Placenta

BACKGROUNDSpontaneous preterm birth (sPTB) remains the foremost cause of neonatal morbidity and mortality worldwide. Although histologic chorioamnionitis (HCA) and placental vascular abnormalities are frequently observed in sPTB, the molecular cascades linking these lesions to labor initiation remain poorly understood. Emerging evidence implicates circadian dysregulation and trophoblast dysfunction as additional drivers of sPTB. OBJECTIVEThis study aims to map placental pathology to distinct transcriptomic functional signatures that may precipitate sPTB, delineate the contribution of circadian regulation - both core-clock genes and circadian transcription-factor target sets (TFTs) - to sPTB, and identify placental cell-type-enriched and developmental pathway signatures that differ between sPTB and term deliveries. STUDY DESIGNWe performed bulk RNA sequencing on 32 formalin fixed, paraffin embedded placental specimens from 12 selected women (9 sPTB and 3 Term) in the POUCH Study cohort. Samples were selected for white ethnicity, maternal age 23-33years, and parity 1-4 to reduce heterogeneity within groups. An extraction-free HTG transcriptome panel assayed 19,398 protein-coding genes. Log2-fold changes of all genes were computed with limma adjusted for maternal age, gestational age, parity, placental region, placental pathology, and POUCHID (a clustering variable) for sPTB vs. Term and HCA/vascular lesion vs. no pathology (no placental pathology adjustment). Gene-set enrichment used 50 Hallmark sets (MSigDB) plus curated placental circadian, circadian TFT, cell-type, and developmental pathways or gene sets. RESULTSsPTB placentas displayed a global suppression of metabolic, secretory, and immune pathways (e.g., protein secretion, oxidative phosphorylation, Interferon responses, Complement, ROS, MYC Targets, TGF {beta}, mTORC1, and Coagulation) while KRAS Signaling Down and EMT were up-regulated. HCA-enriched sets (TNF/NF-{kappa}B, ROS, KRAS Up, IL-2/STAT5, Hypoxia, Interferon-{gamma}) were up-regulated, with EMT and Notch remaining down. Vascular abnormalities alone showed up-regulation of 12 Hallmark sets - including TGF-{beta}, TNF/NF-{kappa}B, ROS, pancreatic {beta}-cell stress, Hypoxia, Oxidative Phosphorylation, EMT, and mTORC1 - while Notch was down-regulated. When HCA co-exists with vascular abnormalities, the Hallmark profile becomes more inflammatory highlighting a synergistic exacerbation of innate immunity, oxidative stress, and programmed cell death with the 12 up-regulated sets (Complement, Interferon /{gamma}, TNF, ROS, Apoptosis, and Heme Metabolism). The exclusive downregulation of DNA Repair suggests compromised genomic integrity. Circadian gene-sets analysis revealed an up-regulated Regulation of Circadian Sleep Wake Cycle in sPTB but down-regulation of core clock pathway and suppressed circadian TF targets. Cell-type enrichment reveals increased trophoblast giant cells and IGFBP1-DKK1 positive fetal cells, with marked suppression of extravillous trophoblasts, syncytiotrophoblasts, villous cytotrophoblasts, and fetal myeloid cells. Placental developmental pathways were downregulated, indicating arrested trophoblast maturation. CONCLUSIONOur pilot analysis demonstrates sPTB placentas exhibit a global suppression of metabolic, secretory, and immune-modulatory programs and maladaptive trophoblast remodeling, whereas HCA and vascular abnormalities drove distinct inflammatory or hypoxic signatures. The shared and opposing Hallmark pathways across phenotypes highlight distinct yet overlapping pathogenic mechanisms. Dysregulated circadian pathways, consistent downregulated transcription factor target gene sets, and trophoblast-specific signatures implicate circadian misalignment and impaired placental maturation as key contributors to preterm parturition. These findings provide a mechanistic atlas linking placental pathology to sPTB and highlight potential targets for chronotherapeutic and cell-type-specific interventions. AJOG at a GlanceO_ST_ABSWhy was this study conducted?C_ST_ABSSpontaneous preterm birth remains a leading cause of neonatal morbidity. Histopathologic lesions of the placenta, particularly chorioamnionitis and vascular abnormalities, are common in preterm deliveries, yet the underlying molecular pathways are poorly understood. We sought to integrate functioning pathway profiles of placental histology, circadian biology, and cell types to identify mechanistic drivers of sPTB. Key findingsO_LIsPTB placentas showed widespread down-regulation of oxidative phosphorylation, mTORC1, hypoxia, interferon, and TNF/NF-{kappa}B pathways. C_LIO_LIHCA placentas up-regulated the same pathways (except androgen response), revealing a reciprocal inflammatory-hypoxic signature. C_LIO_LIVascular abnormalities displayed a distinct mix of up- and down-regulated pathways, suggesting divergent reparative responses. C_LIO_LIPlacentas with co-existing HCA and vascular abnormalities enriched more inflammatory Hallmark pathways: the 12 up-regulated sets (Complement, Interferon /{gamma}, TNF, ROS, Apoptosis, and Heme Metabolism) highlight a synergistic exacerbation of innate immunity, oxidative stress, and programmed cell death and the exclusive down-regulation of DNA Repair suggests compromised genomic integrity, which can contribute to premature placental senescence and preterm labor. C_LIO_LICircadian clock and multiple transcription-factor targets were enriched in sPTB, and trophoblast-specific signatures (giant, extravillous, syncytiotrophoblast) were prominent. C_LI What does this add to what is known?The study demonstrates a clear dichotomy between inflammatory and hypoxic molecular programs in sPTB and HCA, identifies circadian dysregulation as a potential contributor, and highlights trophoblast subpopulations as key players. These insights open avenues for targeted biomarkers and chronotherapy in preterm birth prevention.

Aberrant changes in circulating cell-free mitochondrial DNA (ccf-mtDNA) across gestation are associated with adverse pregnancy outcomes. Given the inflammatory properties of ccf-mtDNA via pattern recognition receptors such as Toll-like receptor 9 (TLR9), we hypothesized that extracellular mtDNA induces placental inflammation via TLR9 signaling and that this response differs by fetal sex. Pregnant Sprague-Dawley rats were treated intravenously with purified mtDNA (300 g/kg), nuclear DNA (nDNA), saline, and/or the TLR9 antagonist ODN2088 across five studies. Placental responses were evaluated 4 h (Studies 1-3) and 24 h (Study 4) post-treatment; pregnancy and neonatal outcomes were assessed at delivery (Study 5). Exposure to mtDNA, but not nDNA, increased placental il1{beta}, tnf, and il10 mRNA (p < 0.05), establishing response specificity. mtDNA-induced placental inflammation was fetal sex-dependent: mtDNA increased il6 and il1{beta} mRNA in male placentas (p [&le;] 0.0004) but not female placentas, whereas ifn{gamma} was selectively induced in female placentas (p = 0.0004). TLR9 and MyD88 abundance increased in female but not male placentas, and TLR9 antagonism modified selected inflammatory responses with sex-specific patterns. The 4 h inflammatory transcriptional signature resolved by 24 h, whereas mtDNA exposure was associated with a sex-specific shift in antioxidant enzyme expression persisting to 24 h. Despite no effects on gestational length or neonatal biometrics, mtDNA exposure was associated with a higher estimated stillbirth count per litter (IRR = 4.23, 95% CI [0.89, 20.1], p = 0.069). These findings establish extracellular mtDNA as an acute, sex-differentiated placental inflammatory stimulus with partial TLR9 dependence and a potential impact on fetal viability. New & NoteworthyThis study demonstrates that acute exposure to extracellular mtDNA induces placental inflammatory responses in vivo. This response is specific to mtDNA, fetal-sex dependent, and partially mediated by TLR9, with male and female placentas engaging distinct inflammatory signals within hours of exposure. The biological effects extend beyond the initial inflammatory window, with mtDNA exposure producing lasting, sex-specific changes in antioxidant enzyme expression. mtDNA-exposed dams had higher expected stillbirth counts, suggesting extracellular mtDNA may affect fetal viability.

BackgroundPrenatal cannabis use is becoming increasingly more commonplace. However, cannabis exposure is linked to adverse pregnancy outcomes, including gestational hypertension, preeclampsia, and preterm birth. The aim of this study was to determine the morphological and molecular effects of prenatal cannabinoid exposure on the placenta. MethodsPregnant Sprague-Dawley rats were exposed daily to vaporized THC (100 mg/mL) starting at gestational day (GD)5 until GD19 when dams were sacrificed and fetuses and placentas collected. Fetuses were genotyped for genetic sex and transcriptomic analysis was performed on male and female THC-exposed and control placentas. ResultsOn GD19, both the fetuses and placentas from the THC group were significantly larger than the control. When separated by sex, both male and female THC fetuses were significantly larger; however, only male THC placentas were significantly larger than male control placentas with no significant difference in placental weight between female control and THC placentas. RNA-sequencing revealed enriched biological processes related to nutrient transport and lipid catabolism, protein-lipid complex formation, and lipoprotein particle remodeling and organization. Further transcriptomic analysis determined that the differentially expressed genes and enriched biological processes related to lipid metabolism were preferentially enriched in the female THC placentas compared to the male, suggesting a sex-specific effect. DiscussionCollectively, these data present sex-specific effects of prenatal cannabinoid exposure on placental growth and global gene expression. These data also suggest that sex influences gene expression of genes related to lipid metabolism in the THC-exposed placentas.

Background The placenta serves a critical role in nutrient uptake and waste elimination for the developing fetus. The placenta is also responsible for the uptake and/or exchange of xenobiotics, including medications, between the maternal and fetal bloodstreams. An estimated 40-80% of women take medications or drugs during pregnancy for a variety of conditions. Very little is understood about fetal drug and nutrient exposure during pregnancy and how it may change over the course of fetal development. Objective This study aimed to characterize the abundance of transport proteins in placental tissue, which are important in modulating fetal nutrient and drug exposure, over the duration of pregnancy. Mass spectrometry-based global proteomic analysis revealed trends in the expression of thousands of proteins throughout gestation. Focusing on the membrane-associated proteome enabled an increased emphasis on the solute carrier and ATP-binding cassette families of transporter proteins that are critical for nutrient and xenobiotic transport across the maternal-fetal barrier. Study Design Using data-independent acquisition proteomics, relative abundance of proteins in placental tissue samples was profiled across all three trimesters of pregnancy (Trimester 1 = 16, Trimester 2 = 9, and Term = 9). Membrane fractions were generated to enrich membrane-associated proteins for proteomic analysis. Placental samples were grouped into randomized batches for membrane fraction generation and mass spectrometry analysis. Proteomic search results from each batch were imported into the R programming environment from Skyline, concatenated, and normalized as one data set for downstream analysis. Results A total of 6,331 proteins were detected across all samples with 4,210 proteins identified in every sample. Pathway analysis revealed that as gestational age increases, membrane-associated proteins involved in more complex metabolic pathways increase in relative abundance while those involved in extracellular remodeling events and simple organic ion transport tended to decrease. A total of 139 solute carrier and ATP-binding cassette transport proteins were identified in all samples, and 80 were identified in every sample. In general, membrane-associated proteins, including solute carrier and ATP-binding cassette transport proteins, were significantly enriched in placental tissue collected during early gestation compared to term placental tissue. Conclusion This study presents a comprehensive profiling of membrane-associated proteomic changes during gestation and identifies significant gestational age associated abundance changes at the protein level in several transport protein families. The application of data-independent acquisition global proteomic techniques enabled in-depth analysis of thousands of proteomic changes across pregnancy in a single experiment. These data provide critical information to support future studies into the understanding of fetal exposure to xenobiotics and nutrients circulating in the maternal bloodstream.

ObjectiveTo develop and validate a cell-free DNA (cfDNA) fragmentomic classifier for the early prediction of spontaneous preterm birth (PTB) using routine first-trimester non-invasive prenatal testing (NIPT) data. MethodsA nested case-control study was conducted within a prospective multicenter Vietnamese cohort comprising 286 pregnancies, including 82 spontaneous PTB cases and 204 term controls. Maternal plasma cfDNA collected during routine first-trimester NIPT (median gestational age, 12 weeks) was sequenced to a depth of approximately 20 million reads per sample. Five fragmentomic feature categories including copy number alterations, end-motif composition, nucleosome distance, fragment length, and joint fragment-lengthxend-motif were evaluated for PTB prediction. Machine learning classifiers were developed in a training cohort (n = 228, 65 PTB vs 163TB) and tested in a validation cohort (n = 58, 17 PTB vs 41 TB). ResultsAmong the five fragmentomic feature classes evaluated, 4-mer end-motif (EM) profiles exhibited the most pronounced differences between PTB and term control samples. Consistent with these findings, the EM-based classifier demonstrated the highest discriminative performance in the validation cohort, achieving an AUC of 0.970 (95% CI, 0.912-1.000). At a specificity >90%, the model achieved a sensitivity of 94% (95% CI, 78-100%). ConclusionThese findings demonstrate that cfDNA EM signatures derived from routine first-trimester NIPT can accurately identify pregnancies at risk of spontaneous preterm birth, without additional blood collection or sequencing, thereby extending the clinical utility of existing prenatal screening infrastructure. KEY POINTSO_ST_ABSWhat is already known about this topic?C_ST_ABSO_LICurrent first-trimester prediction strategies based on maternal characteristics, cervical length, and biochemical markers have limited predictive accuracy, particularly in nulliparous women. C_LIO_LIExisting cfDNA-based approaches have shown only modest performance or require additional assays, limiting clinical applicability. C_LI What does this study add?O_LIExisting NIPT sequencing data can be repurposed (without additional blood sampling or sequencing) for accurate prediction of spontaneous preterm birth (AUC=0.970). C_LIO_LIA classifier employing 4-mer end-motif (EM) profiles achieved an AUC of 0.970. At a specificity >90%, the model achieved a sensitivity of 94%. C_LI

BackgroundThe physical properties of leukocytes, such as cell size and stiffness, are critical for their circulation in microcapillary networks where rapid shape changes are required to squeeze through vascular constrictions. Alterations in the cells physical phenotype can promote venous thromboembolism (VTE), a common cause of death in cancer patients receiving chemotherapy. While biochemical VTE predictors are well studied, physical properties of blood cells receive less attention. MethodsUsing real-time deformability cytometry (RT-DC), we monitored for the first time the physical phenotype of leukocytes in a longitudinal study of a breast cancer patient treated with epirubicin/cyclophosphamide (EC) and paclitaxel (Pax). ResultsThe leukocyte counts extracted from RT-DC were in good agreement with standard clinical leukograms and EC had no immediate effect on leukocyte properties. However, Pax caused a significant softening of granulo/monocytes and a stiffening of lymphocytes immediately after administration. Leukocyte size was constant throughout the therapy, but we observed an overall increase in leukocyte stiffness, which was restored to normal values 45 weeks post treatment. ConclusionTaken together, our data reveal chemotherapy-induced specific alterations of leukocyte stiffness potentially critical for microcirculation. Thus, RT-DC measurements can add important, yet currently not available information to VTE prediction in cancer patients.

Autism development involves multiple genetic and early-life environmental factors. Studying the placenta's gene expression profile may reveal key mechanistic pathways in autism development. Here, using a nested case-cohort design within an Australian population-derived prebirth cohort study (n=1074), we identified 1,644 differentially expressed genes (DEGs; FDR<0.05) in the placenta of children with autism diagnosis (n=43), compared to those without (n=120). The top enriched pathways related to mitochondrial translation, oxidative stress, RNA processing and transcription regulation. CYP1A1, the most important xenobiotic-metabolising enzyme of the placenta, was the top downregulated DEG in the placenta of children with autism, while immuno-regulatory human leukocyte antigen (HLA)-related genes were among the top upregulated DEGs. A machine learning-based approach predicted autism from the transcriptomic data with a median sensitivity of 0.57 (2.5th-97.5th centiles: 0.29, 0.76) and median specificity of 0.92 (2.5th-97.5th centiles: 0.78, 0.98). Weighted Gene Correlation Network Analysis identified eight affected placental gene modules, with the largest five modules being enriched primarily for mitochondrial bioenergetics, oxidative phosphorylation and RNA processing pathways. This placental transcriptomic signature of impaired mitochondrial function and gene transcription regulation among infants subsequently diagnosed with autism has profound implications for understanding both risk factors and prediction, suggesting the possibility of identifying modifiable prenatal pathways to improve autism outcomes.

Background: Endothelial progenitor cells (EPCs) contribute to endothelial repair and neovascularization, and EPC dysfunction is closely associated with oxidative stress-related vascular injury. Forkhead box O3a (FoxO3a) regulates cellular stress responses, whereas miR-34a has been implicated in endothelial dysfunction, senescence, and apoptosis. However, the relationship between FoxO3a and miR-34a-3p in oxidatively injured EPCs remains incompletely defined. Objective: This study investigated the role of FoxO3a in H2O2-induced EPC dysfunction and examined whether miR-34a-3p directly interacts with the FoxO3a 3' untranslated region (3'UTR). Methods: Human umbilical cord blood-derived EPCs were identified by DiI-ac-LDL uptake, FITC-UEA-1 binding, and the expression of EPC-related markers. Oxidative stress was induced by H2O2. Cell viability, apoptosis, and angiogenic capacity were evaluated using CCK-8 assay, Annexin V/7-AAD flow cytometry, and Matrigel tube formation assay, respectively. FoxO3a expression was modulated using adenoviral overexpression or knockdown vectors, and miR-34a was modulated using mimics or antagomir. FoxO3a and miR-34a expression levels were detected by Western blot and qPCR. A dual-luciferase reporter assay was used to verify the interaction between hsa-miR-34a-3p and the FoxO3a 3'UTR. Results: H2O2 reduced EPC viability, increased apoptosis, and impaired tube formation in a concentration-dependent manner. H2O2 increased FoxO3a protein abundance and miR-34a expression, whereas FoxO3a mRNA did not change markedly. FoxO3a overexpression aggravated, whereas FoxO3a knockdown partially alleviated, H2O2-induced EPC dysfunction. Similarly, miR-34a mimics further suppressed EPC viability and tube formation, while miR-34a antagomir exerted a protective effect. Dual-luciferase reporter analysis showed that hsa-miR-34a-3p significantly reduced the activity of the wild-type FoxO3a 3'UTR reporter, while mutation of the predicted binding site abolished this suppression. Conclusion: FoxO3a and miR-34a participate in oxidative stress-induced EPC dysfunction. The dual-luciferase data demonstrate that hsa-miR-34a-3p directly targets the FoxO3a 3'UTR, suggesting the presence of miR-34a-3p-mediated post-transcriptional feedback within the FoxO3a-related stress-response network in EPCs.

Preterm birth is the leading cause of neonatal death. Despite sustained efforts to identify high-risk women in the mid-trimester, accurate prediction remains difficult. Quantitative cervical ultrasound texture has been proposed as a predictor of spontaneous preterm birth. However, earlier models were developed in small single-centre samples and were not externally validated. We developed image-texture (Local Binary Patterns with a Random Forest), deep-learning (Vision Transformer), clinical-variable, and multimodal models to predict spontaneous preterm birth on the prospective GARBH-Ini cohort. We then externally validated our best models on an independent cohort scanned on a different ultrasound machine. Our best overall model reached an internal-test area under the receiver-operating-characteristic curve of 0.71 (95% CI 0.60, 0.82), but performed modestly at 0.52 (95% CI 0.38, 0.64) externally. The deep-learning and multimodal models did not perform better. Discrimination appeared higher in a clinically high-risk subgroup at the 34-week threshold. These estimates were imprecise because of few cases and need to be confirmed in future studies. Among the several likely reasons for the modest external performance is the heterogeneity of preterm birth. Predicting distinct preterm-birth subtypes separately, and integrating additional biomarkers and data domains, might improve model performance. Keywords: preterm birth; cervical ultrasound; prediction model; external validation; deep learning

Proliferation of cytotrophoblasts (CTBs) and their differentiation into invasive extravillous trophoblasts (EVTs) are critical processes in early placental development. Defects in these processes are associated with adverse pregnancy outcomes, including recurrent pregnancy loss (RPL). There is evidence that reduced expression of the maintenance DNA methyltransferase DNMT1 in the placenta occurs in pregnancy loss and that RPL is associated with aberrant DNA methylation patterns. Therefore, we investigated the role of DNMT1 in human trophoblast growth and differentiation. Using human trophoblast stem cells (hTSCs), an in vitro analog to CTBs, we found that shRNA-mediated knockdown of DNMT1 led to decreased hTSC proliferation, genome-wide reductions in methylation, broad changes in gene expression, and impaired EVT differentiation. Transcriptome profiling of DNMT1-deficient hTSCs and hTSC-derived EVTs highlighted aberrant cytokine expression, drawing a connection to prior reports of immunological dysfunction in RPL. Finally, using a catalytic DNMT1 chemical inhibitor, we demonstrate the canonical methyltransferase activity of DNMT1 is essential for EVT differentiation and invasion. This study identifies new roles for DNMT1 in trophoblasts and addresses the molecular basis of the associations between DNMT1 expression, altered DNA methylation profiles, and RPL.

Background: Prospective pregnancy registries and biobanking infrastructures are essential for future translational studies investigating maternal, placental and offspring health. However, circulating nucleic acid analyses are highly sensitive to preanalytical variability, particularly regarding blood-collection tube type and sample processing conditions. We established a prospective pregnancy registry and biobanking workflow at Cruces University Hospital and evaluated the impact of preanalytical variables on circulating cell-free DNA (cfDNA) and cell-free RNA (cfRNA) preservation in maternal plasma collected at delivery. Methods: The Registry of Pregnant Women at Cruces University Hospital was designed as a prospective infrastructure integrating placental sampling, maternal blood collection and ethically controlled future access to maternal and offspring clinical data. Within this framework, peripheral blood samples from 50 women at delivery were simultaneously collected into EDTA, Norgen and Roche tubes. Plasma samples processed within or after 24 hours following collection underwent cfDNA/cfRNA extraction, electrophoretic profiling, fluorometric quantification and RT-qPCR analyses targeting different stress-related genes. Results: By the end of June 2026, 1,127 women had been prospectively recruited into the registry, with 661 plasma samples, 637 serum samples and 858 sets of four placental biopsies collected, processed and stored in the Basque Biobank. In the preanalytical substudy, EDTA tubes yielded higher cfDNA concentrations, likely reflecting reduced cellular preservation and genomic DNA contamination. In contrast, Roche tubes showed superior cfRNA preservation, with higher cfRNA concentrations and more consistent detection of the characteristic 5S rRNA peak compared with EDTA and Norgen tubes. Processing delays beyond 24 hours reduced cfRNA concentration, while associations between circulating transcripts and gestational age were more consistently detectable in preservative-containing tubes. Conclusions: Prospective infrastructures like ours offer strong foundation for large scale, long-term studies in the framework of the Developmental Origins of Health and Disease hypothesis. Technically, Roche tubes provided superior cfRNA preservation and enhanced sensitivity for detecting subtle biological associations, supporting the importance of standardized preanalytical workflows within prospective pregnancy biobanking resource.

Objective To evaluate the feasibility, safety, patient acceptance, and preliminary clinical relevance of automated self-operated transvaginal ultrasound for ovarian stimulation monitoring. Design Prospective observational pilot study. Subjects Ten women undergoing ovarian stimulation for in vitro fertilization or fertility preservation at a single high-volume private IVF center. Exposure Participants performed investigational self-operated transvaginal ultrasound examinations immediately following standard monitoring visits. Patients inserted and stabilized the ultrasound probe while ovarian and endometrial imaging was acquired through controlled motorized probe rotation without real-time anatomical guidance. Main Outcome Measure(s) The primary outcome was feasibility, defined as the generation of evaluable imaging datasets suitable for ovarian stimulation monitoring. Secondary outcomes included bilateral ovarian visualization, procedural safety, patient-reported outcomes, follicular assessment, and agreement of endometrial thickness measurements with standard transvaginal ultrasound. Result(s) Nineteen investigational scan attempts were performed, yielding 18 evaluable datasets (94.7%). Bilateral ovarian visualization was achieved in 16 of 18 evaluable examinations (88.9%), whereas partial ovarian visualization occurred in 2 examinations (11.1%). No adverse events, adverse device effects, vaginal injury, bleeding, or infection were observed. Patient-reported outcomes demonstrated high procedural acceptability, with all participants expressing willingness to reuse the system. Compared with standard transvaginal ultrasound monitoring, investigational self-operated acquisition significantly improved overall examination experience (Wilcoxon p=0.002). Investigational imaging demonstrated clinically relevant agreement with standard transvaginal ultrasound for follicular categorization and endometrial assessment. Counts of follicles [&ge;]14 mm correlated strongly with mature oocyte recovery for both investigational and standard ultrasound measurements (Spearman {rho}=0.83 and {rho}=0.80, respectively). Endometrial thickness measurements also demonstrated strong correlation between modalities (Spearman {rho}=0.91). Conclusion(s) This prospective pilot study demonstrates the feasibility of automated self-operated transvaginal ultrasound during ovarian stimulation monitoring. Investigational imaging generated clinically relevant monitoring information without observed safety concerns and was associated with high patient acceptance. These findings support further investigation of patient-operated acquisition strategies and standardized imaging workflows in reproductive medicine.

Background. Low-birth-weight (LBW) neonates carry a disproportionate share of newborn morbidity and mortality in sub-Saharan Africa. Distinguishing which maternal and neonatal characteristics mark the highest risk supports bedside risk stratification in resource-limited newborn units. This study examined maternal and neonatal predictors of severe adverse outcomes among LBW neonates admitted to a county referral hospital in Kenya. Methods. A facility-based cross-sectional analysis was conducted on 169 LBW neonate-mother pairs admitted to the newborn unit at Kericho County Referral Hospital. The outcome was a severe adverse outcome, defined as a composite of respiratory distress, sepsis, hypothermia, hypoglycaemia, prolonged admission (>= 7 days), or neonatal death. Maternal and neonatal factors were screened using chi-square, Fisher's exact, and independent-samples t tests, with crude odds ratios (ORs) and 95% confidence intervals (CIs). A restricted multivariable logistic regression, limited to maternal and neonatal predictors, produced adjusted odds ratios (AORs). Results. Severe adverse outcomes occurred in 136 of 169 neonates (80.5%). At the bivariate level, pregnancy-induced hypertension (PIH; OR = 6.69, 95% CI 1.53-29.27, p = 0.004) and preterm birth (OR = 3.79, 95% CI 1.50-9.56, p = 0.003) were associated with higher odds of a severe outcome, and severe outcomes clustered at lower birth weight and gestational age and higher neonatal risk-factor counts (all p < 0.001). In the restricted adjusted model, PIH (AOR = 7.66, 95% CI 1.56-37.55, p = 0.012) and birth weight (AOR = 0.997 per gram, p = 0.002) retained independent significance. Conclusion. Neonatal biological vulnerability-particularly lower birth weight-together with maternal pregnancy-induced hypertension were the predictors most strongly and independently associated with severe adverse outcomes. Risk stratification of LBW neonates should prioritise the smallest infants and those born to mothers with hypertensive disease. Keywords: low birth weight; neonatal outcomes; pregnancy-induced hypertension; preterm birth; risk stratification; Kenya

Bovine embryo in vitro production (IVP) is characterised by low efficiency and variable outcomes. Monitoring early embryonic development by videomicroscopy revealed substantial morphokinetic heterogeneity in the first four embryonic cycles (EC, conventionally referred to as the 2-, 4-, 8- and 16-cell stages). Morphokinetic analysis offers a promising approach to characterize divergent developmental trajectories and provide a better understanding of underlying molecular mechanisms. We developed a Random Forest classification system (Bovine Embryo Analyser based on Morphokinetics: BEAM) to predict embryo phenotype. It is based on morphokinetic variables collected from the 1st to the 4th EC and predicts four blastocyst categories (EHB: Early Hatching Blastocyst, HB: Hatching Blastocyst, SSB: Subtle Developmental Shift Blastocyst, ADB: Arrhythmic Development Blastocyst). Classification performance on an independent dataset was good (F1 score = 0.59; Accuracy = 0.78), indicating that the BEAM can be useful for embryo development studies. The BEAM was further applied to embryos submitted to 4.3 days of culture and having completed the 4th EC (16-32 cells). RNA sequencing was performed on sixteen samples (4 x 8 pooled embryos/category). The ADB category was significantly enriched in transcripts involved in the regulation of transcriptional activity compared to the EHB category. In addition, in the ADB category, 22.7% (n = 185/816) of the upregulated genes were of maternal origin while only 6.2% were of embryonic origin (n = 54/816). In conclusion, despite being at a comparable developmental stage and transcriptionally competent, the ADB category showed delayed maternal transcript degradation suggesting delayed transition to embryonic transcriptional autonomy. Illustrated Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=138 SRC="FIGDIR/small/733532v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@13a0958org.highwire.dtl.DTLVardef@13b88a1org.highwire.dtl.DTLVardef@18707c9org.highwire.dtl.DTLVardef@105759_HPS_FORMAT_FIGEXP M_FIG C_FIG Summary SentenceIn vitro produced bovine embryo morphokinetic patterns allow prediction of four blastocyst categories (up to the 4th embryonic cycle) and are associated with distinct transcriptomic profiles at embryonic genome activation in competent embryos.

PurposeComputational models of the uterus during pregnancy enable analysis of electro-chemo-mechanical pathways to predict labor timing and guide treatment planning. We aim to develop a robust image-based modeling pipeline to investigate uterine passive mechanics during late pregnancy. MethodsA parametric model of the uterus and cervix was created using a patients MRI measurements at 38 weeks of gestation. Inspired by advances in cardiac mechanics models, we created Laplace-Dirichlet solutions to inform tissue domains, fiber structure within the uterus and cervix, and spatially varying Robin boundary conditions. Prior imaging and mechanical testing data were used to fit material parameters. Boundary condition parameters were tuned to match the displacements of a previously established approach that employed contact with surrounding tissue. The tissue mechanical response to a physiologic load was assessed across varying material properties and fiber architectures. ResultsDiscrepancies in nodal displacements between the current approach and the contact-based model were limited to 3.4 {+/-} 1.8 mm, yielding nearly 90 % computational savings. Uterine tensile strains were more sensitive to ground substance elastic modulus (E) compared to fiber properties. Reduced E and fiber stiffness increased cervical strains and compression. Fiber dispersion and architecture modulated the opening of the cervical internal ostium but had a reduced impact on compression. ConclusionWe developed a novel workflow for modeling passive uterine mechanics, informed by patient-specific measurements and in vitro mechanical tests. The robust workflow may prove useful for studying labor progression and conducting longitudinal studies to enhance our understanding of normal and pathological pregnancies.

Abstract Background: Acute Lymphoblastic Leukemia (ALL) is a highly heterogeneous pediatric malignancy. Despite high survival rates, relapse and the involvement of central nervous system (CNS) remains a significant clinical challenge. Traditional clinical parameters often lack the precision required for early detection and risk stratification. This study utilizes high-throughput proteomics and machine learning to identify molecular signatures in cerebrospinal fluid (CSF) that characterize disease effect and treatment response. Methods: 82 CSF samples from 41 pediatric ALL patients at diagnosis (VD) and remission (VR) were analyzed. Proteomic profiling of 276 proteins was performed using Olink Proximity Extension Assay. Differentially abundant proteins were identified (q-value< 0.05, |Log_2FC| > 0.5) using the Wilcoxon rank-sum test. Three machine-learning algorithms - Random Forest, LASSO, and SVM-RFE - were integrated to select the differentially abundant proteins in VR and VD and between CNS involvement levels. To validate the data Pan-Cancer Atlas analysis was done using two different platforms. Results: In the remission phase, we observed significant alterations in the expression of key proteins compared to diagnosis, with ADGRG1 and KYNU showing a marked increase, while CCL17, CD5, CD27, CXCL9, CXCL11, FASLG, GZMA, and TNFRSF9 were significantly downregulated. Furthermore, our analysis identified distinct protein signatures associated with CNS involvement: CCL4, CTSC, CXCL10, CXCL9, and MMP7 were differentially abundant at the VD stage, whereas CAIX, CASP-8, HAGH, CXCL9, MMP7, MCP-2, and VWC2 at the VR stage. Conclusion: Integrating Olink proteomics with machine learning identified molecular signatures in ALL that have the potential to be further developed to a biomarker panel for monitoring treatment response and guiding personalized therapeutic strategies shifting the focus toward the Precision One Health approaches.

Background Neural tube defects (NTDs) are preventable congenital malformations that disproportionately affect low and middle income countries and contribute to disability and mortality. Evidence on the long-term outcomes of children and adolescents with NTDs in Eastern Africa has not been comprehensively synthesised. We conducted a systematic review and meta-analysis to assess survival, complications, and functional outcomes among children with NTDs in this region. Methods We searched PubMed, MEDLINE (Ovid), Cochrane Library, Web of Science, and Africa Index Medicus from the earliest records to April 2025. Two reviewers independently screened studies, assessed quality, and extracted data. We included studies reporting outcomes beyond one year of age, except mortality, which was assessed from birth onwards. Random-effects meta-analyses were undertaken when at least two sufficiently studies were available; otherwise, findings were synthesised narratively. Results Of 597 articles screened, 16 studies involving 2,340 children with NTDs met the inclusion criteria. Pooled cumulative mortality was 23% (95% CI: 12 - 36%) in the neonatal period, 9% (95% CI: 2 - 33%) during infancy, 22% (95% CI: 16 - 28%) in toddlers, 37% (95% CI: 30 - 44%) in pre school aged children and 45% (95% CI: 36 - 54%) in school-aged children. The pooled prevalence of hydrocephalus was 41% (95% CI: 34 - 48%) with little variation by age. Neurogenic bladder increased from 53% (95% CI 49 - 81%) in toddlers to 83% (95 % CI 72 - 91%) in adolescents, while impaired mobility affected about 61% (95% CI 48 - 72%) of adolescents. School enrolment was 53% (95% CI: 39 - 67%), with 9% (95% CI: 4% - 19%) in specialized education. Speech, hearing, and bowel dysfunction were understudied (< 2 studies each). Conclusion Many children with NTDs in the Eastern African region survive beyond infancy but frequently experience hydrocephalus, neurogenic bladder and motor impairment. Longitudinal studies, context-specific guidelines, and follow-up systems are urgently needed to improve care and long-term outcomes.

Humans and other vertebrates contain two estrogen receptors (ERs), ER-alpha and ER-beta, which mediate the physiological actions of three estrogens: estrone (E1), estradiol (E2) and estriol (E3). Of these three estrogens, in vivo, E2 is the strongest transcriptional activator of ER-alpha and ER-beta, E1 is next most active, followed by E3. We studied transcriptional activation of human ER-alpha and ER-beta by E2, E1 and E3 in African green monkey kidney (COS-7) cells, which we compared with studies of estrogen stimulation of ER transcription in human em-bryonic kidney (HEK-293) cells. To our surprise, in COS-7 cells, E3 had the lowest half-maximal response (EC50) for human ER-alpha and ER-beta than either E2, which was second most active estrogen, or E1. In contrast, for human ER-alpha and ER-beta transfected into HEK-293 cells, E2 was the most active estrogen, followed by E1 and E3. Similar results were found in COS-7 cells and HEK-293 cells transfected with elephant shark ER-alpha and ER-beta. Thus, under some conditions, E3 is a more active estrogen than either E2 or E1. This suggests that E3 may be a novel physiological ligand for the ER in some mammalian cells.

Background: Postpartum anemia (PPA) is defined as a hemoglobin concentration below 11 g/dL within the first week following delivery, or below 12 g/dL at eight weeks postpartum. It constitutes a major yet under-addressed public health burden in sub-Saharan Africa, contributing substantially to maternal morbidity and mortality. In Ethiopia, anemia prevalence among women of reproductive age has been increasing despite national nutrition strategies, yet aggregated, nationally representative data on PPA remain scarce. This systematic review and meta-analysis aimed to estimate the pooled prevalence of anemia among postpartum mothers attending public health facilities in Ethiopia and to identify significantly associated factors. Methods: A comprehensive literature search was conducted across PubMed/MEDLINE, Cochrane Library, Google Scholar, African Journals Online (AJOL), and HINARI from inception to December 2024. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA 2020) guidelines were followed. Observational studies (cross-sectional, cohort) conducted in public health facilities in Ethiopia, reporting hemoglobin-confirmed PPA prevalence and/or associated factors, were included. Two reviewers independently performed study selection, data extraction, and quality assessment using a modified Newcastle-Ottawa Scale (NOS) adapted for cross-sectional studies. Heterogeneity was quantified using the Cochran Q statistic and I-squared index. Pooled prevalence estimates and odds ratios were computed using a random-effects (DerSimonian-Laird) model with 95% confidence intervals. Subgroup analyses were performed by geographic region (Eastern vs. Western/Central Ethiopia), study period, and facility type. Publication bias was assessed via Egger weighted regression test and funnel plot asymmetry. Results: Six primary studies, encompassing 2,819 postpartum mothers, fulfilled the eligibility criteria and were included in the meta-analysis. The pooled prevalence of anemia among postpartum mothers in Ethiopian public health facilities was 35.4% (95% CI: 27.6-43.6%; I2 = 89.2%), classified as a severe public health problem per WHO thresholds. Subgroup analysis revealed higher prevalence in Eastern Ethiopia (Dire Dawa and Harari regions: 27.5%) compared to Western/Central Ethiopia (Gondar and Debre Markos: 24.3-47.1%), though marked heterogeneity was observed. Factors significantly associated with higher odds of PPA included: fewer than four antenatal care (ANC) visits (pooled OR = 2.72; 95% CI: 2.14-3.30), history of postpartum hemorrhage (OR = 2.49; 95% CI: 1.08-3.98), cesarean section delivery (OR = 4.04; 95% CI: 3.43-4.67), instrumental delivery forceps/vacuum (OR = 3.96; 95% CI: 2.99-4.95), poor adherence to iron and folic acid (IFA) supplementation (OR = 2.80; 95% CI: 2.31-3.30), low pre-delivery hemoglobin < 11 g/dL (OR = 4.20; 95% CI: 1.77-6.67), low dietary diversity (OR = 4.20; 95% CI: 1.77-6.67), and lack of formal education (OR = 3.50; 95% CI: 2.64-4.41). Conclusions: Nearly one in three postpartum mothers attending public health facilities in Ethiopia is anemic, constituting a severe public health emergency. The burden is driven by modifiable clinical and behavioral factors particularly inadequate ANC utilization, poor IFA adherence, hemorrhagic complications, and nutritional deficiencies as well as structural determinants including low educational attainment. Evidence-based, multi-sectoral interventions are urgently needed, including strengthening ANC quality and coverage, universal IFA supplementation monitoring, active postpartum hemorrhage management, and targeted nutritional counseling. Policy frameworks must address regional disparities and integrate anemia screening into routine postpartum care protocols.

Background: Iron deficiency (ID) affects up to 40% of pregnant women in the third trimester, even in highly resourced and iron-supplemented populations, with adverse consequences for maternal health and long-term offspring development. Psychological stress may compromise iron status through hypothalamic-pituitary-adrenocortical (HPA) axis dysregulation and inflammation, but no study has directly examined cortisol in relation to iron status across human pregnancy. Objective: This longitudinal study examined associations between HPA function and maternal iron status across pregnancy and tested whether IL-6 and CRP mediated the relationship between cortisol and ferritin across gestation. Methods: One hundred sixty-eight pregnant Black women with Medicaid insurance completed up to four laboratory assessments across pregnancy. Salivary cortisol was measured before and in response to the Trier Social Stress Test, yielding basal and reactive cortisol indices. Serum ferritin, IL-6, and CRP were collected at each visit. Trimester-specific regression models examined cortisol reactivity in relation to ferritin; linear mixed-effects models with moderated mediation tested whether basal cortisol predicted ferritin via inflammation. Results: Higher cortisol reactivity was associated with lower ferritin specifically in the third trimester (std. {beta} = -0.197, p = .004). Higher basal cortisol predicted a steeper IL-6 rise across gestation (p = .002), and IL-6 was positively associated with ferritin (b = 0.236, p = .006), consistent with inflammatory iron sequestration. The indirect effect of basal cortisol on ferritin via IL-6 was statistically significant, and higher basal cortisol was negatively associated with cortisol reactivity in the third trimester. No pathway was observed through CRP. Conclusion: Greater cortisol reactivity predicted lower third-trimester ferritin, a pattern that suggests cumulative iron depletion, atypically sustained HPA reactivity in late pregnancy, or both. To our knowledge, this is the first prospective study linking cortisol reactivity to iron status across human pregnancy, identifying maternal stress physiology as a novel target for understanding and addressing gestational iron deficiency.