Zebrafish prss59.1 is involved in chorion development.

The prss59.1 gene was identified as one of 11 genes that were highly upregulated during the induction of ovulation in zebrafish by using an in vivo ovulation assay. Previously, we conducted biochemical characterization of Prss59.1 and revealed it to be a trypsin-like proteolytic enzyme. In this study, we established a prss59.1 gene knockout strain using the CRISPR/Cas9 system. Phenotypic analysis of prss59.1 knockout fish showed that prss59.1 is associated with chorion elevation, a prominent event in egg activation during fertilization. The chorions of heterozygous and homozygous prss59.1 mutant zebrafish were smaller than those of the wild type. The results suggested that Prss59.1 is necessary for chorion expansion. The homozygous prss59.1 mutant strain, with a small chorion, showed an extremely low survival rate. Fiber-supported knob-like structures (KS) on the chorion showed an abnormal structure in prss59.1 mutants. Prss59.1 was detected in the KS on the chorion. The pores on the chorion were smaller in the prss59.1 mutants than in the wild type. Transmission electron microscopy (TEM) observations of the cross sections of the chorions showed abnormalities in the chorion structure in prss59.1 mutants. These results demonstrated that Prss59.1 is involved in chorion elevation and in proper formation of the chorion, which is necessary for embryo development.

Twin pregnancies, crown-rump length and birthweight discordancy: the influence of chorionicity / Gestações gemelares, comprimento craniocaudal e discordância de peso ao nascimento: a influência da corionicidade

Abstract Objective The purpose of the present study was to analyze the influence of chorionicity in the biometric parameters crown-rump length (CRL), birthweight (BW), crown-rump length discordancy (CRLD) and birthweight discordancy (BWD), determine the correlation between these latter two in cases of intertwin discordancy, and to analyze the influence of chronicity in the presence of these discordancies with clinical relevance (> 10% and > 15%, respectively). Methods The present study was a retrospective study based on the twin pregnancy database of the Centro Hospitalar S. João (2010-2015), including 486 fetuses among 66 monochorionic (MC) and 177 dichorionic gestations (DC). The inclusion criteria were multiple pregnancies with 2 fetuses and healthy twin gestations. The exclusion criteria were trichorionic gestations and pregnancies with inconclusive chorionicity, multiple pregnancy with ≥ 3 fetuses and pathological twin gestations. Results No statistically significant difference was found in BW (p = 0.09) and in its discordancy (p = 0.06) nor in CRL (p = 0.48) and its discordancy (p = 0.74) between MCs and DCs. Crown-rump length discordancy and birthweight discordancy were correlated by the regression line "BWD = 0.8864 x CRLD + 0.0743," with r2 = 0.1599. Crown-rump length discordancy > 10% was found in 7.58% of monochorionic and in 13.56% of dichorionic twins. Birthweight discordancy > 15% was detected in 16.67% of monochorionic and in 31.64% of dichorionic twins. Conclusion No statistically significant influence of chorionicity was identified in both birthweight and birthweight discordancy, as in crown-rump length and crown-rump length discordancy. Birthweight discordancy was correlated to crown-rump length discordancy in 20% of cases.

Resumo Objetivo O objetivo do presente estudo foi analisar a influência da corionicidade nos parâmetros biométricos comprimento craniocaudal, peso ao nascimento, discordância de comprimento craniocaudal e discordância de peso ao nascimento, determinar a correlação entre estes dois últimos caso haja discordância intergemelar e analisar a influência da corionicidade na presença destas discordâncias com relevância clínica (> 10% e > 15%, respectivamente). Métodos O presente estudo foi um estudo retrospectivo baseado na base de dados de gestações gemelares do Centro Hospitalar S. João (2010-2015), incluindo 486 fetos de 66 gestações monocoriônicas e 177 dicoriônicas. Os critérios de inclusão foram gestações múltiplas de 2 fetos e gestações gemelares saudáveis. Os critérios de exclusão foram gestações tricoriônicas ou de corionicidade inconclusiva, gestações múltiplas com ≥ 3 fetos e gestações gemelares patológicas. Resultados Não se encontrou diferença estatisticamente significativa no peso ao nascimento (p =0,09) e sua discordância (p = 0,06) nem no comprimento craniocaudal (p = 0,48) e sua discordância (p = 0,74) entre gestações monocoriônicas e dicoriônicas. Considerando todas as gestações, as discordâncias de comprimento craniocaudal e peso ao nascimento foram correlacionadas pela reta de regressão "discordância de peso ao nascimento = 0.8864 x discordância de comprimento craniocaudal + 0.0743," com r2 = 0,1599. A discordância de comprimento craniocaudal > 10% descobriu-se em 7.58% das gestações monocoriônicas e em 13.56% das dicoriônicas. A discordância de peso ao nascimento > 15% detectou-se em 16.67% das gestações monocoriônicas e em 31.64% das dicoriônicas. Conclusão Não se identificou influência estatisticamente significativa no peso ao nascimento e sua discordância, bem como no comprimento craniocaudal e sua discordância. A discordância de peso ao nascimento correlacionou-se com a discordância de comprimento craniocaudal em 20% dos casos.

Deficiency of ULK1/ATG1 in the follicle cells disturbs ER homeostasis and causes defective chorion deposition in the vector Rhodnius prolixus.

In insects, synthesis and deposition of the chorion (eggshell) are performed by the professional secretory follicle cells (FCs) that surround the oocytes in the course of oogenesis. Here, we found that ULK1/ATG1, an autophagy-related protein, is highly expressed in the FCs of the Chagas-Disease vector Rhodnius prolixus, and that parental RNAi silencing of ULK1/ATG1 results in oocytes with abnormal chorion ultrastructure and FCs presenting expanded rough ER membranes as well as increased expression of the ER chaperone BiP3, both indicatives of ER stress. Silencing of LC3/ATG8, another essential autophagy protein, did not replicate the ULK1/ATG1 phenotypes, whereas silencing of SEC16A, a known partner of the noncanonical ULK1/ATG1 function in the ER exit sites phenocopied the silencing of ULK1/ATG1. Our findings point to a cooperated function of ULK1/ATG1 and SEC16A in the FCs to complete choriogenesis and provide additional in vivo phenotype-based evidence to the literature of the role of ULK1/ATG1 in the ER in a professional secretory cell.

Twin Pregnancies, Crown-rump Length and Birthweight Discordancy: The Influence of Chorionicity.

OBJECTIVE: The purpose of the present study was to analyze the influence of chorionicity in the biometric parameters crown-rump length (CRL), birthweight (BW), crown-rump length discordancy (CRLD) and birthweight discordancy (BWD), determine the correlation between these latter two in cases of intertwin discordancy, and to analyze the influence of chronicity in the presence of these discordancies with clinical relevance (> 10% and > 15%, respectively). METHODS: The present study was a retrospective study based on the twin pregnancy database of the Centro Hospitalar S. João (2010-2015), including 486 fetuses among 66 monochorionic (MC) and 177 dichorionic gestations (DC). The inclusion criteria were multiple pregnancies with 2 fetuses and healthy twin gestations. The exclusion criteria were trichorionic gestations and pregnancies with inconclusive chorionicity, multiple pregnancy with ≥ 3 fetuses and pathological twin gestations. RESULTS: No statistically significant difference was found in BW (p = 0.09) and in its discordancy (p = 0.06) nor in CRL (p = 0.48) and its discordancy (p = 0.74) between MCs and DCs. Crown-rump length discordancy and birthweight discordancy were correlated by the regression line "BWD = 0.8864 x CRLD + 0.0743," with r2 = 0.1599. Crown-rump length discordancy > 10% was found in 7.58% of monochorionic and in 13.56% of dichorionic twins. Birthweight discordancy > 15% was detected in 16.67% of monochorionic and in 31.64% of dichorionic twins. CONCLUSION: No statistically significant influence of chorionicity was identified in both birthweight and birthweight discordancy, as in crown-rump length and crown-rump length discordancy. Birthweight discordancy was correlated to crown-rump length discordancy in 20% of cases.

OBJETIVO: O objetivo do presente estudo foi analisar a influência da corionicidade nos parâmetros biométricos comprimento craniocaudal, peso ao nascimento, discordância de comprimento craniocaudal e discordância de peso ao nascimento, determinar a correlação entre estes dois últimos caso haja discordância intergemelar e analisar a influência da corionicidade na presença destas discordâncias com relevância clínica (> 10% e > 15%, respectivamente). MéTODOS: O presente estudo foi um estudo retrospectivo baseado na base de dados de gestações gemelares do Centro Hospitalar S. João (2010­2015), incluindo 486 fetos de 66 gestações monocoriônicas e 177 dicoriônicas. Os critérios de inclusão foram gestações múltiplas de 2 fetos e gestações gemelares saudáveis. Os critérios de exclusão foram gestações tricoriônicas ou de corionicidade inconclusiva, gestações múltiplas com ≥ 3 fetos e gestações gemelares patológicas. RESULTADOS: Não se encontrou diferença estatisticamente significativa no peso ao nascimento (p = 0,09) e sua discordância (p = 0,06) nem no comprimento craniocaudal (p = 0,48) e sua discordância (p = 0,74) entre gestações monocoriônicas e dicoriônicas. Considerando todas as gestações, as discordâncias de comprimento craniocaudal e peso ao nascimento foram correlacionadas pela reta de regressão "discordância de peso ao nascimento = 0.8864 x discordância de comprimento craniocaudal + 0.0743," com r2 = 0,1599. A discordância de comprimento craniocaudal > 10% descobriu-se em 7.58% das gestações monocoriônicas e em 13.56% das dicoriônicas. A discordância de peso ao nascimento > 15% detectou-se em 16.67% das gestações monocoriônicas e em 31.64% das dicoriônicas. CONCLUSãO: Não se identificou influência estatisticamente significativa no peso ao nascimento e sua discordância, bem como no comprimento craniocaudal e sua discordância. A discordância de peso ao nascimento correlacionou-se com a discordância de comprimento craniocaudal em 20% dos casos.

Up-to-date role of the dehydrated human amnion/chorion membrane (AMNIOFIX) for wound healing.

INTRODUCTION: Chronic wounds pose a significant burden on patients, society, and the health-care setup. Higher costs, protracted clinical course, and increased risk of complications necessitate identifying novel treatment modalities that hasten healing and wound closure. AREAS COVERED: This article covers newer available treatment modalities for chronic wounds, namely the dehydrated amniotic membrane products, biological skin substitutes, and similar therapies aimed at the healing of chronic non-healing wounds. It presents product description for Amniofix (dehydrated human amniotic/chorionic membrane) and its efficacy, compared to other similar products. EXPERT OPINION: In our experience and review of available literature, we expect Amniofix to offer wound care specialists with a more effective, easy-to-use, and convenient treatment modality for chronic wounds. Amniofix and other dHACM (dehydrated human amniotic/chorionic membrane) therapies reported faster and complete healing with lower complication rates, when compared to other similar products. These features encourage the use of Amniofix in Diabetic foot ulcers and Venous Leg Ulcers, besides other conditions such as plantar fasciitis.

Maternal Larp6 controls oocyte development, chorion formation and elevation.

La-related protein 6 (Larp6) is a conserved RNA-binding protein found across eukaryotes that has been suggested to regulate collagen biogenesis, muscle development, ciliogenesis, and various aspects of cell proliferation and migration. Zebrafish have two Larp6 family genes: larp6a and larp6b Viable and fertile single and double homozygous larp6a and larp6b zygotic mutants revealed no defects in muscle structure, and were indistinguishable from heterozygous or wild-type siblings. However, larp6a mutant females produced eggs with chorions that failed to elevate fully and were fragile. Eggs from larp6b single mutant females showed minor chorion defects, but chorions from eggs laid by larp6a;larp6b double mutant females were more defective than those from larp6a single mutants. Electron microscopy revealed defective chorionogenesis during oocyte development. Despite this, maternal zygotic single and double mutants were viable and fertile. Mass spectrometry analysis provided a description of chorion protein composition and revealed significant reductions in a subset of zona pellucida and lectin-type proteins between wild-type and mutant chorions that paralleled the severity of the phenotype. We conclude that Larp6 proteins are required for normal oocyte development, chorion formation and egg activation.

The effect of placental location identified before delivery on birthweight discordance among diamniotic-dichorionic twin pregnancies: a three-year retrospective cohort study.

This retrospective cohort study aimed to investigate the effect of placental location on birthweight discordance among diamniotic-dichorionic twin pregnancies. Medical records and sonographic reports of 978 diamniotic-dichorionic twin pregnancies delivered at Foshan Maternal and Fetal Health Hospital were reviewed. Pregnancies with congenital malformation, intrauterine death or placenta previa were excluded. The placental location for each twin was determined by last sonographic examination before delivery, and the pregnancies were grouped by different versus same placental location in each pregnancy. Maternal and fetal characteristics were summarized. The primary outcome of interest was birthweight discordance (BWD) ≥20%, and secondary outcomes included small for gestational age (SGA) as a binary outcome and mean value and absolute difference in birthweight as continuous outcomes. Student's t test and the chi-square test were used for univariate analyses, while multivariate regressions were used to adjust for confounders. General estimated equation (GEE) models were used to address the correlation between fetuses when assessing SGA. A total of 866 eligible subjects were included in the analysis. In total, 460 pregnancies had placentas with different locations, and 406 had placentas with same locations. The gestational age at delivery was slightly younger in the same placental location group than in the different placental location group (35.8 ± 0.1 vs. 36.1 ± 0.1 weeks, P = 0.067). Other maternal and fetal characteristics were comparable between the two study groups. There was no significant difference in BWD ≥20% (aOR = 1.06; 95% CI: 0.71-1.59) or SGA (aOR = 1.32; 95% CI: 0.76-2.28) between the same and different placental location groups. Neither the mean value nor the absolute difference in birth weight was associated with placental location combination (P = 0.478 and P = 0.162, respectively). In conclusion, discordant birthweight is not affected by same location of diamniotic-dichorionic placentas.

An investigational study of a dual-layer, chorion-free amnion patch as a protective barrier following lumbar laminectomy in a sheep model.

The inherent properties of the human amniotic membrane (HAM) suggest its potential for use as a physical barrier during surgery to protect neural elements and vessels from the surrounding environment. The objective of this study was to evaluate the effect of a dual-layer, chorion-free amnion patch (DLAM; ViaShield®, Globus Medical Inc., Audubon, PA, USA) processed from HAM as a protective barrier following lumbar laminectomy in a sheep model. A multiplex immunoassay was performed to quantify the inherent cytokines present in the amnion after processing. Twelve skeletally mature female crossbred Suffolk sheep were randomly divided into two equal post-operative periods (4 and 10 weeks). Each sheep underwent a laminectomy at L3 and L5, and one of the surgical sites randomly received the DLAM treatment. At each postsurgical time point, the extent of epidural fibrosis and neurohistopathological responses at the laminectomy sites was assessed based on epidural fibrosis-dura tenacity scores and decalcified histology, respectively. Immunoassay results showed that inflammatory mediators and immunomodulatory cytokines were present in the amnion after processing, but no proangiogenic cytokines were detected. At 10 weeks, tissue tenacity was significantly less in the DLAM treatment group when compared with the operative control (1.2 ± 0.4 vs. 2.8 ± 0.4, p < 0.05), demonstrating the ability of DLAM to act as a barrier and cover the dura. Gross observations showed fewer fibroblasts in the DLAM group in comparison with the control at both post-operative time points. Fibroblast infiltration analysis indicated that at both 4 and 10 weeks, there were significantly more infiltrated fibroblasts in the operative control sites than in the DLAM-treated sites, expressed as a percentage of the total number of fibroblasts present (4 weeks: 72.3 ± 10.2% vs. 10.8 ± 10.1%, p < .05; 10 weeks: 84.9 ± 15.8% vs. 43.1 ± 11.6%, p < .05). Additionally, fibroblasts travelled further into the dura in the operative control group compared with the DLAM-treated group at both time points. In conclusion, this study found that DLAM reduced fibroblast infiltration and tissue tenacity following lumbar laminectomy in a sheep animal model. These findings support the potential use of DLAM in clinical practice as a protective barrier for neural elements and anterior vessels.

DDC plays vital roles in the wing spot formation, egg production, and chorion tanning in the brown planthopper.

The gene dopa decarboxylase (Nlddc) of the brown planthopper (BPH, Nilaparvata lugens) was identified in the genome and transcriptome of the insect. The open reading frame of Nlddc is 1,434 bp in length and, it has the potential to encode a protein of 477 amino acid with a conserved pyridoxal-dependent decarboxylase domain and a typical motif, NFNPHKW. Real-time quantification polymerase chain reaction analyses revealed that this gene was highly expressed in the integument and brain, and transcript level peaked in the late stages of egg period and each nymph instar with periodicity. RNA interference results revealed that Nlddc played essential roles in pigment synthesis, formation of wing spot, egg production, and tanning of the chorion. A rapid accumulation of Nlddc transcripts was detected, and it coincided with the injection of the hormone 20-hydroxyecdysone (20E), suggesting that Nlddc transcription was regulated by 20E.

The Effect of Chorionicity on Maternal Cardiac Adaptation to Uncomplicated Twin Pregnancy: A Prospective Longitudinal Study.

OBJECTIVE: The objective of this study was to longitudinally evaluate maternal echocardiographic findings in uncomplicated twin gestations according to chorionicity. METHODS: Healthy women with twin pregnancy were assessed with transthoracic echocardiography across the first, second, and third trimesters. Cardiac findings were compared within each group and between monochorionic (MC) and dicho-rionic (DC) pregnancies. RESULTS: Overall, 19 MC and 48 DC uncomplicated twin pregnancies were included. In the MC group, no significant maternal haemodynamic changes were documented across gestation, with the exception of a decrease in ejection fraction. Compared to DC pregnancies, in the MC set lower cardiac output (second and third trimester, p = 0.001 and p = 0.006, respectively) and higher total vascular resistance (first trimester, p = 0.032) were observed. Regarding the diastolic function in MC twins, significantly higher values were observed for mitral E/A ratio (third trimester, p = 0.014), septal mitral E1/A1 ratio (third trimester, p = 0.030), lateral mitral E1 (second and third trimester, p = 0.014 and p = 0.029, respectively), and E1/A1 ratio (third trimester, p = 0.006). CONCLUSIONS: Maternal cardiac adaptation in twin pregnancy seems to differ significantly according to chorionicity. In particular, in MC pregnancies the impairment of diastolic function is less pronounced, presumably due to the lower circulating volume.

Postnatal condition of the second twin in respect to mode of delivery, chorionicity and type of fetal growth.

BACKGROUND: The increased use of assisted reproductive techniques (ART) contributes to the increased rate of twin pregnancies, which are burdened with a higher risk of complications. Factors that affect the condition of the second twin are understudied. OBJECTIVES: The objective of this study was to assess the impact of the delivery mode, chorionicity, amnionicity, and the type of fetal growth on the postnatal condition of the second twin. MATERIAL AND METHODS: The study included data from 475 pregnant women with twin pregnancies. Maternal age, parity, chorionicity, amnionicity, type of fetal growth, mode of delivery, gestational weeks at delivery, Apgar score, and umbilical arterial blood pH were retrospectively analyzed. Data normality was checked with the Kolmogorov-Smirnov test. The paired Wilcoxon signed-rank test and χ2 test were used for comparisons between groups. To check predictive value of the analyzed variables multiple linear regression was used. RESULTS: The mean maternal age was 29.22 (standard deviation (SD) ±5.19) years. The maternal age and gestational age at delivery did not differ significantly between women who delivered by cesarean section (CS) and vaginal delivery (VD). In the second twin, the Apgar score and values of arterial umbilical blood pH were lower in infants delivered by VD than in those delivered by CS (6.30 ±2.83 and 7.30 ±0.12; p = 0.0209 and 7.26 ±0.12 and 7.30 ±0.11; p = 0.0236, respectively). In monochorionic diamniotic twins with asymmetric growth, the second twin achieved significantly lower outcome than the first twin. Vaginal delivery was a predictive factor for a lower Apgar score and lower values of umbilical arterial blood pH in second twins, while not in first twins. Symmetrical fetal growth of twins was a predictive factor for better postnatal condition for both twins. CONCLUSIONS: In twin pregnancies, VD, but not CS, is associated with increased risk of worse postnatal condition of the second twin. In monochorionic diamniotic pregnancies complicated by growth discordance, CS seems to be a reasonable mode of delivery.

Short-term outcome of the second twin during vaginal delivery is dependent on delivery time interval but not chorionicity.

The decrease in pH in monochorionic (MC) second twin may exceed that of dichorionic (DC) second twin during vaginal twin delivery. The aim of this study was to investigate the effect of chorionicity on the umbilical artery pH of the second twin in relation to increased twin-to-twin delivery time interval (DTI) from 30 weeks of gestation. A hospital based cohort of all twin deliveries after 30 completed weeks of gestation with the first twin delivered vaginally between 2007 and 2012 in four delivery units in Stockholm county was conducted. The changes of umbilical pH in MC twins compared to DC twins in relation to twin-to-twin DTI was studied using linear regression. The predictors for acidosis (pH ≤ 7.10) in the second twin were studied by logistic regression. The cohort included 681 DC and 186 MC twin pairs. There was a correlation between the umbilical pH and the twin-to-twin DTI but independent of the chorionicity, the pH decreased in average by 0.0012 units/min (p < .001). Twin-to twin DTI was a predictor for acidosis of the second twin (p < .01). IMPACT STATEMENT What is already known on this subject? Vaginal twin deliveries are a major obstetrical challenge because of the increased risk for the second twin after the delivery of the first twin, and are suggested to increase among premature second twins. Previous studies report a continuous decline in the pH of the second twin during the second stage of labour, with an increased risk for acidosis (pH < 7.10) and Apgar scores <7 for twin-to-twin delivery time interval (DTI) as short as 15-30 min. Major studies have not discriminated between MC and DC vaginally delivered twins and the question rise if there is a reason for different delivery guidelines for MC diamniotic second twin compared to DC second twin. What do the results of this study add? The results of the present study show a correlation between umbilical pH and twin-to-twin DTI, but independent of chorionicity, from gestational week 30 to term. A main predictor for acidosis (pH ≤ 7.10) is the twin-to-twin DTI. What are the implications of these findings for clinical practice and/or future research? In cases of uneventful MC pregnancies, we find no support to suggest different vaginal delivery guidelines for MC diamniotic twins compared to DC twins. The risk for acidosis in the second twin increased with 1%/min during the second stage. Active management may be recommended after 15-30 min.

Versican V1 in human endometrial epithelial cells promotes BeWo spheroid adhesion in vitro.

The endometrium extracellular matrix (ECM) is essential for embryo implantation. Versican, a large chondroitin sulfate proteoglycan that binds hyaluronan and forms large ECM aggregates, can influence fundamental physiological phenomena, such as cell proliferation, adhesion and migration. The present study investigated the possible role of versican in human embryo implantation. Versican V1 expression and secretion in human endometrial epithelial cells (EECs) was most prominent in the mid-secretory phase. Versican expression in EECs significantly increased after treatment with estrogen and progesterone, but not by estrogen alone. We also established versican V1-overexpressing Ishikawa (endometrial cancer cell line) cells (ISKW-V1), versican V3-overexpressing (ISKW-V3) and control GFP-overexpressing (ISKW-GFP) Ishikawa cells. By the in vitro implantation model, the attachment ratio of BeWo (choriocarcinoma cell line) spheroids to the monolayer of ISKW-V1, but not of ISKW-V3, was found significantly enhanced compared with attachment to the ISKW-GFP monolayer. The conditioned medium derived from ISKW-V1 (V1-CM) also promoted the attachment of BeWo spheroids to the ISKW monolayer. However, this attachment-promoting effect was abolished when V1-CM was pretreated with chondroitinase ABC, which degrades chondroitin sulfate. Therefore, out of the ECM components, versican V1 may facilitate human embryo implantation.

Differences in the Expression of TLR-2, NOD2, and NF-κB in Placenta Between Twins.

Dizygotic twins share the same type of genetic relationship as non-twin siblings. Whereas monozygotic (MZ) twins are considered to have identical genetic material, they still differ. There is a number of reasons for early MZ twin discordance, including differences in the in utero environment, stochasticity, genetic mosaicism, and epigenetic factors. During gestation, the efficient innate immune system is of utmost importance. Our study was based on immunohistochemical evaluation of the differences in innate immune protein expression (TLR-2, NOD2, and NF-κB) in the 95 placentas between twins. Our study revealed statistical significant differences between diamniotic-dichorionic and monoamniotic-dichorionic twins. Monoamniotic-monochorionic twins exhibited no significant differences in protein expressions. To identify epigenetic factors causing the differences between twins, we made a series of comparisons with clinical data. The study revealed more cases with infections, miscarriages, in vitro fertilization, and premature rupture of membranes within the group with higher differences level of NF-κB, NOD2 and TLR-2 between twins. In case of twin-to-twin transfusion syndrome, there were no significant differences in innate immune protein expressions between twins. These results show that dissimilar genetic material and separate in utero environment promote discordance in innate immune protein expressions between twins. Moreover, additional blood flow between twins may be favorable in life-threatening conditions ensuring similar microenvironment.

The role of indoleamine-2,3-dioxygenase in normal and pathological pregnancies.

The survival of allogeneic fetus during pregnancy contradicts the laws of immune responses. Behind this paradoxical phenomenon, the mechanism is quite complex. Indoleamine-2,3-dioxygenase (IDO) is the first and rate-limiting enzyme of tryptophan catabolism. Emerging evidence shows that IDO is expressed at the maternal-fetal interface, including trophoblast cells, decidual stroma cells, decidual immune cells (eg, natural killer cells, T cells, and macrophages), and vascular endothelial cells of decidua and chorion. Moreover, the expression and activity of IDO are different among non-pregnant, normal pregnant, and pathological pregnant conditions. IDO plays important roles in normal pregnancy through immune suppression and regulation of fetal invasion and circulation. However, the abnormal expression and dysfunction of IDO are associated with some pathological pregnancies (including recurrent spontaneous abortion, preeclampsia, preterm labor, and fetal growth restriction).

Placental weight and birth weight to placental weight ratio in monochorionic and dichorionic growth-restricted and non-growth-restricted twins.

OBJECTIVE:: The aim of the present study was to compare the placental weight and birth weight/placental weight ratio for intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins. METHODS:: This was a retrospective analysis of placentas from twin pregnancies. Placental weight and the birth weight/placental weight ratio were compared in intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins. The association between cord insertion type and placental lesions in intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins was also investigated. RESULTS:: A total of 105 monochorionic (intrauterine growth restriction=40; non-intrauterine growth restriction=65) and 219 dichorionic (intrauterine growth restriction=57; non-intrauterine growth restriction=162) placentas were analyzed. A significantly lower placental weight was observed in intrauterine growth-restricted monochorionic (p=0.022) and dichorionic (p<0.001) twins compared to non-intrauterine growth-restricted twins. There was no difference in the birth weight/placental weight ratio between the intrauterine growth restriction and non-intrauterine growth restriction groups for either monochorionic (p=0.36) or dichorionic (p=0.68) twins. Placental weight and the birth weight/placental weight ratio were not associated with cord insertion type or with placental lesions. CONCLUSION:: Low placental weight, and consequently reduced functional mass, appears to be involved in fetal growth restriction in monochorionic and dichorionic twins. The mechanism by which low placental weight influences the birth weight/placental weight ratio in intrauterine growth-restricted monochorionic and dichorionic twins needs to be determined in larger prospective studies.

Placental weight and birth weight to placental weight ratio in monochorionic and dichorionic growth-restricted and non-growth-restricted twins

OBJECTIVE: The aim of the present study was to compare the placental weight and birth weight/placental weight ratio for intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins. METHODS: This was a retrospective analysis of placentas from twin pregnancies. Placental weight and the birth weight/placental weight ratio were compared in intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins. The association between cord insertion type and placental lesions in intrauterine growth-restricted and non-intrauterine growth-restricted monochorionic and dichorionic twins was also investigated. RESULTS: A total of 105 monochorionic (intrauterine growth restriction=40; non-intrauterine growth restriction=65) and 219 dichorionic (intrauterine growth restriction=57; non-intrauterine growth restriction=162) placentas were analyzed. A significantly lower placental weight was observed in intrauterine growth-restricted monochorionic (p=0.022) and dichorionic (p<0.001) twins compared to non-intrauterine growth-restricted twins. There was no difference in the birth weight/placental weight ratio between the intrauterine growth restriction and non-intrauterine growth restriction groups for either monochorionic (p=0.36) or dichorionic (p=0.68) twins. Placental weight and the birth weight/placental weight ratio were not associated with cord insertion type or with placental lesions. CONCLUSION: Low placental weight, and consequently reduced functional mass, appears to be involved in fetal growth restriction in monochorionic and dichorionic twins. The mechanism by which low placental weight influences the birth weight/placental weight ratio in intrauterine growth-restricted monochorionic and dichorionic twins needs to be determined in larger prospective studies.

An angiogenesis platform using a cubic artificial eggshell with patterned blood vessels on chicken chorioallantoic membrane.

The chorioallantoic membrane (CAM) containing tiny blood vessels is an alternative to large animals for studies involving angiogenesis and tissue engineering. However, there is no technique to design the direction of growing blood vessels on the CAM at the microscale level for tissue engineering experiments. Here, a methodology is provided to direct blood vessel formation on the surface of a three-dimensional egg yolk using a cubic artificial eggshell with six functionalized membranes. A structure on the lateral side of the eggshell containing a straight channel and an interlinked chamber was designed, and the direction and formation area of blood vessels with blood flow was artfully defined by channels with widths of 70-2000 µm, without sharply reducing embryo viability. The relationship between the size of interlinked chamber and the induction of blood vessels was investigated to establish a theory of design. Role of negative and positive pressure in the induction of CAM with blood vessels was investigated, and air pressure change in the culture chamber was measured to demonstrate the mechanism for blood vessel induction. Histological evaluation showed that components of CAM including chorionic membrane and blood vessels were induced into the channels. Based on our design theory, blood vessels were induced into arrayed channels, and channel-specific injection and screening were realized, which demonstrated proposed applications. The platform with position- and space-controlled blood vessels is therefore a powerful tool for biomedical research, which may afford exciting applications in studies involved in local stimulation of blood vessel networks and those necessary to establish a living system with blood flow from a beating heart.

Understanding the Impact of Preservation Methods on the Integrity and Functionality of Placental Allografts.

INTRODUCTION: Human placental membranes (hPMs) have a long history in treating burns and wounds. The composition of hPMs includes structural matrix, growth factors, and neonatal cells, all of which contribute to their regenerative potential. However, most hPM products are devitalized after dehydration and irradiation. We compared the functionality of single-layer viable cryopreserved human amniotic membrane (vCHAM) with multilayer devitalized dehydrated human amnion/chorion membrane (dHACM) in wound-relevant models to determine the effect of different processing methods on hPMs. METHODS: Viable cryopreserved human amniotic membrane and dHACM were compared with fresh hPM for structural integrity and viability. Viable cell persistence in vCHAM over time was evaluated in vitro and in vivo in a diabetic chronic wound mouse model. Proliferation of cells within fresh hPM and vCHAM was evaluated with bromodeoxyuridine and Ki-67 staining, and proliferation of isolated cells in culture was evaluated. Growth factor release over time and in vitro response to chronic wound stimuli (tumor necrosis factor α, lipopolysaccharide, and hypoxia) were used to compare the functionality of vCHAM and dHACM. RESULTS: The structure and thickness of fresh hPM were retained in vCHAM but were compromised in dHACM. Similar to fresh hPM, vCHAM contained viable cells, whereas dHACM did not. Cells in vCHAM remained viable after 4 and 7 days in culture and in an in vitro chronic wound environment and after 4 and 8 days in vivo after application to a mouse chronic wound. Staining for bromodeoxyuridine and Ki-67 did not reveal proliferative cells within fresh hPM and vCHAM. However, isolated cells proliferated in culture. Viable cryopreserved human amniotic membrane increased platelet-derived growth factor BB, hepatocyte growth factor, and epidermal growth factor levels over time and responded to chronic wound stimuli in vitro by significantly increasing levels of vascular endothelial growth factor and prostaglandin E2. Dehydrated human amnion/chorion membrane showed no significant accumulation of growth factors and did not respond to chronic wound stimuli. CONCLUSIONS: These results indicate that vCHAM retains intact, native matrix, and viable, active cells and responds to chronic wound stimuli in vitro. The inclusion of multiple layers of hPM does not compensate for structural degradation and loss of viability caused by dehydration as evidenced by a lack of functional response by dHACM. The clinical significance of these results remains to be answered.

Twin chorionicity and prospective stillbirth risk: experience at a tertiary care hospital.

OBJECTIVE: To determine the prospective risk of stillbirth, perinatal death and neonatal morbidities in twins. METHODS: This retrospective cohort study was conducted at the Aga Khan University Hospital, Karachi, and comprised cases of twin pregnancies from January 2001 to December 2012. . Cases of both monochorionic diamniotic and dichorionic diamniotic twin pregnancies were included. SPSS 19 was used for data analysis.. RESULTS: Of the 394 cases, 84(21.3%) were monochorionic diamniotic twins and 310(78.7%) were dichorionic diamniotic twins. There were no cases of stillbirth beyond 30 and 34 weeks in monochorionic and dichorionic twins, respectively. Neonatal mortality stood reduced beyond 36 weeks of gestation in both groups. Neonatal morbidity reduced beyond 35 weeks of gestation in both groups.. CONCLUSIONS: Perinatal mortality and morbidity in twin gestation decreased as the gestation advanced.