Prevalence of chromosomal alterations in first-trimester spontaneous pregnancy loss.

Pregnancy loss is often caused by chromosomal abnormalities of the conceptus. The prevalence of these abnormalities and the allocation of (ab)normal cells in embryonic and placental lineages during intrauterine development remain elusive. In this study, we analyzed 1,745 spontaneous pregnancy losses and found that roughly half (50.4%) of the products of conception (POCs) were karyotypically abnormal, with maternal and paternal age independently contributing to the increased genomic aberration rate. We applied genome haplarithmisis to a subset of 94 pregnancy losses with normal parental and POC karyotypes. Genotyping of parental DNA as well as POC extra-embryonic mesoderm and chorionic villi DNA, representing embryonic and trophoblastic tissues, enabled characterization of the genomic landscape of both lineages. Of these pregnancy losses, 35.1% had chromosomal aberrations not previously detected by karyotyping, increasing the rate of aberrations of pregnancy losses to 67.8% by extrapolation. In contrast to viable pregnancies where mosaic chromosomal abnormalities are often restricted to chorionic villi, such as confined placental mosaicism, we found a higher degree of mosaic chromosomal imbalances in extra-embryonic mesoderm rather than chorionic villi. Our results stress the importance of scrutinizing the full allelic architecture of genomic abnormalities in pregnancy loss to improve clinical management and basic research of this devastating condition.

The effects of standing in tutorial group meetings on learning: A randomized controlled trial.

BACKGROUND: Standing desks have been brought into the education environment to reduce sedentary behavior among students. The current study explored the effects of standing in tutorial group meetings on learning among undergraduate students. METHODS: Ninety-six participants were randomly allocated to a Sit or Stand group, with 2 h tutorial group meetings scheduled, once or twice per week, for nine weeks. Learning was analyzed using exam grades, concept maps, and tutorial interactions. RESULTS: Overall, the Sit and Stand groups did not differ from each other in terms of learning, measured through their exam, concept map, and the use of learning-oriented interactions. CONCLUSION: Standing in tutorial group meetings neither enhanced nor compromised learning. Considering the health risks associated with prolonged sedentary behavior, offering standing tutorial group meetings to undergraduate students is a recommended solution to break up prolonged sedentary behavior and encourage more physical activity, while maintaining the learning performance of students.

Placental hypoxia-induced alterations in vascular function, morphology, and endothelial barrier integrity.

Preeclampsia (PE) is a pregnancy-related disorder characterized by hypertension and proteinuria that affects 3-10% of all pregnancies. Although its pathophysiology remains obscure, placental hypoxia-induced oxidative stress and alterations in vascular function, morphology, and endothelial barrier integrity are considered to play a key role in the development of preeclampsia. In this study, placental villous explants of noncomplicated placentae and BeWo cells were subjected to hypoxia. The effect of placental hypoxic-conditioned medium (HCM) on intraluminal-induced contraction and endothelial barrier integrity in chorionic arteries was investigated using pressure myography. The impact of BeWo cell HCM on endothelial cell viability, reactive oxygen species formation and inflammation was also determined. Alterations in arterial morphology and contractile responsiveness to the thromboxane A2 analog (U46619) after exposure to placental HCM were examined immunohistochemically and by wire myography, respectively. Intraluminal administration of placental HCM induced vasoconstriction and increased the endothelial permeability for KCl, which was concentration-dependently prevented by quercetin. Placental and BeWo cell HCMs decreased endothelial cell viability, increased the production of reactive oxygen species and enhanced the secretion of IL-6 and IL-8. The cross-sectional area of the arterial media was increased upon exposure to placental HCM, which was associated with increased vascular proliferation and contractile responsiveness to U46619, and all of these effects were prevented by the antioxidants quercetin and RRR-α-tocopherol. This study is the first to comprehensively demonstrate the link between factors secreted by placental cells in response to hypoxia and vascular abnormalities and paves the way for new diagnostic approaches and therapies to better protect the maternal vasculature during and after a preeclampsia-complicated pregnancy.

Histological villous maturation in placentas of complicated pregnancies.

Chorioamnionitis and preeclampsia account for the majority of preterm births worldwide. Thus far, adequate methods for early detection or prevention of these diseases are lacking. In preeclampsia, accelerated villous maturation is believed to compensate placental insufficiency. However, little is known about the effects of placental inflammation in chorioamnionitis on villous maturation. Therefore, we established a set of morphological parameters to evaluate histological villous maturity in pregnancies complicated by chorioamnionitis and preeclampsia. Preterm placentas complicated by chorioamnionitis or preeclampsia were compared to idiopathic preterm placentas and term controls. Histological villous maturation was analyzed by means of 17 histological markers. Fourteen of these markers provided information on absolute and relative numbers of the terminal villi (TV), the extent of their vascularization (using CD31-stained sections) and their exchange capacities. In addition, the numbers of syncytial bridges, syncytial apoptotic knots and shed syncytiotrophoblasts were counted. Accelerated villous maturation in preeclampsia was demonstrated by means of histological villous remodeling and confirmed by 11 relevant markers. Chorioamnionitis, however, only showed increased area of fetal capillaries. In preeclampsia, placentas may transition from growth to maturation earlier than placentas in normal pregnancies, whereas in chorioamnionitis placental changes are more acute and therefore less elaborated at a structural level. Regression analysis suggests the number of all villi and the number of terminal villi as a percentage of all villi as parameters to evaluate histological villous maturity in preeclamptic placentas and to assist diagnosis. However, we would recommend to analyze all 11 relevant parameters to judge placental maturity in detail.

Fetal tolerance in human pregnancy--a crucial balance between acceptance and limitation of trophoblast invasion.

During human pregnancy the semi-allogeneic/allogeneic fetal graft is normally accepted by the mother's immune system. Initially the contact between maternal and fetal cells is restricted to the decidua but during the 2nd trimester it is extended to the entire body. Two contrary requirements influence the extent of invasion of extravillous fetal trophoblast cells (EVT) in the maternal decidua: anchorage of the placenta to ensure fetal nutrition and protection of the uterine wall against over-invasion. To establish the crucial balance between tolerance of the EVT and its limitation, recognition of the semi-allogeneic/allogeneic fetal cell by maternal leukocytes is prerequisite. A key mechanism to limit EVT invasion is induction of EVT apoptosis. Apoptotic bodies are phagocytosed by antigen-presenting cells (APC). Peptides from apoptotic cells are presented by APC cells and induce an antigen-specific tolerance against the foreign antigens on EVT cells. These pathways, including up-regulation of the expression of IDO, IFNgamma and CTLA-4 as well as the induction of T(regulatory) cells, are general immunological mechanisms which have developed to maintain peripheral tolerance to self-antigens. Together these data suggest that the mother extends her "definition of self" for 9 months on the foreign antigens of the fetus.

Tubal abortions but not viable tubal pregnancies are characterized by an increased number of CD8+ T cells.

OBJECTIVE: To examine immune cell phenotypes in viable tubal pregnancies (VTP) and in tubal abortions (TA). METHODS: Paraffin-embedded specimens of VTP (n=7) and ongoing TA (n=6) were double-stained for cytokeratin for trophoblast as well as for CD45, CD3, CD8, CD68 and CD20 for immune cell phenotypes. In all cases, the amniotic sac was detected by ultrasound. Histological examination showed no evidence of necrosis within the tissues included in this study. Quantification of the subpopulations was performed in each slide by two independent examiners in five areas (0.085 mm2 each) of the invasion zone as marked by cytokeratin-positive stromal extravillous trophoblast (EVT) cells. For statistical analysis, the non-parametric two-tailed t-test was used (p<0.05). RESULTS: The differences in the number of CD45(+), CD68(+) and CD20(+) cells was significant (p=0.0423, p=0.0469 and p=0.0494, respectively); however, the number of CD3(+), and among those the number of CD8(+) cells, was approximately eight-fold higher in TA than in VTP (p<0.0001 and p=0.0012, respectively). CONCLUSION: The unequal distribution of CD8(+) cells in VTP and TA suggests a significant role of this immune cell phenotype in the further outcome of a tubal pregnancy either to an abortive or a viable, potentially life-threatening, entity.

Regulation of taurocholate transport in freshly isolated proximal tubular cells of the rat kidney by protein kinases.

BACKGROUND/AIMS: The bile acids filtered through the glomeruli nearly completely escape urinary excretion due to an efficient tubular reabsorption process. Reabsorption is mediated mainly by the sodium-dependent bile acid transporter (ASBT) which is located in the brush border membranes of proximal tubular cells. The present study addresses the question whether this transporter is subject to short-term regulation by protein kinases. METHODS: The effects of specific activators or inhibitors of eight different protein kinases (PKs) on 3H-taurocholate uptake of proximal tubular cells were investigated. The cells were freshly isolated from rat kidneys by nycodenz density gradient centrifugation. RESULTS: Activation of the cAMP/PKA system by forskolin, 8-Br-cAMP, or the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine significantly diminished cellular 3H-taurocholate uptake whereas 8-Br-cGMP had no effect. Also the MEK1/2 inhibitors PD98059 and U0126, and the p38 mitogen-activated protein (MAP) kinase inhibitor SB203580 decreased 3H-taurocholate uptake. Phorbol myristate acetate and dioctanolglycerol, activators of PKC, and chelerythrine, a selective inhibitor of PKC, did not affect 3H-taurocholate uptake. Likewise the phosphatidylinositol-3 kinase inhibitor wortmannin and the tyrosine kinase inhibitor genistein induced no significant change of cellular 3H-taurocholate uptake. In a sodium-free medium forskolin and PD98059 did not affect 3H-taurocholate uptake but SB203580 significantly decreased it. CONCLUSION: It is concluded that PKA and MAP kinases are involved in the regulation of the ASBT-mediated taurocholate uptake into proximal tubular cells. p38 MAP kinase may have an additional effect on a sodium-independent tubular taurocholate transporter.

Cytokine microenvironments in human first trimester decidua are dependent on trophoblast cells.

OBJECTIVE: To compare cytokine expression profiles of decidua basalis (containing trophoblast cells) and decidua parietalis (without trophoblast cells) for determination of microenvironments in human first trimester decidua. DESIGN: Retrospective study. SETTING: School of Medicine, RWTH University of Aachen, Aachen Germany, and Bourgognekliniek Maastricht, Maastricht, The Netherlands. PATIENT(S): Forty-six women who had undergone elective first-trimester termination of viable pregnancy at 5 to 12 weeks. MAIN OUTCOME MEASURE(S): Quantitative cytokine protein analysis in decidual tissues by enzyme-linked immunosorbent assay, qualitative cytokine messenger (m)RNA analysis in isolated decidual cell samples, and comparative mRNA and protein analysis in tissues of decidua basalis compared with decidua parietalis. RESULT(S): Interleukin-2, interferon-gamma (Th-1), interleukin-4 (Th-2), and interleukin-1beta proteins are expressed in the human first-trimester decidua. Interleukin-2, interferon-gamma, and interleukin-4 mRNA mainly derive from the decidual tissue leukocytes. Interleukin-1beta mRNA is expressed by all decidual cell types. Interferon-gamma mRNA and protein is detected predominantly in the decidua basalis, which contains trophoblast cells. CONCLUSION(S): Microenvironments are established topographically by different expression of cytokines in decidua basalis and decidua parietalis. These locally specific patterns are indicative of fetomaternal cross-talk. Higher interferon-gamma concentrations in decidua basalis may influence leukocyte differentiation (e.g., macrophage activation) and trophoblast invasion (e.g., by induction of expression of major histocompatibility complex).

Cell-type specific expression and regulation of apolipoprotein D and E in human endometrium.

OBJECTIVE: To assess the expression and regulation of antilipoprotein D (ApoD) and antilipoprotein E (ApoE) in human endometrium. STUDY DESIGN: Endometrial biopsies from healthy, regularly cycling women were collected during the late proliferative and mid-secretory phase. mRNA gene expression of ApoD and ApoE was determined using real-time PCR in whole tissue, in isolated stromal (ESC), epithelial (EEC) and CD45(+) leukocytes (EIC), as well as after hormonal stimulation of ESC and EEC in vitro. Protein expression was analyzed using immunohistochemistry. RESULTS: ApoD and ApoE mRNA was expressed in all cell types examined. A rise in ApoD mRNA expression was seen in whole endometrium, ESC, and EEC in the secretory phase, as well as after hormonal stimulation of ESC and EEC in vitro. ApoE mRNA was significantly upregulated in whole endometrium of secretory phase biopsies, while its expression was not altered by progesterone in vitro. Immunohistochemistry of whole endometrial tissue localized ApoD mainly in ESC and EEC. While ApoE was localized slightly in ESC, it was particularly noted on the surface of secretory phase endothelial cells. CONCLUSION: We demonstrate for the first time the cell-type and cycle dependent expression of ApoD and ApoE within human endometrium, suggesting their role in endometrial modulation.

Dendritic cells are equally distributed in intrauterine and tubal ectopic pregnancies.

OBJECTIVE: To examine different stages of dendritic cells (DCs) in intrauterine (IUPs) and viable tubal (VTPs) pregnancies to further elucidate mechanisms of fetomaternal tolerance and extravillous trophoblast invasion. DESIGN: Experimental study on patient-controlled material. SETTING: University hospital. PATIENT(S): Seven women with normal IUPs and ten with VTPs in the first trimester. INTERVENTION(S): Suction curettage in IUP, laparoscopy in VTP. MAIN OUTCOME MEASURE(S): Immunohistochemistry for cytokeratin-7 (trophoblast), CD83 (mature DCs), DEC205 (activated but not fully mature DCs), DC-SIGN (immature macrophage-like DCs), and CD14 (macrophages) alone and in double staining. RESULT(S): The numbers of CD83+ and DEC205+ cells were similarly low in IUP and VTP (0.83 and 0.44 cells/mm2; 2.28 and 2.96 cells/mm2). The number of DC-SIGN+ cells was higher, though without significant differences among the entities examined (57.5 and 47.4 cells/mm2). About two-thirds of DC-SIGN+ cells were also CD14+ in IUP and VTP. CONCLUSION(S): The almost equal distribution of CD83+, DEC205+, and DC-SIGN+ cells in IUP and VTP suggests analogue control mechanisms in intrauterine and extrauterine DC differentiation and a comparable role of these DCs for the development of fetomaternal tolerance.

Amniotic fluid adhesion molecules during parturition at term.

AIMS: During term parturition a constant elevation of intercellular adhesion molecule (ICAM-1) as well as increases of endothelial leukocyte adhesion molecule (ELAM-1) and vascular cell adhesion molecule (VCAM-1) in the lower uterine segment and of ICAM-1 in fetal membranes were observed. We examined ELAM-1, ICAM-1 and VCAM-1 during normal term parturition to find out whether amniotic fluid adhesion molecules change accordingly. METHODS: Amniotic fluid specimens of 35 patients undergoing cesarean section at term with various stages of cervical dilatation (<2 cm, n=11; 2-<4 cm, n=10; 4-6 cm, n=6; >6 cm, n=8) and different durations of labor (0 h, n=11; >0 h, n=24) were examined for ELAM-1, ICAM-1 and VCAM-1 by enzyme immunoassay. For statistical analysis one-way ANOVA (cervical dilatation) and unpaired t-test (duration of labor) were used (P<0.05). RESULTS: Neither ELAM-1 nor VCAM-1 correlated with cervical dilatation or with the duration of labor. ICAM-1 showed a tendency to decline with the last phase of cervical dilatation (P=0.06) and a significant decline with labor (P=0.046). CONCLUSIONS: In contrast to adhesion molecules in the lower uterine segment amniotic fluid adhesion molecules ELAM-1 and VCAM-1 do not rise significantly during parturition. The decline of ICAM-1 may be due to a transfer to other compartments (i.e., lower uterine segment or retroplacental blood) thereby contributing to labor and cervical dilatation.

Class I histone deacetylase expression in the human cyclic endometrium and endometrial adenocarcinomas.

BACKGROUND: Class I histone deacetylases (HDACs) and acetylases (HATs) are members of transcriptional pre-initiation complexes assembled by steroid hormone receptors. Recently, HDAC inhibitors were shown to enhance differentiation of endometrial fibroblasts and endometrial adenocarcinomas. However, there is only rare information on HDAC and HAT expression in the human endometrium. METHODS: HDAC-1, -2, -3 and HAT (PCAF and GCN5) mRNA expression was studied in tissue from premenopausal women undergoing hysterectomy by real-time or semiquantitative RT-PCR. HDAC protein expression was assessed by Western Blot and immunohistochemistry. In endometrial adenocarcinomas (n = 17), HDAC-1 expression was studied by immunohistochemistry. RESULTS: In the human endometrium, HDAC-1, -2, -3 and PCAF mRNA are expressed without cyclical changes. Western blot analysis demonstrated that HDAC-2 protein expression was slightly, but significantly elevated in the secretory phase (P < 0.01 versus day 5-8), whereas HDAC-1 and -3 protein expression was constitutive throughout the menstrual cycle. By immunohistochemistry, nuclear expression of HDAC proteins was detected in all endometrial cell types. In the case of HDAC-3, immunostaining was significantly reduced in the endometrial surface epithelium on day 6-10 (P < 0.01 versus days 15-18 and 24-28). Compared to normal endometrium, a high proportion of endometrial adenocarcinomas showed impaired HDAC-1 protein expression in the epithelial and stromal compartment. CONCLUSIONS: Class I HDACs and HATs are expressed in the human endometrium throughout the menstrual cycle, suggesting the cyclic endometrium as a potential target for HDAC inhibitors. We hypothesis that alterations of HDAC and/or HAT expression are potentially involved in impaired endometrial differentiation.

Embryo-maternal signalling: how the embryo starts talking to its mother to accomplish implantation.

The process of implantation and trophoblast invasion is currently considered as the most limiting factor for the establishment of pregnancy. Molecular interactions at the embryo-maternal interface during the time of adhesion and subsequent invasion are crucial to the process of embryonic implantation. Both partners, the mother as well as the embryo, play equal roles in the embryo-maternal dialogue, the embryonic part being the main topic in this study. Investigations of the proteins in the extra-embryonic matrices (i.e. zona pellucida) indicate that the embryo participates intensively in this early embryo-maternal signalling. One unique feature during implantation process of primate embryos is the release of chorionic gonadotrophin, which seems to influence endometrial activity by two different mechanisms: (i) luteotrophic activity with increasing progesterone release and (ii) a direct action on the endometrium. Furthermore, embryonic interleukin-1beta may be involved in embryo-maternal signalling. Other significant signals in this interaction are most likely leukaemia inhibitory factor (LIF) and colony-stimulating factor (CSF), which stimulate matrix metalloproteinase (MMP)/insulin-like growth factor binding protein-1 (IGFBP-1) activity and the insulin-like growth factor (IGF) system, which is modulated by embryonic IGFBP-3. Similar significances are discussed for uteroglobin and haptoglobin. Finally, the phenomenon of maternal immunological tolerance, triggered by the presence of the early embryo, is fundamental to the understanding of implantation and trophoblast invasion. A tightly regulated balance between activated and inactivated T cells at the implantation site may control the beginning of adequate trophoblast invasion and also limit this invasion to a tolerable extent for the maternal system, consequently ensuring a biologically healthy haemo-chorial placenta.