Maternal Samples Research Articles

Maternal vaginal microbiome composition does not affect development of the infant gut microbiome in early life.

Birth mode has been implicated as a major factor influencing neonatal gut microbiome development, and it has been assumed that lack of exposure to the maternal vaginal microbiome is responsible for gut dysbiosis among caesarean-delivered infants. Consequently, practices to correct dysbiotic gut microbiomes, such as vaginal seeding, have arisen while the effect of the maternal vaginal microbiome on that of the infant gut remains unknown. We conducted a longitudinal, prospective cohort study of 621 Canadian pregnant women and their newborn infants and collected pre-delivery maternal vaginal swabs and infant stool samples at 10-days and 3-months of life. Using cpn60-based amplicon sequencing, we defined vaginal and stool microbiome profiles and evaluated the effect of maternal vaginal microbiome composition and various clinical variables on the development of the infant stool microbiome. Infant stool microbiomes showed significant differences in composition by delivery mode at 10-days postpartum; however, this effect could not be explained by maternal vaginal microbiome composition and was vastly reduced by 3 months. Vaginal microbiome clusters were distributed across infant stool clusters in proportion to their frequency in the overall maternal population, indicating independence of the two communities. Intrapartum antibiotic administration was identified as a confounder of infant stool microbiome differences and was associated with lower abundances of Escherichia coli, Bacteroides vulgatus, Bifidobacterium longum and Parabacteroides distasonis. Our findings demonstrate that maternal vaginal microbiome composition at delivery does not affect infant stool microbiome composition and development, suggesting that practices to amend infant stool microbiome composition focus factors other than maternal vaginal microbes.

Hemolytic disease of the newborn due to anti-Jra from a Chinese mother with one novel and one classic heterozygous mutation.

Investigation of a Jr(a-) family samples, identification of the mutant and assessment of the differences of Jr antigen density of the Jr(a-) family members, random adult and newborn individuals' RBCs. The anti-Jra antibody is generated when a Jr(a-) individual pregnant or transfused with Jr(a+) blood unit, which can lead to mild-to-moderate hemolytic disease of the foetus and newborn (HDFN) or hemolytic transfusion reaction (HTR). Several mutations had been identified. The anti-Jra caused HDFN is not rare in East Asia, but due to the lack of antibody and molecular background, it is likely to lead missed detection. One G4P1 woman had been detected as IAT positive during prenatal examination. Suspected as anti-Jra after the laboratory serological testing, the maternal sample was further assessed by molecular analysis. The antigen density was detected by flow cytometry after reacting with anti-Jra serum in family members and the normal individuals. One novel frameshift mutation c.717delC and one previously identified mutation c.706C > T in ABCG2 was identified on proband. The infant haemoglobin(Hb) and bilirubin increased significantly after exchange transfusion and the severe HDFN was relieved. Flow cytometry results showed that the Jra antigens on adult RBCs were significantly less than those on the infant. The c.717delC mutation can lead to the shortening of protein ABCG2 in the site of p.Leu307Stop, result in the loss of Jra antigen. The difference in antigen density between adult and infant RBCs may be a possible reason that leads to severe HDFN but not transfusion reaction. Breastfeeding may lead to slower recovery from HDFN.

The relationship between urinary selenium levels and risk of gestational diabetes mellitus: A nested case-control study.

Selenium (Se) is an essential trace element for the human body. Serum Se and urinary Se are also biomarkers to assess Se exposure status. However, studies focusing on the association between urinary Se and the risk of gestational diabetes mellitus (GDM) are rare. To investigate the association between urinary Se and the risk of GDM. A nested case-control study based on a prospective birth cohort in Wuhan, China, which focuses on the effects of prenatal environmental factors exposure on pregnant women and children's health was conducted. Two hundred and twenty-six cases and 452 controls were included. Maternal urine samples were collected before GDM diagnosis, and the urinary Se levels were determined. We assessed the association of urinary Se with GDM by conditional logistic regression with maternal urinary Se level as a categorical variable, and estimated the association between Se and glucose levels by multiple linear regression. The potential modifier roles of maternal age and fetal sex have also been assessed. Lower urinary level of Se was significantly associated with a higher risk of GDM (OR = 2.35 for the tertile 1, 95% CI:1.36-4.06; adjusted OR = 1.79 for the tertile 2, 95%CI:1.09-2.95; p for trend = 0.01). Fetal sex had an interaction with Se in the association with GDM. The association was more pronounced among pregnant women with female fetuses than with male fetuses. Our study suggested a significant negative association between urinary Se and the risk of GDM, and this association may vary depending on the fetal sex.

Markers of placental function correlate with prevalence and quantity of nucleated fetal cells in maternal circulation in normotensive term pregnancies.

Transplacental fetal cell transfer results in the engraftment of fetal-origin cells in the pregnant woman's body, a phenomenon termed fetal microchimerism. Increased fetal microchimerism measured decades postpartum is implicated in maternal inflammatory disease. Understanding which factors cause increased fetal microchimerism is therefore important. During pregnancy, circulating fetal microchimerism and placental dysfunction increase with increasing gestational age, particularly towards term. Placental dysfunction is reflected by changes in circulating placenta-associated markers, specifically placental growth factor (PlGF), decreased by several 100 pg/mL, soluble fms-like tyrosine kinase-1 (sFlt-1), increased by several 1000 pg/mL, and the sFlt-1/PlGF ratio, increased by several 10 (pg/mL)/(pg/mL). We investigated whether such alterations in placenta-associated markers correlate with an increase in circulating fetal-origin cells. We included 118 normotensive, clinically uncomplicated pregnancies (gestational age 37+1 up to 42+2 weeks' gestation) pre-delivery. PlGF and sFlt-1 (pg/mL) were measured by Elecsys® Immunoassays. We extracted DNA from maternal and fetal samples and genotyped four human leukocyte antigen loci and 17 other autosomal loci. Paternally inherited, unique fetal alleles served as polymerase chain reaction (PCR) targets for detecting fetal-origin cells in maternal buffy coat. Fetal-origin cell prevalence was assessed by logistic regression, and quantity by negative binomial regression. Statistical exposures included gestational age (weeks), PlGF (100 pg/mL), sFlt-1 (1000 pg/mL) and the sFlt-1/PlGF ratio (10 (pg/mL)/(pg/mL)). Regression models were adjusted for clinical confounders and PCR-related competing exposures. Gestational age was positively correlated with fetal-origin cell quantity (DRR = 2.2, P = 0.003) and PlGF was negatively correlated with fetal-origin cell prevalence (odds ratio [OR]100 = 0.6, P = 0.003) and quantity (DRR100 = 0.7, P = 0.001). The sFlt-1 and the sFlt-1/PlGF ratios were positively correlated with fetal-origin cell prevalence (OR1000 = 1.3, P = 0.014 and OR10 = 1.2, P = 0.038, respectively), but not quantity (DRR1000 = 1.1, P = 0.600; DRR10 = 1.1, P = 0.112, respectively). Our results suggest that placental dysfunction as evidenced by placenta-associated marker changes, may increase fetal cell transfer. The magnitudes of change tested were based on ranges in PlGF, sFlt-1 and the sFlt-1/PlGF ratio previously demonstrated in pregnancies near and post-term, lending clinical significance to our findings. Our results were statistically significant after adjusting for confounders including gestational age, supporting our novel hypothesis that underlying placental dysfunction potentially is a driver of increased fetal microchimerism.

Sex-specific effects of prenatal organophosphate ester (OPE) metabolite mixtures and adverse infant birth outcomes in the maternal and developmental risks from environmental and social stressors (MADRES) pregnancy cohort

BackgroundOrganophosphate esters (OPEs) are used as flame retardants and plasticizers in various consumer products. Limited prior research suggests sex-specific effects of prenatal OPE exposures on fetal development. We evaluated overall and sex-specific associations between prenatal OPE exposures and gestational age (GA) at birth and birthweight for gestational age (BW for GA) z-scores among the predominately low-income, Hispanic MADRES cohort. MethodsNine OPE metabolite concentrations were measured in 421 maternal urine samples collected during a third trimester visit (GA = 31.5 ± 2.0 weeks). We examined associations between single urinary OPE metabolites and GA at birth and BW for GA z-scores using linear regression models and Generalized Additive Models (GAMs) and effects from OPE mixtures using Bayesian Kernel Machine Regression (BKMR). We also assessed sex-specific differences in single metabolite analyses by evaluating statistical interactions and stratifying by sex. ResultsWe did not find significant associations between individual OPE metabolites and birth outcomes in the full infant sample; however, we found that higher bis(1,3-dichloro-2-propyl) phosphate (BDCIPP) was associated with earlier GA at birth among male infants (p = 0.04), and a nonlinear, inverted U-shape association between the sum of dibutyl phosphate and di-isobutyl phosphate (DNBP + DIBP) and GA at birth among female infants (p = 0.03). In mixtures analysis, higher OPE metabolite mixture exposures was associated with lower GA at birth, which was primarily driven by female infants. No associations were observed between OPE mixtures and BW for GA z-scores. ConclusionHigher BDCIPP and DNBP + DIBP concentrations were associated with earlier GA at birth among male and female infants, respectively. Higher exposure to OPE mixtures was associated with earlier GA at birth, particularly among female infants. However, we saw no associations between prenatal OPEs and BW for GA. Our results suggest sex-specific impacts of prenatal OPE exposures on GA at birth.

Maternal vaginal fluids play a major role in the colonization of the neonatal intestinal microbiota.

Caesarean section (CS) is associated with newborns' health risks due to the blocking of microbiome transfer. The gut microbiota of CS-born babies was different from those born vaginally, which may be attributed to reduced exposure to maternal vaginal microbes during labour. To understand the microbial transfer and reduce CS disadvantages, the effect of vaginal microbiota exposure on infant gut microbiota composition was evaluated using 16s rDNA sequencing-based techniques. Pregnant women were recruited in the Women and Children's Hospital, School of Medicine, Xiamen University from June 1st to August 15th, 2017. Maternal faeces (n = 26), maternal vaginal fluids (n = 26), and neonatal transitional stools (n = 26) were collected, while the participants underwent natural delivery (ND) (n = 6), CS (n = 4) and CS with the intervention of vaginal seedings (I) (n = 16). 26 mothers with the median age 26.50 (25.00-27.25) years showed no substantial clinical differences. The newborns' gut microbiota altered among ND, CS and I, and clustered into two groups (PERMANOVA P = 0.001). Microbial composition of ND babies shared more features with maternal vaginal samples (PERMANOVA P = 0.065), while the microbiota structure of ND babies was obviously different from that of sample of maternal faeces. The genus Bacteroides in CS-born babies with intervention approached to vaginal-born neonates, compared with CS-born neonates without intervention. Neonatal gut microbiota was dependent on the delivery mode. And the gut microbiota CS newborns with vaginal seeding shared more features with those of ND babies, which hinted the aberrant gut microbiota composition initiated by CS might be partly mitigated by maternal vaginal microbiota exposure.

Correlation analysis between prenatal exposure of per-/polyfluoroalkyl compounds and neonatal outcome

Objective: To investigate the correlation between the prenatal exposure of per-/polyfluoroalkyl substances (PFASs) and the neonatal outcome. Methods: A total of 506 maternal infant cohort samples were collected in Hangzhou, Zhejiang province from 2020 to 2021. The exposure levels of seven PFASs in maternal serum before delivery were detected by solid-phase extraction-ultra performance liquid chromatography tandem mass spectrometry. Multivariable linear regression model was used to analyze the influence of prenatal exposure of PFASs on birth weight, birth length and Apgar score. Results: The maternal age, prenatal body mass index and gestation age were (31.3±4.3) years old, (26.7±3.2) kg/m2 and (265.0±28.3) days, respectively. The birth weight, birth length and scores of Apgar-1 and Apgar-5 were (3.1±0.8) kg, (49.3±2.9) cm, (9.88±0.47) points and (9.99±0.13) points, respectively. PFASs were widely distributed in maternal serum, with the highest concentration of (18.453±19.557) ng/ml, (6.756±9.379) ng/ml and (5.057±8.555) ng/ml for perfluorooctanoic acid (PFOA), perfluorooctane sulfonate (PFOS) and 6∶2 chlorinated polyfluorinated ether sulfonate (Cl-PFESA), respectively. Maternal age, parity and delivery mode were associated with the exposure level of PFASs (P<0.05). Subgroup analysis showed that PFOS had negative effects on birth weight (β=-0.958), birth length (β=-0.073) and Apgar-5 score (β=-0.288) for neonates in the low birth weight (LBW) group. 6∶2 Cl-PFESA and 8∶2 Cl-PFESA inhibited the birth weight (β=-0.926; β=-0.552) and length (β=-0.074; β=-0.045) of newborn in the LBW group. In addition, 4∶2 fluorotelomer sulfonate (FTS) was associated with increased birth weight (β=0.111) and decreased Apgar-5 score (β=-0.030) in the normal weight group. Conclusion: Prenatal exposure to PFASs is associated with birth weight, birth length and Apgar-5 score. It is necessary to continue to pay attention to the impact of PFASs on fetal growth and development through maternal-fetal transmission.

Gestational exposure to organophosphate esters and adiposity measures of children up to 6 years: Effect modification by breastfeeding

Organophosphate esters (OPEs) are synthetic chemicals used in various commercial products. Accumulating evidence has shown that they may act as metabolic disruptors. However, no study has investigated the long-term effects of gestational OPEs exposure on childhood adiposity. Breast milk represents the optimal nutritional form of feeding for infants and may protect against the adverse effects of gestational OPEs exposure on offspring development. Using data from the Shanghai-Minhang birth cohort study, we investigated the associations of gestational OPEs exposure with adiposity measures in children up to 6 years of age, and whether breastfeeding could modify these associations. A total of 733 mother-child pairs with available data on OPE concentrations and child anthropometry were included. Eight OPE metabolites were assessed in maternal urine samples collected at 12-16 weeks of pregnancy. Information on children's weight, height, arm circumference, and waist circumference was collected at birth and 0.5, 1, 4, and 6 years of age. Weight-for-age and body mass index-for-age z scores were calculated. The duration of children's breastfeeding was categorized as ≤4 months or >4 months. The generalized estimate equation and Bayesian Kernel Machine Regression models were used to examine the associations of OPEs exposure with children's adiposity measures. Selected OPEs exposure was associated with higher children's adiposity measures. Particularly, we found stronger associations of bis(1-chloro-2-propyl) phosphate (BCIPP), bis(2-chloroethyl) phosphate (BCEP), bis(1,3-dichloro-2-propyl) phosphate (BDCPP), and di-o-cresyl phosphate and di-p-cresyl phosphate (DCP) with higher adiposity measures in children breastfed for ≤4 months, while little evidence of associations was found among those breastfed for >4 months. Our study suggested that gestational OPEs exposure could alter children's adiposity measures, but the potential effects were attenuated if children were breastfed for >4 months.

Protective role of the placental efflux transporter BCRP/ABCG2 in the relationship between prenatal cadmium exposure, placenta weight, and size at birth

Background and aimPlacental efflux transporter proteins, such as BCRP, reduce the placental and fetal toxicity of environmental contaminants but have received little attention in perinatal environmental epidemiology. Here, we evaluate the potential protective role of BCRP following prenatal exposure to cadmium, a metal that preferentially accumulates in the placenta and adversely impacts fetal growth. We hypothesized that individuals with a reduced function polymorphism in ABCG2, the gene encoding BCRP, would be most vulnerable to the adverse impacts of prenatal cadmium exposure, notably, smaller placental and fetal size. MethodsWe measured cadmium in maternal urine samples at each trimester and in term placentas from UPSIDE-ECHO study participants (NY, USA; n = 269). We fit adjusted multivariable linear regression and generalized estimating equation models to examine log-transformed urinary and placental cadmium concentrations in relation to birthweight, birth length, placental weight, and fetoplacental weight ratio (FPR) and stratified models by ABCG2 Q141K (C421A) genotype. ResultsOverall 17% of participants expressed the reduced-function ABCG2 C421A variant (AA or AC). Placental cadmium concentrations were inversely associated with placental weight (β = −19.55; 95%CI: −37.06, −2.04) and trended towards higher FPR (β = 0.25; 95%CI: −0.01, 0.52) with stronger associations in 421A variant infants. Notably, higher placental cadmium concentrations in 421A variant infants were associated with reduced placental weight (β = −49.42; 95%CI: 98.87, 0.03), and higher FPR (β = 0.85, 95%CI: 0.18, 1.52), while higher urinary cadmium concentration was associated with longer birth length (β = 0.98; 95%CI: 0.37, 1.59), lower ponderal index (β = −0.09; 95%CI: 0.15, −0.03), and higher FPR (β = 0.42; 95%CI: 0.14, 0.71). ConclusionsInfants with reduced function ABCG2 polymorphisms may be particularly vulnerable to the developmental toxicity of cadmium as well as other xenobiotics that are BCRP substrates. Additional work examining the influence of placental transporters in environmental epidemiology cohorts is warranted.

Abstract 64: Prenatal Per- and Polyfluoroalkyl Substance (PFAS) Exposures, Individually and as a Mixture, Are Associated With Obesity Risk at 16-20 Years in the Project Viva Prospective Cohort: Implications for PFAS as Hazardous Substances for Developmental Health

Introduction: PFAS are obesogenic in adults, but findings on the associations between prenatal PFAS exposures and child obesity risk are inconsistent, and it is not clear if such associations extend up to mid- to late adolescence. We investigated the individual and joint associations of prenatal PFAS exposures with obesity risk at 16-20 years. Methods: In the prospective pre-birth cohort Project Viva, we measured PFAS in maternal plasma samples collected in the first trimester and child body mass index (BMI) at the mid-adolescent visit (median: 17.4y; range: 15.9-20.0y). We defined obesity as BMI ≥ 95 th percentile for age and sex based on the CDC Growth Charts. We used Poisson regression with robust variance estimates to examine associations of each PFAS, and Bayesian kernel machine regression (BKMR) to examine PFAS mixtures, with obesity. We adjusted for maternal age, education, pre-pregnancy BMI, race/ethnicity, parity, and smoking status during pregnancy. Results: Among 545 participants, 73 (13%) had a BMI in the obese range. Children with higher prenatal PFOS and PFNA exposures had a greater risk of obesity [relative risk = 1.59 (95% CI: 1.19, 2.11) per doubling of PFOS and 1.48 (95% CI: 1.11, 1.98) per doubling of PFNA] in multivariable-adjusted models. In BKMR models, PFOS and PFNA were similarly associated with greater risk of obesity when accounting for co-exposures of other PFAS (Panel A) . There was an interaction between PFOS and PFOA: the positive association between PFOS and obesity was stronger when PFOA levels were lower (Panel B) , while there was a negative association between PFOA and obesity when PFOS levels were higher (Panel C) . Children with higher concentrations of the mixture PFAS had greater obesity risk in a dose-dependent fashion (Panel D) . Conclusions: Prenatal PFAS exposures may have long-lasting, intergenerational obesogenic effects. Our findings support the U.S. Environmental Protection Agency’s recent designation of PFAS as hazardous substances for human health, including developmental health.

Abstract P169: Prenatal Per- and Polyfluoroalkyl Substance (PFAS) Exposures and Offspring Blood Pressure From Birth to Late Adolescence: A Prospective Study in Project Viva

Introduction: Mice studies showed that prenatal PFAS exposures increase offspring blood pressure (BP), but findings in human studies are inconsistent. Prior studies did not have repeated BP measurements from birth to late adolescence or use a mixture approach to investigate the interactions or overall effects of prenatal PFAS exposures. Methods: Participants are from the prospective pre-birth cohort Project Viva. We measured PFAS in maternal plasma samples collected in the 1 st -trimester (median: 9.6w) and child systolic (SBP) and diastolic BP (DBP) at six follow-up visits: 1) birth; 2) infancy (median: 6m); 3) early childhood (median: 3.2y); 4) mid-childhood (median: 7.7y); 5) early adolescent (median: 13.0y), 6) mid-adolescent (median: 17.5y). At each study visit, BP was measured 4-5 times, and the average value was taken. We used linear regression to examine associations of each individual PFAS, and Bayesian kernel machine regression (BKMR) to examine PFAS mixtures, with BP at each visit, adjusting for maternal age, race/ethnicity, education, pre-pregnancy body mass index, parity, smoking status, and chronic hypertension before pregnancy. Results: Sample sizes at each visit ranged from 549-919 ( Legend ). Prenatal PFAS was not consistently associated with offspring SBP or DBP across all visits. PFOS was associated with higher SBP in mid-childhood ( Panel A1 ), EtFOSAA was associated with higher SBP in infancy and in mid-childhood ( Panel A5 ), and MeFOSAA was associated with higher SBP in infancy ( Panel A6 ). PFOS, PFNA, EtFOSAA, and MeFOSAA were associated with lower DBP only in early childhood but not at other visits ( Panels B1, B4, B5, B6 ). BKMR did not suggest prenatal PFAS interactions or the overall effects of exposure to multiple PFAS on offspring SBP or DBP. Conclusions: There are no consistent associations between prenatal PFAS exposures and BP from birth to late adolescence. Future studies should examine whether there are other critical windows (e.g., mid- or late pregnancy) of susceptibility for PFAS on offspring BP.

Abstract P206: Later Age at Menarche is Associated With Metabolite Signatures for Cardiovascular Diseases: A Metabolome-Wide Association Study of Racially and Ethnically Diverse Women From the Boston Birth Cohort

Introduction: Later menarche onset portends lower risk of cardiometabolic outcomes in adults (e.g., obesity, diabetes, hypertension, and myocardial infarction) and in pregnancy (e.g., preterm birth). However, the underlying mechanisms underlying these associations are unclear. Metabolome-wide association studies afford an opportunity to identify mechanistic pathways. Objective: To identify metabolome signatures of age at menarche in a diverse sample of women. Methods: This analysis includes 1301 women from the Boston Birth Cohort who were enrolled 24-72 hrs. after delivery, when we collected maternal plasma sample for metabolome and information on maternal age at menarche (range 7-18y) via a standard questionnaire interview. We used linear regression and restricted cubic splines to examine the association between age at menarche and metabolites. We adjusted for maternal year of birth, place of birth, education, smoking status, Mediterranean diet score, pre-pregnancy BMI, hypertensive disorders, and diabetes. Results: Of the 1301 study women, 63.7% identified as Black and 21.3% as Hispanic. Maternal age at delivery was 28.3y ±6.6y and age at menarche was 12.87y ±1.95y. Of the 389 metabolites we examined, 63 were associated with late menarche (&gt;15y vs. menarche 12-13y; Fig., A ). A 1-yr increment in menarche age was associated with 7 metabolites ( Fig., B ); of these, imidazole propionate and C18:2 SM were (inverse) linear dose-response ( Fig., C ). Other metabolites, including some implicated in cardiovascular disease (e.g., dimethylguanidino valeric acid), showed a threshold effect starting at menarche &gt;14 years (Fig., C). Conclusion: In this racially and ethnically diverse cohort of women from Boston, later age at menarche is associated with many metabolites, including inverse associations with metabolites that have been previously implicated in cardiovascular disease. Our findings help inform mechanistic pathways and that, if replicated, could be targeted for cardiometabolic prediction and prevention.

Proteomic profile of extracellular vesicles in maternal plasma of women with fetal death

Objectives Fetal death is a complication of pregnancy caused by multiple etiologies rather than being the end-result of a single disease process. Many soluble analytes in the maternal circulation, such as hormones and cytokines, have been implicated in its pathophysiology. However, changes in the protein content of extracellular vesicles (EVs), which could provide additional insight into the disease pathways of this obstetrical syndrome, have not been examined. This study aimed to characterize the proteomic profile of EVs in the plasma of pregnant women who experienced fetal death and to evaluate whether such a profile reflected the pathophysiological mechanisms of this obstetrical complication. Moreover, the proteomic results were compared to and integrated with those obtained from the soluble fraction of maternal plasma. Methods This retrospective case-control study included 47 women who experienced fetal death and 94 matched, healthy, pregnant controls. Proteomic analysis of 82 proteins in the EVs and the soluble fractions of maternal plasma samples was conducted by using a bead-based, multiplexed immunoassay platform. Quantile regression analysis and random forest models were implemented to assess differences in the concentration of proteins in the EV and soluble fractions and to evaluate their combined discriminatory power between clinical groups. Hierarchical cluster analysis was applied to identify subgroups of fetal death cases with similar proteomic profiles. A p-value of <.05 was used to infer significance, unless multiple testing was involved, with the false discovery rate controlled at the 10% level (q < 0.1). All statistical analyses were performed by using the R statistical language and environment-and specialized packages. Results Nineteen proteins (placental growth factor, macrophage migration inhibitory factor, endoglin, regulated upon activation normal T cell expressed and presumably secreted (RANTES), interleukin (IL)-6, macrophage inflammatory protein 1-alpha, urokinase plasminogen activator surface receptor, tissue factor pathway inhibitor, IL-8, E-Selectin, vascular endothelial growth factor receptor 2, pentraxin 3, IL-16, galectin-1, monocyte chemotactic protein 1, disintegrin and metalloproteinase domain-containing protein 12, insulin-like growth factor-binding protein 1, matrix metalloproteinase-1(MMP1), and CD163) were found to have different plasma concentrations (of an EV or a soluble fraction) in women with fetal death compared to controls. There was a similar pattern of change for the dysregulated proteins in the EV and soluble fractions and a positive correlation between the log2-fold changes of proteins significant in either the EV or the soluble fraction (ρ = 0.89, p < .001). The combination of EV and soluble fraction proteins resulted in a good discriminatory model (area under the ROC curve, 82%; sensitivity, 57.5% at a 10% false-positive rate). Unsupervised clustering based on the proteins differentially expressed in either the EV or the soluble fraction of patients with fetal death relative to controls revealed three major clusters of patients. Conclusion Pregnant women with fetal death have different concentrations of 19 proteins in the EV and soluble fractions compared to controls, and the direction of changes in concentration was similar between fractions. The combination of EV and soluble protein concentrations revealed three different clusters of fetal death cases with distinct clinical and placental histopathological characteristics.

Biochemical Profile of African American Women During Three Trimesters of Pregnancy and at Delivery

The data presented are the results from a prospective observational study which was conducted to investigate the effects of nutrition and other related factors on the outcome of pregnancy in nulliparous African American women 16-35 years old. Fasting blood samples were collected from the women during the first, second and third trimesters of pregnancy. At delivery, both maternal and cord samples were collected. Biochemical variables such as, serum folate, vitamin B12, ascorbic acid, vitamin E, ferritin, selected minerals as well as complete blood count (CBC) and red cell folate were analyzed in the blood samples. The concentrations of hematocrit, hemoglobin, white blood cells, red blood cells and vitamin B12 were below the reference non-pregnant ranges throughout gestation. Maternal concentrations of folate and vitamin E increased sequentially with increased gestational age. Serum ferritin, during the third trimester, declined to 58% of the first trimester concentration. Maternal levels of ferritin at delivery were one third of the values found in the infant (cord) sample. Cord levels of folate, ascorbic acid and vitamin B12 were higher than the concentrations in the maternal delivery samples. The data suggest that among this group of pregnant women, major physiological changes, such as plasma volume expansion which alters blood chemistry and maternal to fetal transfer of nutrients, were similar to the findings of other investigators. In this population however, the findings for serum and whole blood folate are contrary to those reported by other researchers, and the sequential increase in the maternal concentration of the vitamin during pregnancy could be attributed to the use of vitamin supplements.

Non-invasive fetal genotyping for maternal alleles with droplet digital PCR: A comparative study of analytical approaches.

To develop a flexible droplet digital PCR (ddPCR) workflow to perform non-invasive prenatal diagnosis via relative mutation dosage (RMD) for maternal pathogenic variants with a range of inheritance patterns, and to compare the accuracy of multiple analytical approaches. Cell free DNA (cfDNA) was tested from 124 archived maternal plasma samples: 88 cases for sickle cell disease and 36 for rare Mendelian conditions. Three analytical methods were compared: sequential probability ratio testing (SPRT), Bayesian and z-score analyses. The SPRT, Bayesian and z-score analyses performed similarly well with correct prediction rates of 96%, 97% and 98%, respectively. However, there were high rates of inconclusive results for each cohort, particularly for z-score analysis which was 31% overall. Two samples were incorrectly classified by all three analytical methods; a false negative result predicted for a fetus affected with sickle cell disease and a false positive result predicting the presence of an X-linked IDS variant in an unaffected fetus. ddPCR can be applied to RMD for diverse conditions and inheritance patterns, but all methods carry a small risk of erroneous results. Further evaluation is required both to reduce the rate of inconclusive results and explore discordant results in more detail.

NanoNIPT: Short-fragment nanopore sequencing of cell-free DNA for non-invasive prenatal testing of fetal aneuploidies and sex chromosome aberrations.

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The investigation of the association of pregnancy weight gain on maternal and neonatal gut microbiota composition and abundance using 16sRNA sequencing

ObjectiveTo investigate the association of pregnancy weight gain on gut microbiota in pregnant women and newborns.MethodsPregnant women who had regular antenatal check-ups and were hospitalised for delivery at Shanxi Maternal and Child Health Hospital from September 2020 to December 2020 were selected as the study subjects. They were divided into the normal pre-pregnancy weight-normal pregnancy weight gain group (N-NG group), the normal pregnancy weight-excessive pregnancy weight gain group (N-EG group), the pre-pregnancy overweight/obese-normal pregnancy weight gain group (O-NG group) and the pre-pregnancy overweight/obese-excessive pregnancy weight gain group (O-EG group). Faecal samples of the pregnant women before delivery (37–41+ 6 weeks of gestation) and the first meconium samples of their newborns were collected, sequenced for 16S rRNA gut microbiota and analysed. The results of different gut microbiota were compared separately. χ2 test, a one-way analysis of variance or the rank sum test were performed according to data type and distribution. The differences in the Alpha diversity between the groups were analysed using the Kruskal–Wallis rank sum test. The differences in the Beta diversity between the groups were analysed using the Adonis method.ResultsA total of 126 pre-delivery faecal samples from pregnant women and the first faecal samples from their newborns were collected. Seven species with significant abundance differences between the maternal O-NG and N-EG groups and 27 species with significant abundance differences in the newborns were analysed by LEfSe. In the Alpha diversity analysis, the differences in the maternal observed species index and the Chao1 index were statistically significant (p < 0.05) when compared between the groups (O-EG group versus the O-NG group, N-EG group and N-NG group), and the differences in the Shannon index and Simpson index were not statistically significant (p > 0.05) when compared between the groups. The neonatal observed species index, Chao1 index, Shannon index and Simpson index showed statistically significant differences in the comparison between the N-EG and O-EG groups (p < 0.05). In the Beta diversity analysis, the maternal samples did not differ Significantly between the four groups (p > 0.05), while the neonatal samples differed Significantly between the N-EG and N-NG, O-NG, and O-EG groups (p < 0.05).ConclusionPregnancy weight gain affects the composition and abundance of maternal and neonatal gut microbiota species as well as the diversity of neonatal gut microbiota.

Exposure to phthalate increases the risk of eczema in children: Findings from a systematic review and meta-analysis

Emerging evidence indicated phthalate exposure might raise the risk of eczema in children. However, these findings were inconsistent. The relation between phthalate exposure and childhood eczema remained debated. Therefore, we performed this meta-analysis to assess their association. PubMed, Web of Science, and Embase were searched for eligible studies. Pooled odds ratio (OR) and 95% confidence interval (CI) were calculated for risk estimate. Thirty studies involving 12,615 participants were included in this meta-analysis. For prenatal phthalate exposure assessed with maternal samples, the pooled results showed gestational exposure to monobenzyl phthalate (MBzP) (OR: 1.17, 95% CI: 1.00–1.36), but not the other phthalates, was correlated with increased risk of eczema in children. For childhood exposure assessed using children's urine sample, our pooled results indicated that postnatal exposure to MBzP (OR: 1.10, 95% CI: 1.02–1.19), mono-2-ethyl-5-hydroxyhexyl phthalate (MEHHP) (OR: 1.32, 95% CI: 1.08–1.61), mono-(2-ethyl-5-carboxypentyl) phthalate (MECPP) (OR: 1.24, 95% CI: 1.06–1.44), and molar summation of di-2-ethylhexyl phthalate (DEHP) (OR: 1.23, 95% CI: 1.06–1.42) were associated with higher risk of eczema. While for studies using household dust to estimate environmental phthalate exposure and eczema risk, the pooled results showed no significant association. Subgroup analyses indicated study country, diagnostic mode, and children's age contributed to the heterogeneity. The results of our meta-analysis demonstrated that phthalate exposure during both prenatal and postnatal periods was associated with elevated risk of eczema in children. However, such association was not strong as the pooled ORs were relatively small. Further studies are warranted to verify these findings and explore the underlying mechanism.

Mutation spectrum analysis of DMD gene in Indonesian Duchenne and Becker muscular dystrophy patients

Background: Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations in the DMD gene. The full mutation spectrum of the DMD gene in Indonesian patients is currently unknown. Mutation-specific therapies are currently being developed, such as exon skipping or stop codon read-through therapy. This study was conducted with the aim of identifying the mutation spectrum of the DMD gene in Indonesia to guide future development and application of feasible therapeutic strategies. Methods: This study is a cross sectional study that enrolled 43 male patients with a clinical suspicion of DMD or BMD. Multiplex ligation-dependent probe amplification (MLPA) reaction was performed to screen for the common mutations in the DMD gene. Results: Out of 43 subjects, deletions accounted for 69.77% (n=30) cases, while duplications were found in 11.63% (n=5) cases. One novel duplication spanning exons 2 to 62 was identified. Deletion mutations clustered around the distal (66.67%) and proximal (26.67%) hot spot regions of the DMD gene while duplication mutations were observed solely at the proximal region. Two false positive cases of single exon deletion detected through MLPA were attributed to sequence mutations affecting primer ligation sites, confirming the need to validate all single exon deletions when using this screening method. Analysis of available maternal DNA samples showed that the rate of de novo mutations (48.15%) appears higher than expected in this population. Out of 31 patients who were classified as DMD based on clinical and genotype characterizations, 60.47% (n=26) of cases were suitable for exon skipping therapy. Conclusion: This is the first comprehensive study showing the feasibility of implementing the MLPA method for routine screening of DMD patients in Indonesia. This is also the first study showing the potential applicability of exon skipping therapy in the majority of DMD cases in the country.

Heterogenous Differences in Cellular Senescent Phenotypes in Pre-Eclampsia and IUGR following Quantitative Assessment of Multiple Biomarkers of Senescence

Premature ageing of the placenta in pregnancy outcomes is associated with the persistent presence of oxidative stress and placental insufficiency reducing its functional capacity. In this study, we investigated cellular senescence phenotypes of pre-eclampsia and IUGR pregnancies by simultaneously measuring several biomarkers of senescence. Maternal plasma and placental samples were collected at term gestation from nulliparous women undergoing pre-labour elective caesarean section with pre-eclampsia without intrauterine growth restriction (PE; n = 5), pre-eclampsia associated with intrauterine growth restriction (n = 8), intrauterine growth restriction (IUGR < 10th centile; n = 6), and age-matched controls (n = 20). Placental absolute telomere length and senescence gene analysis was performed by RTqPCR. The expression of cyclin-dependent kinase inhibitors (p21 and p16) was determined by Western blot. Senescence-associated secretory phenotypes (SASPs) were evaluated in maternal plasma by multiplex ELISA assay. Placental expression of senescence-associated genes showed significant increases in CHEK1, PCNA, PTEN, CDKN2A, and CCNB-1 (p < 0.05) in pre-eclampsia, while TBX-2, PCNA, ATM, and CCNB-1 expression were evident (p < 0.05) and were significantly decreased in IUGR compared with controls. Placental p16 protein expression was significantly decreased in pre-eclampsia only compared with controls (p = 0.028). IL-6 was significantly increased in pre-eclampsia (0.54 pg/mL ± 0.271 vs. 0.3 pg/mL ± 0.102; p = 0.017) while IFN-γ was significantly increased in IUGR (4.6 pg/mL ± 2.2 vs. 2.17 pg/mL ± 0.8; p = 0.002) compared with controls. These results provide evidence of premature senescence in IUGR pregnancies, and while cell cycle checkpoint regulators are activated in pre-eclampsia, the cellular phenotype is one of cell repair and subsequent proliferation rather than progression to senescence. The heterogeneity of these cellular phenotypes highlights the complexity of characterising cellular senescence and may equally be indicative of the differing pathophysiological insults unique to each obstetric complication.