Dynamic prediction of pregnancy outcome after previous stillbirth or perinatal death: pilot study to establish proof-of-concept and explore method feasibility.

OBJECTIVE: To establish proof-of-concept for the dynamic prediction of adverse pregnancy outcome in women with a history of stillbirth or perinatal death, repeatedly throughout the pregnancy. METHODS: A retrospective cohort study of women in a subsequent pregnancy following previous perinatal loss, who received antenatal care at a tertiary hospital between January 2014 and December 2017, was used as the basis for exploratory prognostic model development. Models were developed to repeatedly predict a composite adverse outcome (stillbirth or neonatal death, 5-min Apgar score < 7, umbilical artery pH ≤ 7.05, admission to the neonatal intensive care unit for longer than 24 h, preterm birth (< 37 completed weeks) or birth weight < 10th centile) using the findings of sequential ultrasound scans for fetal biometry and umbilical and uterine artery Doppler. RESULTS: In total, 506 participants were eligible, of whom 504 were included in the analysis. An adverse pregnancy outcome was experienced by 110 (22%) participants. The ability to predict the composite outcome using repeated head circumference and estimated fetal weight measurements improved as the pregnancy progressed (e.g. area under the receiver-operating-characteristics curve improved from 0.59 at 24 weeks' gestation to 0.74 at 36 weeks' gestation), supporting proof-of-concept. Predictors to include in dynamic prediction models were identified, including ultrasound measurements of fetal biometry, umbilical and uterine artery Doppler and placental size and shape. CONCLUSION: The present study supports proof-of-concept for dynamic prediction of adverse outcome in pregnancy following prior stillbirth or perinatal death, which could be used to identify risks earlier in pregnancy, while highlighting methodological challenges and requirements for subsequent large-scale model development studies. © 2024 The Author(s). Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

External validation of prognostic models to predict stillbirth using International Prediction of Pregnancy Complications (IPPIC) Network database: individual participant data meta-analysis.

OBJECTIVE: Stillbirth is a potentially preventable complication of pregnancy. Identifying women at high risk of stillbirth can guide decisions on the need for closer surveillance and timing of delivery in order to prevent fetal death. Prognostic models have been developed to predict the risk of stillbirth, but none has yet been validated externally. In this study, we externally validated published prediction models for stillbirth using individual participant data (IPD) meta-analysis to assess their predictive performance. METHODS: MEDLINE, EMBASE, DH-DATA and AMED databases were searched from inception to December 2020 to identify studies reporting stillbirth prediction models. Studies that developed or updated prediction models for stillbirth for use at any time during pregnancy were included. IPD from cohorts within the International Prediction of Pregnancy Complications (IPPIC) Network were used to validate externally the identified prediction models whose individual variables were available in the IPD. The risk of bias of the models and cohorts was assessed using the Prediction study Risk Of Bias ASsessment Tool (PROBAST). The discriminative performance of the models was evaluated using the C-statistic, and calibration was assessed using calibration plots, calibration slope and calibration-in-the-large. Performance measures were estimated separately in each cohort, as well as summarized across cohorts using random-effects meta-analysis. Clinical utility was assessed using net benefit. RESULTS: Seventeen studies reporting the development of 40 prognostic models for stillbirth were identified. None of the models had been previously validated externally, and the full model equation was reported for only one-fifth (20%, 8/40) of the models. External validation was possible for three of these models, using IPD from 19 cohorts (491 201 pregnant women) within the IPPIC Network database. Based on evaluation of the model development studies, all three models had an overall high risk of bias, according to PROBAST. In the IPD meta-analysis, the models had summary C-statistics ranging from 0.53 to 0.65 and summary calibration slopes ranging from 0.40 to 0.88, with risk predictions that were generally too extreme compared with the observed risks. The models had little to no clinical utility, as assessed by net benefit. However, there remained uncertainty in the performance of some models due to small available sample sizes. CONCLUSIONS: The three validated stillbirth prediction models showed generally poor and uncertain predictive performance in new data, with limited evidence to support their clinical application. The findings suggest methodological shortcomings in their development, including overfitting. Further research is needed to further validate these and other models, identify stronger prognostic factors and develop more robust prediction models. © 2021 The Authors. Ultrasound in Obstetrics & Gynecology published by John Wiley & Sons Ltd on behalf of International Society of Ultrasound in Obstetrics and Gynecology.

Breech presentation is associated with lower adolescent tibial bone strength.

We compared bone outcomes in adolescents with breech and cephalic presentation. Tibia bone mineral content, density, periosteal circumference, and cross-sectional moment of inertia were lower in breech presentation, and females with breech presentation had lower hip CSA. These findings suggest that prenatal loading may exert long-lasting influences on skeletal development. INTRODUCTION: Breech position during pregnancy is associated with reduced range of fetal movement, and with lower limb joint stresses. Breech presentation at birth is associated with lower neonatal bone mineral content (BMC) and area, but it is unknown whether these associations persist into later life. METHODS: We examined associations between presentation at onset of labor, and tibia and hip bone outcomes at age 17 years in 1971 participants (1062 females) from a UK prospective birth cohort that recruited > 15,000 pregnant women in 1991-1992. Cortical BMC, cross-sectional area (CSA) and bone mineral density (BMD), periosteal circumference, and cross-sectional moment of inertia (CSMI) were measured by peripheral quantitative computed tomography (pQCT) at 50% tibia length. Total hip BMC, bone area, BMD, and CSMI were measured by dual-energy X-ray absorptiometry (DXA). RESULTS: In models adjusted for sex, age, maternal education, smoking, parity, and age, singleton/multiple births, breech presentation (n = 102) was associated with lower tibial cortical BMC (- 0.14SD, 95% CI - 0.29 to 0.00), CSA (- 0.12SD, - 0.26 to 0.02), BMD (- 0.16SD, - 0.31 to - 0.01), periosteal circumference (- 0.14SD, - 0.27 to - 0.01), and CSMI (- 0.11SD, - 0.24 to 0.01). In females only, breech presentation was associated with lower hip CSA (- 0.24SD, - 0.43 to 0.00) but not with other hip outcomes. Additional adjustment for potential mediators (delivery method, birthweight, gestational age, childhood motor competence and adolescent height and body composition) did not substantially affect associations with either tibia or hip outcomes. CONCLUSIONS: These findings suggest that prenatal skeletal loading may exert long-lasting influences on skeletal size and strength but require replication.

Breech presentation is associated with lower bone mass and area: findings from the Southampton Women's Survey.

We compared bone outcomes in children with breech and cephalic presentation at delivery. Neonatal whole-body bone mineral content (BMC) and area were lower in children with breech presentation. At 4 years, no differences in whole-body or spine measures were found, but hip BMC and area were lower after breech presentation. INTRODUCTION: Breech presentation is associated with altered joint shape and hip dysplasias, but effects on bone mineral content (BMC), area (BA) and density (BMD) are unknown. METHODS: In the prospective Southampton Women's Survey mother-offspring cohort, whole-body bone outcomes were measured using dual-energy X-ray absorptiometry (DXA) in 1430 offspring, as neonates (mean age 6 days, n = 965, 39 with a breech presentation at birth) and/or at age 4.1 years (n = 999, 39 breech). Hip and spine bone outcomes were also measured at age 4 years. RESULTS: Neonates with breech presentation had 4.2 g lower whole-body BMC (95% CI -7.4 to - 0.9 g, P = 0.012) and 5.9 cm2 lower BA (- 10.8 to - 1.0 cm2, P = 0.019), but BMD was similar between groups (mean difference - 0.007, - 0.016 to 0.002 g/cm2, P = 0.146) adjusting for sex, maternal smoking, gestational diabetes, mode of delivery, social class, parity, ethnicity, age at scan, birthweight, gestational age and crown-heel length. There were no associations between breech presentation and whole-body outcomes at age 4 years, but, in similarly adjusted models, regional DXA (not available in infants) showed that breech presentation was associated with lower hip BMC (- 0.51, - 0.98 to - 0.04 g, P = 0.034) and BA (- 0.67, - 1.28 to - 0.07 cm2, P = 0.03) but not with BMD (- 0.009, - 0.029 to 0.012 g, P = 0.408), or spine outcomes. CONCLUSIONS: These results suggest that breech presentation is associated with lower neonatal whole-body BMC and BA, which may relate to altered prenatal loading in babies occupying a breech position; these differences did not persist into later childhood. Modest differences in 4-year hip BMC and BA require further investigation.

Studies of the dynamics of nuclear clustering in human syncytiotrophoblast.

Syncytial nuclear aggregates (SNAs), clusters of nuclei in the syncytiotrophoblast of the human placenta, are increased as gestation advances and in pregnancy pathologies. The origins of increased SNAs are unclear; however, a better appreciation of the mechanism may give insight into placental ageing and factors underpinning dysfunction. We developed three models to investigate whether SNA formation results from a dynamic process of nuclear movement and to generate alternative hypotheses. SNA count and size were measured in placental explants cultured over 16 days and particles released into culture medium were quantified. Primary trophoblasts were cultured for 6 days. Explants and trophoblasts were cultured with and without cytoskeletal inhibitors. An in silico model was developed to examine the effects of modulating nuclear behaviour on clustering. In explants, neither median SNA number (108 SNA/mm(2) villous area) nor size (283 µm(2)) changed over time. Subcellular particles from conditioned culture medium showed a wide range of sizes that overlapped with those of SNAs. Nuclei in primary trophoblasts did not change position relative to other nuclei; apparent movement was associated with positional changes of the syncytial cell membrane. In both models, SNAs and nuclear clusters were stable despite pharmacological disruption of cytoskeletal activity. In silico, increased nuclear movement, adhesiveness and sites of cytotrophoblast fusion were related to nuclear clustering. The prominence of SNAs in pregnancy disorders may not result from an active process involving cytoskeleton-mediated rearrangement of syncytial nuclei. Further insights into the mechanism(s) of SNA formation will aid understanding of their increased presence in pregnancy pathologies.

Maternal prenatal depression is associated with decreased placental expression of the imprinted gene PEG3.

BACKGROUND: Maternal prenatal stress during pregnancy is associated with fetal growth restriction and adverse neurodevelopmental outcomes, which may be mediated by impaired placental function. Imprinted genes control fetal growth, placental development, adult behaviour (including maternal behaviour) and placental lactogen production. This study examined whether maternal prenatal depression was associated with aberrant placental expression of the imprinted genes paternally expressed gene 3 (PEG3), paternally expressed gene 10 (PEG10), pleckstrin homology-like domain family a member 2 (PHLDA2) and cyclin-dependent kinase inhibitor 1C (CDKN1C), and resulting impaired placental human placental lactogen (hPL) expression. METHOD: A diagnosis of depression during pregnancy was recorded from Manchester cohort participants' medical notes (n = 75). Queen Charlotte's (n = 40) and My Baby and Me study (MBAM) (n = 81) cohort participants completed the Edinburgh Postnatal Depression Scale self-rating psychometric questionnaire. Villous trophoblast tissue samples were analysed for gene expression. RESULTS: In a pilot study, diagnosed depression during pregnancy was associated with a significant reduction in placental PEG3 expression (41%, p = 0.02). In two further independent cohorts, the Queen Charlotte's and MBAM cohorts, placental PEG3 expression was also inversely associated with maternal depression scores, an association that was significant in male but not female placentas. Finally, hPL expression was significantly decreased in women with clinically diagnosed depression (44%, p < 0.05) and in those with high depression scores (31% and 21%, respectively). CONCLUSIONS: This study provides the first evidence that maternal prenatal depression is associated with changes in the placental expression of PEG3, co-incident with decreased expression of hPL. This aberrant placental gene expression could provide a possible mechanistic explanation for the co-occurrence of maternal depression, fetal growth restriction, impaired maternal behaviour and poorer offspring outcomes.

Marvellous to mediocre: findings of national survey of UK practice and provision of care in pregnancies after stillbirth or neonatal death.

BACKGROUND: Pregnancy after stillbirth or neonatal death is an emotionally challenging life-event for women and adequate emotional support during pregnancy should be considered an essential component of quality maternity care. There is a lack of evidence surrounding the role of UK maternity services in meeting womens' emotional and psychological needs in subsequent pregnancies. This study aimed to gain an overview of current UK practice and womens' experiences of care in pregnancy after the death of a baby. METHODS: Online cross-sectional surveys, including open and closed questions, were completed on behalf of 138 United Kingdom (UK) Maternity Units and by 547 women who had experience of UK maternity care in pregnancy after the death of a baby. Quantitative data were analysed descriptively using SPSS software. Open textual responses were managed manually and analysed using the framework method. RESULTS: Variable provision of care and support in subsequent pregnancies was identified from maternity unit responses. A minority had specific written guidance to support care delivery, with a focus on antenatal surveillance and monitoring for complications through increased consultant involvement and technological surveillance (ultrasound/cardiotocography). Availability of specialist services and professionals with specific skills to provide emotional and psychological support was patchy. There was a lack of evaluation/dissemination of developments and innovative practice. Responses across all UK regions demonstrated that women engaged early with maternity care and placed high value on professionals as a source of emotional support. Many women were positive about their care, but a significant minority reported negative experiences. Four common themes summarised womens' perceptions of the most important influences on quality and areas for development: sensitive communication and conduct of staff, appropriate organisation and delivery of services, increased monitoring and surveillance and perception of standard vs. special care. CONCLUSIONS: These findings expose likely inequity in provision of care for UK parents in pregnancy after stillbirth or neonatal death. Many parents do not receive adequate emotional and psychological support increasing the risk of poor health outcomes. There is an urgent need to improve the evidence base and develop specific interventions to enhance appropriate and sensitive care pathways for parents.

Parents' experiences and expectations of care in pregnancy after stillbirth or neonatal death: a metasynthesis.

BACKGROUND: Pregnancy after perinatal death is characterised by elevated stress and anxiety, increasing the risk of adverse short-term and long-term outcomes. OBJECTIVES: This metasynthesis aimed to improve understanding of parents' experiences of maternity care in pregnancy after stillbirth or neonatal death. SEARCH STRATEGY: Six electronic databases were searched using predefined search terms. SELECTION CRITERIA: English language studies using qualitative methods to explore the experiences of parents in pregnancy after perinatal loss, were included subject to quality appraisal framework. DATA COLLECTION AND ANALYSIS: Searches were initiated in December 2011 and finalised in March 2013. Studies were synthesised using an interpretive approach derived from meta-ethnography. MAIN RESULTS: Fourteen studies were included in the synthesis, graded A (no or few flaws, high trustworthiness; n = 5), B (some flaws, unlikely to affect trustworthiness; n = 5) and C (some flaws, possible impact on trustworthiness; n = 4). Three main themes were identified; co-existence of emotions, helpful and unhelpful coping activities and seeking reasssurance through interactions. CONCLUSION: Parents' experiences of pregnancy are profoundly altered by previous perinatal death; conflicted emotions, extreme anxiety, isolation and a lack of trust in a good outcome are commonly reported. Emotional and psychological support improves parents' experiences of subsequent pregnancy, but the absence of an evidence base may limit consistent delivery of optimal care within current services.

A difficult conversation? The views and experiences of parents and professionals on the consent process for perinatal postmortem after stillbirth.

OBJECTIVE: To describe the experiences, knowledge and views of both parents and professionals regarding the consent process for perinatal postmortem. DESIGN: Internet-based survey. SETTING: Obstetricians, midwives and perinatal pathologists currently working in the UK. Parents who have experienced a stillbirth in the UK in the previous 10 years. SAMPLE: Obstetricians, midwives and perinatal pathologists registered with their professional bodies. Parents who accessed the Sands website or online forum. METHODS: Online self-completion questionnaire with both fixed-choice and open-ended questions. RESULTS: Responses were analysed from 2256 midwives, 354 obstetricians, 21 perinatal pathologists and 460 parents. The most common reason for parents to request postmortem examination was to find a cause for their baby's death; the prevention of stillbirths in others also ranked highly. Perinatal pathologists possessed greatest knowledge of the procedure and efficacy of postmortem, but were unlikely to meet bereaved parents. The majority of professionals and parents ranked emotional distress and a lengthy wait for results as barriers to consent. The majority of staff ranked workload, negative publicity, religion and cultural issues as important barriers, whereas most parents did not. Almost twice as many parents who declined postmortem examination later regretted their decision compared with those who accepted the offer (34.4 versus 17.4%). CONCLUSION: Emotional, practical and psychosocial issues can act as real or perceived barriers for staff and bereaved parents. Education is required for midwives and obstetricians, to increase their knowledge to ensure accurate counselling, with due regard for the highly individual responses of bereaved parents. The contribution of perinatal pathologists to staff education and parental decision-making would be invaluable.

Correlation between abnormal placental appearance at routine 2nd trimester ultrasound scan and histological examination of the placenta after birth.

Ultrasound imaging of the placenta and its relationship to poor perinatal outcome has been studied since 1973. Studies have predominantly focussed on placental morphology in the third trimester or in high-risk pregnancies. We aimed to correlate abnormal placental appearances identified in the second trimester with histological appearances. We present four cases of abnormal placental morphology detected at the 20 week routine ultrasound scan in low-risk women and relate them to histological origins, which often involved areas of villous infarction. Abnormal placental appearances at routine second trimester anomaly ultrasound scans might identify women with placental dysfunction who merit increased fetal surveillance in the third trimester.

Fetal blood sampling in early labour: is there an increased risk of operative delivery and fetal morbidity?

OBJECTIVE: To determine whether the rate of caesarean section was increased in women undergoing fetal blood sampling (FBS) in early labour. DESIGN: Retrospective study. SETTING: Secondary and tertiary obstetric units in the UK. POPULATION: A cohort of 381 women undergoing FBS. METHODS: Data relating to demographics, labour and delivery characteristics, and neonatal outcomes were collected on women undergoing FBS in labour. Odds ratios (ORs) for caesarean section compared with vaginal delivery for women who had their first FBS in early labour (≤ 3 cm cervical dilatation) and for women who required multiple samples were calculated. MAIN OUTCOME MEASURES: Mode of delivery. RESULTS: Forty-eight percent of women who required their first FBS at a cervical dilatation of ≤ 3 cm achieved a vaginal delivery; these women were at modestly increased risk of caesarean section (adjusted OR 1.80; 95% CI 1.04-3.13) compared with women who had their first FBS at a cervical dilatation of ≥ 4 cm. The odds ratio for caesarean section in women who required two or more FBS was 1.71 (95% CI 1.37-2.13) compared with those requiring a single sample. There were no differences in instrumental delivery. Infants undergoing three or more FBS were more likely to be admitted to a neonatal intensive care unit (NICU; OR 2.69; 95% CI 1.09-6.64), although this was not associated with increased acidaemia. CONCLUSIONS: Women who require FBS in early labour or multiple FBS are at a modestly increased risk of caesarean section compared with those in established labour. When contemplating FBS at ≤ 3-cm cervical dilatation, practitioners should not be put off by the perceived low chance of vaginal delivery, but repeating FBS on more than three occasions should be considered carefully.

Can post-mortem examination of the placenta help determine the cause of stillbirth?

Some cases of stillbirth are associated with placental abnormality; recent classification systems have included some features of placental pathology. This study aimed to determine whether placental investigations assist in determining the cause of stillbirth. A total of 71 consecutive cases of stillbirth were reviewed. Placental investigations were undertaken in 54% of cases. Women who had placental assessment were significantly less likely to have an unexplained stillbirth (OR = 0.17; 95% CI 0.04-0.70). In 47% of cases, the findings of placental investigation were included in the classification of stillbirth. In 16% of cases the classification was determined primarily by placental examination. Some placental abnormalities found were associated with clinical causes of stillbirth, such as placental infarction and IUGR or leukocyte infiltration and chorioamnionitis (p < 0.05). We conclude that assessment of the placenta can aid classification of stillbirth and recommend that histological analysis of placental tissue be offered in all cases of stillbirth, even when full infant post-mortem is declined.

Predicting poor perinatal outcome in women who present with decreased fetal movements.

Maternal perception of decreased fetal movements (DFM) is associated with increased incidence of stillbirth and intrauterine growth restriction. We hypothesised that clinical assessment of women perceiving DFM may identify patients at highest risk of poor perinatal outcome. This was a retrospective study of 203 patients presenting to the obstetric triage service with DFM. Information on obstetric and past medical history, the current presentation with DFM and the outcome of pregnancy was collected. Using multivariate analysis, odds ratios (OR) and 95% confidence intervals (CI) were calculated for poor pregnancy outcome defined as stillbirth, small for gestational age or pre-term delivery. The rate of stillbirth was increased in women with DFM (OR 2.9). Some 26.6% of women perceiving DFM had a poor perinatal outcome. Women with relevant past obstetric history (OR 2.11), two or more presentations of DFM (OR 1.92), or who measured small-for-dates (OR 19.53) were at increased risk of poor pregnancy outcome. These preliminary data suggest that some features of clinical assessment can identify patients at increased risk of poor perinatal outcome after presentation with DFM. Such patients may be prioritised for detailed assessment of fetal growth and wellbeing.

Live and let die - regulation of villous trophoblast apoptosis in normal and abnormal pregnancies.

Since 1995 the number of publications investigating apoptosis in villous trophoblast has increased exponentially. This scientific interest is in part due to observations that this specialised form of cell death is increased in pregnancy complications such as pre-eclampsia and intra-uterine growth restriction. In addition, apoptosis is described in normal villous trophoblast and elements of the apoptotic machinery are involved in the fusion between cytotrophoblast and the overlying multinucleate syncytiotrophoblast. The increase in descriptions of apoptotic cell death in villous trophoblast has been accompanied by investigations of regulators of apoptosis. It is anticipated that understanding the regulation of apoptosis in villous trophoblast may provide new insights into placental pathologies. This review describes current knowledge regarding the expression and function of these regulators in villous trophoblast, both in normal and complicated pregnancies.

Effects of oxygen on cell turnover and expression of regulators of apoptosis in human placental trophoblast.

Pre-eclampsia (PE) and intrauterine growth restriction (IUGR) are associated with aberrant cell turnover, including increased apoptosis, in placental villous trophoblast. The increased apoptosis is associated with exaggerated expression of p53, which promotes cell cycle arrest or apoptosis via downstream proteins such as p21 or Bax. These changes in apoptosis and p53 expression are purported to result from exposure to altered oxygen tension. Using a model of villous trophoblast turnover, we examined the effect of 20%, 6% and 1% ambient oxygen (O(2)) on apoptosis, necrosis, proliferation and expression of p53 and related regulators of cell turnover, compared to both fresh tissue. Altered O(2) tension exerted an effect on cell turnover in cultured term villous tissue: cytotrophoblast proliferation was increased by culture in 20% O(2) and reduced in 1% O(2) (median proliferative index: fresh tissue=0.32%, 20% O(2)=0.9%, 6% O(2)=0.28%, 1% O(2)=0.07%). Apoptosis was increased in all culture environments, but was significantly enhanced by culture in 1% O(2) (median apoptotic index: fresh tissue=0.64%, 20% O(2)=2.96%, 6% O(2)=3.81%, 1% O(2)=9.2%). Necrotic cell death was also increased by culture in 1% O(2) compared to 6% and 20% O(2). The expression of p53, p21 and Mdm2 in both cytotrophoblast and stromal cells was increased following culture in 1% O(2). There was no alteration in the expression of Bax or Bcl-2. This study provides evidence that p53 is elevated in trophoblast following exposure to hypoxia. The potential role of the p53-pathway in the control of cell turnover in villous trophoblast and the regulation of p53 by altered O(2) tension merits further investigation.

Analysis of the metabolic footprint and tissue metabolome of placental villous explants cultured at different oxygen tensions reveals novel redox biomarkers.

Pre-eclampsia (PE) is a multi-system disorder of pregnancy hypothesised to arise from circulating factors derived from an unhealthy placenta. Some changes in placental phenotype seen in PE can be reproduced by culture in altered oxygen (O2) tension. Currently, these circulating factors are unidentified, partly due to the complexity of maternal plasma. Investigation of factors released from placental tissue provides a potential method to identify bioactive compounds. Experimental strategies to study compounds present in a biological system have expanded greatly in recent years. Metabolomics can detect and identify endogenous and secreted metabolites. We aimed to determine whether metabolites could be identified in placental cultures with acceptable experimental variability and to determine whether altered O2 tension affects the composition of the placental metabolome. In this study we used gas-chromatography-mass spectroscopy to determine the presence of metabolites in conditioned culture medium (CCM) and tissue lysates of placental villous explants cultured in 1, 6 and 20% atmospheric O2 for 96h. This experimental strategy had an intra-assay variation of 6.1-11.6%. Intra and inter-placental variability were 15.7-35.8% and 44.8-46.2% respectively. Metabolic differences were identified between samples cultured in 1, 6 and 20% O2 in both CCM and tissue lysate. Differentially expressed metabolites included: 2-deoxyribose, threitol or erythritol and hexadecanoic acid. We conclude that metabolomic strategies offer a novel approach to investigate placental function. When conducted under carefully controlled conditions, with appropriate statistical analysis, metabolic differences can be identified in placental explants in response to altered O2 tension. Metabolomics could be used to identify changes in conditions associated with placental pathology.

Methods of fetal movement counting and the detection of fetal compromise.

Maternal perception of fetal movements is widely used as a marker of fetal viability and well-being. A reduction in fetal movements is associated with fetal hypoxia, increased incidence of stillbirth and fetal growth restriction (FGR). Therefore, a reduction in fetal movements has been proposed as a screening tool for FGR or fetal compromise. The problem of this approach is that there is no widely accepted definition of reduced fetal activity or 'alarm limits', and pregnant women are currently given a wide range of non-evidence-based advice. We have reviewed the background of published definitions and their potential usefulness in screening. A formal meta-analysis of these studies is not possible due to variation in methodology and definitions of reduced fetal movements. Assessment of fetal movements using formal fetal movement counting has shown equivocal results. Importantly, in all studies, there was a decrease in perinatal mortality suggesting a beneficial role for raising maternal awareness of fetal movements. Most studies implemented limits to define reduced fetal movements based on small groups of high risk pregnancies and obsolete counting methodology. A single case-control study developed 'normal limits' in a low risk population, and successfully implemented it prospectively for screening. At present, there is no evidence that any absolute definition of reduced fetal movements is of greater value than maternal subjective perception of reduced fetal movements in the detection of intrauterine fetal death or fetal compromise. Further investigation is required to determine an effective method of identifying patients with reduced fetal movements and to determine the best subsequent management.

Identification of the functional pathways altered by placental cell exposure to high glucose: lessons from the transcript and metabolite interactome.

The specific consequences of hyperglycaemia on placental metabolism and function are incompletely understood but likely contribute to poor pregnancy outcomes associated with diabetes mellitus (DM). This study aimed to identify the functional biochemical pathways perturbed by placental exposure to high glucose levels through integrative analysis of the trophoblast transcriptome and metabolome. The human trophoblast cell line, BeWo, was cultured in 5 or 25 mM glucose, as a model of the placenta in DM. Transcriptomic analysis using microarrays, demonstrated 5632 differentially expressed gene transcripts (≥± 1.3 fold change (FC)) following exposure to high glucose. These genes were used to generate interactome models of transcript response using BioGRID (non-inferred network: 2500 nodes (genes) and 10541 protein-protein interactions). Ultra performance-liquid chromatography-mass spectrometry (MS) and gas chromatography-MS analysis of intracellular extracts and culture medium were used to assess the response of metabolite profiles to high glucose concentration. The interactions of altered genes and metabolites were assessed using the MetScape interactome database, resulting in an integrated model of systemic transcriptome (2969 genes) and metabolome (41 metabolites) response within placental cells exposed to high glucose. The functional pathways which demonstrated significant change in response to high glucose included fatty acid ß-oxidation, phospholipid metabolism and phosphatidylinositol phosphate signalling.