Tissuepatch is biocompatible and seals iatrogenic membrane defects in a rabbit model.

OBJECTIVE: To evaluate novel sealing techniques for their biocompatibility and sealing capacity of iatrogenic fetal membrane defects in a pregnant rabbit model. METHOD: At day 23 of gestation (term = d31), a standardized fetoscopy was performed through a 14G cannula. The resulting fetal membrane defect was closed with condensed collagen, collagen with fibrinogen, Tissuepatch, Duraseal, or a conventional collagen plug (Lyostypt) as reference. At d30, the fetuses were harvested and full thickness fetal membrane samples were analyzed. The study consisted of 2 consecutive parts: (1) biocompatibility testing by fetal survival, apoptosis, and infiltration of polymorphonuclear cells in the membranes and (2) the efficacy to seal fetal membrane defects. RESULTS: Three sealants (collagen with fibrinogen, Duraseal, or Lyostypt) were associated with a higher fetal mortality compared to control unmanipulated littermates and hence were excluded from further analysis. Tissuepatch was biocompatible, and amniotic fluid levels were comparable to those of control untouched littermates. Compared to the condensed collagen, Tissuepatch was also easier in surgical handling and induced limited cell proliferation. CONCLUSION: Tissuepatch had the best biocompatibility and efficacy in sealing an iatrogenic fetal membrane defect in the pregnant rabbit compared to other readily available sealants.

Diagnosis of umbilical cord entanglement in a monochorionic diamniotic twin pregnancy with spontaneous septostomy of the dividing membranes using dual-gate Doppler imaging.

Umbilical cord entanglement is the leading cause of fetal mortality in monoamniotic twin pregnancies and a pseudo monoamniotic environment. Published methods for detecting this complication include color Doppler and pulsed Doppler sonography; however, no method provides an absolute diagnosis. In this case, we report the diagnosis of umbilical cord entanglement using dual-gate Doppler imaging. A 35-year-old woman was referred to our hospital at 28 weeks of gestation for prenatal management because of diagnosis of a monochorionic diamniotic twin pregnancy with spontaneous septostomy of the dividing membranes. Each fetus displayed normal fetal growth without obvious discordance and anatomical abnormalities. However, the dividing membrane was not detected, and an entangled cord was suspected. Dual-gate Doppler examination was carried out. Two regions of interest were considered at different areas of the umbilical arteries, and when each Doppler image showed two different heart rates at the same time, we considered this to be evidence of umbilical cord entanglement. Cesarean section was performed at 32 weeks of gestation and twins were delivered. The delivered umbilical cords had sixfold entanglement. In this case, dual-gate Doppler seems to have been more accurate than conventional single-gate Doppler for the diagnosis of cord entanglement because we confirmed two different heart rates at the same time with dual-gate Doppler.

Development of bovine embryos derived from reproductive techniques.

Assisted reproduction techniques have improved agricultural breeding in the bovine. However, important development steps may differ from the situation in vivo and there is a high mortality rate during the first trimester of gestation. To better understand these events, we investigated the development of embryos and fetal membranes following fixed-time AI (FTAI), IVF and nuclear transfer (NT). The onset of yolk-sac development was not normal in cloned embryos. Later steps differed from conditions in vivo in all three groups; the yolk-sac was yellowish and juxtaposed with the amniotic membrane. Vascularisation of the chorioallantoic membrane was relatively late and low in NT gestations, but normal in the others. The overall development of the embryos was normal, as indicated by morphology and regression analysis of growth rate. However, NT conceptuses were significantly smaller, with the livers in some embryos occupying the abdominal cavity and others exhibiting heart abnormalities. In conclusion, the yolk-sac and the cardiovascular system seem to be vulnerable to morphogenetic alterations. Future studies will focus on gene expression and early vascularisation processes to investigate whether these changes may be responsible for the high incidence of intrauterine mortality, especially in clones.

A rare case of globosus amorphus in a goat.

We present a case of globosus amorphus delivered from a goat and subjected to radiography and histological examination. Radiography revealed a lack of development of any organ system; histological sections showed evidence of lymphoid aggregations, mononuclear infiltrations, blood capillaries, and dense fibroblasts.

Syncytin-A knockout mice demonstrate the critical role in placentation of a fusogenic, endogenous retrovirus-derived, envelope gene.

In most mammalian species, a key process of placenta development is the fusion of trophoblast cells into a highly specialized, multinucleated syncytiotrophoblast layer, through which most of the maternofetal exchanges take place. Little is known about this process, despite the recent identification of 2 pairs of envelope genes of retroviral origin, independently acquired by the human (syncytin-1 and syncytin-2) and mouse (syncytin-A and syncytin-B) genomes, specifically expressed in the placenta, and with in vitro cell-cell fusion activity. By generating knockout mice, we show here that homozygous syncytin-A null mouse embryos die in utero between 11.5 and 13.5 days of gestation. Refined cellular and subcellular analyses of the syncytin-A-deficient placentae disclose specific disruption of the architecture of the syncytiotrophoblast-containing labyrinth, with the trophoblast cells failing to fuse into an interhemal syncytial layer. Lack of syncytin-A-mediated trophoblast cell fusion is associated with cell overexpansion at the expense of fetal blood vessel spaces and with apoptosis, adding to the observed maternofetal interface structural defects to provoke decreased vascularization, inhibition of placental transport, and fetal growth retardation, ultimately resulting in death of the embryo. These results demonstrate that syncytin-A is essential for trophoblast cell differentiation and syncytiotrophoblast morphogenesis during placenta development, and they provide evidence that genes captured from ancestral retroviruses have been pivotal in the acquisition of new, important functions in mammalian evolution.

E2F4 cooperates with pRB in the development of extra-embryonic tissues.

The retinoblastoma gene, RB-1, was the first identified tumor suppressor. Rb(-/-) mice die in mid-gestation with defects in proliferation, differentiation and apoptosis. The activating E2F transcription factors, E2F1-3, contribute to these embryonic defects, indicating that they are key downstream targets of the retinoblastoma protein, pRB. E2F4 is the major pRB-associated E2F in vivo, yet its role in Rb(-/-) embryos is unknown. Here we establish that E2f4 deficiency reduced the lifespan of Rb(-/-) embryos by exacerbating the Rb mutant placental defect. We further show that this reflects the accumulation of trophectoderm-like cells in both Rb and Rb;E2f4 mutant placentas. Thus, Rb and E2f4 play cooperative roles in placental development. We used a conditional mouse model to allow Rb(-/-);E2f4(-/-) embryos to develop in the presence of Rb wild-type placentas. Under these conditions, Rb(-/-);E2f4(-/-) mutants survived to birth. These Rb(-/-);E2f4(-/-) embryos exhibited all of the defects characteristic of the Rb and E2f4 single mutants and had no novel defects. Taken together, our data show that pRB and E2F4 cooperate in placental development, but play largely non-overlapping roles in the development of many embryonic tissues.

Loss of mouse Ikbkap, a subunit of elongator, leads to transcriptional deficits and embryonic lethality that can be rescued by human IKBKAP.

Familial dysautonomia (FD), a devastating hereditary sensory and autonomic neuropathy, results from an intronic mutation in the IKBKAP gene that disrupts normal mRNA splicing and leads to tissue-specific reduction of IKBKAP protein (IKAP) in the nervous system. To better understand the roles of IKAP in vivo, an Ikbkap knockout mouse model was created. Results from our study show that ablating Ikbkap leads to embryonic lethality, with no homozygous Ikbkap knockout (Ikbkap(-)(/)(-)) embryos surviving beyond 12.5 days postcoitum. Morphological analyses of the Ikbkap(-)(/)(-) conceptus at different stages revealed abnormalities in both the visceral yolk sac and the embryo, including stunted extraembryonic blood vessel formation, delayed entry into midgastrulation, disoriented dorsal primitive neural alignment, and failure to establish the embryonic vascular system. Further, we demonstrate downregulation of several genes that are important for neurulation and vascular development in the Ikbkap(-)(/)(-) embryos and show that this correlates with a defect in transcriptional elongation-coupled histone acetylation. Finally, we show that the embryonic lethality resulting from Ikbkap ablation can be rescued by a human IKBKAP transgene. For the first time, we demonstrate that IKAP is crucial for both vascular and neural development during embryogenesis and that protein function is conserved between mouse and human.

Generation of a conditional Ppap2b/Lpp3 null allele.

Lpp3, formerly known as Pap2b, is a lipid phosphohydrolase enzyme. Some of its substrates and products are lipids with potent biological and signaling activities such as phosphatidic acid, lysophosphatidic acid, sphingosine-1-phosphate, diacylglycerol, sphingosine, and ceramide. Lpp3 is dynamically expressed during development and is widely distributed in adult tissues. Targeted inactivation of Lpp3 gene (Ppap2b) in the mouse results in embryonic lethality because of defects in extraembryonic vascular development and gastrulation. To study the participation of Lpp3 later in development and in specific cell lineages we generated a conditional allele of Ppap2b. This was accomplished by flanking critical exons, responsible for its catalytic activity with loxP sites. A generalized Cre-mediated recombination of this allele yielded a phenotype fundamentally identical to our previously reported Ppap2b null allele.

Ausencia o retraso en la formación de la membrana amniocoriónica: ¿un marcador ecográfico de trisomía 21? / Absence or delay in the amniochorionic membrane formation: a trisomy 21 marker?

Objetivo. Describir la asociación encontrada entre fetos portadores de trisomía 21 y el hallazgo de lo que consideramos como un marcador ecográfico.Material y método. Estudio descriptivo prospectivo de ocho pacientes a quienes se les realizó determinación del cariotipo fetal y que presentaban ausencia o retraso en la formación de la membrana amniocoriónica al realizar la amniocentesis.Resultados. De las ocho pacientes estudiadas, en seis de ellas (75%) se obtuvo un cariotipo fetal aneuploide. Comentarios. La detección de ausencia o retraso en la formación de la membrana amniocoriónica podría constituir un marcador ecográfico de cromosomopatía.Comentarios. La detección de ausencia o retraso en la formación de la membrana amniocoriónica podría constituir un marcador ecográfico de cromosomopatía

Objective. Descibe the possible association of this sonographic marker and fetus with trisomy 21.Material and method. Descriptive study of eigth patients that were subjected to amniocentesis. At the scan, before the amniocentesis was done, all of these patients did not present or presented delay in the amniochorionic membrane formation.Results. In six cases (75 %) there was a fetal aneuploidy.Commentary. Absence or delay in the amniochorionic membrane formation could be a sonographic marker during the second-trimester of pregnancy

Chorioamniotic membrane separation: a potentially lethal finding.

Sonographic detection of chorioamniotic membrane separation (CMS) has been considered a benign incidental finding. We now report 6 cases of CMS identified by prenatal ultrasound; 1 in an otherwise normal pregnancy and 5 following fetal surgery. Following membrane separation, amniotic bands formed and compromised the umbilical cord in 4 cases leading to 2 fetal deaths. In the first case, CMS was detected by ultrasound at 22 weeks' gestation in an otherwise uncomplicated pregnancy. Because CMS was considered benign and umbilical cord blood flow was ample, the mother was followed by intermittent sonographic examinations. Fetal demise occurred 2 weeks later, clearly due to umbilical cord strangulation by an amniotic band. Surprised by this unexpected outcome, we reviewed our experience with CMS after hysterotomy for fetal surgery. Out of more than 40 fetal surgical cases, we have 5 cases in which CMS was recognized after hysterotomy. Three of these fetuses had umbilical cord compromise by a band of amniotic membrane leading to 1 fetal death. This experience demonstrates that membrane separation may be associated with amniotic band formation which can lead to cord strangulation and fetal compromise. Following fetal surgery, serial ultrasound evaluation and close fetal monitoring are indicated. In otherwise unremarkable pregnancies, clinician awareness of the possibility of amniotic band formation following CMS should be heightened. In either situation, knowledge of this potential life-threatening complication may identify cases in which cord compromise requires emergent delivery or fetoscopic release of the strangulating amniotic band.

Fetus in fetu: a case with complete umbilical cord and fetal sac.

The authors present a retroperitoneal fetus in fetus in a 3-month-old girl. A 15-cm cystic mass with a monstrous fetuslike structure surrounded by a complete sac containing serous fluid was removed from the left retroperitoneal space. The draining vessel of the cystic mass was connected to the right renal vein of the host. The included fetus weighed 380 g. It had a well-developed umbilical cord, four extremities, head, buttock, and vertebral bodies with a meningomyelocele. The thoracic cavity of the included fetus had only a saclike foregut structure, but the abdominal cavity revealed a full length of intestine with a Meckel diverticulum, bilateral ovaries, urinary bladder, and cloaca with external opening. The cephalic end was composed of well-developed tooth germs, tongue and buccopharynx, mandible, maxilla, sphenoid bone, and salivary glands. Chromosomal study showed 46,XX with a normal G banding pattern. We report this case as an example of fetus in fetu with a complete umbilical cord and fetal membrane.

Placental pathology for the nineties.

Examination of the placenta in a problem pregnancy is potentially very valuable, yielding information about the nature and duration of processes occurring in gestation. Such evaluation requires thoughtful gross examination, careful sectioning, and an understanding of the basic microscopic changes in a variety of pathologic processes. As there are very few absolute associations in placental pathology, it is most important to appreciate the range of normal in assessing the potential importance of lesions. This chapter has tried to provide the essential relevant material, with emphasis on commonly encountered problem areas, and a variety of topics potentially related to maternal and fetal morbidity and mortality.

Origin of raised maternal serum alpha-fetoprotein levels in second-trimester oligohydramnios.

Concanavalin A (Con A) subtyping of alpha-fetoprotein (AFP) revealed higher concentrations of AFP non-reactive with Con A in sera of 12 pregnant women with second-trimester oligohydramnios and raised total serum AFP levels than in sera of 42 pregnant women with raised total serum AFP levels and a normal amniotic fluid volume. This suggests that in oligohydramnios the origin of excess AFP in the maternal compartment is amniotic fluid. It is proposed that oligohydramnios and the associated raised maternal serum AFP levels are caused by damage of the fetal membranes prior to 16 weeks of gestation resulting in leakage of amniotic fluid to the decidual tissue and resorption in the maternal circulation.

[Case description of a congenital malformation in a calf, characterized by an ingrowth of the fetal membranes into the cranial cavity]. / Fallbeschreibung einer angeborenen Missbildung beim Kalb, charakterisiert durch Einwachsen der Eihäute in die Schädelhöhle.

It is reported on a congenital malformation at the head of a calf born dead, characterized by downgrowth of the fetal membranes into the cranial cavity. The outer appearance and the pathologic-anatomical observations are presented in pictures and the attempt of an interpretation is made.

Frequency of pathologic changes in the young human chorion in therapeutic abortions of normal pregnancies. A report on 500 cases studied histologically.

PIP: 500 chorionic specimens from legal abortions were examined macroscopically and histologically from January 1965 until December 1967. All 500 specimens were obtained between the fifth and the fourteenth weeks of normal pregnancy. Among the developmental defects of the chorion, the most frequent finding was focal micromolar degeneration, which was observed in 21% of all cases. Cases of diffuse involvement of the chorion were found in 2%. Diffuse micromolar degeneration was observed in 2% of all cases. Discrete stromal fibrotic changes were evident in 4.4% of the cases, focal intervillous thromboses in 4.2%, fetal "lymphoid" elements in 3.6%, and acute nonspecific inflammations in 1.8%. Most of the cases in every age group revealed no histopathologic changes at all. The percentage of abnormal cases rose gradually with the duration of pregnancy. The changes that did occur were found in the materials from pregnancies taken as "normal". Since the tissues obtained during therapeutic abortion are not always normal and contain various pathologic changes, the need for systematic microscopic examination of the human chorion is stressed.^ieng