patients presenting for routine elective abortion services. METHODS: Six hundred seventy-four women presenting for routine elective first-trimester abortions were studied. All were 84 or fewer days after the last menstrual period, had no history of bleeding, and had positive urine pregnancy tests. Each woman was scanned initially with an empty-bladder transabdominal technique. If no sac was seen, endovaginal ultrasonography was performed. All terminations had modified gross examination of tissue (3x magnification) as well as staining for microscopic analysis. RESULTS: Six hundred twelve patients (90.8%) demonstrated intrauterine gestations on transabdominal ultrasound, 595 of which were 12 or fewer weeks. Suction and sharp curettage and examination of tissue revealed products of conception in all. Seventeen subjects (2.5%) were found to be 13 or more weeks despite bimanual examinations and last menstrual period suggesting 12 or fewer weeks. Sixty-two patients had no sac seen on transabdominal ultrasound, 34 of whom had definitive intrauterine gestations on endovaginal ultrasound. Curettage revealed chorionic villi in all. Two had unruptured definitive ectopic pregnancies seen on endovaginal ultrasound. Twenty-one women with no sac seen on endovaginal ultrasound underwent curettage as the next step in triage; chorionic villi proved an intrauterine gestation in 17. The additional four had decidua only on pathology. Rising hCG levels in two of these four led to a diagnosis of ectopic pregnancy, whereas falling hCG levels in the other two led to a presumptive diagnosis of complete abortion, possibly tubal pregnancy in light of the lack of vaginal bleeding. CONCLUSION: Pre-abortion sonography eliminates inadvertent second-trimester cases, and immediate postoperative examination of curettage material expedites the diagnosis of ectopic pregnancy when present.

Abortion, Induced↗

Mononuclear phagocytes in the human placenta.

Hofbauer cells have been identified by morphological criteria in human placentae and are considered to be mononuclear phagocytes. An increasing recognition of the morphological heterogeneity of mononuclear phagocytes and of the considerable difficulty in their recognition in histological sections has suggested that they may be considerably more numerous in the placenta than has been appreciated. The use of specific markers for mononuclear phagocytes in the present study facilitated their identification and quantitation in the placenta. Approximately 65 per cent of the cells in cell suspensions from placentae at all stages of gestation exhibited avid Fc receptors and were morphologically indistinguishable from mononuclear phagocytes. Most of these cells also were phagocytic and expressed C3 receptors. Furthermore, when cells expressing alpha-naphthyl butyrate esterase positivity as well as high-affinity receptors for immune complexes were visualized in the stroma of chorionic villi, nearly all stromal cells were positive. The results suggest that Hofbauer cells represent the fully differentiated, morphologically-obvious end-stage members of a large mononuclear phagocyte population which is present in chorionic villi throughout gestation.

Cell Separation↗

Constitutive and induced cytokine production by human placenta and amniotic membrane at term.

Results of our previous study on the immunity of human placenta and amniotic membranes revealed that in majority of cases these organs present constitutive non-specific antiviral immunity in the organ culture (OC) system. It is possible that interferons (IFNs), tumour necrosis factors (TNFs) and interleukin 6 (IL-6) may be responsible for the antiviral effect. Here, the constitutive and lipopolysaccharide (LPS)-induced production of these cytokines and, additionally, interleukin 10 (IL-10) were determined in OC of chorionic villi, decidua and amniotic membranes. Significant amounts of constitutive TNF-alpha (2-64 U/ml), IL-6 (200-12,000 U/ml) and IL-10 (1-70 ng/ml) were detected in the maternal decidua and chorionic villi of placenta. Amniotic membranes produced lower concentrations of the cytokines. LPS increased the production of cytokines from two- to eightfold. In contrast, activity of IFN released spontaneously was found only in four of 50 placentae and amniotic membranes. LPS and Newcastle disease virus (NDV) induced IFN production in the OC system. However, the increase of IFN after induction was also very small (up to 32 U/ml). Individual differentiation in the cytokines production was observed among placentas and amniotic membranes. TNF was identified as type alpha with addition TNF-beta, IFN as type alpha, beta and gamma.

Amnion↗

[Prenatal diagnosis and clinical management of a twin pregnancy consisting of a complete mole and coexisting fetus].

OBJECTIVE: To discuss the differential diagnosis of the hydatidiform mole and a coexisting fetus, to study the prenatal diagnosis and the clinical management of a twin pregnancy consisting of a complete mole and coexisting fetus (CMCF). METHODS: Two cases of CMCF were reported retrospectively. RESULTS: In the first case, the hydatidiform mole and a coexisting fetus was found by B mode ultrasound at the 10th gestational week, the patient asked to terminate the pregnancy. The interphase FISH and karyotype analysis of the normal villi and the mole showed both of them were diploid, thus the CMCF was diagnosed. In the second case, the hydatidiform mole and a coexisting fetus was found by B mode ultrasound at the 21st gestational week. Transabdominal chorionic villi sampling and amniocentesis was performed, interphase FISH and karyotype analysis of the mole and the amniotic fluid showed both of them were diploid, thus the CMCF was diagnosed prenatally. The pregnancy was continued and premature rupture of membrane happened at the 28th gestational week, the cesarean section was performed. The neonate was healthy. The karyotype analysis of the placenta and the neonate was accordant with the prenatal diagnosis. CONCLUSIONS: As long as the hydatidiform mole and a coexisting fetus was found the prenatal diagnosis must be performed in order to differentiate the CMCF and the partial hydatidiform mole (PHM). The transabdominal chorionic villi sampling and the amniocentesis were ideal methods, interphase FISH and karyotype analysis of the mole and the amniotic fluid should be performed. If both of them were diploid, the CMCF could be diagnosed. The clinical management of CMCF should be done individually. If both of them were triploid, the PHM could be diagnosed.

Adult↗

Methodological approaches to immunohistochemical demonstration of beta-hexosaminidase in human placental and renal tissue with monoclonal antibodies.

The lysosomal enzyme, beta-hexosaminidase (Hex), was studied in full-term human placentas and in renal tissue using monoclonal antibodies raised against Hex purified from human placentas. The immunohistochemical reaction for Hex was pronounced in trophoblastic cells and macrophages of the basal plate and the smooth chorion, but was faint or negative in the amnion as well as in the syncytiotrophoblast and Hofbauer cells of the chorionic villi. The maternal decidual cells of the basal plate were negative. Biochemical enzyme analysis showed the highest activity in basal plate cells (containing trophoblast, decidual cells, macrophages and neutrophils) and a low activity in the chorionic villi. Placental tissue was less positive with monoclonal antibodies specific for Hex A, compared with antibodies reacting with both Hex A and Hex B. Epithelial cells of the renal proximal tubules were positive to the same degree with antibodies recognizing both Hex A and Hex B as well as those recognizing only Hex A.

Antibodies, Monoclonal↗