Control Fetuses Research Articles

Fetal magnetic resonance imaging (MRI) enhances the diagnosis of congenital body anomalies

AimsWe sought to assess variability and concordance between fetal MRI and ultrasound (USS) in the evaluation of fetal body abnormalities. MethodsAll fetal body anomalies reported on F-MRI within the iFIND database (http://www.ifindproject.com) were included. Differences in findings regarding anomalies on contemporaneous USS were explored. Three clinical specialists evaluated each case independently, and the anomaly severity was graded: as “insignificant” to “lethal”. The value of MRI in alteration of either antenatal or postnatal care was established. ResultsFifty-four cases were identified consisting of 5 healthy controls, 37 with USS-identified body anomalies, and 12 with known CNS or cardiac anomalies. In fetuses with a known body anomaly, information on the MRI was relevant to change the clinical course in 59% of cases. There was also an incidental detection rate of 7% in fetuses with known cardiac or CNS anomalies, or 1.5% of normal control, although these were rarely clinically relevant.Importantly, fetuses undergoing MRI for cardiac concerns did have major anomalies that were missed (one case of oesophageal atresia and two cases of ARM). ConclusionsIn cases where fetal anomalies are suspected, F-MRI is a valuable means of further characterizing anomalies and may detect additional anomalies in fetuses with recognized cardiac or CNS anomalies. In fetuses with a recognized body anomaly, more than half of those scanned by MRI had information available which changed clinical management. Importantly there were also incidental findings in healthy control fetuses, so the management of these needs to be recognized in fetal MRI research. Level of evidenceII, Prospective cohort study.

Pancreas deficiency modifies bone development in the ovine fetus near term.

Hormones have an important role in the regulation of fetal growth and development, especially in response to nutrient availability in utero. Using micro-CT and an electromagnetic three-point bend test, this study examined the effect of pancreas removal at 0.8 fraction of gestation on the developing bone structure and mechanical strength in fetal sheep. When fetuses were studied at 10 and 25 days after surgery, pancreatectomy caused hypoinsulinaemia, hyperglycaemia and growth retardation which was associated with low plasma concentrations of leptin and a marker of osteoclast activity and collagen degradation. In pancreatectomized fetuses compared to control fetuses, limb lengths were shorter, and trabecular (Tb) bone in the metatarsi showed greater bone volume fraction, Tb thickness, degree of anisotropy and porosity, and lower fractional bone surface area and Tb spacing. Mechanical strength testing showed that pancreas deficiency was associated with increased stiffness and a greater maximal weight load at fracture in a subset of fetuses studied near term. Overall, pancreas deficiency in utero slowed the growth of the fetal skeleton and adapted the developing bone to generate a more compact and connected structure. Maintenance of bone strength in growth-retarded limbs is especially important in a precocial species in preparation for skeletal loading and locomotion at birth.

Serial neurosonography in fetuses with congenital heart defects shows mild delays in cortical development.

IntroductionNeurodevelopmental delay is more common in children born with congenital heart defects (CHD), even with optimal perinatal and peri‐operative care. It is hypothesized that fetuses with CHD are prone to neurological impairment in utero due to their cardiac defect, possibly leading to delayed cortical development.MethodsCerebral cortical maturation was assessed with advanced neurosonographic examinations every 4 weeks in fetuses with CHD and compared to control fetuses. Five different primary fissures and four areas were scored (ranging 0–5) by blinded examiners using a cortical maturation scheme.ResultsCortical staging was assessed in 574 ultrasound examinations in 85 CHD fetuses and 61 controls. Small differences in grading were seen in Sylvian and cingulate fissures. (Sylvian fissure: −0.12 grade, 95% CI (−0.23; −0.01) p = 0.05, cingulate fissure: −0.24 grade, 95% CI (−0.38; −0.10) p = <0.001. Other cortical areas showed normal maturation as compared to control fetuses.ConclusionSmall differences were seen in three of the nine analyzed cortical areas in CHD fetuses, in contrast to previous reports on progressive third‐trimester delay. The clinical implications of the small differences however, remain unknown.

Myocardial deformation analysis in late-onset small-for-gestational-age and growth-restricted fetuses using two-dimensional speckle tracking echocardiography: a prospective cohort study.

An association between fetal growth restriction (FGR) and increased predisposition to cardiovascular disease (CVD) is suggested. The aim of this study was to evaluate subclinical signs of fetal cardiac remodeling in late-onset small-for-gestational-age (SGA) and growth-restricted fetuses using two-dimensional speckle tracking echocardiography (2D-STE). This is a prospective cohort study, including 117 late-onset (≥32weeks) SGA (birthweight≤10th centile) fetuses and 102 gestational age matched controls. A subgroup analysis was performed: FGR was defined based on either BW (<third centile) or the presence of feto-maternal Doppler abnormalities, the remaining cases were called SGA centile 3-10. 2D-STE based myocardial deformation analyses included longitudinal peak systolic strain (LPSS) and strain rate (LSR) of the global left (LV) or right (RV) ventricle and its individual segments. Global and segmental LPSS and LSR values showed no significant difference between late-onset SGA and control fetuses for both ventricles (p>0.05). Regarding global and segmental LPSS and LSR values of LV/RV, subgroup analysis revealed no significant difference between the FGR (n=81), SGA centile 3-10 (n=36) and control group. A mild degree of placental dysfunction seems not to influence myocardial deformation properties measured by 2D-STE.

Redox ratio in the left ventricle of the growth restricted fetus is positively correlated with cardiac output.

Intrauterine growth restriction (IUGR) is a result of limited substrate supply to the developing fetus in utero, and can be caused by either placental, genetic or environmental factors. Babies born IUGR can have poor long-term health outcomes, including being at higher risk of developing cardiovascular disease. Limited substrate supply in the IUGR fetus not only changes the structure of the heart but may also affect metabolism and function of the developing heart. We have utilised two imaging modalities, two-photon microscopy and phase-contrast MRI (PC-MRI), to assess alterations in cardiac metabolism and function using a sheep model of IUGR. Two-photon imaging revealed that the left ventricle of IUGR fetuses (at 140-141 d GA) had a reduced optical redox ratio, suggesting a reliance on glycolysis for ATP production. Concurrently, the use of PC-MRI to measure foetal left ventricular cardiac output (LVCO) revealed a positive correlation between LVCO and redox ratio in IUGR, but not control fetuses. These data suggest that altered heart metabolism in IUGR fetuses is indicative of reduced cardiac output, which may contribute to poor cardiac outcomes in adulthood.

Fetal cardiac geometry and function in pregnancies with well-controlled gestational diabetes mellitus using Fetal HQ

Objective To determine whether abnormal cardiac shape and ventricular global, transverse, and longitudinal contractility are present in fetuses of women with well-controlled GDM. Methods A prospective observational study was performed on 80 fetuses of women with well-controlled GDM and 90 control fetuses. Using Fetal HQ, a new speckle-tracking technique, cardiac shape, global contractility, transverse contractility, and longitudinal contractility were calculated. The number and percentage of fetuses with z score values below 5th or above 95th were computed. Results Compared with controls, there were no significant differences in the frequency of cardiac geometric abnormalities in GDM fetuses. Despite good glycemic control, 60.0% of fetuses in the well-controlled GDM group had one or more types of global, longitudinal, and transverse contractility abnormalities of one or both ventricles, but more frequent on the right ventricle (RV, 50%). The most frequent abnormality of the RV occurred in the transverse contractility (35%), followed by abnormalities of global contractility (25%), and longitudinal contractility (21.3%), compared with controls. The left ventricle (LV) analysis demonstrated that the percentage of study fetuses with only transverse contractility abnormality (18.8%) was significantly higher. Conclusions Despite good glycemic control, abnormal ventricular contractility was present in fetuses of women with GDM, but more frequent in the RV. For both the RV and LV, transverse ventricular contractility abnormality were more prevalent than abnormal global and longitudinal contractility. Fetuses of women with GDM should be evaluated for ventricular contractility abnormality and have more follow-ups despite good glycemic control.

Amniotic fluid metabolomic and lipidomic alterations associated with hemoglobin Bart's diseases.

α-Thalassemia is the most common inherited disease in southern China. The severest form is hemoglobin (Hb) Bart's disease, in which the affected fetuses almost always die in utero or shortly after birth, and the mothers are at high risk for severe morbidity. To investigate the changes in all metabolites in fetuses with Hb Bart's disease and to characterize the metabolomic and lipidomic biomarkers in the development of Hb Bart's fetuses. Amniotic fluid (AF) specimens were selected from 34 pregnant women who underwent interventional prenatal diagnosis from June 2017 to June 2018. Gap-PCR analysis was used to diagnose Hb Bart's disease, and untargeted metabolomic and lipidomic analyses were performed. By analyzing AF samples, 935 differential metabolites were selected between Hb Bart's and control fetuses. The metabolites with significant changes mainly involved D-glutamine and D-glutamate metabolism, histidine metabolism, arginine metabolism, beta-alanine metabolism and alanine, aspartate and glutamate metabolism. Further lipidomics analysis revealed 132 differential lipids, mainly involved phosphatidylcholine and triglyceride metabolism. Through the characterized metabolites in AF, a schematic model of Hb Bart's disease was established. Glutamate and glutathione metabolism, aspartate metabolism, urea metabolism and triglyceride metabolism were significantly changed in the Hb Bart's group compared to the control group. The characterized biomarkers were mainly involved in oxidative stress reaction, iron overload and liver dysfunction. This finding may help improve the treatment options for α-thalassemia as well as diagnosing phenotype of patients.

Characterizing the effects of in utero valproic acid exposure on murine fetoplacental development

IntroductionValproic acid (VPA) is an effective anti-epileptic drug clinically used to treat seizures, bipolar disorders and neuropathic pain in women of reproductive age. Current approval of VPA for psychiatric conditions and migraine has increased the number of VPA exposed pregnancies. VPA crosses the placental barrier and induces birth defects in about 10% of exposed pregnancies. In addition, VPA exposure results in neurodevelopmental disorders in children without any overt birth defects. The current study was designed to investigate the effects of in utero VPA exposure on fetoplacental growth in a mouse model. MethodsPregnant CD-1 dams were exposed to a single teratogenic dose of 400 mg/kg VPA or saline via subcutaneous injection on gestational day (GD) 9 and fetuses were harvested on GD 13, 15, 17 and 19, respectively. Resorptions, gross malformations, fetal weight, fetal head weight, fetal crown-rump length, fetal head transverse and anteroposterior diameters, placental weight and placental diameter were noted. ResultsVPA exposure led to multiple external deformities including exencephaly, open eye defect, subcutaneous hemorrhage and underdevelopment of tail. All fetoplacental growth parameters fetal weight, fetal head weight, fetal crown-rump length, placental weight and placental diameter were significantly reduced in VPA-exposed fetuses with and without congenital malformations such as exencephaly, compared to control fetuses. DiscussionIn conclusion, the effects of in utero VPA exposure on fetal and placental growth persisted throughout pregnancy and our results suggest that the effects of VPA on placental growth may play a role in VPA-induced toxicity.

Decreased tight junction protein intensity in the placenta of porcine reproductive and respiratory syndrome virus-2 infected fetuses

IntroductionExisting strategies to control porcine reproductive and respiratory syndrome (PRRS) are not completely effective and require alternative approaches. Although intrauterine growth restricted (IUGR) fetuses are more resilient to transplacental PRRS virus-2 (PRRSV2) infection compared to normal fetuses, the exact mechanisms are unknown. The objective of this research was to assess abundance and localization of a subset of tight junction (TJ) proteins in the maternal-fetal interface and any alterations that may affect the movement of nutrients or PRRSV2 across the epitheliochorial placenta. MethodsParaffin-embedded samples of placenta from non-infected control (CTRL) and PRRSV2 infected fetuses (IUGR, non(N)-IUGR, meconium-stained (MEC) (n = 6 per group) were randomly selected from a large challenge trial and immunostained for claudins (CLDN) 1, 3, 4, 7 and tight junction protein 1 (TJP1). Immunostaining intensity was semi-subjectively scored by region. ResultsIntensity of CLDN1 was lower in placenta of IUGR, MEC, and N-IUGR fetuses compared to CTRL, mainly in fetal epithelium and maternal endothelial cells (MECL). CLDN4 intensity was lower in MECL of IUGR compared to CTRL and MEC fetuses. TJP1 intensity was lower in maternal and fetal epithelia of placenta within IUGR, MEC, and N-IUGR fetuses versus CTRL. DiscussionDifferences were mainly observed between PRRSV2 infected and non-infected groups indicating TJ integrity was affected by PRRSV2 infection. These results provide insights into the potential mechanisms of transplacental transmission of PRRSV2; however, since only CLDN4 differed amongst the infected groups, PRRSV2 induced changes in TJ integrity do not appear to explain variation in fetal outcomes after infection.

Anemic hypoxemia reduces myoblast proliferation and muscle growth in late-gestation fetal sheep.

Fetal skeletal muscle growth requires myoblast proliferation, differentiation, and fusion into myofibers in addition to protein accretion for fiber hypertrophy. Oxygen is an important regulator of this process. Therefore, we hypothesized that fetal anemic hypoxemia would inhibit skeletal muscle growth. Studies were performed in late-gestation fetal sheep that were bled to anemic and therefore hypoxemic conditions beginning at ∼125 days of gestation (term = 148 days) for 9 ± 0 days (n = 19) and compared with control fetuses (n = 16). A metabolic study was performed on gestational day ∼134 to measure fetal protein kinetic rates. Myoblast proliferation and myofiber area were determined in biceps femoris (BF), tibialis anterior (TA), and flexor digitorum superficialis (FDS) muscles. mRNA expression of muscle regulatory factors was determined in BF. Fetal arterial hematocrit and oxygen content were 28% and 52% lower, respectively, in anemic fetuses. Fetal weight and whole body protein synthesis, breakdown, and accretion rates were not different between groups. Hindlimb length, however, was 7% shorter in anemic fetuses. TA and FDS muscles weighed less, and FDS myofiber area was smaller in anemic fetuses compared with controls. The percentage of Pax7+ myoblasts that expressed Ki67 was lower in BF and tended to be lower in FDS from anemic fetuses indicating reduced myoblast proliferation. There was less MYOD and MYF6 mRNA expression in anemic versus control BF consistent with reduced myoblast differentiation. These results indicate that fetal anemic hypoxemia reduced muscle growth. We speculate that fetal muscle growth may be improved by strategies that increase oxygen availability.

Assessment of fetal cardiac diastolic function of gestational diabetes mellitus using dual-gate Doppler.

Gestational Diabetes Mellitus (GDM), as a common complication of pregnancy, has an increasing trend globally. GDM leads to maternal complications and fetal complications. Fetal cardiac diastolic dysfunction is strongly associated with GDM. This study aims to assess the ventricular diastolic function of fetuses exposed to GDM by looking into the diagnostic parameters using both conventional method and Dual-gate Doppler method (DD). And to investigate the potential of DD method in early detection of fetal cardiac diastolic dysfunction.56 women diagnosed with GDM and 55 non-GDM pregnant women were enrolled in their 24 to 30 weeks of gestation. Conventional method and DD method were applied to measure mitral and tricuspid inflow velocities E-waves, A-waves on pulsed-wave Doppler, and mitral and tricuspid annular velocities e′-waves, a’-waves on Tissue Doppler imaging. E/A, e’/a’ and E/e′ ratio was calculated. The difference between GDM and control groups was statistically tested and analysed using one-sample Kolmogorov-Smirnov test, Student t test, Mann-Whitney U test and Kruskal-Wallis test and Bland–Altman plot analysis.Intraobserver intraclass correlation coefficients of E/A, e’/a’, and E/e’ value of both mitral and tricuspid valve are all greater than 0.80, while interobserver intraclass correlation coefficients are between 0.71 and 0.88. Right (6.35 vs 6.79; P = .001) ventricular function showed significantly lower E/e’ ratios in the GDM group compared with control fetuses by conventional method. Both left (6.16 vs 6.59; P = .036) and right (6.28 vs 6.75; P = .01) ventricular function showed significantly lower E/e’ ratios in the GDM group compared with control fetuses by DD method.Exposure to high level of maternal blood glucose leads to impaired diastolic function in the fetuses. Fetal right ventricular function is a potential key point to study to enable an early detection for fetal diastolic dysfunction since the alteration and damage are more likely to happen in right ventricular. Measurement of E/e’ ratio using DD method is considered as a promising method in fetal cardiac diastolic function assessment. Well or poorly control of the GDM does not have significant influence on the fetal diastolic function thus an early detection of GDM and GDM induced fetal cardiac dysfunction is necessary.

Mapping of the amniotic fluid proteome of fetuses with congenital anomalies of the kidney and urinary tract identifies plastin 3 as a protein involved in glomerular integrity.

Congenital anomalies of the kidney and the urinary tract (CAKUT) are the first cause of chronic kidney disease in childhood. Several genetic and environmental origins are associated with CAKUT, but most pathogenic pathways remain elusive. Considering the amniotic fluid (AF) composition as a proxy for fetal kidney development, we analyzed the AF proteome from non-severe CAKUT (n = 19), severe CAKUT (n = 14), and healthy control (n = 22) fetuses using LC-MS/MS. We identified 471 significant proteins that discriminated the three AF groups with 81% precision. Among them, eight proteins independent of gestational age (CSPG4, LMAN2, ENDOD1, ANGPTL2, PRSS8, NGFR, ROBO4, PLS3) were associated with both the presence and the severity of CAKUT. Among those, five were part of a protein-protein interaction network involving proteins previously identified as being potentially associated with CAKUT. The actin-bundling protein PLS3 (plastin 3) was the only protein displaying a gradually increased AF abundance from control, via non-severe, to severe CAKUT. Immunohistochemistry experiments showed that PLS3 was expressed in the human fetal as well as in both the fetal and the postnatal mouse kidney. In zebrafish embryos, depletion of PLS3 led to a general disruption of embryonic growth including reduced pronephros development. In postnatal Pls3-knockout mice, kidneys were macroscopically normal, but the glomerular ultrastructure showed thickening of the basement membrane and fusion of podocyte foot processes. These structural changes were associated with albuminuria and decreased expression of podocyte markers including Wilms' tumor-1 protein, nephrin, and podocalyxin. In conclusion, we provide the first map of the CAKUT AF proteome that will serve as a reference for future studies. Among the proteins strongly associated with CAKUT, PLS3 did surprisingly not specifically affect nephrogenesis but was found as a new contributor in the maintenance of normal kidney function, at least in part through the control of glomerular integrity. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Perinatal Outcome and its Prediction Using Longitudinal Feto-Maternal Doppler Follow-Up in Late Onset Small for GestationalAge Fetuses - A Prospective Cohort Study.

To describe the perinatal outcome of a prospective cohort of late-onset small-for-gestational-age (SGA) fetuses and to test adverse perinatal outcome (APO) prediction using Doppler measurements. Singleton pregnancies from 32 weeks with suspicion of SGA (followed-up each 2 weeks) and randomly selected healthy controls at a university hospital were included. The whole SGA group was divided into the FGR subgroup or SGA percentile 3-10 subgroup. The following Doppler measurements were evaluated prospectively: umbilical artery (UA) pulsatility index (PI), middle cerebral artery (MCA) PI, cerebro-placental ratio (CPR), and mean uterine artery (mUtA) PI. APO was defined as arterial cord blood pH ≤ 7.15 and/or 5-minute Apgar ≤ 7 and/or emergency operative delivery and/or admission to the neonatal unit. Induction of labor was indicated according to a stage-based protocol. A total of 149 SGA and 143 control fetuses were included. The number of operative deliveries was similar between both groups (control: 29 %, SGA: 28 %), especially the cesarean delivery rate after the onset of labor (11 % vs. 10 %). Most SGA cases ended up in induction of labor (61 % vs. 31 %, p < 0.001). The areas under the curve (AUC) for APO prediction were similar using the last UA PI, MCA PI, CPR, and mUtA PI and barely reached 0.60. The AUC was best for the FGR subgroup, using the minimal CPR or maximum mUtA PI z-score of all longitudinal measurements (AUC = 0.63). SGA fetuses do not have a higher rate of operative delivery if managed according to a risk stratification protocol. Prediction of APO is best for SGA and FGR using the "worst" CPR or mUtA PI but it remains moderate.

Augmented glucose production is not contingent on high catecholamines in fetal sheep with IUGR.

Fetuses with intrauterine growth restriction (IUGR) have high concentrations of catecholamines, which lowers the insulin secretion and glucose uptake. Here, we studied the effect of hypercatecholaminemia on glucose metabolism in sheep fetuses with placental insufficiency-induced IUGR. Norepinephrine concentrations are elevated throughout late gestation in IUGR fetuses but not in IUGR fetuses with a bilateral adrenal demedullation (IAD) at 0.65 of gestation. Euglycemic (EC) and hyperinsulinemic-euglycemic (HEC) clamps were performed in control, intact-IUGR, and IAD fetuses at 0.87 of gestation. Compared to controls, basal oxygen, glucose, and insulin concentrations were lower in IUGR groups. Norepinephrine concentrations were five-fold higher in IUGR fetuses than in IAD fetuses. During the EC, rates of glucose entry (GER, umbilical + exogenous), glucose utilization (GUR), and glucose oxidation (GOR) were greater in IUGR groups than in controls. In IUGR and IAD fetuses with euglycemia and euinsulinemia, glucose production rates (GPR) remained elevated. During the HEC, GER and GOR were not different among groups. In IUGR and IAD fetuses, GURs were 40% greater than in controls, which paralleled the sustained GPR despite hyperinsulinemia. Glucose-stimulated insulin concentrations were augmented in IAD fetuses compared to IUGR fetuses. Fetal weights were not different between IUGR groups but were less than controls. Regardless of norepinephrine concentrations, IUGR fetuses not only develop greater peripheral insulin sensitivity for glucose utilization but also develop hepatic insulin resistance because GPR was maintained and unaffected by euglycemia or hyperinsulinemia. These findings show that adaptation in glucose metabolism of IUGR fetuses are independent of catecholamines, which implicate that hypoxemia and hypoglycemia cause the metabolic responses.

209 Large Offspring Syndrome: Effects of in vitro Production on Embryo Epigenetics and Development

Abstract In cattle, the use of assisted reproductive technologies (ART) can result in a congenital overgrowth condition known as large/abnormal offspring syndrome (LOS/AOS). The phenotypic characteristics of LOS include; somatic overgrowth, abdominal wall defects, large organs, breathing difficulties, skeletal defects, hypoglycemia, abnormal placentas, difficulty suckling, and perinatal death. LOS can have detrimental effects on the offspring and dam and also pose managerial and financial challenges to the producer. Research from the Rivera laboratory has demonstrated that LOS is an epigenetic syndrome. As in cattle, ART can promote the development of congenital overgrowth in humans, a condition known as Beckwith Wiedemann Syndrome (BWS). For the past 13 years, the Rivera laboratory has been characterizing LOS and we have shown that LOS and BWS are phenotypically and epigenotypically similar. In our studies, using gestation day ~105 Bos taurus taurus x Bos taurus indicus F1 hybrids, we showed global misregulation of imprinted and non-imprinted transcripts, micro RNAs and global misregulation of DNA methylation. In brief, LOS fetuses displayed variable loss-of-imprinting in kidney, liver, muscle and brain, when compared to controls. Biallelic expression of imprinted genes in LOS was associated with tissue-specific hypomethylation of the normally methylated parental allele. Not only was there loss of allele-specific expression of imprinted genes in LOS, but we also observed differential transcript amounts of these genes between control and overgrown fetuses. In addition, a positive correlation was observed between bodyweight and the number of biallelically expressed imprinted genes in LOS fetuses. From this work, we concluded that LOS is a multi-locus loss-of-imprinting condition. Current work, aims to determine if LOS is identifiable during pregnancy using day 55 fetal ultrasonography and day 55 and 105 maternal blood. In addition, we aim to determine how serum supplementation of culture medium can program preimplantation embryos to develop LOS. Findings will be discussed.

Forthcoming Issues

Fetuses with an estimated fetal weight below the 10th centile have an increased risk of adverse perinatal and long-term outcomes as well as increased rates of cardiac dysfunction, which often alters cardiac size and shape of the 4-chamber view and the individual ventricles. As a result, a simple method has emerged to screen for potential cardiac dysfunction in fetuses with estimated fetal weights <10th centile by measuring the size and shape of the 4-chamber view and the size of the ventricles.To determine the number of fetuses with an abnormal size and shape of the 4-chamber view and size of the ventricles in fetuses with an estimated fetal weight <10th centile.This was a retrospective study of 50 fetuses between 25 and 37 weeks of gestation with an estimated fetal weight <10th centile. Data from their last examination were analyzed. From an end-diastolic image of the 4-chamber view, the largest basal–apical length and transverse width were measured from their corresponding epicardial borders. This allowed the 4-chamber view area and global sphericity index (4-chamber view length/4-chamber view width) to be computed. In addition, tracing along the endocardial borders with speckle tracking software enabled measurements of the right and left ventricular chamber areas and the right ventricle/left ventricle area ratios to be computed. Doppler waveform pulsatility indices from the umbilical (umbilical artery pulsatility index) and middle cerebral arteries (middle cerebral artery pulsatility index) were analyzed, and the cerebroplacental ratio (middle cerebral artery pulsatility index/umbilical artery pulsatility index) computed. Umbilical artery pulsatility indices >90th and cerebroplacental ratios <10th centile were considered abnormal. Using data from the control fetuses, the centile for each of the cardiac measurements was categorized by whether it was <10th or >90th centile, depending upon the measurement.Of the 50 fetuses with estimated fetal weight <10th centile, 50% (n = 25) had a normal umbilical artery pulsatility index and cerebroplacental ratio. These fetuses had significantly more (P < 0.02 to <0.0001) abnormalities of the size and shape of the 4-chamber view than controls. In all, 44% had a 4-chamber view area >90th centile, 32% had a 4-chamber view global sphericity index <10th centile, 56% had a 4-chamber view width >90th centile, and 80% had 1 or more abnormalities of size and/or shape. The remaining 50% of fetuses (n = 25) had abnormalities of 1 or both for the umbilical artery pulsatility index and/or cerebroplacental ratio. These fetuses had significantly higher rates of abnormalities (P <0.05 to <0.0001) than controls for the following 4-chamber view measurements: 36% had a 4-chamber view area >90th centile; 28% had a 4-chamber view global sphericity index <10th centile; and 68% had a 4-chamber view width >90th centile. Only those fetuses with an abnormal umbilical artery pulsatility index had significant changes in ventricular size; 56% had a left ventricular area <10th centile; 28% had a right ventricular area <10th centile; 36% had right ventricular/left ventricular area ratio >90th centile. One or more of the above abnormal measurements were present in 92% of the fetuses.Higher rates of abnormalities of cardiac size and shape of the 4-chamber view were found in fetuses with an estimated fetal weight <10th centile, regardless of their umbilical artery pulsatility index and cerebroplacental ratio measurements. Those with a normal umbilical artery pulsatility index and an abnormal cerebroplacental ratio had larger and wider measurements of the 4-chamber view. In addition, the shape of the 4-chamber view was more globular or round than in controls. These fetuses may have an increased risk of perinatal complications and childhood and/or adult cardiovascular disease. Screening tools derived from the 4-chamber view, acting as surrogates for ventricular dysfunction, may identify fetuses who could benefit from further comprehensive testing and future preventive interventions.

Assessment of myocardial performance index in late-onset fetal growth restriction.

ABSTRACTThe aim of this study is to determine whether the myocardial performance index (MPI) is increased in fetal growth restriction (FGR) fetuses and if increased MPI is related to adverse outcomes of FGR. This is a prospective cross-sectional study. Seventy-three late-onset FGR fetuses and 97 gestational-age matched control fetuses were enrolled in this study. Fetal blood flow parameters including MPI values were measured and compared between the two groups. For the effect of severity of growth restriction on MPI value, they were also compared with < 3rd and 3rd – 10th centile groups. FGR fetuses were divided into two groups by favorable and adverse outcome and ultrasound parameters were compared between these two groups. Moreover, significant factors related to adverse outcomes by univariate analysis were analyzed by multivariate logistic regression analysis. Pulsatility index of umbilical arterial flow (UA-PI), MPI and amniotic fluid index in the FGR were significantly different from the control fetuses. However, no significant difference between < 3rd and 3rd – 10th centile groups was detected in MPI and UA-PI. The increased levels of MPI and UA-PI were independently related with adverse outcome of late-onset FGR pregnancy. In conclusion, MPI values were increased in late-onset FGR pregnancy, and the higher level of MPI could predict adverse outcome as well as the measurement of UA-PI. Clinicians should consider cardiac dysfunction in FGR through increased MPI.

Transcriptome and DNA Methylation Profiles of Mouse Fetus and Placenta Generated by Round Spermatid Injection.

Low birth efficiency and developmental abnormalities in embryos derived using round spermatid injection (ROSI) limit the clinical application of this method. Further, the underlying molecular mechanisms remain elusive and warrant further in-depth study. In this study, the embryonic day (E) 11.5 mouse fetuses and corresponding placentas derived upon using ROSI, intracytoplasmic sperm injection (ICSI), and natural in vivo fertilized (control) embryos were collected. Transcriptome and DNA methylation profiles were analyzed and compared using RNA-sequencing (RNA-seq) and whole-genome bisulfite sequencing, respectively. RNA-seq results revealed similar gene expression profiles in the ROSI, ICSI, and control fetuses and placentas. Compared with the other two groups, seven differentially expressed genes (DEGs) were identified in ROSI fetuses, and ten DEGs were identified in the corresponding placentas. However, no differences in CpG methylation were observed in fetuses and placentas from the three groups. Imprinting control region methylation and imprinted gene expression were the same between the three fetus and placenta groups. Although 49 repetitive DNA sequences (RS) were abnormally activated in ROSI fetuses, RS DNA methylation did not differ between the three groups. Interestingly, abnormal hypermethylation in promoter regions and low expression of Fggy and Rec8 were correlated with a crown-rump length less than 6 mm in one ROSI fetus. Our study demonstrates that the transcriptome and DNA methylation in ROSI-derived E11.5 mouse fetuses and placentas were comparable with those in the other two groups. However, some abnormally expressed genes in the ROSI fetus and placenta warrant further investigation to elucidate their effect on the development of ROSI-derived embryos.

Blood pressure in the chronically hypoxaemic fetus is regulated by α-adrenergic receptors on the peripheral vasculature.

Blood pressure in the chronically hypoxaemic fetus is regulated by α-adrenergic receptors on the peripheral vasculature.

Transuterine Fetal Tracheal Occlusion Model in Mice

Fetal tracheal occlusion (TO), an established treatment modality, promotes fetal lung growth and survival in severe congenital diaphragmatic hernia (CDH). Following TO, retention of the secreted epithelial fluid increases luminal pressure and induces lung growth. Various animal models have been defined to understand the pathophysiology of CDH and TO. All have their own advantages and disadvantages such as the difficulty of the technique, the size of the animal, cost, high mortality rates, and the availability of genetic tools. Herein, a novel transuterine model of murine fetal TO is described. Pregnant mice were anesthetized, and the uterus exposed via a midline laparotomy. The trachea of selected fetuses were ligated with a single transuterine suture placed behind the trachea, one carotid artery, and one jugular vein. The dam was closed and allowed to recover. Fetuses were collected just before parturition. Lung to body weight ratio in TO fetuses was higher than that in control fetuses. This model provides researchers with a new tool to study the impact of both TO and increased luminal pressure on lung development.