Proteomic Analysis of Maternal Urine for the Early Detection of Preeclampsia and Fetal Growth Restriction

Background: To explore the use of maternal urine proteome for the identification of preeclampsia biomarkers. Methods: Maternal urine samples from women with and without preeclampsia were used for protein discovery followed by a validation study. The targeted proteins of interest were then measured in urine samples collected at 20–24 and 30–34 weeks among nine women who developed preeclampsia, one woman with fetal growth restriction, and 20 women with uncomplicated pregnancies from a longitudinal study. Protein identification and quantification was obtained using liquid chromatography–tandem mass spectrometry (LC–MS/MS). Results: Among the 1108 urine proteins quantified in the discovery study, 21 were upregulated in preeclampsia and selected for validation. Nineteen (90%) proteins were confirmed as upregulated in preeclampsia cases. Among them, two proteins, ceruloplasmin and serpin A7, were upregulated at 20–24 weeks and 30–34 weeks of gestation (p < 0.05) in cases of preeclampsia, and could have served to identify 60% of women who subsequently developed preeclampsia and/or fetal growth restriction at 20–24 weeks of gestation, and 78% at 30–34 weeks, for a false-positive rate of 10%. Conclusions: Proteomic profiling of maternal urine can differentiate women with and without preeclampsia. Several proteins including ceruloplasmin and serpin A7 are upregulated in maternal urine before the diagnosis of preeclampsia and potentially fetal growth restriction.

Preeclampsia prediction –First trimester screening markers

Preeclampsia is a multi-system disorder manifested primarily by hypertension and proteinuria during second half of pregnancy. It is a major cause of maternal morbidity and mortality worldwide. Despite decades of research into the condition, the ability of clinicians to predict preeclampsia prior to the onset of symptoms has not improved significantly. In this review we will look at potential biomarkers for early prediction and diagnosis of preeclampsia. To evaluate the efficacy of different biochemical and biophysical markers in the early weeks of gestation as screening tools for early prediction of preeclampsia. This hospital-based prospective observational study conducted on 52 pregnant women, at less than 13 weeks of gestation were recruited. Maternal urine microalbumin, urinary albumin to creatinine ratio, and USG uterine artery PI levels were analyzed among the pregnant women who subsequently developed PE and compare with those who did not develop PE. Methods used for the detection of markers are: immunoturbidimetric method for urine albumin, modified kinetic Jaffe reaction without deproteinization for Urine creatinine and Uterine artery Doppler velocimetry was done by PHILIPS HD11XE transabdominal ultrasound machine using a 4-6 MHz probe with the same sonographer. In the present study, spot urine microalbumin and spot urine albumin to creatinine ratio (UACR) at 11-13 were significantly higher in women who developed PE subsequently when compared to nonpreeclamptic women.(P&#60;0.0001). The mean levels of 1st and 2nd-trimester uterine artery PI significantly high in women who subsequently developed PE when compared to those who did not develop preeclampsia (P&#60;0.0001). Study results showed a strong association between gestational age at delivery and neonatal outcome (neonatal birth weight and APGAR) with preeclampsia. The maternal urine microalbumin, albumin to creatinine ratio, and uterine artery PI found to have good sensitivity and specificity for early prediction of PE. Study concluded that the women who are prone to develop PE subsequently, had high levels of MAP, UAPI, microalbuminuria and urine albumin to creatinine ratio than the normotensive women. In our setting, MAP, UAPI, microalbuminuria, and UACR markers appeared to be better screening modalities. The combination of biochemical markers with the biophysical markers, demographic characteristics, and other novel markers will establish the effective screening models for early prediction of PE. Early identification of high-risk cases will offer an opportunity for prophylactic therapy, such as Low- dose Aspirin in selected groups of high-risk women screened in the first trimester, thus improving the maternal and perinatal outcome.

Prenatal Exposure to Di-Ethyl Phthalate (DEP) Is Related to Increasing Neonatal IgE Levels and the Altering of the Immune Polarization of Helper-T Cells

Introduction: Phthalates are substances that are added to plastic products to increase their plasticity. These substances are released easily into the environment and can act as endocrine disruptors. Epidemiological studies in children have showed inconsistent findings regarding the relationship between prenatal or postnatal exposure to phthalates and the risk of allergic disease. Our hypothesis is that prenatal exposure to phthalates may contribute to the development of allergies in children. Material and methods: The objective of this study was to determine the associations between urinary phthalate metabolite concentrations in pregnant women, maternal atopic diathesis, maternal lifestyle, and cord blood IgE. Pregnant mothers and paired newborns (n = 101) were enrolled from an antenatal clinic. The epidemiologic data and the clinical information were collected using standard questionnaires and medical records. The maternal blood and urine samples were collected at 24–28 weeks gestation, and cord blood IgE, IL-12p70, IL-4, and IL-10 levels were determined from the newborns at birth. The link between phthalates and maternal IgE was also assessed. To investigate the effects of phthalates on neonatal immunity, cord blood mononuclear cells (MNCs) were used for cytokine induction in another in vitro experiment. Results: We found that maternal urine monoethyl phthalate (MEP) (a metabolite of di-ethyl phthalate (DEP)) concentrations are positively correlated with the cord blood IgE of the corresponding newborns. The cord blood IL-12p70 levels of mothers with higher maternal urine MEP groups (high DEP exposure) were lower than mothers with low DEP exposure. In vitro experiments demonstrated that DEP could enhance IL-4 production of cord blood MNCs rather than adult MNCs. Conclusion: Prenatal DEP exposure is related to neonatal IgE level and alternation of cytokines relevant to Th1/Th2 polarization. This suggests the existence of a link between prenatal exposure to specific plasticizers and the future development of allergies.

Prenatal exposure to butyl paraben is associated with fat percentage in 7-year old boys

Context Parabens are used as preservatives in consumer products but are suspected of having endocrine-disrupting properties. A recent study reported an association between in utero exposure to butyl paraben and overweight in childhood, with a stronger trend in girls. Objective We therefore studied the association between parabens in maternal urine in third trimester and fat percentage in children aged 7 years. Design, setting & Participants We used data from the Odense Child Cohort, a mother-child cohort with enrollment from 2010-2012, in which the children are followed. Paraben concentration was assessed in maternal urine at median gestational week 28.7 and body composition measured as total, gynoid, and android fat percentages assessed by Dual Xray Absorptiometry in their children at age 7 years. Main Outcome Measurements Total, Gynoid, and Android fat percentages and z-score for BMI. Interventions None Results Paraben exposure was low. In multivariate linear regressions, detection of butylparaben in maternal urine was associated with an increase of 17% (95% confidence intervals CI 3.0%;32%) in total body fat percentage and an increase of 23% (95% CI 5.1%;43%) in android fat percentage in boys, compared to boys whose mother had no detectable butylparaben in urine. No significant associations between in utero exposure to methyl-, ethyl- or propyl parabens and body composition were found, and no significant associations were seen in girls. Conclusion Our findings suggest that parabens, which are believed to have low toxicity, may affect obesity development at vulnerable time periods during development

Diethylhexyl phthalate induces teratogenic effects through oxidative stress response in a chick embryo model

Diethylhexyl phthalate (DEHP) is known as a persistent environmental pollutant. However, the possible effects of DEHP on human neural tube defects (NTDs) remain elusive. We set out to investigate the exposure of DEHP in human and explore the association of DEHP and NTDs. The level of DEHP in maternal urine was measured and analyzed by GC-MS. To further validate the results in human NTDs, chick embryos were used as animal models. Viability, reactive oxygen species (ROS) level, oxidative stress indicators and apoptosis were detected in DEHP-treated chick embryos. Our research revealed that the detection ratio of positive DEHP and its metabolites in maternal urine were observed dramatically higher in NTDs population than that in normal controls (P &lt; 0.01, P &lt; 0.05, respectively). Moreover, DEHP treatment (10−6 M) led to developmental toxicity in chick embryos via accelerating oxidative stress response and cell apoptosis, and changing the level of oxidative stress-related indicators. Moreover, high dose choline (100 μg/μl) could partially restrain the toxicity effects induced by DEHP. Our data collectively imply that the incidence of NTDs may closely associate with DEHP exposure, which disturbs the development of neural tubes by enhancing oxidative stress.