Membrane proteins are fundamental to many crucial cellular processes but removing them from their native environment for structural and functional studies creates experimental challenges. Numerous strategies have been developed to replicate native-like membrane environments in vitro for membrane protein research, however, most studies have focused on systems for either structural or functional characterisation, not both together. Here, we apply an in-vivo split intein strategy to produce stable circularised nanodiscs for combined structural and functional analysis of respiratory complex I, using its highly hydrophobic native ubiquinone-10 substrate and an auxiliary ubiquinol oxidase from Trypanosoma brucei brucei. We successfully reconstituted Paracoccus denitrificans complex I into circularised nanodiscs, determined its cryo-EM structure at 3.1 \u00c5 resolution and conducted biophysical and biochemical analyses to demonstrate how the \u2018oversized\u2019 nanodiscs have space to accommodate both enzymes and substrates to sustain steady-state catalysis. Our work establishes a proof-of-principle for using oversized nanodiscs as an integrated platform for structural and functional interrogation of complex membrane proteins in near-native membrane environments.
\n \n\n \n \nReducing maternal mortality remains a global challenge. Weaknesses in health systems are an important contributor to deaths around the world. Digital technologies could accelerate progress and support maternity services worldwide. Many digital innovations have been developed and implemented in maternity services. These target: the end users themselves (i.e., the pregnant woman and her family) to improve health knowledge and health-seeking behaviors; the primary and secondary care health workers in the community to support decision-making, scheduling, and referrals; and the health system and health management supporting the tracking of births and deaths. This chapter outlines the case for digital health innovations to support pregnancy and maternity services through a series of case studies, considering the potential advantages and challenges of scale-up. While the examples presented are not specific to Muslim populations, they have potential relevance for the challenges faced in Muslim countries worldwide.
\n \n\n \n \nINTRODUCTION: Congenital heart defect (CHD) is a significant, rapidly emerging global problem in child health and a leading cause of neonatal and childhood death. Prenatal detection of CHDs with the help of ultrasound allows better perinatal management of such pregnancies, leading to reduced neonatal mortality, morbidity and developmental complications. However, there is a wide variation in reported fetal heart problem detection rates from 34% to 85%, with some low- and middle-income countries detecting as low as 9.3% of cases before birth. Research has shown that deep learning-based or more general artificial intelligence (AI) models can support the detection of fetal CHDs more rapidly than humans performing ultrasound scan. Progress in this AI-based research depends on the availability of large, well-curated and diverse data of ultrasound images and videos of normal and abnormal fetal hearts. Currently, CHD detection based on AI models is not accurate enough for practical clinical use, in part due to the lack of ultrasound data available for machine learning as CHDs are rare and heterogeneous, the retrospective nature of published studies, the lack of multicentre and multidisciplinary collaboration, and utilisation of mostly standard planes still images of the fetal heart for AI models. Our aim is to develop AI models that could support clinicians in detecting fetal CHDs in real time, particularly in nonspecialist or low-resource settings where fetal echocardiography expertise is not readily available. METHODS AND ANALYSIS: We have designed the Clinical Artificial Intelligence Fetal Echocardiography (CAIFE) study as an international multicentre multidisciplinary collaboration led by a clinical and an engineering team at the University of Oxford. This study involves five multicountry hospital sites for data collection (Oxford, UK (n=1), London, UK (n=3) and Southport, Australia (n=1)). We plan to curate 14\u2009000 retrospective ultrasound scans of fetuses with normal hearts (n=13\u2009000) and fetuses with CHDs (n=1000), as well as 2400 prospective ultrasound cardiac scans, including the proposed research-specific CAIFE 10 s video sweeps, from fetuses with normal hearts (n=2000) and fetuses diagnosed with major CHDs (n=400). This gives a total of 16\u2009400 retrospective and prospective ultrasound scans from the participating hospital sites. We will build, train and validate computational models capable of differentiating between normal fetal hearts and those diagnosed with CHDs and recognise specific types of CHDs. Data will be analysed using statistical metrics, namely, sensitivity, specificity and accuracy, which include calculating positive and negative predictive values for each outcome, compared with manual assessment. ETHICS AND DISSEMINATION: We will disseminate the findings through regional, national and international conferences and through peer-reviewed journals. The study was approved by the Health Research Authority, Care Research Wales and the Research Ethics Committee (Ref: 23/EM/0023; IRAS Project ID: 317510) on 8 March 2023. All collaborating hospitals have obtained the local trust research and development approvals.
\n \n\n \n \nBackground Uterine sparing surgery has become an option for patients with placenta accreta spectrum disorders. The decision to perform a cesarean hysterectomy versus uterine sparing surgery is made intraoperatively. This study was undertaken to assess the value of ultrasound markers in predicting hysterectomy. Objective To describe ultrasound markers associated with the need for cesarean hysterectomy in patients at risk of placenta accreta spectrum. Study design This was an analysis of a prospectively collected data of high risk placenta accreta spectrum patients between September 2023 and August 2024. Ultrasound examination was performed by an expert focusing on the diagnosis of placenta accreta spectrum. All patients were counselled regarding the management options available at our center, namely uterine-sparing surgery and hysterectomy. All patients opted for a uterine sparing surgery if safe and technically feasible. The final choice of surgical management approach was solely based on the intraoperative topography which describes the size and location of the abnormally adhered placenta. The primary outcome was the need for hysterectomy despite a preoperative plan for uterine-sparing surgery. Results A total of 123 participants were enrolled: 93 placenta accreta spectrum cases and 30 non-placenta accreta scar dehiscence cases. Uterine sparing surgery was successful in 74 out of 93(79.6%) placenta accreta spectrum cases and 100% non-placenta accreta scar dehiscence cases. Least Absolute Shrinkage and Selection Operator penalised regression revealed intracervical hypervascularity >50%, distorted urinary bladder wall, and parametrial hypervascularity as the most influential predictors for hysterectomy. This best-fitted model achieved accuracy of 94% (95% CI: 81.3% \u2013 99.3%) after model cross-validation. The combination of intracervical hypervascularity >50% and distorted bladder wall had the highest predictive probability for hysterectomy, with a value of 0.87 (95% CI: 0.81 \u2013 0.93), sensitivity of 96.0% (95% CI 89.0 \u2013 99.0%) and specificity of 92.0%(95% CI 62.0 \u2013 100.0). Conclusion Comprehensive preoperative ultrasound can reasonably predict the appropriate surgical approach to placenta accreta spectrum. This can be achieved by assessing intracervical hypervascularity and a distorted urinary bladder wall using a combination of transabdominal, transvaginal, and color Doppler ultrasound techniques, as these signs have a strong correlation with the need for hysterectomy in a cohort where the intention to treat was uterine sparing surgery.
\n \n\n \n \nBACKGROUND: Each year 182,000 babies are stillborn in Nigeria, representing nearly 10% of the annual global stillbirth burden. Imo state in south-eastern Nigeria has one of the highest levels of maternal health service access in Nigeria, yet this has not translated into good pregnancy outcomes. Many stillbirth prevention initiatives in Nigeria focus on maternal health education but empirical evidence suggests that sociocultural factors impact healthcare choices and outcomes. This study aims to explore women's and health workers' perspectives of the sociocultural barriers to following medical advice during pregnancy and childbirth, and specifically how these barriers may contribute to an increased risk of stillbirth. This study is part of a broader community-based stillbirth prevention mixed-methods research in Imo State, Nigeria. METHODS: A qualitative descriptive study was conducted using in-depth interviews and focus group discussions. 38 participants were purposively recruited; 20 women and 18 health workers. Audio recordings were transcribed, translated and analysed using inductive thematic analysis. RESULTS: Four themes were identified: (1) trust, where scepticism about health worker motives or competence and trust in community informal networks were highlighted (2) power dynamics within families, with husbands and older female relatives influencing health decisions; (3) personal and community beliefs that undermine confidence in medical interventions, including a pervasive stigma associated with caesarean section; and (4) grassroots proposals for solutions, emphasising the importance of a whole-community approach to maternal health education, mobilising peer voices, engaging traditional leaders and training of traditional birth attendants. CONCLUSION: This study provides insights into the sociocultural barriers to following medical advice during pregnancy in Nigeria, which include a lack of trust in health professionals, power dynamics within a woman's family, and entrenched cultural and religious beliefs that oppose medical intervention. Women's decisions about pregnancy and childbirth are heavily influenced by family and cultural norms. Culturally sensitive, community-wide interventions which aim to rebuild trust in the health system, involve women as decision-makers in antenatal care, and engage religious and traditional leaders would be beneficial for improving outcomes.
\n \n\n \n \nDespite Nigeria's stillbirth rate reducing from 28.6 to 22.5 per 1,000 births from 2000-2021, progress trails comparable indicators and regional variations persist. We assessed stillbirth incidences and associated risk factors in 20 secondary hospitals in Imo state, to generate essential local evidence to inform policymaking to reduce mortality. The total numbers of births and their outcomes were determined through hospital maternity registers. An unmatched case-control study was conducted. We collected retrospective data about 157 antepartum and 193 intrapartum stillbirths, and from 381 livebirths (controls). Potential risk factors were categorised into sociodemographic, obstetric and maternity care and biological determinants using a theoretical framework. Independent multivariable logistic regression models were used to investigate the association of risk factors with each stillbirth type. The overall stillbirth rate was 38 per 1,000 total births. The rate of antepartum and intrapartum stillbirths were 16 and 19 per 1,000 respectively. The risk factors independently associated with antepartum stillbirths were nulliparity (adjusted odds ratio (aOR) 1.87, 95%CI 1.04-3.36); preterm birth (aOR 14.29, 95%CI 6.31-32.38); being referred from another facility (aOR 3.75, 95%CI 1.96-7.17); unbooked pregnancy (aOR 2.58, 95%CI 1.37-4.85); and obstetric complications (aOR 4.04, 95%CI 2.35-6.94). For intrapartum stillbirths, associated factors were preterm birth (aOR 11.28, 95%CI 4.66-27.24); referral (aOR 2.50, 95%CI 1.19-5.24); not using a partogram (aOR 2.92, 95%CI 1.23-6.95) and obstetric complications (aOR 10.71, 95%CI 5.92-19.37). The findings highlight specific risk factors associated with antepartum and intrapartum stillbirths, shedding light on potential areas for targeted interventions.
\n \n\n \n \nBACKGROUND: Better geographical accessibility to comprehensive emergency obstetric care (CEmOC) facilities can significantly improve pregnancy outcomes. However, with other factors, such as affordability critical for care access, it is important to explore accessibility across groups. We assessed CEmOC geographical accessibility by wealth status in the 15 most-populated Nigerian cities. METHODS: We mapped city boundaries, verified and geocoded functional CEmOC facilities, and assembled population distribution for women of childbearing age and Meta's Relative Wealth Index (RWI). We used the Google Maps Platform's internal Directions Application Programming Interface to obtain driving times to public and private facilities. City-level median travel time (MTT) and number of CEmOC facilities reachable within 60\u2009min were summarised for peak and non-peak hours per wealth quintile. The correlation between RWI and MTT to the nearest public CEmOC was calculated. RESULTS: We show that MTT to the nearest public CEmOC facility is lowest in the wealthiest 20% in all cities, with the largest difference in MTT between the wealthiest 20% and least wealthy 20% seen in Onitsha (26 vs 81\u2009min) and the smallest in Warri (20 vs 30\u2009min). Similarly, the average number of public CEmOC facilities reachable within 60\u2009min varies (11 among the wealthiest 20% and six among the least wealthy in Kano). In five cities, zero facilities are reachable under 60\u2009min for the least wealthy 20%. Those who live in the suburbs particularly have poor accessibility to CEmOC facilities. CONCLUSIONS: Our findings show that the least wealthy mostly have poor accessibility to care. Interventions addressing CEmOC geographical accessibility targeting poor people are needed to address inequities in urban settings.
\n \n\n \n \nBACKGROUND: Sub-Saharan Africa (SSA) alone contributed to 42% of global stillbirths in 2019, and the rate of stillbirth reduction has remained slow. There has been an increased uptake of community-based interventions to combat stillbirth in the region, but the effects of these interventions have been poorly assessed. Our objectives were to examine the effect of community-based interventions on stillbirth in SSA. METHODS: In this systematic review and meta-analysis, we searched eight databases (MEDLINE [OvidSP], Embase [OvidSP], Cochrane Central Register of Controlled Trials, Global Health, Science Citation Index and Social Science Citation index [Web of Science Core Collection], CINAHL [EBSCOhost] and Global Index Medicus) and four grey literature sources from January 1, 2000 to July 7, 2023 for relevant studies from SSA. Community-based interventions targeting stillbirths solely or as part of complex interventions, with or without hospital interventions were included, while hospital-only interventions, microcredit schemes and maternity waiting home interventions were excluded. Study quality was assessed using the Cochrane risk of bias and National Heart, Lung and Blood Institute's tools. The study outcome was odds of stillbirth in intervention versus control communities. Pooled odds ratios (ORs) were estimated using random-effects models, and subgroup analyses were performed by intervention type and strategies. Publication bias was evaluated by funnel plot and Egger's test. This study is registered with PROSPERO, CRD42021296623. FINDINGS: Of the 4223 records identified, seventeen studies from fifteen SSA countries were eligible for inclusion. One study had four arms (community only, hospital only, community and hospital, and control arms), so information was extracted from each arm. Analysis of 13 of the 17 studies which had community-only intervention showed that the odds of stillbirth did not vary significantly between community-based intervention and control groups (OR 0.96; 95% CI 0.78-1.17, I2\u00a0=\u00a057%, p\u00a0\u2264\u00a00.01, n\u00a0=\u00a063,884). However, analysis of four (out of five) studies that included both community and health facility components found that in comparison with community only interventions, this combination strategy significantly reduced the odds of stillbirth by 17% (OR 0.83; 95% CI 0.79-0.87, I2\u00a0=\u00a011%, p\u00a0=\u00a00.37, n\u00a0=\u00a0244,868), after excluding a study with high risk of bias. The quality of the 17 studies were graded as poor (n\u00a0=\u00a02), fair (n\u00a0=\u00a09) and good (n\u00a0=\u00a06). INTERPRETATION: Community-based interventions alone, without strengthening the quality and capacity of health facilities, are unlikely to have a substantial effect on reducing stillbirths in SSA. FUNDING: Nuffield Department of Population Health, Balliol College, the Clarendon Fund, Medical Research Council.
\n \n\n \n \nBackground: Dashboards are increasingly being used in sub-Saharan Africa (SSA) to support health policymaking and governance. However, their use has been mostly limited to routine care, not emergency services like emergency obstetric care (EmOC). To ensure a fit-for-purpose dashboard, we conducted an online survey with policymakers and researchers to understand key considerations needed for developing a policy-ready dashboard of geospatial access to EmOC in SSA. Methods: Questionnaires targeting both stakeholder groups were pre-tested and disseminated in English, French, and Portuguese across SSA. We collected data on participants\u2019 awareness of concern areas for geographic accessibility of EmOC and existing technological resources used for planning of EmOC services, the dynamic dashboard features preferences, and the dashboard's potential to tackle lack of geographic access to EmOC. Questions were asked as multiple-choice, Likert-scale, or open-ended. Descriptive statistics were used to summarise findings using frequencies or proportions. Free-text responses were recoded into themes where applicable. Results: Among the 206 participants (88 policymakers and 118 researchers), 90% reported that rural areas and 23% that urban areas in their countries were affected by issues of geographic accessibility to EmOC. Five percent of policymakers and 38% of researchers were aware of the use of maps of EmOC facilities to guide planning of EmOC facility location. Regarding dashboard design, most visual components such as location of EmOC facilities had almost universal desirability; however, there were some exceptions. Nearly 70% of policymakers considered the socio-economic status of the population and households relevant to the dashboard. The desirability for a heatmap showing travel time to care was lower among policymakers (53%) than researchers (72%). Nearly 90% of participants considered three to four data updates per year or less frequent updates adequate for the dashboard. The potential usability of a dynamic dashboard was high amongst both policymakers (60%) and researchers (82%). Conclusion: This study provides key considerations for developing a policy-ready dashboard for EmOC geographical accessibility in SSA. Efforts should now be targeted at establishing robust estimation of geographical accessibility metrics, integrated with existing health system data, and developing and maintaining the dashboard with up-to-date data to maximise impact in these settings.
\n \n\n \n \nINTRODUCTION: Access to skilled birth attendance has been prioritised as an intervention to minimise burden of maternal deaths in sub-Saharan Africa (SSA). However, poor experience of care (EoC) is impeding progress. We conducted a systematic review to holistically explore EoC patterns of facility-based childbirth in SSA. METHODS: PubMed, Embase and Scopus databases were searched to identify SSA EoC studies conducted between January 2000 and December 2019. Studies meeting our pre-defined inclusion criteria were quality assessed and relevant data extracted. We utilised the EoC quality standards (defined by the World Health Organization) to summarise and analyse findings while highlighting patterns. RESULTS: Twenty-two studies of varying quality from 11 SSA countries were included for review. Overall, at least one study from all included countries reported negative EoC in one or more domains of the WHO framework. Across SSA, 'respect and preservation of dignity' was the most reported domain of EoC. While most women deemed the pervasive disrespect as unacceptable, studies in West Africa suggest a \"normalisation\" of disrespect, if the intent is to save their lives. Women often experienced sub-optimal communication and emotional support with providers in public facilities compared to non-public ones in the region. These experiences had an influence on future institutional deliveries. DISCUSSION: Sub-optimal EoC is widespread in SSA, more so in public facilities. As SSA heath systems explore approaches make progress towards the Sustainable Development Goal 3, emphasis needs to be placed on ensuring women in the region have access to both high-quality provision and experience of care.
\n \n\n \n \nBACKGROUND: Follicular aspiration under transvaginal ultrasound guidance is routinely performed as part of assisted reproductive technology (ART) to retrieve oocytes for in vitro fertilisation (IVF). The process involves aspiration of the follicular fluid followed by the introduction of flush, typically culture media, back into the follicle followed by re-aspiration. However, there is a degree of controversy as to whether this intervention yields a larger number of oocytes and is hence associated with greater potential for pregnancy than aspiration only. OBJECTIVES: To assess the safety and efficacy of follicular flushing as compared with aspiration only performed in women undergoing ART. SEARCH METHODS: We searched the following electronic databases up to 13 July 2021: the Cochrane Gynaecology and Fertility Specialised Register of Controlled Trials, CENTRAL (containing output from two trial registries and CINAHL), MEDLINE, Embase, and PsycINFO. We also searched LILACS, Google Scholar, and Epistemonikos. We reviewed the reference lists of relevant papers and contacted experts in the field to identify further relevant studies. SELECTION CRITERIA: We included randomised controlled trials (RCTs) that compared follicular aspiration and flushing with aspiration alone in women undergoing ART using their own gametes. Primary outcomes were live birth rate and miscarriage rate per woman randomised. DATA COLLECTION AND ANALYSIS: Two review authors independently assessed studies identified by search against the inclusion criteria, extracted data, and assessed risk of bias. A third review author was consulted if required. We contacted study authors as needed. We analysed dichotomous outcomes using Mantel-Haenszel odds ratios (ORs), 95% confidence intervals (CIs), and a fixed-effect model, and we analysed continuous outcomes using mean differences (MDs) between groups presented with 95% CIs. We examined the heterogeneity of studies via the I2 statistic. We assessed the certainty of evidence using the GRADE approach. MAIN RESULTS: We included 15 studies with a total of 1643 women. Fourteen studies reported outcomes per woman randomised, and one study reported outcomes per ovary. No studies were at low risk of bias across all domains; the main limitation was lack of blinding. The certainty of the evidence ranged from moderate to very low, and was downgraded for risk of bias, imprecision, and inconsistency. We are uncertain of the effect of follicular flushing on live birth rate compared to aspiration alone (OR 0.93, 95% CI 0.59 to 1.46; 4 RCTs; n = 467; I2 = 0%; moderate-certainty evidence). This suggests that with a live birth rate of approximately 30% with aspiration alone, the equivalent live birth rate with follicular flushing lies between 20% and 39%. \u00a0We are uncertain of the effect of follicular flushing on miscarriage rate compared to aspiration alone (OR 1.98, 95% CI 0.18 to 22.22; 1 RCT; n = 164; low-certainty evidence). This suggests that with a miscarriage rate of approximately 1% with aspiration alone, the equivalent miscarriage rate with follicular flushing lies between 0% and 22%. We are uncertain of the effect of follicular flushing on oocyte yield (MD -0.47 oocytes, 95% CI -0.72 to -0.22; 9 RCTs; n = 1239; I2 = 61%; very low-certainty evidence); total number of embryos (MD -0.10 embryos, 95% CI -0.34 to 0.15; 2 RCTs; n = 160; I2 = 58%; low-certainty evidence); and clinical pregnancy rate (OR 1.12, 95% CI 0.85 to 1.51; 7 RCTs; n = 939; I2 = 46%; low-certainty evidence). The duration of the retrieval process may be longer with flushing (MD 175.44 seconds, 95% CI 152.57 to 198.30; 7 RCTs; n = 785; I2 = 87%; low-certainty evidence). It was not possible to perform a meta-analysis for adverse events, although individual studies reported on outcomes ranging from depression and anxiety to pain and pelvic organ injury. AUTHORS' CONCLUSIONS: The effect of follicular flushing on both live birth and miscarriage rates compared with aspiration alone is uncertain. Although the evidence does not permit any firm conclusions on the impact of follicular flushing on oocyte yield, total number of embryos, number of cryopreserved embryos, or clinical pregnancy rate, it may be that the procedure itself takes longer than aspiration alone. The evidence was insufficient to permit any firm conclusions with respect to adverse events or safety.
\n \n\n \n \nSTUDY QUESTION: What are the similarities and differences in endometrial B cells in the normal human endometrium and benign reproductive pathologies? SUMMARY ANSWER: Endometrial B cells typically constitute <5% of total endometrial CD45+ lymphocytes, and no more than 2% of total cells in the normal endometrium, and while their relative abundance and phenotypes vary in benign gynaecological conditions, current evidence is inconsistent. WHAT IS KNOWN ALREADY: B cells are vitally important in the mucosal immune environment and have been extensively characterized in secondary lymphoid organs and tertiary lymphoid structures (TLSs), with the associated microenvironment germinal centre. However, in the endometrium, B cells are largely overlooked, despite the crucial link between autoimmunity and reproductive pathologies and the fact that B cells are present in normal endometrium and benign female reproductive pathologies, scattered or in the form of lymphoid aggregates (LAs). A comprehensive summary of current data investigating B cells will facilitate our understanding of endometrial B cells in the endometrial mucosal immune environment. STUDY DESIGN SIZE DURATION: This systematic review retrieved relevant studies from four databases (MEDLINE, EMBASE, Web of Science Core Collection and CINAHL) from database inception until November 2021. PARTICIPANTS/MATERIALS SETTING METHODS: The search strategy combined the use of subject headings and relevant text words related to 'endometrium', 'B cells' and B-cell derivatives, such as 'antibody' and 'immunoglobulin'. Non-benign diseases were excluded using cancer-related free-text terms, and searches were limited to the English language and human subjects. Only peer-reviewed research papers were included. Each paper was graded as 'Good', 'Fair' or 'Poor' quality based on the NEWCASTLE-OTTAWA quality assessment scale. Only 'Good' quality papers were included. MAIN RESULTS AND THE ROLE OF CHANCE: Twenty-seven studies met the selection criteria and were included in this review: 10 cross-sectional studies investigated B cells in the normal endometrium; and 17 case-control studies compared the characteristics of endometrial B cells in control and benign female reproductive pathologies including endometritis, endometriosis, infertility, abnormal uterine bleeding, endometrial polyps and uterine fibroids. In all studies, B cells were present in the endometrium, scattered or in the form of LAs. CD20+ B cells were more abundant in patients with endometritis, but the data were inconsistent as to whether B-cell numbers were increased in endometriosis and patients with reproductive pathologies. LIMITATIONS REASONS FOR CAUTION: Although only 'good' quality papers were included in this systematic review, there were variations in patients' age, diagnostic criteria for different diseases and sample collection time among included studies. Additionally, a large number of the included studies only used immunohistochemistry as the identification method for endometrial B cells, which may fail to provide an accurate representation of the numbers of endometrial B cells. WIDER IMPLICATIONS OF THE FINDINGS: Histological studies found that endometrial B cells are either scattered or surrounded by T cells in LAs: the latter structure seems to be under hormonal control throughout the menstrual cycle and resembles TLSs that have been observed in other tissues. Further characterization of endometrial B cells and LAs could offer insights to endometrial B-cell function, particularly in the context of autoimmune-associated pathologies, such as endometriosis. Additionally, clinicians should be aware of the limited value of diagnosing plasma cell infiltration using only CD138. STUDY FUNDING/COMPETING INTERESTS: This study was funded by Finox Biotech. The authors have no conflicts of interest to declare. PROSPERO REGISTRATION NUMBER: This systematic review was registered in PROSPERO in January 2020 (PROSPERO ID: CRD42020152915).
\n \n\n \n \nPhospholipase C zeta (PLC\u03b6) is a sperm-specific protein that triggers oocyte activation. The analysis of PLC\u03b6 expression in human spermatozoa can be used as a diagnostic marker for oocyte activation deficiency. Our laboratory has previously optimized a standard \"in-house\" assay to determine PLC\u03b6 expression in human spermatozoa. However, one study has suggested that an antigen unmasking method (AUM) would be more efficient in visualizing PLC\u03b6 in human sperm. This study aimed to compare our established assay and AUM (involving HCl, acidic Tyrode's solution [AT], and heat). The mean relative fluorescence (RF) intensity of PLC\u03b6 in frozen-thawed spermatozoa from fourteen fertile donors stained with the in-house method was significantly higher than three other AUM groups (in-house [mean \u00b1 standard error of mean]: 18.87 \u00b1 2.39 arbitrary units [a.u.] vs non-AUM: 11.44 \u00b1 1.61 a.u., AT-AUM: 12.38 \u00b1 1.89 a.u., and HCl-AUM: 12.51 \u00b1 2.16 a.u., P < 0.05, one-way analysis of variance). The mean RF intensity of PLC\u03b6 in AT- and HCl-treated spermatozoa from 12 infertile males was not significantly different from that of the non-AUM group. However, the in-house method resulted in the highest RF intensity (12.11 \u00b1 1.36 a.u., P < 0.01). Furthermore, specificity testing of antibody-antigen binding indicated that the in-house method showed more specific binding than spermatozoa treated by the AUM. In conclusion, our in-house method showed superior visualization and reliability than the AUM, thus supporting the continued use of our in-house assay for clinical research screening.
\n \n\n \n \nPlatelets are anucleate blood cells vital for hemostasis and immunity. During cell death and aberrant mitosis, nucleated cells release DNA, resulting in \"cell-free\" DNA in plasma (cfDNA). An excess of cfDNA is deleterious. Given their ability to internalize pathogen-derived nucleic acids, we hypothesized that platelets may also clear endogenous cfDNA. We found that, despite lacking a nucleus, platelets contained a repertoire of DNA fragments mapping across the nuclear genome. We detected fetal DNA in maternal platelets and cancer-derived DNA in platelets from patients with premalignant and cancerous lesions. As current liquid biopsy approaches utilize platelet-depleted plasma, important genetic information contained within platelets is being missed. This study establishes a physiological role for platelets that has not previously been highlighted, with broad translational relevance.
\n \n\n \n \nObstetric care in Japan is organized in 3 levels and half of all deliveries are conducted at primary facilities without neonatal intensive care unit. We evaluated the predictive accuracy of the sFlt-1/PlGF ratio for the onset of preeclampsia and adverse outcomes in pregnant women with suspected preeclampsia at multiple facilities in Japan, including primary facilities. 356 pregnant women from 18\u2009+\u20090 to 36\u2009+\u20096 weeks of pregnancy were enrolled. 303 women were included in the final analysis. The negative predictive value for ruling out preeclampsia within 1 week using the cut-off value 38 was 98.4% (95% CI, 96.2-99.3) with a negative likelihood ratio of 0.22 (95% CI, 0.09-0.53). The positive predictive value for ruling in preeclampsia within 4 weeks using the cut-off value 38 was 48.2% (95% CI, 38.0-58.5). The positive predictive value using a cut-off value of 85 was 65.0% (95% CI, 44.1-81.4) with a positive likelihood ratio of 12.21 (95% CI, 5.20-28.80). The positive predictive value for prediction of adverse outcomes within 4 weeks using the cut-off value 38 was 64.8% (95% CI, 53.1-75.0). To clarify the accuracy of the test currently covered by insurance in Japan, we additionally conducted an analysis focusing on pregnant women between 18\u2009+\u20090 and 35\u2009+\u20096 weeks, yielding even higher accuracy. Pregnant women with sFlt-1/PlGF ratio >38 should be referred to a higher-level medical institution. The appropriate use of sFlt-1/PlGF supported by a robust collaboration between primary and tertiary care institutions may help to improve perinatal outcomes in Japan.
\n \n\n \n \nRetained products of conception (RPOC) is a common complication following first-trimester pregnancy loss. However, there are no formal recommendations regarding the diagnosis of RPOC. This systematic review aimed to synthesise and critically appraise the existing evidence on the definition and diagnostic criteria for RPOC. We registered this systematic review prospectively with PROSPERO (CRD42023444456). A comprehensive literature search was conducted in October 2024 using the following databases: Embase (OVID), Medline (OVID), Global Health, CINAHL on EBSCOhost, Cochrane Central and Web of Science. Databases were searched using free-text keywords and subject headings for the key concepts of 'early', 'miscarriage' and 'retained'. Data were extracted and 2 by 2 tables populated. Risk of bias and quality assessments were performed using the QUADAS-2 tool. The literature search yielded 2,014 articles that were screened for eligibility, resulting in the inclusion of 17 studies in the final analysis. Ultrasound scan was the primary diagnostic tool, used in 16 of the 17 included studies. Ultrasound diagnostic markers included: endometrial thickness (ET), the presence of hyperechoic or echogenic material, and colour flow Doppler. One study used persistent bleeding for more than 14\u2009days as the primary diagnostic marker. There was significant variation in the diagnostic thresholds used and no single ultrasound marker demonstrated consistent reliability in diagnosing RPOC. The findings of this review highlight the limitations of ultrasound as a standalone diagnostic tool for RPOC. Given the lack of clear diagnostic criteria, clinicians should integrate ultrasound findings with clinical symptoms to improve diagnostic accuracy. RPOC appears to be a distinct pathology within the spectrum of early pregnancy loss, characterised by the persistence of pregnancy tissue within the uterine cavity despite initial management which distinguishes it from incomplete miscarriage. This review provides a foundation for future research and calls for a Delphi consensus to refine the diagnosis and management of RPOC.
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