Characterizing Chemical Exposure Trends from NHANES Urinary Biomonitoring Data.

BACKGROUND: Xenobiotic metabolites are widely present in human urine and can indicate recent exposure to environmental chemicals. Proper inference of which chemicals contribute to these metabolites can inform human exposure and risk. Furthermore, longitudinal biomonitoring studies provide insight into how chemical exposures change over time. OBJECTIVES: We constructed an exposure landscape for as many human-exposure relevant chemicals over as large a time span as possible to characterize exposure trends across demographic groups and chemical types. METHODS: We analyzed urine data of nine 2-y cohorts (1999-2016) from the National Health and Nutrition Examination Survey (NHANES). Chemical daily intake rates (in milligrams per kilogram bodyweight per day) were inferred, using the R package bayesmarker, from metabolite concentrations in each cohort individually to identify exposure trends. Trends for metabolites and parents were clustered to find chemicals with similar exposure patterns. Exposure variation by age, gender, and body mass index were also assessed. RESULTS: Intake rates for 179 parent chemicals were inferred from 151 metabolites (96 measured in five or more cohorts). Seventeen metabolites and 44 parent chemicals exhibited fold-changes ≥10 between any two cohorts (deltamethrin, di-n-octyl phthalate, and di-isononyl phthalate had the greatest exposure increases). Di-2-ethylhexyl phthalate intake began decreasing in 2007, whereas both di-isobutyl and di-isononyl phthalate began increasing shortly before. Intake for four parabens was markedly higher in females, especially reproductive-age females, compared with males and children. Cadmium and arsenobetaine exhibited higher exposure for individuals >65 years of age and lower for individuals <20 years of age. DISCUSSION: With appropriate analysis, NHANES indicates trends in chemical exposures over the past two decades. Decreases in exposure are observable as the result of regulatory action, with some being accompanied by increases in replacement chemicals. Age- and gender-specific variations in exposure were observed for multiple chemicals. Continued estimation of demographic-specific exposures is needed to both monitor and identify potential vulnerable populations. https://doi.org/10.1289/EHP12188.

Reproducibility of organ-level effects in repeat dose animal studies.

This work estimates benchmarks for new approach method (NAM) performance in predicting organ-level effects in repeat dose studies of adult animals based on variability in replicate animal studies. Treatment-related effect values from the Toxicity Reference database (v2.1) for weight, gross, or histopathological changes in the adrenal gland, liver, kidney, spleen, stomach, and thyroid were used. Rates of chemical concordance among organ-level findings in replicate studies, defined by repeated chemical only, chemical and species, or chemical and study type, were calculated. Concordance was 39 - 88%, depending on organ, and was highest within species. Variance in treatment-related effect values, including lowest effect level (LEL) values and benchmark dose (BMD) values when available, was calculated by organ. Multilinear regression modeling, using study descriptors of organ-level effect values as covariates, was used to estimate total variance, mean square error (MSE), and root residual mean square error (RMSE). MSE values, interpreted as estimates of unexplained variance, suggest study descriptors accounted for 52-69% of total variance in organ-level LELs. RMSE ranged from 0.41 - 0.68 log10-mg/kg/day. Differences between organ-level effects from chronic (CHR) and subchronic (SUB) dosing regimens were also quantified. Odds ratios indicated CHR organ effects were unlikely if the SUB study was negative. Mean differences of CHR - SUB organ-level LELs ranged from -0.38 to -0.19 log10 mg/kg/day; the magnitudes of these mean differences were less than RMSE for replicate studies. Finally, in vitro to in vivo extrapolation (IVIVE) was employed to compare bioactive concentrations from in vitro NAMs for kidney and liver to LELs. The observed mean difference between LELs and mean IVIVE dose predictions approached 0.5 log10-mg/kg/day, but differences by chemical ranged widely. Overall, variability in repeat dose organ-level effects suggests expectations for quantitative accuracy of NAM prediction of LELs should be at least ± 1 log10-mg/kg/day, with qualitative accuracy not exceeding 70%.

Characterizing surface water concentrations of hundreds of organic chemicals in United States for environmental risk prioritization.

BACKGROUND: Thousands of chemicals are observed in freshwater, typically at trace levels. Measurements are collected for different purposes, so sample characteristics vary. Due to inconsistent data availability for exposure and hazard, it is complex to prioritize which chemicals may pose risks. OBJECTIVE: We evaluated the influence of data curation and statistical practices aggregating surface water measurements of organic chemicals into exposure distributions intended for prioritizing based on nation-scale potential risk. METHODS: The Water Quality Portal includes millions of observations describing over 1700 chemicals in 93% of hydrologic subbasins across the United States. After filtering to maintain quality and applicability while including all possible samples, we compared concentrations across sample types. We evaluated statistical methods to estimate per-chemical distributions for chosen samples. Overlaps between resulting exposure ranges and distributions representing no-effect concentrations for multiple freshwater species were used to rank estimated chemical risks for further assessment. RESULTS: When we apply explicit data quality and statistical assumptions, we find that there are 186 organic chemicals for which we can make screening-level estimates of surface water chemical concentration. Of the original 1700 observed chemicals, this number decreased primarily due to a predominance of censored values (that is, observations indicating concentrations too low to be measured). We further identify 423 chemicals where all measurements were censored but, through consideration of detection limits, risk might still be prioritized based on the detection limits themselves. In the final set of 1.5 million samples, the median environmental concentration of one chemical (acetic acid) exceeded the 5th percentile of no-effect concentrations for the most delicate freshwater species (the highest priority risk condition identified here), and a further 29 chemicals were identified for possible further evaluation based on a small margin between occurrence and toxicity values. SIGNIFICANCE: This method shows the broad range of chemical concentrations seen for organic chemicals across the country and identifies methods of determining their central tendency, allowing for researchers to characterize higher-than-normal or lower-than-normal surface water conditions as well as providing an overall indication of the presence of organic chemicals in the United States. The highest chemical concentrations did not always indicate the highest-risk conditions. Even when accounting for the high level of uncertainty in these data due to differences in data collection and reporting across the set, some chemicals may still be categorized as higher environmental risk than others using this method, providing value to chemical safety decision makers and researchers by suggesting avenues for more focused investigation.

Bayesian inference of chemical exposures from NHANES urine biomonitoring data.

BACKGROUND: Knowing which environmental chemicals contribute to metabolites observed in humans is necessary for meaningful estimates of exposure and risk from biomonitoring data. OBJECTIVE: Employ a modeling approach that combines biomonitoring data with chemical metabolism information to produce chemical exposure intake rate estimates with well-quantified uncertainty. METHODS: Bayesian methodology was used to infer ranges of exposure for parent chemicals of biomarkers measured in urine samples from the U.S population by the National Health and Nutrition Examination Survey (NHANES). Metabolites were probabilistically linked to parent chemicals using the NHANES reports and text mining of PubMed abstracts. RESULTS: Chemical exposures were estimated for various population groups and translated to risk-based prioritization using toxicokinetic (TK) modeling and experimental data. Exposure estimates were investigated more closely for children aged 3 to 5 years, a population group that debuted with the 2015-2016 NHANES cohort. SIGNIFICANCE: The methods described here have been compiled into an R package, bayesmarker, and made publicly available on GitHub. These inferred exposures, when coupled with predicted toxic doses via high throughput TK, can help aid in the identification of public health priority chemicals via risk-based bioactivity-to-exposure ratios.

Influence of living in the same home on biomonitored levels of consumer product chemicals.

BACKGROUND: Individuals living in the same home may share exposures from direct contact with sources or indirectly through contamination of the home environment. OBJECTIVE: We investigated the influence of sharing a home on urine levels of ten phenolic chemicals present in some consumer products. METHODS: We used data from Silent Spring Institute's Detox Me Action Kit (DMAK), a crowdsourced biomonitoring program in the US. Of the 726 DMAK participants, 185 lived in the same home with one or more other DMAK participants (n = 137 pairs, up to six participants in a home). The concentration distributions included values below the detection limit so we used statistical methods that account for left-censored data, including non-parametric correlation estimation and hierarchical Bayesian regression models. RESULTS: Concentrations were significantly positively correlated between pair-members sharing a home for nine of the ten chemicals. Concentrations of 2,5-dichlorophenol were the most strongly correlated between pair-members (tau = 0.46), followed by benzophenone-3 (tau = 0.31) and bisphenol A (tau = 0.21). The relative contribution of personal product use reported product use of other household members (up to 5 others), and the residual contribution from a shared household, including exposures not asked about, varied by chemical. Paraben concentrations were largely influenced by personal behaviors whereas dichlorophenol and bisphenol concentrations were largely influenced by shared home exposures not related to reported behaviors. SIGNIFICANCE: Measuring the influence of personal and household practices on biomonitoring exposures helps pinpoint major sources of exposure and highlights chemical-specific intervention strategies to reduce them.

Comparison of Approaches for Determining Bioactivity Hits from High-Dimensional Profiling Data.

Phenotypic profiling assays are untargeted screening assays that measure a large number (hundreds to thousands) of cellular features in response to a stimulus and often yield diverse and unanticipated profiles of phenotypic effects, leading to challenges in distinguishing active from inactive treatments. Here, we compare a variety of different strategies for hit identification in imaging-based phenotypic profiling assays using a previously published Cell Painting data set. Hit identification strategies based on multiconcentration analysis involve curve fitting at several levels of data aggregation (e.g., individual feature level, aggregation of similarly derived features into categories, and global modeling of all features) and on computed metrics (e.g., Euclidean and Mahalanobis distance metrics and eigenfeatures). Hit identification strategies based on single-concentration analysis included measurement of signal strength (e.g., total effect magnitude) and correlation of profiles among biological replicates. Modeling parameters for each approach were optimized to retain the ability to detect a reference chemical with subtle phenotypic effects while limiting the false-positive rate to 10%. The percentage of test chemicals identified as hits was highest for feature-level and category-based approaches, followed by global fitting, whereas signal strength and profile correlation approaches detected the fewest number of active hits at the fixed false-positive rate. Approaches involving fitting of distance metrics had the lowest likelihood for identifying high-potency false-positive hits that may be associated with assay noise. Most of the methods achieved a 100% hit rate for the reference chemical and high concordance for 82% of test chemicals, indicating that hit calls are robust across different analysis approaches.

Variability in in vivo studies: Defining the upper limit of performance for predictions of systemic effect levels.

New approach methodologies (NAMs) for chemical hazard assessment are often evaluated via comparison to animal studies; however, variability in animal study data limits NAM accuracy. The US EPA Toxicity Reference Database (ToxRefDB) enables consideration of variability in effect levels, including the lowest effect level (LEL) for a treatment-related effect and the lowest observable adverse effect level (LOAEL) defined by expert review, from subacute, subchronic, chronic, multi-generation reproductive, and developmental toxicity studies. The objectives of this work were to quantify the variance within systemic LEL and LOAEL values, defined as potency values for effects in adult or parental animals only, and to estimate the upper limit of NAM prediction accuracy. Multiple linear regression (MLR) and augmented cell means (ACM) models were used to quantify the total variance, and the fraction of variance in systemic LEL and LOAEL values explained by available study descriptors (e.g., administration route, study type). The MLR approach considered each study descriptor as an independent contributor to variance, whereas the ACM approach combined categorical descriptors into cells to define replicates. Using these approaches, total variance in systemic LEL and LOAEL values (in log10-mg/kg/day units) ranged from 0.74 to 0.92. Unexplained variance in LEL and LOAEL values, approximated by the residual mean square error (MSE), ranged from 0.20-0.39. Considering subchronic, chronic, or developmental study designs separately resulted in similar values. Based on the relationship between MSE and R-squared for goodness-of-fit, the maximal R-squared may approach 55 to 73% for a NAM-based predictive model of systemic toxicity using these data as reference. The root mean square error (RMSE) ranged from 0.47 to 0.63 log10-mg/kg/day, depending on dataset and regression approach, suggesting that a two-sided minimum prediction interval for systemic effect levels may have a width of 58 to 284-fold. These findings suggest quantitative considerations for building scientific confidence in NAM-based systemic toxicity predictions.

Simulating exposure-related behaviors using agent-based models embedded with needs-based artificial intelligence.

Exposure to a chemical is a critical consideration in the assessment of risk, as it adds real-world context to toxicological information. Descriptions of where and how individuals spend their time are important for characterizing exposures to chemicals in consumer products and in indoor environments. Herein we create an agent-based model (ABM) that simulates longitudinal patterns in human behavior. By basing the ABM upon an artificial intelligence (AI) system, we create agents that mimic human decisions on performing behaviors relevant for determining exposures to chemicals and other stressors. We implement the ABM in a computer program called the Agent-Based Model of Human Activity Patterns (ABMHAP) that predicts the longitudinal patterns for sleeping, eating, commuting, and working. We then show that ABMHAP is capable of simulating behavior over extended periods of time. We propose that this framework, and models based on it, can generate longitudinal human behavior data for use in exposure assessments.

Development of a prioritization method for chemical-mediated effects on steroidogenesis using an integrated statistical analysis of high-throughput H295R data.

Synthesis of 11 steroid hormones in human adrenocortical carcinoma cells (H295R) was measured in a high-throughput steroidogenesis assay (HT-H295R) for 656 chemicals in concentration-response as part of the US Environmental Protection Agency's ToxCast program. This work extends previous analysis of the HT-H295R dataset and model by examining the utility of a novel prioritization metric based on the Mahalanobis distance that reduced these 11-dimensional data to 1-dimension via calculation of a mean Mahalanobis distance (mMd) at each chemical concentration screened for all hormone measures available. Herein, we evaluated the robustness of mMd values, and demonstrate that covariance and variance of the hormones measured appear independent of the chemicals screened and are inherent to the assay; the Type I error rate of the mMd method is less than 1%; and, absolute fold changes (up or down) of 1.5 to 2-fold have sufficient power for statistical significance. As a case study, we examined hormone responses for aromatase inhibitors in the HT-H295R assay and found high concordance with other ToxCast assays for known aromatase inhibitors. Finally, we used mMd and other ToxCast cytotoxicity data to demonstrate prioritization of the most selective and active chemicals as candidates for further in vitro or in silico screening.

Assessing Toxicokinetic Uncertainty and Variability in Risk Prioritization.

High(er) throughput toxicokinetics (HTTK) encompasses in vitro measures of key determinants of chemical toxicokinetics and reverse dosimetry approaches for in vitro-in vivo extrapolation (IVIVE). With HTTK, the bioactivity identified by any in vitro assay can be converted to human equivalent doses and compared with chemical intake estimates. Biological variability in HTTK has been previously considered, but the relative impact of measurement uncertainty has not. Bayesian methods were developed to provide chemical-specific uncertainty estimates for 2 in vitro toxicokinetic parameters: unbound fraction in plasma (fup) and intrinsic hepatic clearance (Clint). New experimental measurements of fup and Clint are reported for 418 and 467 chemicals, respectively. These data raise the HTTK chemical coverage of the ToxCast Phase I and II libraries to 57%. Although the standard protocol for Clint was followed, a revised protocol for fup measured unbound chemical at 10%, 30%, and 100% of physiologic plasma protein concentrations, allowing estimation of protein binding affinity. This protocol reduced the occurrence of chemicals with fup too low to measure from 44% to 9.1%. Uncertainty in fup was also reduced, with the median coefficient of variation dropping from 0.4 to 0.1. Monte Carlo simulation was used to propagate both measurement uncertainty and biological variability into IVIVE. The uncertainty propagation techniques used here also allow incorporation of other sources of uncertainty such as in silico predictors of HTTK parameters. These methods have the potential to inform risk-based prioritization based on the relationship between in vitro bioactivities and exposures.

Considerations for Strategic Use of High-Throughput Transcriptomics Chemical Screening Data in Regulatory Decisions.

Recently, numerous organizations, including governmental regulatory agencies in the U.S. and abroad, have proposed using data from New Approach Methodologies (NAMs) for augmenting and increasing the pace of chemical assessments. NAMs are broadly defined as any technology, methodology, approach or combination thereof that can be used to provide information on chemical hazard and risk assessment that avoids the use of intact animals. High-throughput transcriptomics (HTTr) is a type of NAM that uses gene expression profiling as an endpoint for rapidly evaluating the effects of large numbers of chemicals on in vitro cell culture systems. As compared to targeted high-throughput screening (HTS) approaches that measure the effect of chemical X on target Y, HTTr is a non-targeted approach that allows researchers to more broadly characterize the integrated response of an intact biological system to chemicals that may affect a specific biological target or many biological targets under a defined set of treatment conditions (time, concentration, etc.). HTTr screening performed in concentration-response mode can provide potency estimates for the concentrations of chemicals that produce perturbations in cellular response pathways. Here, we discuss study design considerations for HTTr concentration-response screening and present a framework for the use of HTTr-based biological pathway-altering concentrations (BPACs) in a screening-level, risk-based chemical prioritization approach. The framework involves concentration-response modeling of HTTr data, mapping gene level responses to biological pathways, determination of BPACs, in vitro-to-in vivo extrapolation (IVIVE) and comparison to human exposure predictions.

Methods for evaluating variability in human health dose-response characterization.

The Reference Dose (RfD) and Reference Concentration (RfC) are human health reference values (RfVs) representing exposure concentrations at or below which there is presumed to be little risk of adverse effects in the general human population. The 2009 National Research Council report Science and Decisions recommended redefining RfVs as "a risk-specific dose (for example, the dose associated with a 1 in 100,000 risk of a particular end point)." Distributions representing variability in human response to environmental contaminant exposures are critical for deriving risk-specific doses. Existing distributions estimating the extent of human toxicokinetic and toxicodynamic variability are based largely on controlled human exposure studies of pharmaceuticals. New data and methods have been developed that are designed to improve estimation of the quantitative variability in human response to environmental chemical exposures. Categories of research with potential to provide new database useful for developing updated human variability distributions include controlled human experiments, human epidemiology, animal models of genetic variability, in vitro estimates of toxicodynamic variability, and in vitro-based models of toxicokinetic variability. In vitro approaches, with further development including studies of different cell types and endpoints, and approaches to incorporate non-genetic sources of variability, appear to provide the greatest opportunity for substantial near-term advances.

Empirical models for anatomical and physiological changes in a human mother and fetus during pregnancy and gestation.

Many parameters treated as constants in traditional physiologically based pharmacokinetic models must be formulated as time-varying quantities when modeling pregnancy and gestation due to the dramatic physiological and anatomical changes that occur during this period. While several collections of empirical models for such parameters have been published, each has shortcomings. We sought to create a repository of empirical models for tissue volumes, blood flow rates, and other quantities that undergo substantial changes in a human mother and her fetus during the time between conception and birth, and to address deficiencies with similar, previously published repositories. We used maximum likelihood estimation to calibrate various models for the time-varying quantities of interest, and then used the Akaike information criterion to select an optimal model for each quantity. For quantities of interest for which time-course data were not available, we constructed composite models using percentages and/or models describing related quantities. In this way, we developed a comprehensive collection of formulae describing parameters essential for constructing a PBPK model of a human mother and her fetus throughout the approximately 40 weeks of pregnancy and gestation. We included models describing blood flow rates through various fetal blood routes that have no counterparts in adults. Our repository of mathematical models for anatomical and physiological quantities of interest provides a basis for PBPK models of human pregnancy and gestation, and as such, it can ultimately be used to support decision-making with respect to optimal pharmacological dosing and risk assessment for pregnant women and their developing fetuses. The views expressed in this article are those of the authors and do not necessarily represent the views or policies of the U.S. Environmental Protection Agency.

The Next Generation Blueprint of Computational Toxicology at the U.S. Environmental Protection Agency.

The U.S. Environmental Protection Agency (EPA) is faced with the challenge of efficiently and credibly evaluating chemical safety often with limited or no available toxicity data. The expanding number of chemicals found in commerce and the environment, coupled with time and resource requirements for traditional toxicity testing and exposure characterization, continue to underscore the need for new approaches. In 2005, EPA charted a new course to address this challenge by embracing computational toxicology (CompTox) and investing in the technologies and capabilities to push the field forward. The return on this investment has been demonstrated through results and applications across a range of human and environmental health problems, as well as initial application to regulatory decision-making within programs such as the EPA's Endocrine Disruptor Screening Program. The CompTox initiative at EPA is more than a decade old. This manuscript presents a blueprint to guide the strategic and operational direction over the next 5 years. The primary goal is to obtain broader acceptance of the CompTox approaches for application to higher tier regulatory decisions, such as chemical assessments. To achieve this goal, the blueprint expands and refines the use of high-throughput and computational modeling approaches to transform the components in chemical risk assessment, while systematically addressing key challenges that have hindered progress. In addition, the blueprint outlines additional investments in cross-cutting efforts to characterize uncertainty and variability, develop software and information technology tools, provide outreach and training, and establish scientific confidence for application to different public health and environmental regulatory decisions.

Consensus Modeling of Median Chemical Intake for the U.S. Population Based on Predictions of Exposure Pathways.

Prioritizing the potential risk posed to human health by chemicals requires tools that can estimate exposure from limited information. In this study, chemical structure and physicochemical properties were used to predict the probability that a chemical might be associated with any of four exposure pathways leading from sources-consumer (near-field), dietary, far-field industrial, and far-field pesticide-to the general population. The balanced accuracies of these source-based exposure pathway models range from 73 to 81%, with the error rate for identifying positive chemicals ranging from 17 to 36%. We then used exposure pathways to organize predictions from 13 different exposure models as well as other predictors of human intake rates. We created a consensus, meta-model using the Systematic Empirical Evaluation of Models framework in which the predictors of exposure were combined by pathway and weighted according to predictive ability for chemical intake rates inferred from human biomonitoring data for 114 chemicals. The consensus model yields an R2 of ∼0.8. We extrapolate to predict relevant pathway(s), median intake rate, and credible interval for 479 926 chemicals, mostly with minimal exposure information. This approach identifies 1880 chemicals for which the median population intake rates may exceed 0.1 mg/kg bodyweight/day, while there is 95% confidence that the median intake rate is below 1 µg/kg BW/day for 474572 compounds.

Evaluating In Vitro-In Vivo Extrapolation of Toxicokinetics.

Prioritizing the risk posed by thousands of chemicals potentially present in the environment requires exposure, toxicity, and toxicokinetic (TK) data, which are often unavailable. Relatively high throughput, in vitro TK (HTTK) assays and in vitro-to-in vivo extrapolation (IVIVE) methods have been developed to predict TK, but most of the in vivo TK data available to benchmark these methods are from pharmaceuticals. Here we report on new, in vivo rat TK experiments for 26 non-pharmaceutical chemicals with environmental relevance. Both intravenous and oral dosing were used to calculate bioavailability. These chemicals, and an additional 19 chemicals (including some pharmaceuticals) from previously published in vivo rat studies, were systematically analyzed to estimate in vivo TK parameters (e.g., volume of distribution [Vd], elimination rate). For each of the chemicals, rat-specific HTTK data were available and key TK predictions were examined: oral bioavailability, clearance, Vd, and uncertainty. For the non-pharmaceutical chemicals, predictions for bioavailability were not effective. While no pharmaceutical was absorbed at less than 10%, the fraction bioavailable for non-pharmaceutical chemicals was as low as 0.3%. Total clearance was generally more under-estimated for nonpharmaceuticals and Vd methods calibrated to pharmaceuticals may not be appropriate for other chemicals. However, the steady-state, peak, and time-integrated plasma concentrations of nonpharmaceuticals were predicted with reasonable accuracy. The plasma concentration predictions improved when experimental measurements of bioavailability were incorporated. In summary, HTTK and IVIVE methods are adequately robust to be applied to high throughput in vitro toxicity screening data of environmentally relevant chemicals for prioritizing based on human health risks.

High-Throughput H295R Steroidogenesis Assay: Utility as an Alternative and a Statistical Approach to Characterize Effects on Steroidogenesis.

The U.S. Environmental Protection Agency Endocrine Disruptor Screening Program and the Organization for Economic Co-operation and Development (OECD) have used the human adrenocarcinoma (H295R) cell-based assay to predict chemical perturbation of androgen and estrogen production. Recently, a high-throughput H295R (HT-H295R) assay was developed as part of the ToxCast program that includes measurement of 11 hormones, including progestagens, corticosteroids, androgens, and estrogens. To date, 2012 chemicals have been screened at 1 concentration; of these, 656 chemicals have been screened in concentration-response. The objectives of this work were to: (1) develop an integrated analysis of chemical-mediated effects on steroidogenesis in the HT-H295R assay and (2) evaluate whether the HT-H295R assay predicts estrogen and androgen production specifically via comparison with the OECD-validated H295R assay. To support application of HT-H295R assay data to weight-of-evidence and prioritization tasks, a single numeric value based on Mahalanobis distances was computed for 654 chemicals to indicate the magnitude of effects on the synthesis of 11 hormones. The maximum mean Mahalanobis distance (maxmMd) values were high for strong modulators (prochloraz, mifepristone) and lower for moderate modulators (atrazine, molinate). Twenty-five of 28 reference chemicals used for OECD validation were screened in the HT-H295R assay, and produced qualitatively similar results, with accuracies of 0.90/0.75 and 0.81/0.91 for increased/decreased testosterone and estradiol production, respectively. The HT-H295R assay provides robust information regarding estrogen and androgen production, as well as additional hormones. The maxmMd from this integrated analysis may provide a data-driven approach to prioritizing lists of chemicals for putative effects on steroidogenesis.

Evaluation and calibration of high-throughput predictions of chemical distribution to tissues.

Toxicokinetics (TK) provides critical information for integrating chemical toxicity and exposure assessments in order to determine potential chemical risk (i.e., the margin between toxic doses and plausible exposures). For thousands of chemicals that are present in our environment, in vivo TK data are lacking. The publicly available R package "httk" (version 1.8, named for "high throughput TK") draws from a database of in vitro data and physico-chemical properties in order to run physiologically-based TK (PBTK) models for 553 compounds. The PBTK model parameters include tissue:plasma partition coefficients (Kp) which the httk software predicts using the model of Schmitt (Toxicol In Vitro 22 (2):457-467, 2008). In this paper we evaluated and modified httk predictions, and quantified confidence using in vivo literature data. We used 964 rat Kp measured by in vivo experiments for 143 compounds. Initially, predicted Kp were significantly larger than measured Kp for many lipophilic compounds (log10 octanol:water partition coefficient > 3). Hence the approach for predicting Kp was revised to account for possible deficiencies in the in vitro protein binding assay, and the method for predicting membrane affinity was revised. These changes yielded improvements ranging from a factor of 10 to nearly a factor of 10,000 for 83 Kp across 23 compounds with only 3 Kp worsening by more than a factor of 10. The vast majority (92%) of Kp were predicted within a factor of 10 of the measured value (overall root mean squared error of 0.59 on log10-transformed scale). After applying the adjustments, regressions were performed to calibrate and evaluate the predictions for 12 tissues. Predictions for some tissues (e.g., spleen, bone, gut, lung) were observed to be better than predictions for other tissues (e.g., skin, brain, fat), indicating that confidence in the application of in silico tools to predict chemical partitioning varies depending upon the tissues involved. Our calibrated model was then evaluated using a second data set of human in vivo measurements of volume of distribution (Vss) for 498 compounds reviewed by Obach et al. (Drug Metab Dispos 36(7):1385-1405, 2008). We found that calibration of the model improved performance: a regression of the measured values as a function of the predictions has a slope of 1.03, intercept of - 0.04, and R2 of 0.43. Through careful evaluation of predictive methods for chemical partitioning into tissues, we have improved and calibrated these methods and quantified confidence for TK predictions in humans and rats.

A Method for Identifying Prevalent Chemical Combinations in the U.S. Population.

BACKGROUND: Through the food and water they ingest, the air they breathe, and the consumer products with which they interact at home and at work, humans are exposed to tens of thousands of chemicals, many of which have not been evaluated to determine their potential toxicities. Furthermore, while current chemical testing tends to focus on individual chemicals, the exposures that people actually experience involve mixtures of chemicals. Unfortunately, the number of mixtures that can be formed from the thousands of environmental chemicals is enormous, and testing all of them would be impossible. OBJECTIVES: We seek to develop and demonstrate a method for identifying those mixtures that are most prevalent in humans. METHODS: We applied frequent itemset mining, a technique traditionally used for market basket analysis, to biomonitoring data from the 2009-2010 cycle of the continuous National Health and Nutrition Examination Survey (NHANES) to identify combinations of chemicals that frequently co-occur in people. RESULTS: We identified 90 chemical combinations consisting of relatively few chemicals that occur in at least 30% of the U.S. population, as well as three supercombinations consisting of relatively many chemicals that occur in a small but nonnegligible proportion of the U.S. population. CONCLUSIONS: We demonstrated how FIM can be used in conjunction with biomonitoring data to narrow a large number of possible chemical combinations down to a smaller set of prevalent chemical combinations. https://doi.org/10.1289/EHP1265.