Stroma Of Villi Research Articles

The interplay of inflammation and placenta in maternal diabetes: insights into Hofbauer cell expression patterns.

Inflammation of the placenta is harmful to both the fetus and the mother. Inflammation is strongly associated with diabetes, a common complication of pregnancy. Hofbauer cells (HBCs), unique immune system cells of fetal origin in the placenta, play complex roles, including growth of placental villi and their branching, stromal remodelling, and angiogenesis. Our study investigated the expression of IL-1β, IL-10, CYP2C8, CYP2C9, CYP2J2 and sEH in HBCs from patients with type 1 diabetes mellitus (T1DM) and gestational diabetes mellitus (GDM) compared to healthy controls using immunohistochemistry. We also assessed the structure of the villus stroma using Masson´s trichrome. In T1DM, HBCs showed inflammatory activation characterised by increased IL-1β and decreased CYP epoxygenase expression compared to normal placentas. Conversely, significant inflammation in HBCs appeared less likely in GDM, as levels of IL-1β and CYP epoxygenases remained stable compared to normal placentas. However, GDM showed a significant increase in sEH expression. Both types of diabetes showed delayed placental villous maturation and hypovascularisation, with GDM showing a more pronounced effect. The expression profiles of IL-1β, CYP epoxygenases and sEH significantlly differ between controls and diabetic placentas and between T1DM and GDM. These facts suggest an association of the CYP epoxygenase-EETs-sEH axis with IL-1β expression as well as villous stromal hypovascularisation. Given the stable high expression of IL-10 in both controls and both types of diabetes, it appears that immune tolerance is maintained in HBCs.

Don't sugar coat it: The independent and synergistic impacts of obesity and gestational diabetes on placental parameters.

Don't sugar coat it: The independent and synergistic impacts of obesity and gestational diabetes on placental parameters.

CD68 expression in the placenta of gestational diabetic mothers: A case-control study.

Gestational diabetes mellitus (GDM) is defined as any degree of glucose intolerance with the onset or first recognition during pregnancy and is the most common metabolic complication of pregnancy. Significant maternal and fetal complications can result from undiagnosed or inadequately treated GDM. To investigate the difference in the expression of the CD-68 marker in the Hofbauer cells (HCs) and their distribution within the villi in the placentas of diabetic and non-diabetic mothers. Sixty placentas were included in the study, 30 as controls and 30 from mothers with diagnosed GDM as cases. Full-thickness cross sections of placentas were obtained. Tissue processing was done, followed by haematoxylin and eosin (H&E). A study of CD68 markers (placental macrophages) was done using standard protocols of immunohistochemistry. Frequencies and percentages of Hofbauer cells (HCs) found in case and control placental tissue were calculated. Student's t-test was used to compare two groups using SPSS 13.0 software. When P is 0.0001, differences were considered statistically significant. We studied the distribution and number of fetal macrophages (CD68+) in diabetic and non-diabetic placentas. The immunostained CD68+ cell count was identified to be significantly higher in the GDM placenta. In relation to fetal blood vessels in the villus stroma of the GDM placenta in comparison to control, CD68+ cells were found more frequently. This study shows a significant increase in the number of Hofbauer cells in the placenta of mothers with GDM in comparison to control (P < 0.0001). An increase in macrophages in these placentae might be related to the protective mechanism against inflammation. Further studies are required to investigate the mechanism in detail.

ADAMTS18+ villus tip telocytes maintain a polarized VEGFA signaling domain and fenestrations in nutrient-absorbing intestinal blood vessels

The small intestinal villus tip is the first point of contact for lumen-derived substances including nutrients and microbial products. Electron microscopy studies from the early 1970s uncovered unusual spatial organization of small intestinal villus tip blood vessels: their exterior, epithelial-facing side is fenestrated, while the side facing the villus stroma is non-fenestrated, covered by pericytes and harbors endothelial nuclei. Such organization optimizes the absorption process, however the molecular mechanisms maintaining this highly specialized structure remain unclear. Here we report that perivascular LGR5+ villus tip telocytes (VTTs) are necessary for maintenance of villus tip endothelial cell polarization and fenestration by sequestering VEGFA signaling. Mechanistically, unique VTT expression of the protease ADAMTS18 is necessary for VEGFA signaling sequestration through limiting fibronectin accumulation. Therefore, we propose a model in which LGR5+ ADAMTS18+ telocytes are necessary to maintain a “just-right” level and location of VEGFA signaling in intestinal villus blood vasculature to ensure on one hand the presence of sufficient endothelial fenestrae, while avoiding excessive leakiness of the vessels and destabilization of villus tip epithelial structures.

MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL SIGNS OF PLACENTAL DISORDERS OF WOMEN AFTER SYPHILITIC INFECTION.

The aim: To define morphological and immunohistochemical signs of placental disorders of women after syphilitic infection. Materials and methods: The prospective study of 60 pregnant women with history of syphilitic infection (main group) and 57 pregnant patients without syphilis (control group) was conducted. The morphological and immunohistochemical study of the afterbirth was performed. Results: In the placentas of women of the main group the following phenomena were found out: circulatory disorders in the form of hemorrhages into the intervillous space and the stroma of villi; accumulation of fibrinoid around villi with dystrophically altered stroma, compensatory-adaptive reactions resulted in hyperplasia of terminal villi and vessels in them, which provoked narrowing of the intervillous space and disruption of blood supply in it. Pathogenic immune complexes containing Ig G, M and C3 of the complement fraction were located in the central part of the placenta - 45.00% of cases, 16.67% - in the regional, 8.33% - in both parts. Immune complexes with Ig M content occurred in 38.33% of cases. The content of pathogenic immune complexes was the most concentrated in the placentas of women with latent forms and secondary recurrent syphilis - 60.00% of cases. Conclusions: changes in morphohistological and immunohistochemical examination of the placenta of this group of women confirmed the detrimental effect of syphilitic infection in the anamnesis on the structure of placenta during the next pregnancies.

The pathology of natural and experimentally induced Campylobacter jejuni abortion in sheep.

We describe here the gross and microscopic lesions in 18 experimentally induced and 120 natural Campylobacter abortions. In natural Campylobacter abortions, gross lesions were reported infrequently; placentitis was recorded in 6% and hepatic lesions in 4% of our field cases. Placentitis was the microscopic lesion identified most consistently in natural abortions (93%) and was often observed in association with abundant bacterial colonies in chorionic villi (54%) and less often with placental vasculitis (13%). In natural abortions, suppurative fetal pneumonia (48%), necrosuppurative hepatitis (16%), and purulent meningitis (7%) were also observed. The better-preserved specimens from experimentally induced abortions were utilized to define placental changes more precisely. Placentitis was identified in all 18 experimentally induced abortions and was observed most consistently in the chorionic villus stroma (100%), often accompanied by suppurative surface exudate (89%). An inflammatory infiltrate was less commonly identified in the cotyledonary hilus (39%) and intercotyledonary placenta (22%). Bacteria were visualized in H&E-stained sections in 89% of placentas from experimentally infected ewes, primarily as well-demarcated bacterial colonies within subtrophoblastic, sinusoidal capillaries (89%), in the cotyledonary villus stroma (89%), and within the cytoplasm of trophoblasts (22%). Transmission electron microscopy and immunohistochemistry confirmed that the vast majority of the well-demarcated bacterial colonies characteristic of Campylobacter abortion were within subtrophoblastic sinusoidal capillaries. The most characteristic microscopic lesions identified in cases of Campylobacter abortion in sheep were placentitis with placental bacterial colonies, placental vasculitis, and fetal pneumonia.

Intrinsic antibody mediated enhancement of ZIKV infection in placental macrophages

Abstract Zika virus (ZIKV) vertical transmission from mother to child in utero is associated with a number of neonatal abnormalities, including congenital microcephaly. Yet, the mechanism by which ZIKV infects the placenta and subsequently the fetus remains poorly understood. ZIKV and Dengue virus (DENV) endemic regions are commonly overlapping, and DENV specific antibodies can cross-react with ZIKV. We and others have previously demonstrated that cross-reactive DENV IgG can mediate ZIKV infection of placental cells though increased viral entry via Fc receptors. Specifically, we found that infection with ZIKV immune complexes (ZIKV-IC) generated from cross-reactive anti-DENV monoclonal IgG was enhanced in human placental macrophages called Hofbauer cells, which are found within the chorionic villus stroma near the fetal vasculature. In this study, we aimed to characterize the intrinsic cellular changes that occur in Hofbauer cells as a result of ZIKV-IC infection. We performed RNA sequencing on Hofbauer cells infected with ZIKV complexed with different concentrations of anti-DENV monoclonal IgG. Principal component analysis revealed that while ZIKV incubated with anti-influenza IgG had a similar gene expression profile to ZIKV alone, ZIKV-IC infection with increasing anti-DENV IgG skewed gene expression towards a unique phenotype. We identified a number of transcriptional profiles related to anti-DENV IgG dose. Notably, we observed a pattern of decreased interferon stimulated genes with increasing anti-DENV IgG dose. Yet, increasing anti-DENV IgG also increased ZIKV viral RNA. These results may indicate that ZIKV-IC infection makes Hofbauer cells more permissive to ZIKV replication by modulating the innate antiviral response.

INVESTIGATION OF SFLT-1 AND VEGF EXPRESSION IN NORMOTENSIVE AND PREECLAMPTIC PLACENTA. AN IMMUNOHISTOCHEMICAL STUDY

Objectives: The pathogenesis of preeclampsia is still not clear, but endothelial dysfunction believed to be one of the most encountered problems during placenta development in preeclamptic patients. Both vascular endothelial growth factor (VEGF) and its antagonist, soluble Fms-Like tyrosine kinase-1 (sFlt-1), have roles in vascular function. In this study, we have investigated immunohistochemical expression of VEGF and sFlt-1 in term placenta of normotensive and preeclampsia patients. Methods: Totally twenty term placentas were obtained from pregnant women of which 10 preeclampsia patient and 10 normotensive. Placentas were dissected and tissue samples were subjected to routine tissue processing protocol, then embedded in paraffin blocks. Serial sections were obtained from paraffin blocks and stained with H&amp;amp;E and PAS for routine histopathology. VEGF and sFlt-1 immunohistochemistry was performed to the sections. Results: When compared to control group, severe pathological changes were observed in preeclamptic placentas. Increase in number of syncytial knots and intervillous bridges, hemorrhage in interstitium, dilatation and congestion in villous capillaries, increase in fibrin accumulation in villus stroma, and increase in thickening of basement membrane were very clear. VEGF expression was significantly higher in normotensive placentas compared to preeclamptic. On the other hand, sFlt-1 expression was significantly increased in preeclamptic placentas. Conclusions: When the VEGF and sFlt-1 expression was considered, higher expression of sFlt-1 at preeclampsia, but decrease in VEGF expression, might be related to endothelial dysfunction in preeclampsia. Overall, this study demonstrates, imbalance between VEGF and sFlt-1 is one of the major reason of endothelial dysfunction in preeclamptic placenta.

Peculiarities of the inflammatory reactions development in placenta at delivery in different gestational terms

Objective of the study: to study peculiarities of the inflammatory reactions development in placenta according to results of a general histological and immunohistochemical study of the prevalence and intensity of reactions caused by CD45 and tumor necrosis factor α (TNF-α) during childbirth at different gestational periods.Materials and methods. Morphological and immunohistochemical studies of various placental structures and CD45 and TNF-α expression level in 142 women with preterm delivery at different gestational periods were performed: group I – at 23–29 weeks, group II – at 30–34 weeks; group III – at 35–36 weeks + 6 days, group IV (control) – 40 women at 38–40 weeks.Results. Inflammatory infiltration and the most expressive CD45 expression prevailed in the decidual membrane, TNF-α expression was predominantly negative in group I. In group II, inflammatory changes were evidenced by collagenization and stromal fibrosis, a decrease in fetal vessels, fibroblast proliferation, an increase in intervillous fibrinoid, and CD45 spread was expressed in decreasing order of intensity in decidual membrane, in stroma of intermediate villi and in stem villi. In group III, foci of inflammatory infiltration spread from the decidual membrane to the intervillous space and villi stroma, vessels plethora of all types of villi, dystrophic changes, and fibrinoid volume increased. CD45 expression was more common and expressive than in the groups I and II, and TNF-α expression was weakly positive only in the decidual membrane. The control group determined the widespread expressive CD45 expression in all placental structures and positive moderate TNF-α expression.Conclusions. Preterm delivery and delivery on term occur when there is an activation of inflammatory reactions in placenta. Inflammation in the placental structures is a necessary condition for development of the mother-fetal interphase, however, the features of local immune response prove the need to determine genetic mechanisms of the preterm labor.

Characterization of Placental Infection by Zika Virus in Humans: A Review of the Literature.

The aim of the current review is to present a systematic evaluation of reported human placental findings in cases of zika virus (ZIKV) infection. We reviewed the EMBASE, PUBMED, and SCIELO databases until June 2019, without language restrictions. The search terms placenta AND zika virus were used. The inclusion criteria of the studies were studies that reported placental findings in humans. Experimental studies, reviews, notes or editorials were excluded. A total of 436 studies were retrieved; after duplicate exclusion, 243 articles had their titles screened, and 128 had their abstract read; of those, 32 were included in the final analysis (18 case reports, 10 case series, and 4 cohorts) DATA COLLECTION: We collected data concerning the author, year of publication, study design, number of participants, number of placental samples, onset of symptoms, perinatal outcomes, and main findings on histological analysis. The placental pathologic findings were described as mild and nonspecific, similar to those of other placental infections, including chronic placentitis, chronic villitis, increased Hofbauer cells, irregular fibrin deposits, increased mononuclear cells in the villus stroma, villous immaturity, edema, hypervascularization, stromal fibrosis, calcification, and focal necrosis of syncytiotrophoblasts. Zika infection presents unspecific placental findings, similar to other infections in the toxoplasmosis, other agents, rubella, cytomegalovirus, and herpes (TORCH)group. Characterizing and standardizing placental findings after zika virus infection is key to understanding the mechanisms of congenital diseases.

Molecular and morphological peculiarities of chronic placental insufficiency formation caused by different types of diabetes mellitus

Рathogenesis of the chronic placental insufficiency is largely determined by the type of diabetes mellitus and the degree of its compensation. Trophic function failure of placenta changes its hormonal activity, formation of respiratory disorders and development of oxidative stress. The histological structure of placentas among patients with type 1 diabetes mellitus (T1D) is represented by reduced chorionic villi dimensions of all levels. Stromal edema and increased number of mesenchymal stromal cells are found in the stem and intermediate villi, and hypervascularisation and thickening of syncytiocapillary membranes are detected in terminal villi. In type 2 diabetes mellitus (T2D), the histological structure of the placenta may be represented as a premature maturation and abnormal immaturity of the villous tree with focal fibrosis of villi stroma, hypervascularisation of villi, abundance of syncytial nodules and infarction in the subchorial space. The peculiarity of the placenta structure in gestational diabetes mellitus is predominantly an intermediate immature type of development with angiogenesis abnormality. Angiogenesis processes failure and endothelial dysfunction in chorionic villi associated with hyperglycaemia change the permeability of cell membranes, transferring cells to anaerobic respiration. Metabolic imbalance in the placenta causes the development of diabetic micro-angiopathy in the fetal-placental complex, antenatal hypoxia and negative perinatal outcomes.

Placental Expression of the Forelimb Patterning Transcription Factor MEIS2 in Trisomy 15

Background Observations of first trimester human trisomy 15 (T15) embryos have identified meromelic changes in the upper limbs. These changes are similar to those observed in animal studies investigating the effects of overexpression of Meis2, a signaling transcription factor expressed during forelimb development. Although it would be exceedingly difficult to assess MEIS2 expression in the human embryonic arm, MEIS2 has been reported as consistently expressed in first trimester placental villus stroma. Methods: This study addresses whether gene dosage effect might underlie meromelia in T15 by comparing MEIS2 expression in placentas from T15 and euploid spontaneous abortions employing manual and automated assessment of MEIS2 immunohistochemical scoring. Results: Average MEIS2 expression is increased in T15 first trimester placental tissue compared to euploid controls but that the difference is marginal. Manual and automated scoring showed moderately strong correlation. Conclusion: Extrapolation of these results suggests MEIS2 overexpression may not be required for meromelia in T15.

The human placenta and aneuploidy — trisomies 21, 18, 13

The purpose of this research was to investigation the morphological features of human placenta with karyotyped trisomy 21, 18 and 13 chromosomes. The study included 50 placenta of fetuses in miscarriages pregnancy at 19–20 weeks, among them 10 placentas obtained as a result of induced abortions due to karyotyped trisomy 21 of the fetuses; 10 placentas obtained as a result of induced abortions due to karyotyped trisomy 18 of the fetuses; 10 placentas obtained as a result of induced abortions due to karyotyped trisomy 13 in the fetuses; 20 placentas obtained as a result of spontaneous miscarriages without any congenital defects and abnormal karyotype of the fetuses. As a result of the investigation, in placentas with karyotyped trisomies, a violation of villi branching was noted. With karyotyped trisomy 21, inhibition of angiogenesis and sclerosis of villi with proliferation of fibroblasts were revealed. In cells of chorionic villi with karyotyped trisomy 18, increased apoptosis was noted. In placentas with karyotyped trisomy 13 hydropic changes in villus stroma and inhibition of angiogenesis were noted. As a result of the study, in placentas with karyotyped trisomies, a violation of villi branching was noted. With karyotyped trisomy 21, inhibition of angiogenesis and sclerosis of villi with proliferation of fibroblasts were revealed. In cells of chorionic villi with karyotyped trisomy 18, increased apoptosis was noted. In placentas with karyotyped trisomy 13 hydropic changes in villus stroma and inhibition of angiogenesis were noted.

MORPHOLOGICAL AND IMMUNOLOGICAL CHANGES IN STRUCTURES FETUS-PLACENTAL BARRIER IN WOMEN WITH TORCH - INFECTIONS IN ANAMNESIS

Formation of chronic placental insufficiency in the stage of decompensation in pregnant women promotes intrauterine infection of the child.Today, histologic, morphological and immunohistochemical changes of the placenta in women with TORCH infections in the anamnesis. At TORCH-infection there are considerable infringements of mechanisms of functioning of a feto-placental barrier. Identifying these features should be an additional criterion for early diagnosis of perinatal infections in newborn risk groups.The aims. To evaluate macroscopic, histological, morphological and immunohistochemical changes in the placenta and to prove violation of the feto-placental barrier of women with TORCH-infections in the anamnesis.Materials and methods of research. 40 placens of women were investigated. The I group - 20 placenta of women with transmitted TORCH - infections in the anamnesis (confirmed immunologically during pregnancy transmitted of CMV, herpesvirus and toxoplasmic infections); The II group of control - 20 placets of healthy women with the physiological couse of pregnancy. In the placenta study, organometric, macroscopic, and general histological methods were used. Immunohistochemical - indirect streptavidin-peroxidase method of detecting the level of expression of the marker of proliferation and regeneration of Ki-67. Histochemical method - DNA according to Felgen with the following definition of apoptosis index.Results of the research. In the micro and macroscopic study placenta from mothers with transmitted TORCH infections revealed significant changes: an increase in the mass of the placenta (50%), thickening of the umbilical cord (75%), moderate swelling of precious candles (75%), vasculitis (30%), amniotic edema (30%), inflammatory infiltration in the decidua (25%), chorionic plate (35%), intervertebral space (30%), stroma villi (25%), vessels (30%); impaired blood circulation and degenerative changes in the decidua (50%). Distribution of calcifications (30%) in the regional and paracentral zones of the placenta. The uneven intensity of the DNA reaction in the placental barrier structures and a significant increase in the apoptotic index indicate an injury and a violation of their development. Immunohistochemically increased increase in expression levels of the proliferative marker KI-67 and the index of the proliferative index.Conclusions. Identified macroscopic and histological changes in the state structures feto-placental barrier in women with TORCH-infections indicate that they placental insufficiency due to productive vascular-cell response in the stroma of villi and decidua membrane and possibly the result the effects of viruses and toxoplasma. The findings should be taken into account by neonatologists for more careful monitoring of newborns at risk for perinatal infection from mothers with TORCH-infected history.

323. Peculiarities of placental expression of erythropoietin in preeclampsia

Introduction The role of erythropoietin (EPO) in the pathogenesis of preeclampsia (PE) is important and associated with the anti-apoptotic and angiogenic effects of the hormone. Objectives The aim was to study the peculiarities of placental expression of erythropoietin in preeclampsia. Methods In the cross-sectional cohort study we included 55 pregnant women: group 1 (control) – 11 women without PE; group 2–14 women with mild PE; group 3–30 women with severe PE. Immunohistochemical studies of the placenta (syncytium, capillary endothelium, macrophages of villi stroma) were performed to evaluate the expression of EPO (primary specific anti-Epo (N-19): sc-1310-R (Santa Cruz Biotechnology, Inc., California, USA) in 10 randomly selected areas using an objective micrometer under light microscope at a magnification of 400 high-power field. The mean for 10 fields was used as the cell count for that section. Results In group 1 EPO expression was more often detected in the villous syncytiotrophoblast (19.73 ± 0.54, ps−e =0.005) compared with the capillaries endothelium (9.46 ± 0.60, ps−e =0.003) and stromal macrophages (12.27 ± 0.51, ps−m =0.003, pe−m=0.005). In mild PE expression of EPO in the syncytium (26.20 ± 0.46, p1−2 Discussion It has been suggested that in severe PE, placental hypoxia could induced EPO expression in placenta (syncytium, capillary endothelium, macrophages of villi stroma).

SPECIAL ASPECTS OF MISCARRIAGE IN WOMEN WITH RECCURENT MISCARRIAGE

Missing pregnancy is a consequence of the simultaneous or sequential action of several factors. The main causes of miscarriage and spontaneous interruption of pregnancy include: genetic factors, endocrine disorders, immune and infectious factors, congenital and acquired diseases of female genital organs. In almost 50% of women, it is not impossible to determine the reason of miscarriage, so the question of early diagnosis and prevention of this condition is quite acute.Materials and methods. In this study, we performed a pathohistological study of the deciduum in order to determine the etiological factor of the pathology of implantation of the embryo and placentation. The study included 88 women with a diagnosis of "recurrent miscarriage" that met the criteria for inclusion and exclusion.Results of research. In the study group, the age of women was from 19 to 35 years old (mean age was 27.6±4.1 years old). The abortion was observed at differentst ages of pregnancy, more often during the period of 4-9 weeks of gestation (67 cases – 76.14%). According to the results of the histopathological study of decidouum lymphohistiocytic infiltration was revealed in the stroma of villi in 62 cases (70.45%), other changes were less common.Conclusions and perspectives of further research. The obtained data indicate that the determination of the etiologic factor of miscarriage of the pregnancy, especially in women with a diagnosis "Reccurent miscarriage of obscure etiology", will allow to predict the development of the pathology of implantation and placentation in subsequent pregnancies.Prospects for further research are to develop adequate preparation before pregnancy and prevention of the pathology of implantation and placentation.

P 24 Cell renovation markers, apoptotic markers and placental expression of erythropoietin in preeclampsia

Introduction The role of erythropoietin (EPO) in trophoblast invasion, placental insufficiency and preeclampsia (PE) are associated with angiogenesis-stimulated and antiapoptotic effects. Objectives To study cell renovation markers, apoptotic markers and erythropoietin placental expression in early-onset and late onset preeclampsia. Materials and methods In a case-control study, we included 15 pregnant women with early-onset PE and 13 women – with late-onset PE. We have studied cell proliferation marker ki-67, markers of antiapoptosis bcl-2 and proapoptosis p53, placental expression of EPO in syncytium, capillaries endothelium, villi stroma macrophages. Results In early-onset PE bcl-2 expression was lower in syncytium (5.22 ± 0.78 vs 10.21 ± 0.75, p Conclusion Despite the high expression of EPO in PE, the degree of adaptive changes, particularly in syncytium, was insufficient and that was accompanied by dystrophic changes in placenta (ischemic infarcts, immature chorionic villi) with the realization of proapoptosis and cell proliferation mechanisms.

50 Erythropoietin production adequacy and its placental expression in preeclampsia

Introduction The erythropoietin (EPO) participation in preeclampsia (PE) may be associated with its angiogenesis-stimulating and antiapoptotic effects in hypoxia conditions, including placental genesis. Objectives To study serum EPO-level and its placental expression level in preeclampsia. Methods In case-control study there were 11 women without preeclampsia (the 1st group), 14 – with moderate PE (the 2nd group) and 14 – with severe PE (the 3rd group). EPO-level of mother's blood serum was determined by means of solid-phase chemiluminescent enzyme immunoassay (sandwich-type) with coefficient estimation of EPO-production adequacy (CAepo – relation of observed serum EPO logarithm to tentative EPO logarithm, in normal limits 0.8–1.2). Immune histochemical test was used to estimate EPO-expression in symplastotrophoblast, capillary endothelium, placenta villi stroma macrophages (monoclonal antibodies to Epo (N-19): sc-1310-R, Santa Cruz Biotechnology, Inc., California, USA). Quantitative investigation was performed in 10 randomly selected fields of vision by magnification 400. We used statistical software programs: SPSS 12.1, Statistica for Windows 6.0, STADIA 6.3 prof. (Mann–Whitney test, Pearson's chi-squared test, nonparametric correlation analysis with calculation of Spearman's correlation coefficient). Results Blood serum EPO-level in moderate PE (59.25±13.54mIU/ml; 95% CI 30.34–88.11) appeared to be a bit higher and in severe PE (39.88±10.60mIU/ml; 95% CI 18.78–64.97) – lower that in the 1st group (43.02±8.24mIU/ml, 95% CI 25.21–60.83). The cases of EPO inadequate production ( p =0.005) were registered only in PE, the detection rate was the highest in severe PE (50.0% of cases). CAepo in severe PE (0.87±0.04; 0.75–0.98) was less (p1-3=0.021) compared with the 1st group (1.03±0.03; 95% CI 0.95–1.11). EPO-expression in placenta tissues during the pregnancy not complicated by PE was oftener determined in syncytiotrophoblast (19.73±0.54; 95% CI 18.52–20.93) in comparison with vascular endothelium (9.46±0.60; 95% CI 8.13–10.78; p =0.003) and villi stroma macrophages (12.27±0.51; 95% CI 11.15–13.40; p =0.003). In moderate PE erythropoietin amount in indicated placental structures (26.20±0.46, 95% CI 25.21–27.19; 18.87±0.50, 95% CI 17.80–19.93; 19.87±0.48, 95% CI 18.84–20.89 respectively) was notably higher that analogic indices in the 1st group (p1-2 p =0.001). EPO-representation in placenta of women with severe PE in symplast (33.06±0.76; 95% CI 31.45–34.67), in vascular endothelium (26.00±0.40; 95% CI 25.15–26.85) and in villi stroma macrophages (27.29±0.52; 95% CI 26.19–28.40) was higher (p1-3 p r =−0.377, p =0.014; r =−0.429, p =0.005; r =0.444; p =0.003 respectively) and negative – with CAepo ( r =−0.377, p =0.014; r =−0.429, p =0.005; r =−0.398; p =0.009 respectively). Conclusions Considerable increase of placenta EPO-expression was registered in association with its inadequate production in PE, especially in severe PE. We suppose that the decreasing of mother's serum EPO-level is probably connected with its renal and perhaps placental production disturbance.

Pathological and molecular study of mycotic abortion in ewes

The aims of the present study is to isolate the fungal species associated with ewe abortion and to examine the histopathological changes in the placenta associated with fungal isolates .In addition to study the genetic information of TNF alpha gene, intensity and score of TNF-alpha protein in aborted and normal placenta by PCR and IHC respectivly. Frozen placental tissue and paraffin embedded tissue belong to aborted and normal groups were used for assessment of TNF alpha gene by studying the genetic information of TNF-alpha gene in aborted and normal placental sample by PCR .immmunohistochemical study was carried out to determine TNF protein in the placental sample. The same frozen placenta tissue samples were also used for DNA extraction to detect TNF alpha gene in the two groups. A fragment of (238 bp ) of the TNF alpha gene was amplified by PCR using primers ,the sequence of sense and antisense primer for TNF alpha gene was: GAA TAC CTG GAC TAT GCC GA,CC TCA CTT CCC TAC ATC CCT (bp 238) Gene Bank: X56756. The study showed that 10 fungal species were isolated from aborted placenta with high percentage of these isolates was recorded in March ,.The fungal isolates were variable according to region of sample collecting ,high percentage in the AL-Manathera(36%),followed by Al-Kuzweenia (34%) and Al-Shabaka(30%).The fungal isolates include Coccidioides immitis (16%). Asperigllus.fumigitus (14%), Trichophyton, Rhodetella rubra, and Aspergillus.ochraus ,(10%) for each one . Candida kruzei,(8%). Aspergillus.flavus and Geotrichum ,6%) for each one . Prototheca..zopfi, Saccharomyces.cerevisia, Blast.capitatuss, Cryptococcus uniguttultus and Candida zeylanoides,(4%) for each one . This study revealed the presence of ~238bp band in addition to the ~300 ,400 and 600 bp desired fragment in PCR products that related to aborted samples compared to normal one .,this may be due to polymorphism within the TNF gene itself. We found a significant elevated TNF alpha protein in inflammatory cell (macrophage) of aborted sheep placenta (increasing of intensity and score), while no changes could be observed for pro-inflammatory molecules in the control sample (normal delivery) The current study expressed correlation between polymorphisms in TNF- alpha gene ,immunohistochimistry and the fungal isolated species. The histopathological examination showed acute suppurative placentitis, necrosis of chorion villi, congestion of blood vessels with fibrin deposition in the villus stroma and inter villus space, in addition to calcification in the villus stroma and sloughing and desquamation of cytotrophoblast .numerous yeast and spheriols were detected in the placental tissue by periodic acid Schiff reaction(PAS).

Diverse patterns of cell-specific gene expression in response to glucocorticoid in the developing small intestine

Although glucocorticoids are known to elicit functional maturation of the gastrointestinal tract, the molecular mechanisms of glucocorticoid action on the developing intestine have not been fully elucidated. Our previous microarray studies identified 66 transcripts as being rapidly induced in the jejunum following dexamethasone (Dex) administration to suckling mice. Now we report the specific cellular location of a subset of these transcripts. Mouse pups at P8 received Dex or vehicle and intestinal segments were collected 3-4 h later. Robotic-based in situ hybridization (ISH) was performed with digoxygenin-labeled riboprobes. Transcripts studied included Ndrg1, Sgk1, Fos, and two unknown genes (Gene 9 and Gene 36). As predicted, ISH revealed marked diversity of cellular expression. In small intestinal segments, Sgk1 mRNA was in all epithelial cells; Fos mRNA was confined to epithelial cells at the villus tip; and Ndrg1 and Gene 36 mRNAs were localized to epithelial cells of the upper crypt and villus base. The remaining transcript (Gene 9) was induced modestly in villus stroma and strongly in the muscle layers. In the colon, Ndrg1, Sgk1, and Gene 36 were induced in all epithelial cells; Gene 9 was in muscle layers only; and Fos was not detectable. For jejunal segments, quantitation of ISH signals in tissue from Dex-treated and vehicle-treated mice demonstrated mRNA increases very similar to those measured by Northern blotting. We conclude that glucocorticoid action in the intestine reflects diverse molecular mechanisms operating in different cell types and that quantitative ISH is a valuable tool for studying hormone action in this tissue.