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Per- and polyfluoroalkyl substances (PFAS) are a family of environmental contaminants that have been detected in human placenta and cord blood. The mechanisms driving PFAS-induced adverse pregnancy outcomes and effects on the placenta are not well understood. We investigated the impact of perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid (PFOA), and a replacement PFAS known as hexafluoropropylene oxide dimer acid (HFPO-DA, tradename GenX) on placental trophoblasts in vitro. First, we assessed factors that can influence data quality including quantification of PFAS in common cell culture reagents. Second, we investigated the role of supplemental media serum in intracellular accumulation of PFAS in a human trophoblast (JEG3) cell line. Finally, we evaluated the impact of PFAS on mRNA expression of 96 genes involved in proper placental function in JEG3 cells. Analytical results revealed that, at baseline, serum-free media (SFM) contained no detectable PFAS. Fetal bovine serum-supplemented media (SSM) contained PFNA, PFUdA, PFTrDA, and 6:2 FTS, but these PFAS were not detected internally in cells. Intracellular accumulation following 24 hr treatments was significantly higher when cultured in SFM compared to SSM for PFOS and PFOA, but not HFPO-DA. Finally, PFAS exposure was associated with modest gene expression changes in pathways vital to placental function, including viability, syncytialization, inflammation, transport, and invasion/mesenchymal transition. Among the most robust PFAS-associated changes were those observed in BAD and BAX, apoptosis-related genes. These results suggest a complex relationship between PFAS and culture conditions in vitro, and that PFAS alter key genes necessary for trophoblast involvement in placentation.

The investigation of per- and polyfluorinated alkyl substances (PFAS) in environmental and biological samples relies on both high- and low-resolution mass spectrometry (MS) techniques. While high-resolution MS (HRMS) can be used for identification and quantification of novel compounds, low-resolution MS is the more commonly used and affordable approach for studies examining previously identified PFAS. Of note, perfluorobutanoic acid (PFBA) is one of the smaller PFAS observed in biological and environmental samples and has only one major MS/MS transition, preventing the use of qualitative transitions for verification. Recently, our laboratories undertook a targeted investigation of PFAS in the human placenta from high-risk pregnancies utilizing low-resolution, targeted MS/MS. Examination of placental samples revealed a widespread (n = 93/122 (76%)) chemical interferent in the quantitative ion channel for PFBA (213 → 169). PFBA concentrations were influenced by up to ∼3 ng/g. Therefore, additional chromatographic and HRMS/MS instrumentation was utilized to investigate the suspect peak and putatively assign the identity of the interfering compound as the saturated oxo-fatty acid (SOFA) 3-oxo-dodecanoic acid.

Pre-pregnancy BMI is associated with adverse pregnancy and neonatal health outcomes, with differences in risk observed between sexes. Given that the placenta is a sexually dimorphic organ and critical regulator of development, examining differences in placental mRNA and miRNA expression according to pre-pregnancy BMI may provide insight into responses to maternal BMI in utero. Here, genome-wide mRNA and miRNA expression levels were assessed in the placentas of infants born extremely preterm. Differences in expression were evaluated according to pre-pregnancy BMI status (1) overall and (2) in male and female placentas separately. Overall, N = 719 mRNAs were differentially expressed in relation to underweight status. Unexpectedly, no genes were differentially expressed in relation to overweight or obese status. In male placentas, N = 572 mRNAs were associated with underweight status, with N = 503 (70%) overlapping genes identified overall. Notably, 43/572 (8%) of the mRNAs associated with underweight status in male placentas were also gene targets of two miRNAs (miR-4057 and miR-128-1-5p) associated with underweight status in male placentas. Pathways regulating placental nutrient metabolism and angiogenesis were among those enriched in mRNAs associated with underweight status in males. This study is among the first to highlight a sexually dimorphic response to low pre-pregnancy BMI in the placenta.

Mice have been frequently used to study the adverse effects of inorganic arsenic (iAs) exposure in laboratory settings. Like humans, mice metabolize iAs to monomethyl-As (MAs) and dimethyl-As (DMAs) metabolites. However, mice metabolize iAs more efficiently than humans, which may explain why some of the effects of iAs reported in humans have been difficult to reproduce in mice. In the present study, we searched for mouse strains in which iAs metabolism resembles that in humans. We examined iAs metabolism in male mice from 12 genetically diverse Collaborative Cross (CC) strains that were exposed to arsenite in drinking water (0.1 or 50 ppm) for 2 weeks. Concentrations of iAs and its metabolites were measured in urine and livers. Significant differences in total As concentration and in proportions of total As represented by iAs, MAs, and DMAs were observed between the strains. These differences were more pronounced in livers, particularly in mice exposed to 50 ppm iAs. In livers, large variations among the strains were found in percentage of iAs (15-48%), MAs (11-29%), and DMAs (29-66%). In contrast, DMAs represented 96-99% of total As in urine in all strains regardless of exposure. Notably, the percentages of As species in urine did not correlate with total As concentration in liver, suggesting that the urinary profiles were not representative of the internal exposure. In livers of mice exposed to 50 ppm, but not to 0.1 ppm iAs, As3mt expression correlated with percent of iAs and DMAs. No correlations were found between As3mt expression and the proportions of As species in urine regardless of exposure level. Although we did not find yet a CC strain in which proportions of As species in urine would match those reported in humans (typically 10-30% iAs, 10-20% MAs, 60-70% DMAs), CC strains characterized by low %DMAs in livers after exposure to 50 ppm iAs (suggesting inefficient iAs methylation) could be better models for studies aiming to reproduce effects of iAs described in humans.

BACKGROUND To identify modifiable antecedents during pre-pregnancy and pregnancy windows associated with a positive child health at 10 years of age. METHODS Data on 889 children enrolled in the Extremely Low Gestational Age Newborn (ELGAN) study in 2002-2004 were analyzed for associations between potentially modifiable maternal antecedents during pre-pregnancy and pregnancy time windows and a previously described positive child health index (PCHI) score at 10 years of age. Stratification by race was also investigated for associations with investigated antecedents. RESULTS Factors associated with higher PCHI (more positive health) included greater gestational age, birth weight, multiple gestation, and medical interventions, including assisted reproduction and cervical cerclage. Factors associated with lower PCHI included correlates of lower socioeconomic status, pre-pregnancy chronic medical disorders in the mother such as pre-pregnancy body mass index (BMI), and maternal asthma. When stratified by race, variation in significant results was observed. CONCLUSIONS Among children born extremely preterm, medical interventions and higher socioeconomic status were associated with improved PCHI, while chronic illness and high BMI in the mother is associated with lower PCHI at 10 years of age. Knowledge of such antecedent factors could inform efforts to develop interventions that promote positive child health outcomes in future pregnancies.

Birth weight (BW) represents an important clinical and toxicological measure, indicative of the overall health of the newborn as well as potential risk for later-in-life outcomes. BW can be influenced by endogenous and exogenous factors and is known to be heavily impacted in utero by the health and function of the placenta. An aspect that remains understudied is the influence of genomic and epigenomic programming within the placenta on infant BW. To address this gap, we set out to test the hypothesis that genes involved in critical placental cell signaling are associated with infant BW, and are likely regulated, in part, through epigenetic mechanisms based on microRNA (miRNA) mediation. This study leveraged a robust dataset based on 390 infants born at low gestational age (ranged 23 to 27 weeks) to evaluate genome-wide expression profiles of both mRNAs and miRNAs in placenta tissues and relate these to infant BW. A total of 254 mRNAs and 268 miRNAs were identified as associated with BW, the majority of which showed consistent associations across placentas derived from both males and females. BW-associated mRNAs were found to be enriched for important biological pathways, including glycoprotein VI (the major receptor for collagen), human growth, and hepatocyte growth factor signaling, a portion of which were predicted to be regulated by BW-associated miRNAs. These miRNA-regulated pathways highlight key mechanisms potentially linking endogenous/exogenous factors to changes in birth outcomes that may be deleterious to infant and later-in-life health.

Prenatal exposure to per- and polyfluoroalkyl substances (PFAS), a ubiquitous class of chemicals, is associated with adverse outcomes such as pre-eclampsia, low infant birth weight, and later-life adiposity. The objectives of this study were to examine PFAS levels in the placenta and identify sociodemographic risk factors in a high-risk pregnancy cohort (n = 122) in Chapel Hill, North Carolina. Of concern, PFOS, PFHxS, PFHpS, and PFUnA were detected above the reporting limit in 99, 75, 55, and 49% of placentas, respectively. Maternal race/ethnicity was associated with significant differences in PFUnA levels. While the data from this high-risk cohort did not provide evidence for an association with hypertensive disorders of pregnancy, fetal growth, or gestational age, the prevalence of detectable PFAS in the placenta suggests a need to biomonitor for exposure to PFAS during pregnancy. Future research should investigate factors underlying the differences in PFAS levels in association with a mother's race/ethnicity, as well as potential effects on pregnancy and child health.

Environmental chemicals comprise a major portion of the human exposome, with some shown to impact the health of susceptible populations, including pregnant women and developing fetuses. The placenta and cord blood serve as important biological windows into the maternal and fetal environments. In this article we review how environmental chemicals (defined here to include man-made chemicals [e.g., flame retardants, pesticides/ herbicides, per- and polyfluoroalkyl substances], toxins, metals, and other xenobiotic compounds) contribute to the prenatal exposome and highlight future directions to advance this research field. Our findings from a survey of recent literature indicate the need to better understand the breadth of environmental chemicals that reach the placenta and cord blood, as well as the linkages between prenatal exposures, mechanisms of toxicity, and subsequent health outcomes. Research efforts tailored towards addressing these needs will provide a more comprehensive understanding of how environmental chemicals impact maternal and fetal health.

During pregnancy, the placenta is critical for the regulation of maternal homeostasis and fetal growth and development. Exposures to environmental chemicals during pregnancy can be detrimental to the health of the placenta and therefore adversely impact maternal and fetal health. Though research on placental-derived developmental toxicity is expanding, testing is limited by the resources required for traditional test methods based on whole animal experimentation. Alternative strategies utilizing in vitro methods are well suited to contribute to more efficient screening of chemical toxicity and identification of biological mechanisms underlying toxicity outcomes. This review aims to summarize methods that can be used to evaluate toxicity resulting from exposures during the prenatal period, with a focus on newer in vitro methods centered on placental toxicity. The following key aspects are reviewed: (i) traditional test methods based on animal developmental toxicity testing, (ii) in vitro methods using monocultures and explant models, as well as more recently developed methods, including co-cultures, placenta-on-a-chip, and 3-dimensional (3D) cell models, (iii) endpoints that are commonly measured using in vitro designs, and (iv) the translation of in vitro methods into chemical evaluations and risk assessment applications. We conclude that findings from in vitro placental models can contribute to the screening of potentially hazardous chemicals, elucidation of chemical mechanism of action, incorporation into adverse outcome pathways, estimation of doses eliciting toxicity, derivation of extrapolation factors, and characterization of overall risk of adverse outcomes, representing key components of chemical regulation in the 21(st) century.

Per- and polyfluoroalkyl substances (PFAS) are ubiquitous, synthetic anthropogenic chemicals known to infiltrate and persist in biological systems as a result of their stability and bioaccumulation potential. This study investigated 15 PFAS, including short-chain carboxylic and sulfonic acids, and their presence in a threatened herbivore, the West Indian manatee (Trichechus manatus). Seven of the 15 PFAS examined were detected in manatee plasma. Perfluorooctanesulfonic acid (PFOS) (ranging from 0.13 to 166 ng/g ww) and perfluorononanoic acid (PFNA) (ranging from 0.038 to 3.52 ng/g ww) were detected in every manatee plasma sample examined (n = 69), with differing medians across sampling sites in Florida, Crystal River (n = 39), Brevard County (n = 18), Everglades National Park (n = 8), and four samples (n = 4) from Puerto Rico. With an herbivorous diet and long life-span, the manatee provides a new perspective to monitoring PFAS contamination.

This study focused on an estuarine wildlife species exhibiting high site fidelity and ubiquitous distribution in coastal environments along the Atlantic and Gulf coasts of the United States to monitor per- and polyfluoroalkyl substances (PFAS). A total of 75 diamondback terrapin (Malaclemys terrapin) plasma samples were collected from five creeks associated with Kiawah (Oyster Creek, Fiddler Creek, Sandy Creek, Gnat Creek) and Edisto (Townsend Creek) islands in Charleston County, South Carolina and investigated for 15 legacy PFAS. Of those, PFHxS was the only PFAS found in all terrapin plasma samples. Four additional PFAS were routinely detected (greater than 90% of the samples) and were included in statistical analyses: PFOS, PFNA, PFDA, and PFUnA. Sex-differences were observed for two creeks with male plasma containing higher PFAS than female plasma (PFHxS at Townsend Creek, PFOS at Oyster Creek). Sex-specific site differences in PFAS concentrations were observed primarily for males, suggesting male terrapins may be more sensitive indicators of localized contaminant profiles than females. Three PFAS were observed to have negative correlations with body mass: PFOS in males (p = 0.045, tau = −0.220), PFNA in males (p = 0.016, tau = −0.269), and PFHxS in both males (p = 0.007, tau = −0.302) and females (p = 0.001, tau = −0.379). No relationships for body mass and PFDA and PFUnA were observed.

OBJECTIVE. To assess the development of a positive child health index (PCHI) based on 11 adverse outcomes and evaluate PCHI association with quality of life (QoL) scores in a preterm cohort. STUDY DESIGN. A total of 889 children enrolled in the Extremely Low Gestational Age Newborn (ELGAN) study in 2002–2004 were followed up at 10 years of age. A parent/caregiver completed questionnaires for child QoL, asthma, visual or hearing impairment, gross motor function impairment, epilepsy, attention deficit/hyperactivity disorder, anxiety, and depression. The child was assessed for cognitive impairment, autism, and obesity. PCHI scores were computed, and linear regression models were used to evaluate the relationship between QoL categories (psychosocial, physical, emotional, social, school, and total) and the PCHI (dichotomized and coded as a multi-level categorical predictor) and to assess sex differences. RESULTS. Among ELGAN children, higher PCHI scores were associated with higher reported QoL scores for all QoL categories. Children with no disorders and a PCHI of 100% had PedsQL total scores that were 11 points higher than children with one or more adverse outcomes (PCHI < 100%). Boys had lower QoL scores for categories total, psychosocial, social, and school. CONCLUSIONS. Positive child health assessed using a quantitative PCHI was associated with QoL across the ELGAN cohort at school age. In the current study, PCHI encompassed 11 outcomes assessed in ELGANs. Future research could include an enhanced panel of child health outcomes to support the use of PCHI as an indicator of positive child health.

This study investigated wild caught striped mullet (Mugil cephalus) at Merritt Island National Wildlife Refuge (MINWR) for levels of 15 perfluoroalkyl acids (PFAA) in tandem with individual fecundity measurements (Oocyte sub-stage 2 late, n = 42) and oocyte reproductive stages (Stages 1–5, n = 128). PFAA measurements were quantified in striped mullet liver (n = 128), muscle (n = 49), and gonad (n = 10). No significant negative impacts of liver PFAA burden on wild-caught, mullet fecundity endpoints were observed in this study; however, changes in PFAA were observed in the liver as mullet progressed through different sub-stages of oocyte development. Of the PFAA with significant changes by sub-stage of oocyte development, the carboxylic acids (perfluorooctanoic acid, perfluorononanoic acid, and perfluorotridecanoic acid) increased in the liver with increasing sub-stage while the sulfonic acid and its precursor (perfluorooctanesulfonic acid (PFOS) and perfluorooctanesulfonamide, respectively) decreased in the liver with increasing sub-stage of oocyte development. This is a unique find and suggests PFAA change location of compartmentalization as mullet progress towards spawning. Investigations also revealed higher than expected median muscle and gonad levels of PFOS in striped mullet collected at MINWR (9.01 ng/g and 80.2 ng/g, respectively).

This study examined concentrations of 15 perfluoroalkyl acids (PFAAs) in tissues from male Mozambique tilapia (Oreochromis mossambicus) collected at Loskop Dam, Mpumalanga, South Africa in 2014 and 2016. Nine of the 15 PFAAs were detected frequently and were included in statistical analysis and included two of the most commonly known PFAAs, perfluorooctanesulfonic acid (PFOS) (median, 41.6 ng/g) and perfluorooctanoic acid (PFOA) (median, 0.0825 ng/g). Of the tissues measured, plasma (2016 and 2014 median, 22.2 ng/g) contained the highest PFAA burden followed by (in descending order): liver (median, 11.6 ng/g), kidney (median, 9.04 ng/g), spleen (median, 5.92 ng/g), adipose (median, 2.54 ng/ g), and muscle (median, 1.11 ng/g). Loskop Dam tilapia have been affected by an inflammatory disease of the adipose tissue known as pansteatitis, so this study also aimed to investigate relationships between PFAA tissue concentrations and incidence of pansteatitis or fish health status. Results revealed that healthy tilapia exhibited an overall higher (p-value < 0.05) PFAA burden than pansteatitis-affected tilapia across all tissues. Further analysis showed that organs previously noted in the literature to contain the highest PFAA concentrations, such as kidney, liver, and plasma, were the organs driving the difference in PFAA burden between the two tilapia groups. Care must be taken in the interpretations we draw from not only the results of our study, but also other PFAA measurements made on populations (human and wildlife alike) under differing health status.

This study aimed to quantify concentrations of fifteen perfluoroalkyl acids (PFAAs) in the plasma of American alligators (Alligator mississippiensis) inhabiting wetlands surrounding the Kennedy Space Center (KSC) in Florida, USA located at Merritt Island National Wildlife Refuge (MINWR). Approximately 10 male and 10 female alligators (ntotal = 229) were sampled each month during 2008 and 2009 to determine if seasonal or spatial trends existed with PFAA burden. PFOS represented the highest plasma burden (median 185 ng/g) and PFHxS the second highest (median 7.96 ng/g). While no significant seasonal trends were observed, unique spatial trends emerged. Many of the measured PFAAs co-varied strongly together and similar trends were observed for PFOS, PFDA, PFUnA, and PFDoA, as well as for PFOA, PFHxS, PFNA, PFTriA, and PFTA, suggesting more than one source of PFAAs at MINWR. Higher concentrations of PFOS and the PFAAs that co-varied with PFOS were collected from animals around sites that included the Shuttle Landing Facility (SLF) fire house and the Neil Armstrong Operations and Checkout (O&C) retention pond, while higher concentrations of PFOA and the PFAA that co-varied with PFOA were sampled from animals near the gun range and the old fire training facility. Sex-based differences and snout-vent length (SVL) correlations with PFAA burden were also investigated.

The present study aimed to quantitate 15 perfluoroalkyl acids (PFAAs) in 125 adult American alligators at 12 sites across the southeastern United States. Of those 15 PFAAs, 9 were detected in 65% to 100% of samples: perfluorooctanoic acid, perfluorononanoic acid, perfluorodecanoic acid (PFDA), perfluoroundecanoic acid (PFUnA), perfluorododecanoic acid, perfluorotridecanoic acid (PFTriA), perfluorotetradecanoic acid, perfluorohexanesulfonic acid (PFHxS), and perfluorooctane sulfonate (PFOS). Males (across all sites) showed significantly higher concentrations of 4 PFAAs: PFOS (p = 0.01), PFDA (p = 0.0003), PFUnA (p = 0.021), and PFTriA (p = 0.021). Concentrations of PFOS, PFHxS, and PFDA in plasma were significantly different among the sites in each sex. Alligators at both Merritt Island National Wildlife Refuge (FL, USA) and Kiawah Nature Conservancy (SC, USA) exhibited some of the highest PFOS concentrations (medians of 99.5 ng/g and 55.8 ng/g, respectively) in plasma measured to date in a crocodilian species. A number of positive correlations between PFAAs and snout-vent length were observed in both sexes, suggesting that PFAA body burdens increase with increasing size. In addition, several significant correlations among PFAAs in alligator plasma may suggest conserved sources of PFAAs at each site throughout the greater study area. The present study is the first to report PFAAs in American alligators, to reveal potential PFAA hot spots in Florida and South Carolina, and to provide a contaminant of concern when assessing anthropogenic impacts on ecosystem health. Environ Toxicol Chem 2017;36:917-925. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US government work and, as such, is in the public domain in the United States of America.