Placenta

Vertical transmission of SARS-CoV-2 is controversial. Owing to the low expression of ACE2 along with TMPRSS2 in placenta, role of secondary receptors becomes significant. In our study we predicted twelve interactors of spike protein expressed in placenta through interactome analysis followed by docking and molecular dynamic simulation. Three interactors of SARS-CoV-2 spike protein were found to be CD147, EGFR and LDLR. Expression at transcriptomic and translational level by real-time PCR and immunohistochemistry respectively was observed in placenta samples obtained from women with COVID exposure risk and compared to control. Elevated protein levels of CD147 (OR=14.667), ACE2 (OR=8.25) and LDLR (OR=1.9) was associated with higher exposure risk. However, the variation in mRNA expression was not statistically significant. Computational study showed that CD147/basigin, EGFR and LDLR stably interact with spike protein outside its primary receptor binding domain. In-vitro binding assay confirmed the physical interaction between spike protein and the placental receptors. In-silico site-directed mutagenesis further identified key potential binding hotspots of spike protein- Tyr28 and Trp64 for CD147, Phe643, Val642, Thr632 for LDLR and Leu8, Phe59, Phe32, Tyr28, Trp64, Trwp152, His245, and Val213 for EGFR. Finally, concluded that CD147, EGFR and LDLR might act as secondary non-canonical cofactors for SARS-CoV-2 spike protein in placenta, thereby increasing its susceptibility and tissue tropism. The study overall gives a better insight into potential placental suscpetibility providing targeted therapeutic opportunities.

Preeclampsia is a pregnancy complication characterized by hypertension, proteinuria, and end-organ dysfunction. Abnormal placentation leading to reduced placental perfusion may contribute to its development. Previous studies demonstrated that the activation of the apelin receptor (APJ) system has hypotensive, renoprotective, and antioxidant effects in preeclamptic rat models. Apelin and elabela (ELA) can stimulate the proliferation of trophoblast cells, suggesting a role in embryonic development. However, the mechanisms underlying the actions of apelin or ELA in trophoblast cells are not well understood, particularly in hypoxic settings. The immortalized HTR-8/SVneo trophoblastic cells were treated with cobalt chloride (CoCl2) at 0.2 mM for 24 hours to mimic hypoxic conditions. RT-qPCR, ELISA or Western blotting was used to measure mRNA or protein levels of apelin, elabela, and the components of IL-6 signaling in cell lysates or conditioned media. The exposure to CoCl2 increased total apelin and elabela content approximately 2-fold in the conditioned media but did not affect APJ levels. CoCl2 upregulated proinflammatory cytokine concentrations: soluble fms-like tyrosine kinase 1 (sFlt-1), soluble gp130 (sgp130), interleukin-6 (IL-6), and sIL-6 receptor (IL-s6R). Both apelin and elabela downregulated IL-6 mRNA but had no effect on sFlt-1 mRNA. Apelin attenuated sgp130, while ELA decreased the membrane form of IL-s6R. Apelin also decreased the pSTAT3/STAT3 ratio. CoCl2-induced hypoxia upregulated the pro-inflammatory milieu in HTR-8/SVneo cells. Local activation of this peptidergic system may be a compensatory response of the trophoblast cells to hypoxia as exogenous apelin and elabela treatment ameliorated the hypoxia-induced pro-inflammatory milieu.

IntroductionPreeclampsia is characterized by a pro-inflammatory, anti-migratory and anti-angiogenic placental phenotype. Impaired spiral artery remodeling stemming from trophoblast dysfunction is a key pathogenic mechanism. Little is known about the processes that govern trophoblast function normally and in preeclampsia. In preeclampsia, placental Let-7b-5p is reduced. The objectives of this study were to determine the normal function of Let-7b-5p in human trophoblast cells, to examine whether the ssRNA sensors, Toll-like receptor (TLR) 7 and/or TLR8 are mediators of trophoblast Let-7b-5p function, and whether disruption of this pathway promotes a preeclampsia-like phenotype in the trophoblast. MethodsThe human first trimester trophoblast cell line, Sw.71, was transfected with a Let-7b- 5p mimic, a Let-7b-5p inhibitor, or scramble control. Cells were treated with or without the TLR7 inhibitor IRS661 or the TLR8 inhibitor CUCPT9a. Trophoblast migration was measured using a two-chamber assay and interactions with human endometrial endothelial cells (HEECs) was measured using a 3D matrigel model. Trophoblast anti-angiogenic sFlt-1 release was measured by ELISA and sFLT1 mRNA measured by RT-qPCR. ResultsTransfection of trophoblast cells with a Let-7b-5p mimic elevated migration through activation of TLR7 and TLR8, while in a TLR7-dependent manner, the Let-7b-5p mimic negatively regulated sFlt-1 production. Furthermore, inhibition of trophoblast Let-7b-5p reduced migration, elevated FLT1 mRNA expression and sFlt-1 release, and reduced trophoblast-endometrial endothelial cell interactions. ConclusionsThis study highlights a role for TLR7/TLR8-activating Let-7b-5p in promoting normal trophoblast function and endothelial interactions and that disruption in this miR-driven signaling pathway may be relevant to processes driving a pre-eclamptic placental phenotype. HighlightsTrophoblast migration is positively driven by Let-7b-5p activating TLR7 and TLR8 Let-7b-5p, via TLR7, negatively regulates trophoblast anti-angiogenic sFlt-1 production. Inhibition of trophoblast Let-7b-5p reduces trophoblast migration and normal interactions with endometrial endothelial cells, while sFlt-1 production is elevated. TLR7/TLR8-activating Let-7b-5p promotes normal trophoblast function and endothelial interactions and disruption in this miR-driven signaling pathway may promote a preeclamptic placental phenotype.

Abstract Background: Placental insufficiency (PI), characterized by impaired placental function, is a leading cause of stillbirth. Although rare coding variants are known to cause a subset of stillbirths, their contribution to PI is poorly understood. Methods: We conducted a secondary analysis of stillbirth cases (N=292) of the Stillbirth Collaborative Research Network study with exome sequencing available on postmortem placental or fetal samples. PI was defined by standardized placental pathological lesions (e.g., maternal vascular malperfusion) and prespecified clinical criteria for placental dysfunction. Candidate gene-based rare-variant burden analysis was performed using the Sequence Kernel Association Test-Optimal framework, focusing on evolutionarily constrained developmental and placental-function genes. Primary analysis compared stillbirths with and without placental vascular malperfusion. Similar comparisons by clinical PI and other placental pathological lesions were conducted in secondary analysis. Results: Stillbirths with PI phenotypes demonstrated different patterns of genetic variation than stillbirths without PI. Specifically, we observed nominal differential enrichment of constrained variants in biologically prioritized placental-function genes, regulating extravillous trophoblast invasion and spiral artery remodeling (CTNND2, PHACTR1, SLIT1, VEGFA), placental angiogenesis (IGF2R, TEK), mitochondrial homeostasis (OPA1), and fetal growth (DLK1, GNAS). Secondary analyses on specific placental pathological lesions suggested recurrent enrichment of genes within similar placental developmental pathways. Conclusions: We observed nominal differential enrichment of constrained developmental and placental-function genes in stillbirths with PI compared to stillbirths without PI. The observed genetic signals across PI phenotypes suggest shared biological pathways underlie placental dysfunction. Our findings advance genotype-phenotype characterization and help prioritize candidate pathways for larger collaborative genomic and functional studies to improve stillbirth risk stratification and prevention.

Background: The placenta has a unique transcriptomic profile, including microRNAs that are secreted into maternal circulation throughout pregnancy. MicroRNAs are small, non-coding RNA that post-transcriptionally regulate gene expression. Spontaneous preterm birth (sPTB) is associated with substantial differences in both placental pathophysiology and placental gene expression compared to term birth. We aimed to generate microRNA signatures of sPTB and map them to target genes using a microRNA-mRNA network. Methods: This study was conducted within the Conditions Affecting Neurocognitive Development and Learning in Early childhood (CANDLE) study. Placental samples were collected at delivery, and RNA was isolated for mRNA and microRNA sequencing. To investigate sPTB, this study excluded placental samples of participants with iatrogenic indications for PTB or induced labor. We examined differences in microRNA expression in participants who delivered before 37 weeks (N=35) compared to term participants (N=404) in a series of covariate-adjusted linear regression models. We used paired placental microRNA and mRNA expression data from this cohort to validate associations between computationally predicted microRNA-mRNA pairs and establish a microRNA-mRNA network. Results: Expression of 7 microRNAs were increased in sPTB (FDR<0.05) and were inversely correlated with sPTB-associated genes involved in immune signaling. Expression of 12 microRNAs were decreased in sPTB, including 4 members of the maternally expressed chromosome 14 microRNA cluster (miR-376a-3p, miR-376c-3p, miR-377-3p, and miR-381-3p). These microRNAs were predicted to negatively regulate oxidative phosphorylation genes that were increased in sPTB. The associations between miR-376c-3p and miR-377-3p and oxidative phosphorylation were confirmed in microRNA knockdown experiments. Conclusions: This study highlights potential biological mechanisms by which placental microRNA dysfunction might contribute to sPTB and highlights putative sPTB biomarkers that may be detectable in maternal circulation.

Opioid and cannabis co-use during pregnancy is increasingly common, yet the placental mechanisms linking combined exposure to adverse fetal outcomes remain poorly defined. Using a controlled mouse model of gestational drug exposure, we tested how fentanyl, {Delta}9-tetrahydrocannabinol (THC), or their combination altered placental structure, immune signaling, and gene expression and how these changes related to fetal growth. Drug exposure produced fetal growth restriction and reduced placental efficiency, with the greatest impairment in the combined fentanyl+THC group. Placental alterations were detectable by mid-gestation, when fentanyl exposure increased nucleated red blood cells within the labyrinth, consistent with hypoxic stress. By term, placentas showed compartment-specific remodeling, with THC selectively expanding the labyrinth and combined fentanyl+THC exposure increasing decidual area. Labyrinth composition and proliferative activity were altered, ultrastructural analysis revealed disruption of the maternal-fetal exchange interface, and placental interleukin-10 and interferon-{beta} levels were reduced across exposure groups. Transcriptomic analyses identified suppression of innate immune and antiviral defense pathways together with treatment-specific stress responses, and integration of placental gene expression with fetal weight showed coordinated repression of vascular and developmental regulators and activation of hypoxia- and metabolic stress-associated genes. These findings identify the placenta as a key mediator of adverse fetal outcomes associated with prenatal polysubstance exposure.

Background: Prenatal psychosocial distress is a risk factor for adverse birth outcomes such as preterm birth, but the biological pathways remain incompletely understood. Allopregnanolone (ALLO), a stress-responsive, progesterone-derived neuroactive steroid, may contribute to pregnancy maintenance by inhibiting uterine activation and modulating inflammatory and neuroendocrine pathways involved in parturition. Few human studies have examined ALLO during pregnancy in relation to birth timing and related outcomes, and no study to our knowledge has examined the entire pathway of prenatal psychosocial distress, ALLO, and birth outcomes. Here, we evaluated whether maternal ALLO concentrations and the ALLO-to-progesterone ratio mediate associations between prenatal psychosocial distress and birth outcomes. Methods: This study included 237 pregnant participants from the Healthy Start Cohort, enriched for psychosocial distress (n=57 high-distress; n=180 low-distress) assessed using the Edinburgh Perinatal Depression Scale (EPDS) and EPDS-3A anxiety subscale. We measured maternal serum ALLO and related steroid hormones at approximately 17 and 27 weeks gestation (range: 10-34 weeks) using a validated HPLC-MS/MS assay and evaluated natural log (ln)-transformed ALLO and the ALLO-to-progesterone ratio as potential mediators. The primary outcome was gestational age at birth; secondary outcomes included birthweight-for-gestational-age z-score (BW/GA), percent fat mass (%FM), and birth length-for-gestational-age z-score (BL/GA). We conducted regression-based mediation analyses by comparing the total and direct effects of prenatal psychosocial distress after adjustment for each proposed mediator. Results: Participants had a mean +- SD age of 29+-6 years; 38% were nulliparous, and 60% identified as non-Hispanic White. Of the birth outcomes assessed, prenatal psychosocial distress was only associated with BL/GA ({beta} = -0.49; 95% CI: -0.84, -0.14). Adjustment for ALLO at ~27 weeks as a mediator minimally attenuated the association between prenatal distress and BL/GA ({beta} = -0.46; 95% CI: -0.81, -0.10, 6.1% attenuation), whereas adjustment for the ALLO-to-progesterone ratio resulted in 20.8% attenuation ({beta} = -0.38; 95% CI: -0.74, -0.03). Conclusions: Maternal prenatal distress was associated with shorter birth length. The ALLO-to-progesterone ratio, but not ALLO itself, may partially mediate this relationship. These findings support neurosteroid metabolism as a biological pathway linking prenatal distress to fetal length accrual. Future mechanistic studies are needed to confirm these findings.

IntroductionWhile placental perfusion and pathology jointly affect pregnancy outcomes, cotyledon-specific perfusion across gestation and its correlation with local injury is not yet well understood. Ferumoxytol dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) offers a promising way to noninvasively identify cotyledons across gestation and quantify longitudinal cotyledon-specific perfusion changes. Additionally, intraplacental injection of bioactive fibrin sealant allows us to model thrombotic placental injury and further assess cotyledon-level relationships between perfusion and significant injury. MethodsPregnant rhesus macaques (N=13) received intrauterine saline or fibrin sealant injections at gestational day (GD) [~]101 and underwent ferumoxytol DCE-MRI at GDs [~]100, 115, and 145. Placental perfusion domains derived from contrast arrival time were segmented at each imaging time point and matched to cotyledons identified following tissue collection by cesarean section, with cotyledon perfusion quantified longitudinally and correlated with cotyledon-specific quantitative histopathology. ResultsAll pregnancies were successfully carried to term. Fibrin sealant injections induced significantly higher levels of placental pathology compared to saline controls. MRI-derived perfusion domains were largely consistent across gestation and showed predominantly one-to-one correspondence with term cotyledons, with successful perfusion-pathology pairing achieved in 153 cotyledons. Longitudinal cotyledon perfusion changes showed significant positive correlations with villous agglutination injuries. ConclusionsFeasibility of noninvasively tracking placental cotyledon perfusion using ferumoxytol DCE-MRI was demonstrated, and the efficacy of the rhesus macaque thrombotic injury model was confirmed. The positive perfusion-pathology correlations suggested intrinsic placental regulatory mechanisms and functional plasticity. This new framework is promising for future translational studies and validation of ex vivo cotyledon perfusion models. HighlightsO_LILongitudinal tracking of placental perfusion domains with ferumoxytol MRI C_LIO_LISuccessful matching of cotyledons and MRI-derived perfusion domains C_LIO_LIConfirmed thrombotic injury-model induced cotyledon pathology C_LIO_LIMaternal perfusion compensation in presence of villous pathology C_LI

Males experience elevated mortality and morbidity during the perinatal period and infancy; however, the biological origins of this vulnerability remain unclear. Emerging evidence suggests that sex-dependent placental development may contribute to differences in fetal growth and outcomes. Vascular networks are central to placental function, established early in the first trimester to support growth and mitigate harmful exposures, and maturing by term to sustain late fetal development. Although these processes are tightly regulated by gene-expression programmes, the role of sex-specific vascular signalling in placental development remains poorly understood. To address this, we analysed raw, publicly available RNA-sequencing transcriptomics data from first trimester and term placental samples. We conducted parallel differential gene expression and functional enrichment analyses across the datasets to ensure direct comparability and to disentangle the specific contribution of sex chromosomes. We identified multiple differentially expressed genes and enriched pathways between male and female placentas. In the first trimester, male placentas exhibited significant enrichment of vascular signalling pathways, particularly Notch signalling, cell adhesion, and migration, consistent with heightened proliferative and remodelling activity. At the end of gestation, sex differences remained detectable; however differentially expressed genes was clearly sex-chromosome driven, with autosomal single-gene sex differences and autosomal-sex-specific pathways markedly reduced. This reflected a mature placental vascular system with minimal sex-based differences outside of the sex chromosome genes. These results reveal gestational stage-dependent sex differences in placental vascular development, with early male fetuses exhibiting increased angiogenic activity and immature vascular signalling, potentially contributing to accelerated growth and increased vulnerability in male neonates.

Progesterone is essential for the establishment and maintenance of pregnancy, primarily through the classical nuclear progesterone receptor (PGR). However, progesterone also exerts rapid, non-genomic effects through membrane progesterone receptors, including members of the progestin and adipoQ receptor (PAQR) family, whose cellular distribution and potential roles at the human maternal-fetal interface remain poorly understood. We therefore sought to define the cell-type-specific expression landscape of classical and membrane progesterone receptors during early human pregnancy. We analyzed publicly available single-cell RNA-sequencing datasets of the first-trimester human maternal-fetal interface to characterize the expression of PGR and PAQR family members across trophoblast, immune, stromal, and vascular cell populations. Major expression patterns were examined across independent datasets and further assessed using spatial transcriptomic data. PGR expression was predominantly localized to decidual stromal and perivascular populations, whereas individual PAQR family members showed distinct cellular distributions. PAQR6 was enriched in decidual natural killer cells, PAQR7 was broadly expressed in trophoblast populations, PAQR8 was prominently expressed in fetal stromal and macrophage populations, including Hofbauer cells, and PAQR9 showed prominent enrichment in syncytiotrophoblasts. Selected cell-type-specific patterns were further supported by spatial transcriptomic analysis. These findings provide a cell-type-resolved framework for investigating how distinct progesterone receptor systems mediate progesterone signaling across maternal and fetal cellular compartments during early human placentation.

Background: Prenatal cannabis use is increasing, yet the effects of inhaled cannabis smoke on placental molecular function remain poorly defined. Objective: To characterize placental transcriptomic alterations following prenatal cannabis smoke exposure using a physiologically relevant murine inhalation model. Methods: Timed-pregnant CD-1 mice were exposed to cannabis cigarette smoke or room air from embryonic day (E)6.5 to 18.5. Placentae were collected at E18.5 for microarray-based transcriptomic profiling. Differential gene expression was assessed using fetal sex adjusted linear models and false discovery rate correction. Gene set variation and ranked pathway enrichment were performed to evaluate compartment-associated and biological transcriptional programs. Selected genes were validated by quantitative PCR. Results: Large-scale transcriptomic profiles revealed no clear visual clustering by exposure; however, 23 genes were differentially expressed (FDR <0.05, |log2 fold change| > 0.5). Labyrinth-associated transcriptional signatures were reduced, whereas junctional-associated signatures were increased in exposed placentae. Pathway analyses revealed negative enrichment of fatty acid metabolism, bioenergetic, and transmembrane transport pathways, alongside positive enrichment of RNA processing and transcriptional-regulatory pathways. Quantitative PCR confirmed altered expression of multiple transport-related genes. Electron transport chain-associated gene expression was altered despite preserved placental respiratory measurements. Conclusions: Prenatal cannabis smoke exposure is associated with subtle yet coordinated placental transcriptional alterations involving metabolic, transport-associated, and gene-regulatory pathways, alongside compartment-associated transcriptional shifts. These findings provide insight into placental responses to whole cannabis smoke exposure and identify pathways that may contribute to altered placental function during pregnancy.

Background: A major hallmark of necrotizing enterocolitis (NEC) is intestinal hypoxia, but detection of this feature is difficult. Photoacoustic imaging (PAI) is an emerging imaging method to noninvasively visualize intestinal oxygenation and microvasculature and is promising for improved NEC detection. Prior to clinical translation of PAI for NEC detection, initial studies must be performed in a pediatric population to assess safety and feasibility. Methods: Under IRB approval, eighteen healthy pediatric participants underwent PAI scans before and after eating a lipid-rich snack. PAI was acquired in each abdominal quadrant with intestinal regions-of-interest (ROI) manually drawn on each image series to measure averaged values of deoxygenated hemoglobin (Hb), oxygenated hemoglobin (HbO2), total hemoglobin (HbT), and oxygen saturation (sO2). Results: Minor temporary erythema and pressure from safety goggles in some participants were the only adverse events reported. Pre-/post-prandial changes for HbT, HbO2, Hb, and sO2 were assessed for each abdominal region and age cohort. Results showed significant post-prandial hyperemia in the intestines with no changes in bowel oxygenation. Conclusions: PAI is a safe and feasible imaging modality for evaluating intestinal blood flow in pediatric patients that detects physiological changes in bowel microvasculature and shows strong potential as a quantitative biomarker for assessment and monitoring of intestinal vascular health.

Preeclampsia (PE) is a hypertensive pregnancy-specific disorder and a leading cause of maternal and fetal mortality. A common feature of PE placentas and maternal plasma is dyslipidemia, or abnormal lipid levels, which can increase oxidative stress and endothelial dysfunction. However, the precise transcriptional, post-transcriptional, and epigenetic mechanisms underlying these abnormalities remain poorly characterized. Identifying such changes may clarify disease mechanisms and identify lipid-related PE biomarkers. We conducted a large-scale meta-analysis integrating public placental datasets from NCBI GEO, comprising four DNA methylation (DNAm) datasets (n = 172), three RNA-sequencing datasets (n = 92), and an independent RNA microarray validation cohort (n =146). We evaluated differential DNAm (limma), gene expression (DESeq2), transcript-level shifts (Swish), and alternative splicing (rMATS) in PE versus control placentas, with all analyses stratified by fetal sex via an interaction term model. We also performed placental cell-type deconvolution to quantify PE-associated cell-type proportion changes. Our results demonstrated that lipid-related regulation changes in PE placentas occur primarily at the gene and transcript level, with DNAm showing no changes. We also identified significant isoform switching in PE that were undetected by differential gene expression analysis, and primarily driven by alternative transcription initiation and termination sites rather than alternative splicing. A subset of these isoform switches mapped to pathways dysregulated in PE and were predicted to cause functional protein changes. An interaction term model identified several sex-specific differentially expressed genes (DEGs) in PE, including a subset of male-specific downregulated genes involved in oxidative metabolism. However, many of the remaining sex-specific DEGs across both sexes were previously uncharacterized in the literature. These findings suggest that transcriptional and isoform-level regulation play a role in PE-associated dyslipidemia, with certain regulatory pathways displaying fetal sex-specific patterns. Highlights- Preeclampsia-associated dyslipidemia manifests at the gene and transcript level - Reciprocal isoform switches were missed by standard gene-level analyses - Alternative transcript initiation and termination drove isoform switching - Sex-interaction modeling identified sex-specific transcriptional shifts in PE

BACKGROUND Twin pregnancies have a two- to three-fold higher risk of preeclampsia (PE), but early prediction remains poor because current biomarkers and prediction models are largely derived from singleton pregnancies. Defining the biological changes that precede PE in twins may facilitate the development of more effective risk assessment approaches. OBJECTIVE We aimed to characterize maternal plasma protein changes preceding PE in twin pregnancies and to explore whether early circulating proteins could serve as improved risk stratification for twin PE. STUDY DESIGN We performed a nested case-control study within a prospective twin pregnancy cohort at Peking University Third Hospital. A total of 84 women with twin pregnancies were included, comprising 40 pregnancies complicated by PE and 44 normotensive controls. Longitudinal maternal plasma samples were collected in the first, second, and third trimesters. In the discovery cohort, maternal plasma proteins were profiled by mass spectrometry to identify trimester-specific protein changes associated with PE. Candidate proteins were then validated in an independent validation set. We further assessed the functional relevance of selected proteins in trophoblast and endothelial cell models and constructed prediction models integrating biomarkers with maternal clinical characteristics. RESULTS Twin pregnancies complicated by PE showed distinct trimester-dependent protein alterations, with first- and second-trimester changes mainly involving immune regulation and glycolipid metabolism, and later changes enriched in cardiovascular-related pathways. Several proteins demonstrated early discriminatory patterns, including increased annexin A3, chitotriosidase-1, and ferritin light chain (FTL), together with decreased annexin A2 in the first trimester, whereas the conventional singleton markers (sFlt-1 and PAPP-A) did not differ. ELISA validation of these candidate biomarkers yielded an AUC of 0.93 for twin preeclampsia prediction; integration with standard maternal clinical factors further improved discriminatory performance to an AUC of 0.97. Accordingly, a multivariable model combining annexin A2, annexin A3, chitotriosidase-1, FTL, and maternal clinical factors achieved strong internal performance for early discrimination of PE in twin pregnancies (AUC, 0.97; 95% CI, 0.93-1.00). Moreover, annexin A2 knockdown impaired trophoblast migration and invasion and reduced inflammatory cytokine secretion in endothelial cells, supporting a role in placental development and maternal vascular adaptation. CONCLUSION Twin pregnancies that later develop PE exhibit early and progressive maternal plasma protein alterations that can be detected before clinical disease onset. These findings identify candidate biomarkers and biologically relevant pathways that may help explain why PE in twins differs from singleton disease and may support future twin-specific risk stratification strategies. These results require external validation before clinical application.

Maternal thermal strain is associated with adverse pregnancy outcomes, yet fetal temperatures cannot currently be directly measured, limiting quantification of fetal thermal strain. Here, we develop two steady-state models for estimating internal temperatures in a near-term fetus. First, we improve the only previously published human fetal thermoregulation model, deriving a closed-form solution within its simplified uniform-cylinder representation. Second, we introduce a multilayer, anatomically segmented model that resolves tissue-specific temperatures. Both couple the fetal body to central blood pool and amniotic fluid compartments and incorporate a new placenta-umbilical cord heat-exchanger representation. Predictions agree with available intrauterine scalp measurements, with fetal core and head-center temperatures approximately 0.5{degrees}C and 0.8{degrees}C above maternal core, respectively. Physiologically plausible changes in umbilical cord heat-exchanger effectiveness or blood flow increased fetal temperatures by approximately 0.3{degrees}C. These models enable estimation of otherwise inaccessible temperatures, while the multilayer formulation lays a foundation for transient, coupled maternal-fetal thermoregulation modeling.

Background: Early detection of preeclampsia with noninvasive and reliable biomarkers is the foremost step for minimizing adverse effects during pregnancy. However, none maternal serum analyte evaluated so far is sufficiently accurate to allow recommending their routine use. Methods: Microarray was used to first identify differentially expressed LncRNA and mRNA. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to determine bio-functions and signaling pathways. Quantitative real-time polymerase chain reaction was used to validate the results of microarray. Finally, the lncRNA-mRNA co-expression network was constructed to find the interaction of genes. Result: A total of 1476 differentially expressed lncRNAs and 594 mRNAs were identified. Both RNA-seq and RT-qPCR showed the expression of T241171,T338586, uc002ywy.3 was up-regulated, and the expression of ENST00000524858, T131416, T357032, uc.335+ was down regulated in the plasma of patients with preeclampsia. Pathway analysis showed the targeted mRNAs were enriched in apoptosis, sulfur metabolism, starch and sucrose metabolism. ceRNA network found that LncRNA-uc002ywy.3 might be the upstream regulator of miRNA-4498. LncRNA-uc002ywy.3/miRNA-4498 was predicted to interplay with genes involved in programmed cell death-1and its ligand (PD-1/PD-L1) pathway by KEGG analysis. Conclusion s PD-1/PD-L1 signaling pathway may be involved in the development of preeclampsia. The dysregulated LncRNA-uc002ywy.3/miRNA-4498 shed light on a new layer involved in the regulatory network of preeclampsia.

Environmental exposure to endocrine disrupting chemicals, like bisphenol-A (BPA), can impart detrimental effects on developing feto-placental unit, during pregnancy. Placenta remains a central player maintaining this feto-placental homeostasis for sustenance of a healthy pregnancy. Thus, the bisphenol-A mediated endocrine disruption affects the healthy functioning of placenta by altering key processes, such as tissue remodelling, angiogenesis, and metabolism. However, the underlying mechanism of BPA-altered ECM remodelling remains elusive. Therefore, in this study we investigated the BPA mediated changes in placental tissue remodelling using a bisphenol-A exposed murine model during pregnancy. The results reveal that, the phenotypic changes in feto-placental interface correlates with perturbed placental proteome in response to BPA. Further investigation highlights a S100a10-Annexin A2 axis mediated upregulation of tissue plasminogen activator (tPA), which drives altered extracellular matrix (ECM) degradation in placental decidua. This culminates into functional dysregulation in feto-placental axis, leading to reduced size of fetus and placenta. Therefore, this study provides novel insights of a S100a10-Annexin A2 axis associated mechanism for alteration of ECM remodelling in placental decidua due to BPA exposure, which may lead to toxicity related adverse pregnancy outcome.

Background: Preeclampsia (PE) is a hypertensive pregnancy disorder with contributions from both maternal and fetal genetic factors, but how both genomes regulate placental transcription remains incompletely understood. Results: We analyzed placental 5'-end RNA sequencing data from 59 pregnancies in the FINNPEC cohort and performed separate maternal and fetal cis-eQTL analyses using gene-level expression and promoter-level expression defined as Transcript Far 5' Ends (TFE). Genotype-by-PE interaction models were used to identify genetic effects that differ according to PE status. No gene-level interaction eQTLs remained significant after multiple-testing correction, although suggestive signals included LILRB2 in the maternal and LPCAT3 in the fetal analyses. In contrast, we identified two significant syntenic promoter-level interaction eQTLs: rs12504642-TFE61820 at the FHDC1 locus in the maternal analysis and rs1557150-TFE9032 at the PPIAP31 locus in the fetal analysis. Conclusion: These findings suggest that PE-dependent genetic interaction effects may be detectable at specific transcript promoter regions even when they are not captured at annotated gene levels.

Stigmatizing language in medical documentation may reflect and perpetuate bias, but its prevalence in obstetrics has not been systematically quantified. We applied a keyword-guided BERT classifier to 640,345 obstetric notes from 26,178 pregnancies at an academic medical center. Stigmatizing language was detected in 47% of 26,178 pregnancies. Black pregnancies had significantly higher odds of stigmatizing language compared with Asian (aOR=1.5, p=3x10-8) or White (aOR=1.4, p=6x10-6). Indicated and spontaneous preterm births were also significantly associated with stigmatizing language compared to term (aORs=1.5, 1.2; p=7x10-12, 0.01). Pregnant individuals with only 12th-grade maternal education were more likely to experience stigma than those with college (aOR=1.5; p=4x10-14). These findings provide evidence of differences in clinical documentation across race, education levels, and clinical conditions. They also demonstrate how automated natural language processing can enable systematic monitoring of bias in healthcare language at scale.

Background: Pregnancies complicated by fetal growth restriction are at increased risk of fetal demise and adverse neonatal outcomes. Distinguishing growth-restricted fetuses from constitutionally small ones remains challenging. Given the variability in current diagnostic criteria and the importance of identifying at-risk fetuses, fetal growth velocity has emerged as a predictor of adverse neonatal outcomes. Objective: To evaluate whether percentile-based fetal growth velocity, defined as change in estimated fetal weight percentile per week, predicts adverse neonatal outcomes in pregnancies affected by fetal growth restriction or small-for-gestational-age neonates. The primary aim was to determine the relationship between growth velocity and a composite of adverse neonatal outcomes. Study Design: This was a retrospective cohort study of pregnant patients 18 - 45 years old who delivered between August 2017 to December 2022 in a single healthcare system. Patients were excluded who had fewer than 2 ultrasounds after 16 weeks gestation, genetic or anatomic abnormalities, or a multiple gestation. Results: 300 patients met all inclusion criteria, and most patients (n=199) delivered at the tertiary care center. Three had an intrauterine fetal demise at a mean gestational age of 35w3d. 74 neonates were admitted to the NICU with a mean length of stay of 8.5 days; 29 required respiratory support. No neonatal deaths occurred. In fetuses with initial estimated fetal weight <3rd percentile (n=33), the probability of composite outcome was increased (50%, 95% CI 44.8-55.2) compared to those >50th percentile (5%, 95% CI 3.7-6.6). In the 3-10th and 10 - 50th percentile subgroups with decelerated growth, composite outcome rates were also increased (56.2% and 44.1%) compared to those with neutral or increased growth velocity. Conclusion: Percentile-based fetal growth velocity is a simple calculation that correlates with adverse neonatal outcomes regardless of initial estimated fetal weight. As fetal growth velocity decreased, our cohort saw increased rates of adverse outcomes. Change in EFW percentile normalizes for gestational age and allows ease of clinical interpretation. Decelerated growth identified fetuses at highest risk, suggesting growth velocity as a useful metric in routine surveillance.