Hurricane Maria was the most powerful hurricane to hit Puerto Rico in over 80 years, and recovery and clean-up has been painfully slow. Two months after Maria, 60% of the island is still without electricity and many Puerto Ricans are still struggling to find adequate housing, clean water, and food. Prior to Maria, environmental contamination in Puerto Rico was already extensive, and flooding and damage caused by Maria likely exacerbated these conditions. This has potentially resulted in considerable exposure to a range of hazardous substances for many Puerto Ricans. The extended recovery period has also likely increased exposures to environmental contaminants due to extensive use of gas powered generators, use of water from uncertain sources for drinking, cooking, and washing, and increased consumption of canned and packaged foods, all with potential impacts on health. Experiencing a traumatic natural disaster can itself also have long-term physical and psychological health effects. Exposure to such events during pregnancy has been associated with increased risk of preterm birth, low birth weight, and other adverse birth outcomes. However, no previous studies have evaluated measures of both chemical exposure and stress both before and after the hurricane in the same population, so identification of specific, modifiable, hurricane-related risk factors for adverse birth outcomes has not been possible. The proposed research will leverage an ongoing longitudinal birth cohort, the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT), in which 93 participants were pregnant when Hurricane Maria made landfall. The goals of this research are to 1) assess hurricane-related exposure to a range of contaminants (metals, polycyclic aromatic hydrocarbons (PAHs), phthalates, phenols, parabens), and psychological stress, among PROTECT participants who were pregnant when Hurricane Maria struck the island, 2) identify sources of hurricane-related exposures and stress, and 3) evaluate the effects of these exposures on birth outcomes, such as preterm birth, gestational age, and birth weight. Existing biological samples and prenatal data, including urinary biomarkers of exposure and questionnaires on maternal stress and depression, will be utilized in the proposed work. In addition, post-hurricane urine samples, stress questionnaires, and an additional questionnaire on hurricane-related experiences will be collected. Prior to Hurricane Maria, Puerto Rico?s preterm birth rate was already among the highest within the U.S. and the world, so identifying individuals in need of help, as well as modifiable risk factors within this highly vulnerable population in a timely manner is critical.

Our study will provide new evidence of the impacts of air pollution on neonatal and early childhood development for infants and children living in Puerto Rico and of factors that affect susceptibility to air pollution’s harmful impacts. We will do so by leveraging the success of PROTECT, our cohort study of 1800 pregnant women in Puerto Rico for whom a rich dataset of environmental, health, social, demographic, and behavioral factors are being collected. To these data, we will add measurements of air pollution exposures, early childhood development, and non-nutritive suck (NNS), a measure of newborn central nervous system function that has not yet been used to assess neonatal development in environmental epidemiology. We will use these data to achieve three aims: to (1) assess the association of air pollution on adverse birth outcomes; (2) determine the impact of air pollution on child development; and (3) examine effect modification of the air pollution-adverse health relationship by social and personal factors, including socio-economic status, maternal stress, birth outcomes, other pollutant exposures, and housing characteristics. 2) Experimental Approach: We will obtain air pollution exposure and health measurements for a cohort of 600 children living in Puerto Rico, whom we will follow from gestation through age four. We will do so using numerous measures of adverse birth outcomes and development, including preterm birth, annual respiratory and neurodevelopmental evaluations, and NNS. We will link these health measures to measurements of air pollution exposures, metal biomarkers, in utero exposures to contaminants of emerging concern (CECs), and maternal and child characteristics for each child. We will use multiple logistic regression, multi-level mixed models, and cluster analysis techniques to assess the effects of air pollution on adverse birth and developmental outcomes and effect modification of these effects by socioeconomic factors, maternal stress, birth outcomes, CEC mixtures, and home characteristics. Primary analyses will be performed for particulate pollution, with secondary analyses examining impacts of ozone, NO2, PM2.5 sources, air pollutant mixtures, and exposure biomarkers. All models will control for confounders. 3) Expected Results: We will provide evidence of air pollution-mediated risks of developmental delays and adverse birth outcomes for infants and children living in Puerto Rico, who are highly exposed to environmental pollution yet understudied. The insights gained from our research will inform strategies to reduce the frequency of adverse birth outcomes and developmental delays and to improve the cognitive and respiratory health of Puerto Rican children. In so doing, our Project will advance understanding of (1) the impact of air pollution exposures on the developing brain, a susceptible organ that is difficult to access in clinical evaluations, (2) how air pollution exposures and birth outcomes impact neurocognitive development of Puerto Rican children, and (3) the ability of personal factors and chemical mixtures to modify the impact of air pollution on development.

Children in the northern coast of Puerto Rico are exposed to a complex mix of environmental contaminants. The region has over 200 hazardous waste sites, including 16 active Superfund sites, resulting in contaminated water resources. Air pollution from refineries, power plants, motor vehicles, and large ships at ports is also very high, and made worse by Saharan dust storms crossing the Atlantic which have increased in frequency in recent decades. Children in highly polluted areas like Puerto Rico rarely experience any of these exposures in isolation, yet health effect studies associated with exposure to chemical mixtures are rare, with most research instead focusing on single chemicals. Significant challenges remain in understanding the complex risks that pollutant mixtures and their metabolites pose for human health. Novel approaches to examining the impacts of pollutant mixtures are critical to developing preventative strategies to reduce the burden of disease in children in Puerto Rico and the U.S. These challenges motivate this project, which will employ novel in vitro, high throughput screening (HTS) assays to yield new toxicity information needed to reveal the impacts of exposure to complex pollutant mixtures on neonatal and early childhood development in Puerto Rico. These HTS assays will specifically measure translational changes in oxidative stress, DNA damage and inflammation pathways occurring in response to pollutants and pollutant mixtures. Importantly, these pollutants and mixtures are drawn directly from environmental sources, such as water and air, and also from biological matrices like urine, providing information about exposures to chemical mixtures actually occurring in the real-world. Through this approach, we can provide pathway- and sample-specific fingerprints of pollutant exposures and their effects that can inform epidemiological studies, including CRECE Projects 1 and 3. These cross-project collaborations will enable CRECE to perform health effect analyses for pollutant mixtures identified as toxic by the HTS and to examine the association of exposure biomarkers with neonatal and early childhood health outcomes. We will use this approach to achieve four specific aims: (1) assess the toxicity of individual pollutants and pollutant mixtures found in tap water collected from 40 participants' homes; (2) quantify the toxicity associated with metal and organic carbon extracts from airborne fine particle (PM2.5) samples collected as part of Project 1; (3) analyze mixture toxicity effects of CECs, metals, and other chemicals and chemical metabolites in urine samples of 40 pregnant mothers, and (4) use toxicity measures from Aims 2 and 3 and pollutant concentrations measured in Projects 1 and 3 to identify pollutants most responsible for eliciting the observed pathway-specific toxicity impacts. The outcomes of this project will help to inform strategies to minimize health impacts from multiple exposures on Puerto Rico children, a highly exposed population that experiences disproportionate levels of childhood illness, as well as children in the U.S.

Incidence rates for a range of adverse child developmental conditions have increased in recent years in the U.S. and elsewhere. There is emerging evidence and concern that exposure to environmental chemicals such as phenols, parabens, and others may contribute to this recent rise in developmental disorders. Additionally, disparities exist in both incidence rates of adverse health conditions and level of pollutant exposure. There are particularly high rates of a number of developmental conditions, as well as elevated exposures to phenols and parabens, in Puerto Rico. Environmental phenols and parabens disrupt endocrine function, induce oxidative stress, and cause other alterations that may result in reduced fetal or child growth, preterm birth, reproductive tract anomalies, neurodevelopmental delays, obesity, allergies/asthma, and other effects. While near-ubiquitous exposures to these chemicals have been documented among pregnant women, well-designed human studies are greatly needed to determine whether developmental impacts are related to early life in utero exposure. In addition, few studies have addressed the real life situation of considering multiple exposures and susceptibility factors individually and together. Our preliminary data show that pregnant women in Puerto Rico may have higher exposures to several of these chemicals than women in the US and elsewhere, significant associations between these chemicals and markers of hypothesized mechanistic pathways, and that certain modifiable behaviors, such as use of specific products during pregnancy, are associated with elevated chemical exposure and suggest effective intervention strategies may be possible. The proposed project is designed to fill these research gaps by leveraging an ongoing NIH-funded pregnancy cohort study in Puerto Rico (P42ES017198) that is building a rich dataset on environmental, clinical, social, demographic, behavioral, dietary and other factors among 1800 pregnant women. The proposed study will follow 600 children born into the cohort through the age of 4 years. Our specific aims are to 1) Investigate the association between prenatal exposure to environmental phenols and parabens, estimated via urinary biomarkers at multiple times during pregnancy, and pregnancy outcomes and child development (including preterm birth, measures of infant feeding, child growth, reproductive development, neurodevelopment, and lung function); 2) Provide evidence for biological mechanisms that may mediate the relationship between early life exposure to environmental phenols or parabens and fetal/child development, including endocrine disruption, inflammation, and oxidative stress; 3) Test effect modification of the relationship between phenols or parabens and fetal/child development by a range of susceptibility factors, including socioeconomic status, maternal stress, and preterm birth; and 4) develop and apply innovative statistical approaches to explore fetal/child development effects of early-life exposure to multi-pollutant mixtures when also incorporating air pollution data that will be collected in Project 1 of this Center, and biomarker data on phthalates and metals being measured as part of the parent study.