Search

Introduction: Opioid use disorder (OUD) is implicated in major obstetrical diseases such as fetal growth restriction. Whether or not opioids directly impact placental trophoblast development and function remains unclear. We sought to examine the expression of opioid receptors (OPRs) in villous trophoblasts and the effect of opioids on placental transcriptomics., Methods: Trophoblast stem (TS) cells and primary human trophoblast (PHT) cells from healthy term placentas were used to assess OPR expression in conditions that enhance trophoblast stemness vs differentiation. Placental RNAseq was conducted using our retrospective cohorts of pregnant people with OUD vs controls, both without major obstetrical complications. RT-qPCR was used to determine the effect of fentanyl on the expression of putative opioid targets and stemness or differentiation-associated genes in TS and PHT cells., Results: Three main OPRs, including OPRM1, OPRD1, and OPRK1 were expressed in term PHT cells cultured in the stemness medium, whereas only OPRD1 and OPRK1 were expressed in TS cells. Interestingly, upon induction of differentiation, the expressed OPR mRNAs in TS or in PHT cells were downregulated. We found 286 differentially expressed long RNAs in placentas from the OUD participants vs controls. While three putative opioid targets differed their expression in stemness vs differentiation states of trophoblasts, fentanyl had no effect on their expression or the expression of major stemness or differentiation-relevant genes in TS and PHT cells., Discussion: Trophoblastic expression of OPRs and opioid RNA targets is impacted by cell differentiation, suggesting differential susceptibility of villous trophoblasts to the effect of opioids., Competing Interests: Declaration of competing interest The authors certify that there are no financial interests that influence the results and findings obtained through this study. Yoel Sadovsky is a consultant to Bio-Rad Laboratories, Inc., (Copyright © 2023. Published by Elsevier Ltd.)

Trophoblast injury is central to clinically relevant placenta dysfunction. We hypothesized that the mRNA of primary human trophoblasts, exposed to distinct injuries in vitro, capture transcriptome patterns of placental biopsies obtained from common obstetrical syndromes. We deployed a CIBERSORTx deconvolution method to correlate trophoblastic RNAseq-based expression matrices with the transcriptome of omics-defined placental dysfunction patterns in vivo. We found distinct trophoblast injury patterns in placental biopsies from women with fetal growth restriction and a hypertensive disorder, or in biopsies clustered by their omics analysis. Our RNAseq data are useful for defining the contribution of trophoblast injuries to placental dysfunction syndromes., Competing Interests: Declaration of competing interest The authors certify that there are no financial interests that influence the results and findings obtained through this study. As stated on the title page, Yoel Sadovsky is a consultant to Bio-Rad Laboratories, Inc., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Background: Placental dysfunction, a root cause of common syndromes affecting human pregnancy, such as preeclampsia (PE), fetal growth restriction (FGR), and spontaneous preterm delivery (sPTD), remains poorly defined. These common, yet clinically disparate obstetrical syndromes share similar placental histopathologic patterns, while individuals within each syndrome present distinct molecular changes, challenging our understanding and hindering our ability to prevent and treat these syndromes., Methods: Using our extensive biobank, we identified women with severe PE (n = 75), FGR (n = 40), FGR with a hypertensive disorder (FGR + HDP; n = 33), sPTD (n = 72), and two uncomplicated control groups, term (n = 113), and preterm without PE, FGR, or sPTD (n = 16). We used placental biopsies for transcriptomics, proteomics, metabolomics data, and histological evaluation. After conventional pairwise comparison, we deployed an unbiased, AI-based similarity network fusion (SNF) to integrate the datatypes and identify omics-defined placental clusters. We used Bayesian model selection to compare the association between the histopathological features and disease conditions vs SNF clusters., Results: Pairwise, disease-based comparisons exhibited relatively few differences, likely reflecting the heterogeneity of the clinical syndromes. Therefore, we deployed the unbiased, omics-based SNF method. Our analysis resulted in four distinct clusters, which were mostly dominated by a specific syndrome. Notably, the cluster dominated by early-onset PE exhibited strong placental dysfunction patterns, with weaker injury patterns in the cluster dominated by sPTD. The SNF-defined clusters exhibited better correlation with the histopathology than the predefined disease groups., Conclusions: Our results demonstrate that integrated omics-based SNF distinctively reclassifies placental dysfunction patterns underlying the common obstetrical syndromes, improves our understanding of the pathological processes, and could promote a search for more personalized interventions., (© 2023. BioMed Central Ltd., part of Springer Nature.)

In eutherians, the placenta plays a critical role in the uptake, storage, and metabolism of lipids. These processes govern the availability of fatty acids to the developing fetus, where inadequate supply has been associated with substandard fetal growth. Whereas lipid droplets are essential for the storage of neutral lipids in the placenta and many other tissues, the processes that regulate placental lipid droplet lipolysis remain largely unknown. To assess the role of triglyceride lipases and their cofactors in determining placental lipid droplet and lipid accumulation, we assessed the role of patatin like phospholipase domain containing 2 (PNPLA2) and comparative gene identification-58 (CGI58) in lipid droplet dynamics in the human and mouse placenta. While both proteins are expressed in the placenta, the absence of CGI58, not PNPLA2, markedly increased placental lipid and lipid droplet accumulation. These changes were reversed upon restoration of CGI58 levels selectively in the CGI58-deficient mouse placenta. Using co-immunoprecipitation, we found that, in addition to PNPLA2, PNPLA9 interacts with CGI58. PNPLA9 was dispensable for lipolysis in the mouse placenta yet contributed to lipolysis in human placental trophoblasts. Our findings establish a crucial role for CGI58 in placental lipid droplet dynamics and, by extension, in nutrient supply to the developing fetus.

The coronavirus disease 2019 (COVID-19) pandemic affected people at all ages. Whereas pregnant women seemed to have a worse course of disease than age-matched non-pregnant women, the risk of feto-placental infection is low. Using a cohort of 66 COVID-19-positive women in late pregnancy, we correlated clinical parameters with disease severity, placental histopathology, and the expression of viral entry and Interferon-induced transmembrane (IFITM) antiviral transcripts. All newborns were negative for SARS-CoV-2. None of the demographic parameters or placental histopathological characteristics were associated with disease severity. The fetal-maternal transfer ratio for IgG against the N or S viral proteins was commonly less than one, as recently reported. We found that the expression level of placental ACE2, but not TMPRSS2 or Furin, was higher in women with severe COVID-19. Placental expression of IFITM1 and IFITM3, which have been implicated in antiviral response, was higher in participants with severe disease. We also showed that IFITM3 protein expression, which localized to early and late endosomes, was enhanced in severe COVID-19. Our data suggest an association between disease severity and placental SARS-CoV-2 processing and antiviral pathways, implying a role for these proteins in placental response to SARS-CoV-2., (© 2021. The Author(s).)

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has had a massive impact on human lives worldwide. While the airborne SARS-CoV-2 primarily affects the lungs, viremia is not uncommon. As placental trophoblasts are directly bathed in maternal blood, they are vulnerable to SARS-CoV-2. Intriguingly, the human fetus is largely spared from SARS-CoV-2 infection. We tested whether the human placenta expresses the main SARS-CoV-2 entry factors angiotensin-converting enzyme 2 (ACE2), transmembrane protease serine 2 (TMPRSS2), and furin and showed that ACE2 and TMPRSS2 are expressed in the trophoblast rather than in other placental villous cells. While furin is expressed in the main placental villous cell types, we surveyed, trophoblasts exhibit the highest expression. In line with the expression of these entry factors, we demonstrated that a SARS-CoV-2 pseudovirus could enter primary human trophoblasts. Mechanisms underlying placental defense against SARS-CoV-2 infection likely involve postentry processing, which may be germane for mitigating interventions against SARS-CoV-2. IMPORTANCE Pregnant women worldwide have been affected by COVID-19. As the virus is commonly spread to various organs via the bloodstream and because human placental trophoblasts are directly bathed in maternal blood, feto-placental infection by SARS-CoV-2 seems likely. However, despite the heightened risk to pregnant women, thus far the transmission risk of COVID-19 to the feto-placental unit seems extremely low. This has been recently attributed to a negligible expression of SARS-CoV-2 entry factors in the human placenta. We therefore sought to explore the expression of the entry factors ACE2 and TMPRSS2 in the different cell types of human placental villi. Using a combination of transcriptome sequencing (RNA-seq), real-time quantitative PCR (RT-qPCR), in situ hybridization, and immunofluorescence, we found that trophoblasts, but not the other main villous cell types, express ACE2 and TMPRSS2, with a broad expression of furin. Correspondingly, we also showed that primary human trophoblasts are permissive to entry of SARS-CoV-2 pseudovirus particles., (Copyright © 2021 Ouyang et al.)

Introduction: The chromosome 19 miRNA cluster (C19MC) encodes a large family of microRNAs (miRNAs) that are abundantly expressed in the placenta of higher primates and also in certain cancers. In the placenta, miRNAs from this cluster account for nearly 40% of all miRNAs present in trophoblasts. However, the function of these miRNAs in the placenta remains poorly understood. Recent observations reveal a role for these miRNAs in cell migration, and suggest that they are involved in the development and function of the human placenta. Here, we examine the placenta in transgenic mice expressing the human C19MC miRNAs., Methods: We produced transgenic mice using pronuclear microinjection of a bacterial artificial chromosome plasmid carrying the entire human C19MC locus and derived a homozygous line using crossbreeding. We performed morphological characterization and profiled gene expression changes in the placentas of the transgenic mice., Results: C19MC transgenic mice delivered on time with no gross malformations. The placentas of transgenic mice expressed C19MC miRNAs and were larger than wild type placentas. Histologically, we found that the transgenic placenta exhibited projections of spongiotrophoblasts that penetrated deep into the labyrinth. Gene expression analysis revealed alterations in the expression of several genes involved in cell migration, with evidence of enhanced cell proliferation., Discussion: Mice that were humanized for transgenically overexpressed C19MC miRNAs exhibit enlarged placentas with aberrant delineation of cell layers. The observed phenotype and the related gene expression changes suggest disrupted migration of placental cell subpopulations., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Pregnancy is a unique situation, in which placenta-derived small extracellular vesicles (sEVs) may communicate with maternal and foetal tissues. While relevant to homoeostatic and pathological functions, the mechanisms underlying sEV entry and cargo handling in target cells remain largely unknown. Using fluorescently or luminescently labelled sEVs, derived from primary human placental trophoblasts or from a placental cell line, we interrogated the endocytic pathways used by these sEVs to enter relevant target cells, including the neighbouring primary placental fibroblasts and human uterine microvascular endothelial cells. We found that trophoblastic sEVs can enter target cells, where they retain biological activity. Importantly, using a broad series of pharmacological inhibitors and siRNA-dependent silencing approaches, we showed that trophoblastic sEVs enter target cells using macropinocytosis and clathrin-mediated endocytosis pathways, but not caveolin-dependent endocytosis. Tracking their intracellular course, we localized the sEVs to early endosomes, late endosomes, and lysosomes. Finally, we used coimmunoprecipitation to demonstrate the association of the sEV microRNA (miRNA) with the P-body proteins AGO2 and GW182. Together, our data systematically detail endocytic pathways used by placental sEVs to enter relevant fibroblastic and endothelial target cells, and provide support for "endocytic escape" of sEV miRNA to P-bodies, a key site for cytoplasmic RNA regulation., (© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles.)

Although preeclampsia is a common and serious complication of pregnancy, insight into its pathobiology and diagnosis is lacking. Circulating plasma exosomes, which contain RNA and other molecules and have recently become accessible for diagnostics, may be informative in this regard. We tested the hypothesis that preeclampsia may affect the miRNA cargo within circulating maternal blood exosomes. We collected plasma from 60 pregnant women at term, including 20 women with pregnancy complicated by preeclampsia, and 20 women with fetal growth restriction and 20 with healthy pregnancy, serving as controls. We isolated exosomes from the maternal plasma by continuous density gradient ultracentrifugation. Our main outcome variable was exosomal miRNA cargo, analyzed by quantitative polymerase chain reaction-based TaqMan advanced miRNA assay in a card format and the expression of differentially expressed exosomal miRNA in whole plasma from the same participants. We found that 7 miRNA species were differentially expressed in exosomes from women with preeclampsia and those from controls. In contrast, there was no significant difference in exosomal miRNA expression between women with fetal growth restriction and controls. The results were not affected by fetal sex. Only one of the preeclampsia-related, differentially expressed exosomal miRNAs was significantly different in whole plasma miRNA analysis. We concluded that unlike whole plasma miRNA, exosomes extracted from the plasma of women with preeclampsia exhibit a unique miRNA profile, suggesting that plasma exosomal miRNA could provide insight into the pathophysiology of preeclampsia, and may play a role in disease diagnostics.

Introduction: Imprinted genes are preferentially expressed from one parentally inherited allele, and many are crucial to the regulation of placental function and fetal growth. Murine Krüppel-like factor 14 (Klf14) is a maternally expressed imprinted transcription factor that is a component of the Mest imprinted gene cluster on mouse chromosome 6. We sought to determine if loss of Klf14 expression alters the course of normal mouse extraembryonic development. We also used high-throughput RNA sequencing (RNAseq) to identify a set of differentially expressed genes (DEGs) in placentas with loss of Klf14., Methods: We generated a Klf14 knockout (Klf14 null ) mouse using recombineering and transgenic approaches. To identify DEGs in the mouse placenta we compared mRNA transcriptomes derived from 17.5dpc Klf14 matKO and wild-type littermate placentas by RNAseq. Candidate DEGs were confirmed with quantitative reverse transcription PCR (qPCR) on an independent cohort of male and female gestational age matched Klf14 matKO placentas., Results: We found that 17.5dpc placentas inheriting a maternal null allele (Klf14 matKO ) had a modest overgrowth phenotype and a near complete ablation of Klf14 expression. However, there was no effect on fetal growth. We identified 20 DEGs differentially expressed in Klf14 matKO placentas by RNAseq, and subsequently validated five that are highly upregulated (Begain, Col26a1, Fbln5, Gdf10, and Nell1) by qPCR. The most enriched functional gene-networks included those classified as regulating cellular development and metabolism., Conclusion: These results suggest that loss of the maternal Klf14 locus in the mouse placenta acts results in changes in gene expression patterns that modulate placental growth., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Introduction: Cultured primary human trophoblasts (PHT), derived from term placentas, are relatively resistant to infection by diverse viruses. The resistance can be conferred to non-trophoblastic cells by pre-exposing them to medium that was conditioned by PHT cells. This antiviral effect is mediated, at least in part, by microRNAs (miRNA) expressed from the chromosome 19 microRNA cluster (C19MC). Recently we showed that PHT cells and cells pre-exposed to PHT medium are also resistant to infection by Zika virus (ZIKV), an effect mediated by the constitutive release of the type III interferons (IFN) IFN lambda-1 and IFN lambda-2 in trophoblastic medium. We hypothesized that trophoblastic C19MC miRNA are active against ZIKV, and assessed the interaction of this pathway with IFN lambda-1 - mediated resistance., Methods: Term PHT cells were cultured using standard techniques. An osteosarcoma cell line (U2OS) was used as non-trophoblastic cells, which were infected with either ZIKV or vesicular stomatitis virus (VSV). Trophoblastic extracellular vesicles (EVs) were produced by gradient ultracentrifugation. RT-qPCR was used to determine viral infection, cellular or medium miRNA levels and the expression of interferon-stimulated genes., Results: We showed that C19MC miRNA attenuate infection of U2OS cells by ZIKV, and that C19MC miRNA or exosomes that contain C19MC miRNA did not influence the type III IFN pathway. Similarly, cell exposure to recombinant IFN lambda-1 had no effect on miRNA expression, and these pathways did not exhibit synergistic interaction., Discussion: PHT cells exert antiviral activity by at least two independent mechanisms, mediated by C19MC miRNA and by type III IFNs., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

During pregnancy, placental trophoblasts at the feto-maternal interface produce a broad repertoire of microRNA (miRNA) species. These species include miRNA from the primate-specific chromosome 19 miRNA cluster (C19MC), which is expressed nearly exclusively in the placenta. Trafficking of these miRNAs among the maternal, placental, and fetal compartments is unknown. To determine miRNA expression and trafficking patterns during pregnancy, we sequenced miRNAs in triads of human placenta and of maternal and fetal blood and found large subject-to-subject variability, with C19MC exhibiting compartment-specific expression. We therefore created humanized mice that transgenically express the entire 160-kb human C19MC locus or lentivirally express C19MC miRNA members selectively in the placenta. C19MC transgenic mice expressed a low level of C19MC miRNAs in diverse organs. When pregnant, female C19MC mice exhibited a strikingly elevated (>40-fold) expression of C19MC miRNA in the placenta, compared with other organs, that resembled C19MC miRNAs patterns in humans. Our mouse models showed that placental miRNA traffic primarily to the maternal circulation and that maternal miRNA can traffic to the placenta and even into the fetal compartment. These findings define an extraordinary means of nonhormonal, miRNA-based communication between the placenta and feto-maternal compartments.-Chang, G., Mouillet, J.-F., Mishima, T., Chu, T., Sadovsky, E., Coyne, C. B., Parks, W. T., Surti, U., Sadovsky, Y. Expression and trafficking of placental microRNAs at the feto-maternal interface., (© FASEB.)

Manipulation of microRNA (miRNA) levels, including overexpression of mature species, has become an important biological tool, even motivating miRNA-based therapeutics. To assess key determinants of miRNA overexpression in a mammalian system in vivo, we sought to bypass the laborious generation of a transgenic animal by exploiting placental trophoblast-specific gene manipulation using lentiviral vectors, which has been instrumental in elucidating trophoblast biology. We examined the impact of several key components of miRNA stem loops and their flanking sequences on the efficiency of mature miRNA expression in vivo. By combining established and novel approaches for miRNA expression, we engineered lentivirus-driven miRNA expression plasmids, which we tested in the mouse placenta. We found that reverse sense inserts minimized single-strand splicing and degradation, and that maintaining longer, poly-A-containing arms flanking the miRNA stem-loop markedly enhanced transgenic miRNA expression. Additionally, we accomplished overexpression of diverse mammalian, drosophila, or C. elegans miRNAs, either based on native context or using a "cassette" replacement of the mature miRNA sequence. Together, we have identified primary miRNA sequences that are paramount for effective expression of mature miRNAs, and validated their role in mice. Principles established by our findings may guide the design of efficient miRNA vectors for in vivo use.

Early in pregnancy, trophoblast invasion into the decidua and inner myometrium is essential for establishment of proper implantation, maternal-fetal exchange, and immunological tolerance of the feto-placental allograft. Unlike villous trophoblasts (VTs), extravillous trophoblasts (EVTs) are unique in their capacity to invade the maternal decidua and myometrium. The largest human microRNA (miRNA) gene cluster, the chromosome 19 miRNA cluster (C19MC), is expressed almost exclusively in the placenta and, rarely, in certain tumors and undifferentiated cells. In the work reported here, we found that the expression of C19MC miRNAs is higher in VTs than in EVTs. Using a bacterial artificial chromosome (BAC)-mediated overexpression of C19MC miRNAs in an EVT-derived cell line, which does not naturally express these miRNAs, we found that C19MC miRNAs selectively attenuate cell migration without affecting cell proliferation or apoptosis. A microarray analysis revealed that C19MC miRNAs regulate target transcripts related to cellular movement. Our data also implicated a specific C19MC member, miR-519d, indirectly regulating the EVT invasive phenotype by targeting CXCL6, NR4A2 and FOXL2 transcripts through a 3'UTR miRNA-responsive element. Together, our data suggest a role for C19MC miRNAs in modulating the migration of EVTs.

We performed comparative sequence analysis of 3 blaKPC-2 encoding plasmids to examine evolution of these plasmids and their dissemination. We found that all of them have an IncN replicon with a newly determined IncN plasmid sequence type (ST), ST15. The 2 Klebsiella pneumoniae (KPN) plasmids also harbor an IncF2A1-B1- replicon. The blaKPC-2 is located in the Tn4401c transposon with a newly discovered mutation in the P2 promoter. Screening of the 27 additional blaKPC-2 carrying plasmids from Enterobacter cloacae, Escherichia coli (EC), and K. pneumoniae showed that: all KPN and EC plasmids are IncN plasmids belonging to ST15; 4/7 KPN and 1/6 EC plasmids contain an additional IncF2A1-B1- replicon; all Enterobacter plasmids belong to neither IncN nor IncF2A1-B1- replicon plasmids; 6/7 KPN and 2/5 EC plasmids carry the mutated P2 promoter. Study of the blaKPC-2 environment, transposon, pMLST, and Inc group suggests transposon and plasmid inter- and intra-species dissemination and evolution., (Copyright © 2014 Elsevier Inc. All rights reserved.)

The information flow between distal elements of a protein may rely on allosteric communication trajectories lying along the protein's tertiary or quaternary structure. To unravel the underlying features of energy parsing along allosteric pathways in voltage-gated K(+) channels, high-order thermodynamic coupling analysis was performed. We report that such allosteric trajectories are functionally conserved and delineated by well defined boundaries. Moreover, allosteric trajectories assume a hierarchical organization whereby increasingly stronger layers of cooperative residue interactions act to ensure efficient and cooperative long-range coupling between distal channel regions. Such long-range communication is brought about by a coupling of local and global conformational changes, suggesting that the allosteric trajectory also corresponds to a pathway of physical deformation. Supported by theoretical analyses and analogy to studies analyzing the contribution of long-range residue coupling to protein stability, we propose that such experimentally derived trajectory features are a general property of allosterically regulated proteins.

Gold-tipped CdSe rods (nanodumbbells) were solubilized in an aqueous phase and self-assembled in a head-to-tail manner using biotin disulfide and avidin. The disulfide end of the biotin molecule attaches to the gold tip of the nanodumbbell, and the biotin end of the molecule is able to conjugate to an avidin protein. The avidin can strongly conjugate up to four biotin molecules. Changing the ratios of biotin to nanodumbbells leads to the formation of dimers, trimers, and flowerlike structures. To further improve the distribution of chain lengths, a separation method based upon weight was applied using a concentration gradient. The gold tips provide effective anchor points for constructing complex nanorod structures by self-assembly.

Sum frequency generation (SFG) vibrational spectroscopy and atomic force microscopy (AFM) have been used to study the surface structure and surface mechanical behavior of biologically-relevant polymer systems. These techniques have emerged as powerful surface analytical tools to deduce structure/property relationships in situ, at both air/solid and air/liquid interfaces. SFG and AFM studies have been performed to understand how the surface properties of polymers are linked to polymer bulk compositions, changes in the ambient environment, or the degree of mechanical strain. Specifically, this review discusses (1) the macroscopic- and molecular-level tracking of small end groups attached to polyurethane blends, engineered to reduce blood clotting; (2) the role of ambient humidity on the surface mechanics of soft contact lenses possessing different water content in the bulk; (3) the affect of cyclic stretch on the molecular surface structure of polyurethane films, designed to mimic the mechanical deformation caused by heartbeat; and (4) the molecular ordering of functional groups at the polystyrene-protein interface. The correlation of spectroscopic and mechanical data by SFG and AFM is a powerful methodology to study and design materials with tailored surface properties.

Since the introduction of different ovulation induction regimens the incidence of triplet pregnancies increased from a spontaneous rate of about 1:10,000 to the frequency of nearly 1:1000. Sex ratio of trigemini and male/female proportion of the three newborns within the same triplet set is the amazing issue that we attempted to elucidate in the present study. Data on 36 women with triplet pregnancies delivered in our hospital and information regarding 2717 triplet pregnancies reported by 16 relevant papers were included in this study. 36 women delivered 63 (58%) male and 45 (42%) female babies. Triplet sets of same gender comprised 33% of all trigemini. Sex ratios (male/female) for the spontaneous, menotropin and clomiphene group were 2.00, 1.57 and 0.94 respectively. Homogenous male/female set ratio was 5.0, 2.0 and 0.5 in menotropin, spontaneous and clomiphene groups respectively. As the percentage of ovulation induction triplet pregnancies increased constantly from 0% to 100%, a decline in the fraction of homogenous gender triplet sets was observed--from 73% to 23%. It may be concluded that sex ratio in triplet newborns is divergent according to different publications with most of the authors reporting a male/female ratio of less than 1.00. Ovulation induction results in a lower proportion of homogenous sex triplets sets. Superovulation by menotropins cause a higher percentage of male triplet newborns when compared to ovulation induction by clomiphene.

It is commonly accepted that cesarean section is the preferred mode of delivery of triplets. The present study was performed to evaluate maternal complications and neonatal outcome in 21 women operated on by a transverse lower segment incision as compared to 8 parturients who underwent surgery by a longitudinal uterine incision. There was no significant difference in mean gestational age at delivery, mean weight of the newborns, Apgar scores, perinatal mortality rates, or intraoperative or postpartum hemorrhage incidence between the two groups. Thus, a transverse lower segment uterine incision should be used in triplet deliveries to increase the chances of a normal vaginal delivery in the future.

Sonographic growth curves of 47 normal triplet pregnancies were compared with those of 71 uncomplicated twin gestations. Starting from the 25th week, the biparietal diameter in triplets was found to lag progressively compared with that of twins, reaching a mean difference of 2 weeks at 36 weeks gestation. Similarly, a significant difference was found between the femur length and head to abdomen circumference ratio growth curves of triplets and twins. It is concluded that as pregnancy continues from the 25th to the 36th gestational week, there is a significant delay in the growth patterns of normal triplet pregnancies compared with twin gestations.

Discordancy in twins is generally described as a weight difference between the fetuses of 15 to 25% and its prevalence in normal twin gestations is estimated at 15 to 29%. The present study was performed in order to determine the frequency of discordant newborns in uncomplicated triplet gestations. One hundred and fourteen normal triplet newborns were delivered during a period of 11 years. Their birthweight ranged from 493 to 2533 gm, with a mean of 1894.2 +/- 468.5 gm (+/- sd). In 58% of these 38 sets of triplets the difference in weight between the largest and smallest newborn was more than 15%. About a quarter of the newborns had a greater than 25% dissimilarity in their birthweights. There was no substantial fetal-fetal transfusion or difference in the Apgar scores between the discordant fetuses. It may be concluded that triplet newborns have a rate of discordancy double that of twins. Discordance per se does not necessarily mark an underlying complication of pregnancy or a bad perinatal outcome.

The diagnosis of inappropriate intrauterine fetal growth in triplet pregnancies requires normal standard sonographically determined growth curves. The aim of the present study was to establish such nomograms. The biparietal diameter (BPD), femur length, head and abdominal circumferences (HC, AC) were once in 3 weeks sonographically evaluated in 108 normal triplet fetuses. The resulting growth curves were compared with those of normal singleton fetuses. The data showed that, compared with singletons, the mean triplet fetal BPD progressively lags to a maximum of 2 1/2 weeks as pregnancy continuous from the 25th to the 36th gestational week. Similarly, mean femur length of triplet fetuses gradually shortfalls to the greatest of 2 weeks as gestation advances from the 25th to the 36th week. HC to AC ratio of the triplet fetuses does not differ from the singletons curve. It may be concluded that a normal fetal growth curve in triplet pregnancies demonstrates a 1- to 3-week delay compared with singleton gestations.

A generalized phenomenological model is presented for stereospecific recognition between biological receptors and their ligands. We ask what is the distribution of binding constants psi(K) between an arbitrary ligand and members of a large receptor repertoire, such as immunoglobulins or olfactory receptors. For binding surfaces with B potential subsite and S different types of subsite configurations, the number of successful elementary interactions obeys a binomial distribution. The discrete probability function psi(K) is then derived with assumptions on alpha, the free energy contribution per elementary interaction. The functional form of psi(K) may be universal, although the parameter values could vary for different ligand types. An estimate of the parameter values of psi(K) for iodovanillin, an analog of odorants and immunological haptens, is obtained by equilibrium dialysis experiments with nonimmune antibodies. Based on a simple relationship, predicted by the model, between the size of a receptor repertoire and its average maximal affinity toward an arbitrary ligand, the size of the olfactory receptor repertoire (Nolf) is calculated as 300-1000, in very good agreement with recent molecular biological studies. A very similar estimate, Nolf = 500, is independently derived by relating a theoretical distribution of maxima for psi(K) with published human olfactory threshold variations. The present model also has implications to the question of olfactory coding and to the analysis of specific anosmias, genetic deficits in perceiving particular odorants. More generally, the proposed model provides a better understanding of ligand specificity in biological receptors and could help in understanding their evolution.

Pregnancy during familial Mediterranean fever complicated with amyloidosis and severe nephrotic syndrome is rare and may cause several maternal and fetal complications. Asymmetrical intrauterine growth retardation, superimposed preeclampsia, thromboembolic phenomena, resistant anemia and renal failure only partially represent the possible complications. A successful outcome of a full-term pregnancy is presented and the efficiency of colchicine, a high protein diet, acetylsalicylic acid and dipyridamole is discussed.

The incidence of triplet pregnancies has increased several times due to the widespread use of ovulation induction agents. Premature delivery is the main complication of triplet gestations. In twin conceptions it was shown definitely that elective cervical suture does not prolong these pregnancies. However, in higher order multifetal gestations the uterus is extremely overdistended and the application of cerclage is equivocal. This work intended to assess the value of elective cervical sutures in prolonging triplet pregnancies. Twelve of 35 women with triplet conceptions hospitalized at our high-risk pregnancy unit underwent an elective cerclage at 12 to 14 weeks and 23 women served as control subjects. None of them had any history or physical findings of cervical incompetence prior to the procedure. The gestational age (weeks) at delivery was 33.0 +/- 5.1 and 34.7 +/- 2.8 (mean +/- SD) for the sutured and noncerclage groups, respectively. The birthweight (grams) of the neonates was 1833.0 +/- 524.5 and 1884 +/- 455.5 (mean +/- SD) for the cerclage and nonsutured women, respectively. It seems that triplet gestations as such do not benefit from an elective cervical suture.

Triplet gestations present a considerable management challenge due to a high rate of antenatal, intrapartum and neonatal complications. The introduction of ovulation inducing agents resulted in a marked increase of these pregnancies. Consequently, triplet conceptions are not considered a rare phenomenon and have become almost routine high risk pregnancies. In our department the triplet gestation frequency increased 2-4 times in a 10 year period from a rate of 0.07-0.14% during 1978-84 to 0.28% of all deliveries in 1987. This high incidence of triplet deliveries results in a threefold increase in the frequency of triplet neonates and their associated complications. It is reasonable to assume that the wide application of various assisted reproductive techniques will result in continuous increase in the incidence of triplets which will in turn require coordinated management of a high risk pregnancy unit, delivery room and neonatal intensive care unit.

A case is reported of a severe postpartum maternal hemorrhagic complication, which was related to prophylactic antithrombotic therapy with daily low-dose aspirin (75 mg) combined with dipyridamole (225 mg) for the prevention of preeclampsia. The postpartum course was complicated by recurrent episiotomy site hematomas of nonclotted blood and prolonged bleeding time. Transfusion of platelet concentrates was necessary to control the bleeding. This case report draws attention to maternal hemorrhagic complications which may be associated with prophylactic low-dose aspirin and dipyridamole in pregnancy.

A case report of uterine rupture in labor with epidural anesthesia is presented. The woman had good analgesia on the left side, but complained of severe labor pais on her right side. Uterine rupture occurred which was manifested by sudden vaginal bleeding, fainting, low blood pressure and fetal distress. She did not feel any pains typical of uterine rupture. Rupture of the left uterine wall, with a large hematoma in the left parametrium was seen at surgery. It seems the unilateral anesthesia of the left side concealed the early signs of rupture.

This study was undertaken in order to evaluate pregnancy outcome in three groups of triplet conceptions. Of 36 delivered triplet gestations, seven were spontaneous, 12 achieved using clomiphene citrate and 17 induced by menotropins. The mean gestational age (weeks) at delivery was 36.5 +/- 1.0 SD, for spontaneous and clomiphene citrate pregnancies it was 35.9 +/- 1.9 SD which differed significantly from that in pregnancies induced by menotropins which was 32.9 +/- 3.7 SD. Similarly, triplets born following menotropin treatment had a significantly lower gestational weight (g) of 1656 +/- 516 SD vs. 2015 +/- 198 SD and 2176 +/- 315 SD for spontaneous and clomiphene citrate gestations, respectively. It is concluded that triplets following menotropins are at an increased risk for premature delivery and low birth weight.

Fetal movements (FM) may be spontaneous, originating in the fetus itself, or evoked, caused by external stimuli. The FM in 21 normal twin pregnancies at 10 and 21 weeks of gestation were studied. The percentage rate of evoked FM which occur simultaneously in both twins was 4.96% of all observed FM. The rate of the spontaneous FM which occur independently in each fetus was 95.04% of all FM. The low incidence of evoked FM, at an early gestational age (10-21 weeks) emphasizes the ability of these young fetuses to perceive external stimuli and to react to them by FM.

During the past 30 years there has been great progress in perinatology. However, perinatal mortality has remained relatively high. To reduce perinatal mortality and morbidity, high risk pregnancy units have been established throughout the world. Our unit was established in 1976 for women with diseases complicating pregnancy, and in whom evaluation of fetal respiratory and metabolic function is indicated. The cesarean section rate during these 12 years was 33%. There was no maternal mortality. The intrauterine fetal death rate was 10.3 per 1,000 (very low for this unique population) and the corrected perinatal mortality was 3.4 per 1,000. The monitoring and treatment protocols, developed and applied in this unit, resulted in a significant decrease in fetal morbidity and mortality. These dramatically improved results demonstrate and justify the need for high risk pregnancy units.

The study evaluates the benefit of elective hospitalization in preventing premature deliveries of twin gestations. Three groups of women with twin gestations, having no other complications of pregnancy which could cause premature delivery, were evaluated. The study group was comprised of 43 women who were electively hospitalized between 30-32 and 36 weeks of gestation. Control group 1 was comprised of 55 women who were not hospitalized but were instructed to rest at home. Control group 2 was comprised of 53 women who were not hospitalized and were not instructed to rest at home. Our results showed that elective hospitalization did not significantly affect the gestational duration or the prematurity rate. However the mean birthweight difference between the study group and the two control groups were 143 +/- 83 g and 205 +/- 84 g, respectively. This result was more significant in multiparous women. The slight increase in birthweight of the hospitalized women compared to the controls, does not seem to justify the cost of hospitalization.

Sinusoidal fetal heart rate pattern (FHR) is regarded by most authors as signifying a compromised fetus. In most cases, this is secondary to fetal anemia of different causes, usually Rh isoimmunization, more rarely to fetomaternal transfusion, bleeding vasa previa, placental chorioangioma, or traumatic amniocentesis. A case is described which to our knowledge is the first reported of a sinusoidal FHR pattern occurring in the relatively anemic donor twin in a case of the twin to twin transfusion syndrome. The importance of separate antepartum biophysical evaluation of each member of a twin pregnancy is emphasized.

High levels of serotonin and 5-HIAA were found in the urine of a group of 102 oligospermic and azoospermic men. These levels were significantly higher than those of normal fertile men.

Fetal movement (FM) counts by mothers with premature rupture of membranes (PROM) may be helpful in estimating fetal well being, provided FM assessment is not altered by the reduced amniotic fluid. It is possible that the decreased uterine volume restricts fetal movements, though the closer contact of the fetus to the uterine wall may enhance maternal perception of even weak FM. The present study compared maternally perceived FM in 41 cases of PROM to 120 uncomplicated pregnancies. No significant differences in FM counts were detected between the two groups at any of the gestational ages. Fetal movement counts by mothers with PROM, therefore, can assist fetal surveillance as is the case in pregnancies with intact membranes.

Fetal activity in multiple pregnancies, as expressed by maternal daily fetal movement recordings, was compared with that in singleton pregnancies. It was found that the mean fetal activity in 33 cases of twins, six cases of triplets, one case of quadruplets, and one case of quintuplets was generally higher than that obtained during singleton pregnancies. In most cases, fetal movements of triplets, quadruplets, and quintuplets were even higher than the movement of twins.

A case of maternal pulmonary edema occurring in a patient in the 32nd week of gestation is presented. This was our first case of pulmonary edema seen during a period of five years' usage of isoxsuprine in the treatment of premature labor. The patient presented was 28 years old, gravida 2, para 1, admitted to the maternity ward with premature uterine contractions. Her past history eliminate cardiac or pulmonary disease. Isoxsuprine therapy was begun with initial dose of 0.04 mg/min. and increased to 0.32 mg/min., the total dose administered was 560 mg during 48 hours. During this period she was given in dexamethasone 24 mg. Fluid balance on the first day of the treatment was +1.7 liters and on the second day +5.2 liters. Forty-eight hours from the commencement of the treatment, the patient experienced shortness of breath and chest pain. Physical examination disclosed wet rales over both lungs, sinus tachycardia and tachypnea. Laboratory examination disclosed hypopotassemia of 3 mEq/liter, hypoxemia (PO2 of 80 torr on 0.5 FiO2 face mask) with mild hyperventilation 28 torr PCO2 with normal ph 7.43. Recognition of the early signs of pulmonary edema enable swift clinical diagnosis and steps to be taken to prevent disasterous condition due to progressive hypoxemia. The prompt treatment in this complication includes discontinuation of isoxsuprine and fluid administration, placement of the patient in an erect position, intravenous furosemid 40 mg, oxygen supplement by face mask and 25 mg of meperidine. The patient's condition dramatically improved though the lung fields became completely clear from wet rales only eight hours from the start of dyspneic attack.(ABSTRACT TRUNCATED AT 250 WORDS)

Assessment of fetal movements by the pregnant woman is a useful screening test in high and low risk pregnancies. Decreased fetal movements to less than 10 during 12 hours is an expression of fetal distress, and may be the first alert of impending fetal death. Pregnancies with decreased fetal activity comprise a very high risk group, however some normal pregnancies are still included. With the object of excluding false positive observations, fetal heart rate monitoring was added as a secondary screening procedure. There were seventy patients with diminished fetal movements, 28 had two or more fetal heart rate accelerations in twenty minutes, while 42 had one or no accelerations in twenty minutes. In the former group there was no perinatal mortality, while in the latter, 18 (42.8%) succumbed either pre- (16.6%) or post-natally (26.2%). It is suggested that daily fetal movement recording should be the primary screening test for pregnant women. Whenever reduced fetal activity to less than 10 in 12 hours is perceived, fetal heart rate should be monitored. The existence of at least two accelerations in 20 minutes excludes all the fetuses prone to disaster. One or no accelerations in 20 minutes when coexisting with decreased fetal movements, have a very grave prognosis, and interruption of pregnancy should be considered.

The danger to the newborn inherent in a postmature pregnancy has gained recognition. However, the diagnosis of postmaturity remains a difficult problem. The thromboplastic activity of amniotic fluid (TAAF) increases with the gestational age. Based on this phenomenon, the TAAF of 45 women in the 41st to 43rd weeks of gestation was established, using their amniotic fluid as a source of thromboplastin. It was found that in all cases in which the TAAF value was less than 42 seconds, the newborns exhibited clinical evidence of postmaturity.

Leukocyte alkaline phosphatase (LAP) activity was studied in 50 women at delivery and in the newborn after delivery. The following findings were observed: (1) the LAP activity of newborns is lower than that of the mothers; (2) significant decrease in the LAP activity has been observed in newborns on the 4th day after delivery; (3) there are no significant differences between newborns from normal and prolonged labor; (4) immediately after delivery the LAP scores of male newborns are lower than those of females, but 4 days later these scores were reversed, and (5) 4 days after delivery, the LAP scores do correlate to the weight of the newborn.

Various fetal scalp lesions are related to the use of the vacuum extractor. Blood sequestered in these lesions could result in an increased bilirubin load on the functionally limited neonatal liver, leading to the development of hyperbilirubinemia. In the present study bilirubin levels of vacuum extracted neonates were compared with those of non-instrumentally delivered babies during the first 72 hours of life. Sixty-nine vacuum extracted neonates had higher bilirubin levels than 56 non-instrumentally delivered babies at 24 (114 mumol/l vs. 96 mumol/l), 48 (163 vs. 141) and 72 (194 vs. 144) hours of age. The p values were 0.05, less than 0.025 and less than 0.001 respectively. This trend was apparent in both oxytocin induced and non-induced deliveries and whether or not phototherapy cases were included in the analysis. The incidence of hyperbilirubinemia requiring phototherapy was higher after vacuum extraction than after non instrumental delivery (27.5% vs. 12.5%; p less than 0.04). Analysis of our results unexpectedly indicated that oxytocin induction was generally associated with an attenuation of bilirubin levels after both vacuum extraction and spontaneous delivery. The clinician attending newborn babies should be aware of the higher incidence of neonatal hyperbilirubinemia associated with vacuum extraction.

In three of 1,094 cases of cord complications the mothers experienced reduction of fetal movments until cessation. In these three instances the fetal heart beat was audible but changes appeared on the fetal heart rate monitor. The course of loss of fetal movements resembled that seen in cases of placental insufficiency. It is suggested that the reduced fetal movements and the changes in fetal heart rate were due to a diminished blood flow in the cord vessels as a result of gradual cord compression.

Severe cardiac disease in pregnancy may be complicated by reduced placental perfusion and subsequent fetal deprivation. In the present study we have evaluated one fetal parameter that may thus be affected, namely fetal activity. The study group included patients with rheumatic heart disease: 36 women with mild and 5 with severe disease. Each counted fetal movements 3 times a day and from this the daily fetal movement recording was calculated. One hundred and twenty women with normal pregnancies were the controls. The mild cases had fetal activity which was similar to that of the controls. The severe cases had reduced fetal activity, which was significant statistically at 29 to 32 weeks of gestation. Three in the severe disease group had marked decrease of fetal movements on admission to hospital. A return to normal fetal activity was observed following improvement of the maternal cardiac state. As was previously suggested for other high and low risk pregnancies, the maternal perception and counting of fetal movements may aid the fetal surveillance of patients with rheumatic heart disease.

A dietary history was obtained from 180 patients with preeclampsia and eclampsia, representing 92% of all cases occurring in the pregnant population of West Jerusalem over a defined period. The intake of nutrients and 78 items of diet was compared for each patient and two healthy control subjects who were matched for country of origin, parity, month of delivery, age, year of immigration and years of schooling. All diagnoses were made by the same research team and dietary histories were elicited by one of two experienced dieticians. Comparisons between the 180 patients and their respective control pairs showed a significantly reduced intake of calories, proteins and fats and an increase in consumption of sugar and sweets by the patients. The food items involved, including cheese, eggs, olives and nuts, did not appear consistently, however, in population subgroups defined by age, period of immigration and country of origin. Sixty-six patients reported not having changed their diets during pregnancy and these showed no significant differences in consumption of any of the foods or nutrients at the 1% level, when compared with their matched control subjects. All differences observed were confined to the 114 patients who, for one reason or another, had changed their diets. It is concluded that the differences in diet between patients and control subjects are the result, and not the cause, of the toxemia of pregnancy. Supporting and contradictory evidence in the literature is analyzed, and possible sources of the discrepancies are pointed out.