Aging promotes accumulation of senescent and multiciliated cells in human endometrial epithelium.

STUDY QUESTION: What changes occur in the endometrium during aging, and do they impact fertility? SUMMARY ANSWER: Both the transcriptome and cellular composition of endometrial samples from women of advanced maternal age (AMA) are significantly different from that of samples from young women, suggesting specific changes in epithelial cells that may affect endometrial receptivity. WHAT IS KNOWN ALREADY: Aging is associated with the accumulation of senescent cells in aging tissues. Reproductive aging is mostly attributed to the decline in ovarian reserve and oocyte quality, whereas the endometrium is a unique complex tissue that is monthly renewed under hormonal regulation. Several clinical studies have reported lower implantation and pregnancy rates in oocyte recipients of AMA during IVF. Molecular studies have indicated the presence of specific mutations within the epithelial cells of AMA endometrium, along with altered gene expression of bulk endometrial tissue. STUDY DESIGN SIZE DURATION: Endometrial transcriptome profiling was performed for 44 women undergoing HRT during the assessment of endometrial receptivity before IVF. Patients younger than 28 years were considered as the young maternal age (YMA) group (age 23-27 years) and women older than 45 years were considered as the AMA group (age 47-50 years). Endometrial biopsies were obtained on Day 5 of progesterone treatment and RNA was extracted. All endometrial samples were evaluated as being receptive based on the expression of 68 common endometrial receptivity markers. Endometrial samples from another 24 women classified into four age groups (YMA, intermediate age group 1 (IMA1, age 29-35), intermediate age group 2 (IMA2, age 36-44), and AMA) were obtained in the mid-secretory stage of a natural cycle (NC) and used for validation studies across the reproductive lifespan. PARTICIPANTS/MATERIALS SETTING METHODS: A total of 24 HRT samples (12 YMA and 12 AMA) were subject to RNA sequencing (RNA-seq) and differential gene expression analysis, 20 samples (10 YMA and 10 AMA) were used for qPCR validation, and 24 NC samples (6 YMA, 6 IMA1, 6 IMA2 and 6AMA) were used for RNA-seq validation of AMA genes across the woman's reproductive lifespan. Immunohistochemistry (IHC) was used to confirm some expression changes at the protein level. Computational deconvolution using six endometrial cell type-specific transcriptomic profiles was conducted to compare the cellular composition between the groups. MAIN RESULTS AND THE ROLE OF CHANCE: Comparisons between YMA and AMA samples identified a lower proportion of receptive endometria in the AMA group (P = 0.007). Gene expression profiling identified 491 differentially expressed age-sensitive genes (P adj < 0.05) that revealed the effects of age on endometrial epithelial growth and receptivity, likely contributing to decreased reproductive performance. Our results indicate that changes in the expression of the cellular senescence marker p16INK4a and genes associated with metabolism, inflammation, and hormone response are involved in endometrial aging. Importantly, we demonstrate that the proportion of multi-ciliated cells, as discovered based on RNA-seq data deconvolution and tissue IHC results, is affected by endometrial aging, and propose a putative onset of age-related changes. Furthermore, we propose that aging has an impact on the transcriptomic profile of endometrial tissue in the context of endometrial receptivity. LARGE SCALE DATA: The raw sequencing data reported in this article are deposited at the Gene Expression Omnibus under accession code GSE236128. LIMITATIONS REASONS FOR CAUTION: This retrospective study identified changes in the endometrium of patients undergoing hormonal replacement and validated these changes using samples obtained during a NC. However, future studies must clarify the importance of these findings on the clinical outcomes of assisted reproduction. WIDER IMPLICATIONS OF THE FINDINGS: The findings reported in this study have important implications for devising future strategies aimed at improving fertility management in women of advanced reproductive age. STUDY FUNDING/COMPETING INTERESTS: This research was funded by the Estonian Research Council (grant no. PRG1076), Horizon 2020 innovation grant (ERIN, grant no. EU952516), Enterprise Estonia (grant no. EU48695), MSCA-RISE-2020 project TRENDO (grant no. 101008193), EU 874867 project HUTER, the Horizon Europe NESTOR grant (grant no. 101120075) of the European Commission, the EVA specialty program (grant no. KP111513) of the Maastricht University Medical Center (MUMC+), MICIU/AEI/10.13039/501100011033 and FEDER, EU projects Endo-Map (grant no. PID2021-12728OB-100), ROSY (grant no. CNS2022-135999), and the National Science Fund of Bulgaria (grant no. KII-06 H31/2). The authors declare no competing interests.

Giant urinary bladder stone: A rare case report.

INTRODUCTION: Giant urinary bladder stones are rare phenomenon which is associated with chronic urinary infections, intravesical foreign bodies, urethral strictures, bladder diverticula etc. CASE REPORT: A 52-year-old man presented with complaints of severe dysuria, urgency, frequency, suprapubic pain, and pollakuria for the last ten years. Physical examination revealed a palpable suprapubic mass with no obvious flank masses. Ultrasonography of the abdomen and pelvis revealed right-sided gross hydroureteronephrosis and thinning of renal parenchyma along with a hyperechoic structure with posterior acoustic shadowing was noted in the region of the urinary bladder, suggesting a vesical calculus. Plain CT of the kidneys, ureters, and bladder (KUB) confirmed right nephrolithiasis in lower poly calyx and a large vesical calculus (10.6 cm × 8.6 cm x8.8 cm, +1110 HU). Open cystolithotomy with a right-sided double-J "DJ" stent insertion was performed. Post-operative period went uneventful. DISCUSSION: Giant bladder stones are extremely rare, often resulting from neglected symptoms in otherwise normal individuals. They typically develop over several years and present with symptoms like severe dysuria, urgency, frequency, supra-pubic pain, and hematuria. Diagnosis is made by cystoscopy, ultrasonography and CT-KUB. Treatment includes intracorporeal cystolithotripsy using a laser, ultrasonic lithotripter, or pneumatic lithotripter, depending on availability. Endourologic procedures have been safer and more cost-effective for bladder stones, however, open removal is the treatment of choice for giant bladder stones. CONCLUSION: Open cystolithotomy can be performed to remove giant bladder stone with near 100 % stone removal rate.

Exploring the dynamics of situational interest in team-based learning in undergraduate medical education.

BACKGROUND: Team-based learning (TBL) is a widely recognized instructional approach in medical education blending direct instruction with active-cooperative learning in small groups. While TBL is known to enhance knowledge acquisition, its impact on student motivation, particularly through situational interest, remains underexplored. This study aimed to investigate the development of situational interest across the distinct phases of TBL, focusing on how each phase (individual readiness assurance test; iRAT, team readiness assurance test; tRAT, and application exercise; AE) influences students' situational interest. The study sought to provide insights into the motivational dynamics underpinning TBL in a medical education setting. METHODS: A total of 88 medical students participated in a TBL session on "Bleeding during Pregnancy." Situational interest was measured after each TBL phase. A one-way repeated-measures analysis of variance (ANOVA) was conducted to assess the fluctuation of situational interest throughout the session. RESULTS: The analysis revealed significant variations in situational interest across different TBL stages. There was a significant increase in situational interest following the tRAT (p = .001). Post-tRAT, situational interest significantly decreased after the AE (p = .007), returning to levels observed at the session's start. Post hoc correlation analysis suggested a negative association between tRAT performance and situational interest, indicating heightened interest in response to awareness of knowledge gaps during the tRAT. CONCLUSIONS: The findings of this study may challenge the traditional view of TBL, suggesting a more integrated and dynamic interplay between knowledge acquisition and application phases. The results highlight the importance of the AE phase in clinical education and suggest that situational interest is one key driver in the learning process within TBL. Future research should focus on replicating these findings and comparing situational interest development between pre-clinical and clinical student cohorts to further understand the effects of situational interest on TBL in medical education.

Citalopram exposure of hESCs during neuronal differentiation identifies dysregulated genes involved in neurodevelopment and depression.

Selective serotonin reuptake inhibitors (SSRIs), including citalopram, are widely used antidepressants during pregnancy. However, the effects of prenatal exposure to citalopram on neurodevelopment remain poorly understood. We aimed to investigate the impact of citalopram exposure on early neuronal differentiation of human embryonic stem cells using a multi-omics approach. Citalopram induced time- and dose-dependent effects on gene expression and DNA methylation of genes involved in neurodevelopmental processes or linked to depression, such as BDNF, GDF11, CCL2, STC1, DDIT4 and GAD2. Single-cell RNA-sequencing analysis revealed distinct clusters of stem cells, neuronal progenitors and neuroblasts, where exposure to citalopram subtly influenced progenitor subtypes. Pseudotemporal analysis showed enhanced neuronal differentiation. Our findings suggest that citalopram exposure during early neuronal differentiation influences gene expression patterns associated with neurodevelopment and depression, providing insights into its potential neurodevelopmental impact and highlighting the importance of further research to understand the long-term consequences of prenatal SSRI exposure.

CHK1 inhibitor induced PARylation by targeting PARG causes excessive replication and metabolic stress and overcomes chemoresistance in ovarian cancer.

Chemoresistance contributes to the majority of deaths in women with ovarian cancer (OC). Altered DNA repair and metabolic signaling is implicated in mediating therapeutic resistance. DNA damage checkpoint kinase 1 (CHK1) integrates cell cycle and DNA repair in replicating cells, and its inhibition causes replication stress, repair deficiency and cell cycle dysregulation. We observed elevated Poly-ADP-ribosylation (PAR) of proteins (PARylation) and subsequent decrease in cellular NAD+ levels in OC cells treated with the CHK1 inhibitor prexasertib, indicating activation of NAD+ dependent DNA repair enzymes poly-ADP-ribose polymerases (PARP1/2). While multiple PARP inhibitors are in clinical use in treating OC, tumor resistance to these drugs is highly imminent. We reasoned that inhibition of dePARylation by targeting Poly (ADP-ribose) glycohydrolase (PARG) would disrupt metabolic and DNA repair crosstalk to overcome chemoresistance. Although PARG inhibition (PARGi) trapped PARylation of the proteins and activated CHK1, it did not cause any significant OC cell death. However, OC cells deficient in CHK1 were hypersensitive to PARGi, suggesting a role for metabolic and DNA repair crosstalk in protection of OC cells. Correspondingly, OC cells treated with a combination of CHK1 and PARG inhibitors exhibited excessive replication stress-mediated DNA lesions, cell cycle dysregulation, and mitotic catastrophe compared to individual drugs. Interestingly, increased PARylation observed in combination treatment resulted in depletion of NAD+ levels. These decreased NAD+ levels were also paralleled with reduced aldehyde dehydrogenase (ALDH) activity, which requires NAD+ to maintain cancer stem cells. Furthermore, prexasertib and PARGi combinations exhibited synergistic cell death in OC cells, including an isogenic chemoresistant cell line and 3D organoid models of primary patient-derived OC cell lines. Collectively, our data highlight a novel crosstalk between metabolism and DNA repair involving replication stress and NAD+-dependent PARylation, and suggest a novel combination therapy of CHK1 and PARG inhibitors to overcome chemoresistance in OC.

Generative artificial intelligence to produce high-fidelity blastocyst-stage embryo images.

STUDY QUESTION: Can generative artificial intelligence (AI) models produce high-fidelity images of human blastocysts? SUMMARY ANSWER: Generative AI models exhibit the capability to generate high-fidelity human blastocyst images, thereby providing substantial training datasets crucial for the development of robust AI models. WHAT IS KNOWN ALREADY: The integration of AI into IVF procedures holds the potential to enhance objectivity and automate embryo selection for transfer. However, the effectiveness of AI is limited by data scarcity and ethical concerns related to patient data privacy. Generative adversarial networks (GAN) have emerged as a promising approach to alleviate data limitations by generating synthetic data that closely approximate real images. STUDY DESIGN, SIZE, DURATION: Blastocyst images were included as training data from a public dataset of time-lapse microscopy (TLM) videos (n = 136). A style-based GAN was fine-tuned as the generative model. PARTICIPANTS/MATERIALS, SETTING, METHODS: We curated a total of 972 blastocyst images as training data, where frames were captured within the time window of 110-120 h post-insemination at 1-h intervals from TLM videos. We configured the style-based GAN model with data augmentation (AUG) and pretrained weights (Pretrained-T: with translation equivariance; Pretrained-R: with translation and rotation equivariance) to compare their optimization on image synthesis. We then applied quantitative metrics including Fréchet Inception Distance (FID) and Kernel Inception Distance (KID) to assess the quality and fidelity of the generated images. Subsequently, we evaluated qualitative performance by measuring the intelligence behavior of the model through the visual Turing test. To this end, 60 individuals with diverse backgrounds and expertise in clinical embryology and IVF evaluated the quality of synthetic embryo images. MAIN RESULTS AND THE ROLE OF CHANCE: During the training process, we observed consistent improvement of image quality that was measured by FID and KID scores. Pretrained and AUG + Pretrained initiated with remarkably lower FID and KID values compared to both Baseline and AUG + Baseline models. Following 5000 training iterations, the AUG + Pretrained-R model showed the highest performance of the evaluated five configurations with FID and KID scores of 15.2 and 0.004, respectively. Subsequently, we carried out the visual Turing test, such that IVF embryologists, IVF laboratory technicians, and non-experts evaluated the synthetic blastocyst-stage embryo images and obtained similar performance in specificity with marginal differences in accuracy and sensitivity. LIMITATIONS, REASONS FOR CAUTION: In this study, we primarily focused the training data on blastocyst images as IVF embryos are primarily assessed in blastocyst stage. However, generation of an array of images in different preimplantation stages offers further insights into the development of preimplantation embryos and IVF success. In addition, we resized training images to a resolution of 256 × 256 pixels to moderate the computational costs of training the style-based GAN models. Further research is needed to involve a more extensive and diverse dataset from the formation of the zygote to the blastocyst stage, e.g. video generation, and the use of improved image resolution to facilitate the development of comprehensive AI algorithms and to produce higher-quality images. WIDER IMPLICATIONS OF THE FINDINGS: Generative AI models hold promising potential in generating high-fidelity human blastocyst images, which allows the development of robust AI models as it can provide sufficient training datasets while safeguarding patient data privacy. Additionally, this may help to produce sufficient embryo imaging training data with different (rare) abnormal features, such as embryonic arrest, tripolar cell division to avoid class imbalances and reach to even datasets. Thus, generative models may offer a compelling opportunity to transform embryo selection procedures and substantially enhance IVF outcomes. STUDY FUNDING/COMPETING INTEREST(S): This study was supported by a Horizon 2020 innovation grant (ERIN, grant no. EU952516) and a Horizon Europe grant (NESTOR, grant no. 101120075) of the European Commission to A.S. and M.Z.E., the Estonian Research Council (grant no. PRG1076) to A.S., and the EVA (Erfelijkheid Voortplanting & Aanleg) specialty program (grant no. KP111513) of Maastricht University Medical Centre (MUMC+) to M.Z.E. TRIAL REGISTRATION NUMBER: Not applicable.

Towards ultrafast quantitative phase imaging via differentiable microscopy [Invited].

With applications ranging from metabolomics to histopathology, quantitative phase microscopy (QPM) is a powerful label-free imaging modality. Despite significant advances in fast multiplexed imaging sensors and deep-learning-based inverse solvers, the throughput of QPM is currently limited by the pixel-rate of the image sensors. Complementarily, to improve throughput further, here we propose to acquire images in a compressed form so that more information can be transferred beyond the existing hardware bottleneck of the image sensor. To this end, we present a numerical simulation of a learnable optical compression-decompression framework that learns content-specific features. The proposed differentiable quantitative phase microscopy (∂-QPM) first uses learnable optical processors as image compressors. The intensity representations produced by these optical processors are then captured by the imaging sensor. Finally, a reconstruction network running on a computer decompresses the QPM images post aquisition. In numerical experiments, the proposed system achieves compression of × 64 while maintaining the SSIM of ∼0.90 and PSNR of ∼30 dB on cells. The results demonstrated by our experiments open up a new pathway to QPM systems that may provide unprecedented throughput improvements.

The outcomes of team-based learning vs small group interactive learning in the obstetrics and gynecology course for undergraduate students.

INTRODUCTION: Team-based learning (TBL) is a well-established active teaching method which has been shown to have pedagogical advantages in some areas such as business education and preclinical disciplines in undergraduate medical education. Increasingly, it has been adapted to clinical disciplines. However, its superiority over conventional learning methods used in clinical years of medical school remains unclear. The aim of this study was to compare TBL with traditional seminars delivered in small group interactive learning (SIL) format in terms of knowledge acquisition and retention, satisfaction and engagement of undergraduate medical students during the 6-week obstetrics and gynecology clerkship. MATERIAL AND METHODS: The study was conducted at Karolinska Institutet, a medical university in Sweden, and had a prospective, crossover design. All fifth-year medical students attending the obstetrics and gynecology clerkship, at four different teaching hospitals in Stockholm (approximately 40 students per site), in the Autumn semester of 2022 were invited to participate. Two seminars (one in obstetrics and one in gynecology) were designed and delivered in two different formats, ie TBL and SIL. The student:teacher ratio was approximately 10:1 in the traditional SIL seminars and 20:1 in the TBL. All TBL seminars were facilitated by a single teacher who had been trained and certified in TBL. Student knowledge acquisition and retention were assessed by final examination scores, and the engagement and satisfaction were assessed by questionnaires. For the TBL seminars, individual and team readiness assurance tests were also performed and evaluated. RESULTS: Of 148 students participating in the classrooms, 132 answered the questionnaires. No statistically significant differences were observed between TBL and SIL methods with regard to student knowledge acquisition and retention, engagement and satisfaction. CONCLUSIONS: We found no differences in student learning outcomes or satisfaction using TBL or SIL methods. However, as TBL had a double the student to teacher ratio as compared with SIL, in settings where teachers are scarce and suitable rooms are available for TBL sessions, the method may be beneficial in reducing faculty workload without compromising students' learning outcomes.

Outcomes of SARS-CoV-2 infection in early pregnancy-A systematic review and meta-analysis.

INTRODUCTION: Available data on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and pregnancy outcomes mostly refer to women contracting the infection during advanced pregnancy or close to delivery. There is limited information on the association between SARS-CoV-2 infection in early pregnancy and outcomes thereof. MATERIAL AND METHODS: We aimed to systematically review the maternal, fetal and neonatal outcomes following SARS-CoV-2 infection in early pregnancy, defined as <20 weeks of gestation (PROSPERO Registration 2020 CRD42020177673). Searches were carried out in PubMed, Medline, EMBASE, and Scopus databases from January 2020 until April 2023 and the WHO database of publications on coronavirus disease 2019 (COVID-19) from December 2019 to April 2023. Cohort and case-control studies on COVID-19 occurring in early pregnancy that reported data on maternal, fetal, and neonatal outcomes were included. Case reports and studies reporting only exposure to SARS-CoV-2 or not stratifying outcomes based on gestational age were excluded. Data were extracted in duplicate. Meta-analyses were conducted when appropriate, using R meta (R version 4.0.5). RESULTS: A total of 18 studies, 12 retrospective and six prospective, were included in this review, reporting on 10 147 SARS-CoV-2-positive women infected in early pregnancy, 9533 neonates, and 180 882 SARS-CoV-2 negative women. The studies had low to moderate risk of bias according to the Newcastle-Ottawa quality assessment Scale. The studies showed significant clinical and methodological heterogeneity. A meta-analysis could be performed only on the outcome miscarriage rate, with a pooled random effect odds ratio of 1.44 (95% confidence interval 0.96-2.18), showing no statistical difference in miscarriage in SARS-CoV-2-infected women. Individual studies reported increased incidences of stillbirth, low birthweight and preterm birth among neonates born to mothers affected by COVID-19 in early pregnancy; however, these results were not consistent among all studies. CONCLUSIONS: In this comprehensive systematic review of available evidence, we identified no statistically significant adverse association between SARS-CoV-2 infection in early pregnancy (before 20 weeks of gestation) and fetal, neonatal, or maternal outcomes. However, a 44% increase in miscarriage rate is concerning and further studies of larger sample size are needed to confirm or refute our findings.

Team-based learning (TBL) in clinical disciplines for undergraduate medical students-a scoping review.

BACKGROUND: Team-based learning (TBL) is an evidence-based pedagogical method that has been used in undergraduate medical education since 2001. However, its use in clinical disciplines is rarely reported, and the impact of its implementation is not known. The aim of this study was to explore and map the published literature on the impact of implementing TBL in clinical disciplines in undergraduate medical education. METHODS: A comprehensive search of Medline, Education Resources Information Center (ERIC), and Web of Science databases was performed on November 24, 2021 and updated April 6, 2023, using relevant Medical Subject Headings (MeSH) and free-text terms. Original research studies reporting on the implementation of TBL in clinical disciplines in undergraduate medical education published in peer-reviewed English language journals were included irrespective of their methodological design. RESULTS: The initial search identified 2,383 records. Of these, 49 met the inclusion criteria. Most of the studies (n = 44, 90%) described the implementation of a modified version of TBL in which one or more TBL steps were missing, and one study had undefined protocol for the implementation. The most reported outcomes were knowledge acquisition (n = 38, 78%) and students' satisfaction or attitudes toward TBL (n = 34, 69%). Despite some differences in their results, the studies found that implementing TBL is associated with increased knowledge acquisition (n = 19, 39%), student engagement (n = 6, 12%), and student satisfaction (n = 31, 63%). CONCLUSIONS: Most of the studies reported positive results in students' satisfaction and students' engagement, whilst the results on knowledge acquisition and retention were more contradictory. In most of the studies, TBL was implemented in a modified form and diverse comparators were used. The methodological quality also varied. Thus, no unequivocal conclusions could be drawn regarding the value of implementing TBL in clinical disciplines. More studies with rigorous methodologies are needed in this field.

Association of maternal SARS-CoV-2 infection at the time of admission for delivery with labor process and outcomes of vaginal birth: A cohort study.

INTRODUCTION: This study aimed to investigate the impact of maternal SARS-CoV-2 infection at the time of admission for delivery on labor process and outcomes of vaginal birth. MATERIAL AND METHODS: A cohort study was carried out at the Obstetrics Department of Anhui Provincial Hospital, China, where universal reverse transcriptase polymerase chain reaction (RT-PCR) testing for SARS-CoV-2 infection was introduced for all women admitted for labor and delivery from December 1-31, 2022. Women were divided into positive and negative groups based on the test result. All women having a singleton vaginal birth were included in final analysis. The effect of SARS-CoV-2 positivity on labor process and outcomes of vaginal birth was estimated by regression analyses. RESULTS: Among a total of 360 women included, 87 had a positive SARS-CoV-2 test and 273 a negative test. Women in the positive group had an increased likelihood of having longer labor (median 9.3 vs 8.3 hours; sB [log-transformed] 0.19; 95% confidence interval [CI] 0.09-0.28), episiotomy (39.1% vs 23.8%; adjusted odds ratio [aOR] 2.31; 95% CI 1.27-4.21), grade III meconium-stained amniotic fluid (19.5% vs 7.0%; aOR 2.52; 95% CI 1.15-5.54) and postpartum hospital stay exceeding 37 hours (58.6% vs 46.5%; aOR 1.71; 95% CI 1.00-2.91). They had reduced rates exclusive breastfeeding (26.7% vs 39%; aOR 0.21; 95% CI 0.09-0.46) as well as mixed feeding (46.5% vs 52.2%; aOR 0.28; 95% CI 0.13-0.60) at 1 week postpartum. No significant differences were observed in other aspects of labor process and birth outcomes, including the uptake of labor analgesia, postpartum hemorrhage (>500 mL) or neonatal outcomes. CONCLUSIONS: A positive maternal SARS-CoV-2 test in labor among women having vaginal birth was associated with a slightly longer duration of labor, increased likelihood of episiotomy, increased incidence of grade III meconium-stained amniotic fluid, a longer postpartum hospital stay and a lower rate of breastfeeding 1 week postpartum. However, it did not have an adverse impact on other birth outcomes.

Multi-omics approach reveals dysregulated genes during hESCs neuronal differentiation exposure to paracetamol.

Prenatal paracetamol exposure has been associated with neurodevelopmental outcomes in childhood. Pharmacoepigenetic studies show differences in cord blood DNA methylation between unexposed and paracetamol-exposed neonates, however, causality and impact of long-term prenatal paracetamol exposure on brain development remain unclear. Using a multi-omics approach, we investigated the effects of paracetamol on an in vitro model of early human neurodevelopment. We exposed human embryonic stem cells undergoing neuronal differentiation with paracetamol concentrations corresponding to maternal therapeutic doses. Single-cell RNA-seq and ATAC-seq integration identified paracetamol-induced chromatin opening changes linked to gene expression. Differentially methylated and/or expressed genes were involved in neurotransmission and cell fate determination trajectories. Some genes involved in neuronal injury and development-specific pathways, such as KCNE3, overlapped with differentially methylated genes previously identified in cord blood associated with prenatal paracetamol exposure. Our data suggest that paracetamol may play a causal role in impaired neurodevelopment.

Immune cell profiling of vaginal blood from patients with early pregnancy bleeding.

PROBLEM: Vaginal bleeding during early pregnancy is estimated to occur in 20% of all pregnancies and it is often difficult to predict who will ultimately miscarry. The role of immune cells in early pregnancy loss is poorly understood. METHOD OF STUDY: In this prospective cohort study, 28 pregnant women presenting with first-trimester vaginal bleeding donated vaginal blood, peripheral venous blood, and saliva during their initial emergency room visit, and at a follow-up. The composition, frequency, and phenotype of immune cells in the vaginal blood were determined using flow cytometry. The proteome of serum and saliva was analyzed with OLINK proximity extension assay and correlated to vaginal immune cell phenotype and outcome of pregnancy. The course and outcome of pregnancies were followed and recorded. RESULTS: Vaginal blood contained all main immune cell lineages including B cells, NK cells, T cells, and monocytes/macrophages. Notably, vaginal blood immune cells expressed tissue residency markers including CD49a. Women who subsequently miscarried had a higher frequency of vaginal blood CD49a+ NK cells compared to those who did not miscarry, and this correlated with serum levels of granzyme A and H, as well as CSF1, CAIX, and TWEAK. Women in the miscarriage group also had a higher frequency of peripheral blood T cells expressing CD49a. CONCLUSIONS: Our study provides novel insight into human reproductive immunology in relation to miscarriage. Tissue-resident NK cells in vaginal blood alone or in combination with serological biomarkers hold potential as prognostic factors in the prediction of pregnancy outcome in women with early pregnancy bleedings.

Endometrial receptivity in women of advanced age: an underrated factor in infertility.

BACKGROUND: Modern lifestyle has led to an increase in the age at conception. Advanced age is one of the critical risk factors for female-related infertility. It is well known that maternal age positively correlates with the deterioration of oocyte quality and chromosomal abnormalities in oocytes and embryos. The effect of age on endometrial function may be an equally important factor influencing implantation rate, pregnancy rate, and overall female fertility. However, there are only a few published studies on this topic, suggesting that this area has been under-explored. Improving our knowledge of endometrial aging from the biological (cellular, molecular, histological) and clinical perspectives would broaden our understanding of the risks of age-related female infertility. OBJECTIVE AND RATIONALE: The objective of this narrative review is to critically evaluate the existing literature on endometrial aging with a focus on synthesizing the evidence for the impact of endometrial aging on conception and pregnancy success. This would provide insights into existing gaps in the clinical application of research findings and promote the development of treatment options in this field. SEARCH METHODS: The review was prepared using PubMed (Medline) until February 2023 with the keywords such as 'endometrial aging', 'receptivity', 'decidualization', 'hormone', 'senescence', 'cellular', 'molecular', 'methylation', 'biological age', 'epigenetic', 'oocyte recipient', 'oocyte donation', 'embryo transfer', and 'pregnancy rate'. Articles in a language other than English were excluded. OUTCOMES: In the aging endometrium, alterations occur at the molecular, cellular, and histological levels suggesting that aging has a negative effect on endometrial biology and may impair endometrial receptivity. Additionally, advanced age influences cellular senescence, which plays an important role during the initial phase of implantation and is a major obstacle in the development of suitable senolytic agents for endometrial aging. Aging is also accountable for chronic conditions associated with inflammaging, which eventually can lead to increased pro-inflammation and tissue fibrosis. Furthermore, advanced age influences epigenetic regulation in the endometrium, thus altering the relation between its epigenetic and chronological age. The studies in oocyte donation cycles to determine the effect of age on endometrial receptivity with respect to the rates of implantation, clinical pregnancy, miscarriage, and live birth have revealed contradictory inferences indicating the need for future research on the mechanisms and corresponding causal effects of women's age on endometrial receptivity. WIDER IMPLICATIONS: Increasing age can be accountable for female infertility and IVF failures. Based on the complied observations and synthesized conclusions in this review, advanced age has been shown to have a negative impact on endometrial functioning. This information can provide recommendations for future research focusing on molecular mechanisms of age-related cellular senescence, cellular composition, and transcriptomic changes in relation to endometrial aging. Additionally, further prospective research is needed to explore newly emerging therapeutic options, such as the senolytic agents that can target endometrial aging without affecting decidualization. Moreover, clinical trial protocols, focusing on oocyte donation cycles, would be beneficial in understanding the direct clinical implications of endometrial aging on pregnancy outcomes.

Racial differences in RAD51 expression are regulated by miRNA-214-5P and its inhibition synergizes with olaparib in triple-negative breast cancer.

BACKGROUND: Triple-negative breast cancer (TNBC) affects young women and is the most aggressive subtype of breast cancer (BC). TNBCs disproportionally affect women of African-American (AA) descent compared to other ethnicities. We have identified DNA repair gene RAD51 as a poor prognosis marker in TNBC and its posttranscriptional regulation through microRNAs (miRNAs). This study aims to delineate the mechanisms leading to RAD51 upregulation and develop novel therapeutic combinations to effectively treat TNBCs and reduce disparity in clinical outcomes. METHODS: Analysis of TCGA data for BC cohorts using the UALCAN portal and PrognoScan identified the overexpression of RAD51 in TNBCs. miRNA sequencing identified significant downregulation of RAD51-targeting miRNAs miR-214-5P and miR-142-3P. RT-PCR assays were used to validate the levels of miRNAs and RAD51, and immunohistochemical and immunoblotting techniques were used similarly for RAD51 protein levels in TNBC tissues and cell lines. Luciferase assays were performed under the control of RAD51 3'-UTR to confirm that miR-214-5P regulates RAD51 expression. To examine the effect of miR-214-5P-mediated downregulation of RAD51 on homologous recombination (HR) in TNBC cells, Dr-GFP reporter assays were performed. To assess the levels of olaparib-induced DNA damage responses in miR-214-5P, transfected cells, immunoblots, and immunofluorescence assays were used. Furthermore, COMET assays were used to measure DNA lesions and colony assays were performed to assess the sensitivity of BRCA-proficient TNBC cells to olaparib. RESULTS: In-silico analysis identified upregulation of RAD51 as a poor prognostic marker in TNBCs. miRNA-seq data showed significant downregulation of miR-214-5P and miR-142-3P in TNBC cell lines derived from AA women compared to Caucasian-American (CA) women. miR-214-5P mimics downregulated RAD51 expression and induces HR deficiency as measured by Dr-GFP assays in these cell lines. Based on these results, we designed a combination treatment of miR-214-5P and olaparib in HR-proficient AA TNBC cell lines using clonogenic survival assays. The combination of miR-214-5P and olaparib showed synergistic lethality compared to individual treatments in these cell lines. CONCLUSIONS: Our studies identified a novel epigenetic regulation of RAD51 in TNBCs by miR-214-5P suggesting a novel combination therapies involving miR-214-5P and olaparib to treat HR-proficient TNBCs and to reduce racial disparity in therapeutic outcomes.

A systematic review and critical evaluation of quality of clinical practice guidelines on fetal growth restriction.

INTRODUCTION: To systematically identify and critically assess the quality of clinical practice guidelines (CPGs) on management fetal growth restriction (FGR). CONTENT: Medline, Embase, Google Scholar, Scopus and ISI Web of Science databases were searched to identify all relevant CPGs on FGR. SUMMARY: Diagnostic criteria of FGR, recommended growth charts, recommendation for detailed anatomical assessment and invasive testing, frequency of fetal growth scans, fetal monitoring, hospital admission, drugs administrations, timing at delivery, induction of labor, postnatal assessment and placental histopathological were assessed. Quality assessment was evaluated by AGREE II tool. Twelve CPGs were included. Twenty-five percent (3/12) of CPS adopted the recently published Delphi consensus, 58.3% (7/12) an estimated fetal weight (EFW)/abdominal circumference (AC) EFW/AC <10th percentile, 8.3% (1/12) an EFW/AC <5th percentile while one CPG defined FGR as an arrest of growth or a shift in its rate measured longitudinally. Fifty percent (6/12) of CPGs recommended the use of customized growth charts to assess fetal growth. Regarding the frequency of Doppler assessment, in case of absent or reversed end-diastolic flow in the umbilical artery 8.3% (1/12) CPGs recommended assessment every 24-48, 16.7% (2/12) every 48-72 h, 1 CPG generically recommended assessment 1-2 times per week, while 25 (3/12) did not specifically report the frequency of assessment. Only 3 CPGs reported recommendation on the type of Induction of Labor to adopt. The AGREE II standardized domain scores for the first overall assessment (OA1) had a mean of 50%. OUTLOOK: There is significant heterogeneity in the management of pregnancies complicated by FGR in published CPGs.

Impact of the COVID-19 pandemic on women's perinatal mental health and its association with personality traits: An observational study.

INTRODUCTION: The burden of perinatal mental health problems was expected to increase during the COVID-19 pandemic. We prospectively investigated the impact of the COVID-19 pandemic on the mental health of pregnant and postpartum women in Norway and explored associations with their sociodemographic characteristics and personality traits. MATERIAL AND METHODS: Sociodemographic information and the self-reported impact of pandemic on wellbeing of pregnant women was collected using an online survey. To assess women's mental health, two validated questionnaires, the Edinburgh Postpartum Depression Scale (EPDS) and the Generalized Anxiety Disorder-7 item Scale (GAD-7), were used prenatally and postnatally. Personality traits were evaluated using HumanGuide, a web-based ipsative psychological evaluation instrument. RESULTS: 772 women were included prenatally, of which 526 also responded to the survey 4-6 weeks postnatally. The median age was 29 years, 53.6% of the women were nulliparous when enrolled, and 35.1% worked in the healthcare sector. The median EPDS (6.0; interquartile range [IQR] 3.0-10.0 vs 6.0; IQR: 3.0-10.0) and the median GAD-7 (5.0; IQR 2.0-9.0 vs 5.0; IQR 2.0-9.0) were similar pre-and postnatally. Prenatally, the proportion of women scoring ≥13 on EPDS and ≥10 on GAD-7 was 14.5% (112/772) and 21.5% (166/772), whereas the postnatal figures were 15.6% (82/526) and 21.5% (113/526), respectively. The differences were not significant (P = 0.59 and P = 0.99). Being <25 years of age, being on pre-pregnancy psychotherapy or psychotropic medication, frequent voluntary isolation, perception of maternity care not proceeding normally, avoiding seeking medical assistance due to fear of infection and having negative economic consequences during the COVID19 pandemic significantly increased the risk of both anxiety (GAD-7 ≥10) and depression (EPDS ≥13). Nullipara had a higher risk of anxiety, whereas being a healthcare worker had a lower risk. The personality trait factors Power (P = 0.008), Quality (P = 0.008), Stability (P < 0.001) and Contacts (P < 0.001) were significant predictors of depression among pregnant women, whereas the Quality (P = 0.005) and Contacts (P = 0.003) were significant predictors of anxiety. CONCLUSIONS: During the initial phase of the COVID-19 pandemic, the prevalence of depression (EPDS ≥ 13) and anxiety (GAD-7 ≥ 10) was 14.5% and 21.5%, respectively, among Norwegian pregnant women. Certain sociodemographic characteristics and personality traits were significant predictors of depression and anxiety.

The acceptability of implementing patient-reported measures in routine maternity care: A systematic review.

INTRODUCTION: Patient-reported measures (PRMs) are becoming popular as they might influence clinical decisions, help to deliver patient-centered care, and improve health care quality. However, the limited knowledge and consensus about the acceptability of implementing PRMs in maternity care hinder their widespread use in clinical practice, and evidence-based recommendations are lacking. This systematic review aims to synthesize available evidence on the acceptability of implementing PRMs in routine maternity care. MATERIAL AND METHODS: Literature on the implementation of PRMs in maternity care was electronically searched in six databases (PsycARTICLES, PubMed, Scopus, Web of Science, Cochrane Database of Systematic Reviews, and CINAHL), screened and selected for the topic of "acceptability". Theoretical Framework of Acceptability was used as the basic framework guiding data analysis and synthesis. Evidence was thematically analyzed and synthesized. Mixed Method Appraisal Tool and GRADE-CERQual approach were used to assess the quality of studies and evaluate the confidence in the review findings. RESULTS: Overall, 4971 articles were screened. From 24 studies, we identified five themes regarding the acceptability of implementing PRMs in routine maternity care: (1) user's action and behavior, (2) stakeholders' attitudes, (3) perceived benefits, (4) perceived challenges and risks, and (5) stakeholders' preferences and suggestions on implementation. While pregnant and postpartum women, health professionals and other stakeholders involved in maternity care were generally positive about the implementation of PRMs in routine care and recognized the potential benefits (eg health improvement, women empowerment, care and services improvement and healthcare system advancement), they pointed out possible challenges and risks in answering PRMs questions, responding to answers, and setting up integrated information systems as well as suggested solutions in the aspects of PRMs data collection, follow-up care, and system-level management. The confidence in the review findings was moderate due to methodological limitations of included studies. CONCLUSIONS: Available empirical evidence suggested that the use of PRMs in routine maternity care is acceptable among stakeholders involved in maternity care and the potential benefits of its integration in routine clinical practice to healthcare improvement has been recognized. However, possible challenges in data collection, follow-up care arrangement and system-level integration should be appropriately addressed.