An in vitro-in silico workflow for predicting renal clearance of PFAS.

Per- and poly-fluoroalkyl substances (PFAS) have a wide range of elimination half-lives (days to years) in humans, thought to be in part due to variation in proximal tubule reabsorption. While human biomonitoring studies provide important data for some PFAS, renal clearance (CLrenal) predictions for hundreds of PFAS in commerce requires experimental studies with in vitro models and physiologically-based in vitro-to-in vivo extrapolation (IVIVE). Options for studying renal proximal tubule pharmacokinetics include cultures of renal proximal tubule epithelial cells (RPTECs) and/or microphysiological systems. This study aimed to compare CLrenal predictions for PFAS using in vitro models of varying complexity (96-well plates, static 24-well Transwells and a fluidic microphysiological model, all using human telomerase reverse transcriptase-immortalized and OAT1-overexpressing RPTECs combined with in silico physiologically-based IVIVE. Three PFAS were tested: one with a long half-life (PFOS) and two with shorter half-lives (PFHxA and PFBS). PFAS were added either individually (5 µM) or as a mixture (2 µM of each substance) for 48 h. Bayesian methods were used to fit concentrations measured in media and cells to a three-compartmental model to obtain the in vitro permeability rates, which were then used as inputs for a physiologically-based IVIVE model to estimate in vivo CLrenal. Our predictions for human CLrenal of PFAS were highly concordant with available values from in vivo human studies. The relative values of CLrenal between slow- and faster-clearance PFAS were most highly concordant between predictions from 2D culture and corresponding in vivo values. However, the predictions from the more complex model (with or without flow) exhibited greater concordance with absolute CLrenal. Overall, we conclude that a combined in vitro-in silico workflow can predict absolute CLrenal values, and effectively distinguish between PFAS with slow and faster clearance, thereby allowing prioritization of PFAS with a greater potential for bioaccumulation in humans.

Hazard and risk characterization of 56 structurally diverse PFAS using a targeted battery of broad coverage assays using six human cell types.

Per- and poly-fluoroalkyl substances (PFAS) are extensively used in commerce leading to their prevalence in the environment. Due to their chemical stability, PFAS are considered to be persistent and bioaccumulative; they are frequently detected in both the environment and humans. Because of this, PFAS as a class (composed of hundreds to thousands of chemicals) are contaminants of very high concern. Little information is available for the vast majority of PFAS, and regulatory agencies lack safety data to determine whether exposure limits or restrictions are needed. Cell-based assays are a pragmatic approach to inform decision-makers on potential health hazards; therefore, we hypothesized that a targeted battery of human in vitro assays can be used to determine whether there are structure-bioactivity relationships for PFAS, and to characterize potential risks by comparing bioactivity (points of departure) to exposure estimates. We tested 56 PFAS from 8 structure-based subclasses in concentration response (0.1-100 µM) using six human cell types selected from target organs with suggested adverse effects of PFAS - human induced pluripotent stem cell (iPSC)-derived hepatocytes, neurons, and cardiomyocytes, primary human hepatocytes, endothelial and HepG2 cells. While many compounds were without effect; certain PFAS demonstrated cell-specific activity highlighting the necessity of using a compendium of in vitro models to identify potential hazards. No class-specific groupings were evident except for some chain length- and structure-related trends. In addition, margins of exposure (MOE) were derived using empirical and predicted exposure data. Conservative MOE calculations showed that most tested PFAS had a MOE in the 1-100 range; ∼20% of PFAS had MOE<1, providing tiered priorities for further studies. Overall, we show that a compendium of human cell-based models can be used to derive bioactivity estimates for a range of PFAS, enabling comparisons with human biomonitoring data. Furthermore, we emphasize that establishing structure-bioactivity relationships may be challenging for the tested PFAS.

Assessing proarrhythmic potential of environmental chemicals using a high throughput in vitro-in silico model with human induced pluripotent stem cell-derived cardiomyocytes.

QT prolongation and the potentially fatal arrhythmia Torsades de Pointes are common causes for withdrawing or restricting drugs; however, little is known about similar liabilities of environmental chemicals. Current in vitro-in silico models for testing proarrhythmic liabilities, using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM), provide an opportunity to address this data gap. These methods are still low- to medium-throughput and not suitable for testing the tens of thousands of chemicals in commerce. We hypothesized that combining high-throughput population- based in vitro testing in hiPSC-CMs with a fully in silico data analysis workflow can offer sensitive and specific predictions of proarrhythmic potential. We calibrated the model with a published hiPSC-CM dataset of drugs known to be positive or negative for proarrhythmia and tested its performance using internal cross-validation and external validation. Additionally, we used computational down-sampling to examine three study designs for hiPSC-CM data: one replicate of one donor, five replicates of one donor, and one replicate of a population of five donors. We found that the population of five donors had the best performance for predicting proarrhythmic potential. The resulting model was then applied to predict the proarrhythmic potential of environmental chemicals, additionally characterizing risk through margin of exposure (MOE) calculations. Out of over 900 environmental chemicals tested, over 150 were predicted to have proarrhythmic potential, but only seven chemicals had a MOE < 1. We conclude that a high-throughput in vitro-in silico approach using population-based hiPSC-CM testing provides a reasonable strategy to screen environmental chemicals for proarrhythmic potential.

This article discusses a new method for testing the potential harmful effects of environmental chemicals on the heart. We used human heart cells grown in a lab to test the chemicals and developed a computer model to predict their potential to cause dangerous heart rhythms. This method could help identify harmful chemicals more quickly and accurately than current testing methods. The study has the potential to improve evaluation of chemical risks and protect public health without the use of animals.

Development of physiologically-based gut absorption model for probabilistic prediction of environmental chemical bioavailability.

Absorption in the gastrointestinal tract is a key factor determining the bioavailability of chemicals after oral exposure but is frequently assumed to have a conservative value of 100% for environmental chemicals, particularly in the context of high-throughput toxicokinetics for in vitro-to-in vivo extrapolation (IVIVE). For pharmaceutical compounds, the physiologically based advanced compartmental absorption and transit (ACAT) model has been used extensively to predict gut absorption but has not generally been applied to environmental chemicals. Here we develop a probabilistic environmental compart­mental absorption and transit (PECAT) model, adapting the ACAT model to environmental chemicals. We calibrated the model parameters to human in vivo, ex vivo, and in vitro datasets of drug permeability and fractional absorption by con­sidering two key factors: (1) differences between permeability in Caco-2 cells and in vivo permeability in the jejunum, and (2) differences in in vivo permeability across different gut segments. Incorporating these factors probabilistically, we found that given Caco-2 permeability measurements, predictions of the PECAT model are consistent with the (limited) available gut absorption data for environmental chemicals. However, the substantial chemical-to-chemical variability observed in the cal­ibration data often led to wide probabilistic confidence bounds in the predicted fraction absorbed and resulting steady state blood concentration. Thus, while the PECAT model provides a statistically rigorous, physiologically based approach for incor­porating in vitro data on gut absorption into toxicokinetic modeling and IVIVE, it also highlights the need for more accurate in vitro models and data for measuring gut segment-specific in vivo permeability of environmental chemicals.

Bayesian population physiologically-based pharmacokinetic model for robustness evaluation of withdrawal time in tilapia aquaculture administrated to florfenicol.

The antimicrobial residues of aquacultural production is a growing public concern, leading to reexamine the method for establishing robust withdrawal time and ensuring food safety. Our study aims to develop the optimizing population physiologically-based pharmacokinetic (PBPK) model for assessing florfenicol residues in the tilapia tissues, and for evaluating the robustness of the withdrawal time (WT). Fitting with published pharmacokinetic profiles that experimented under temperatures of 22 and 28 °C, a PBPK model was constructed by applying with the Bayesian Markov chain Monte Carol (MCMC) algorithm to estimate WTs under different physiological, environmental and dosing scenarios. Results show that the MCMC algorithm improves the estimates of uncertainty and variability of PBPK-related parameters, and optimizes the simulation of the PBPK model. It is noteworthy that posterior sets generated from temperature-associated datasets to be respectively used for simulating residues under corresponding temperature conditions. Simulating the residues under regulated regimen and overdosing scenarios for Taiwan, the estimated WTs were 12-16 days at 22 °C and 9-12 days at 28 °C, while for the USA, the estimated WTs were 14-18 and 11-14 days, respectively. Comparison with the regulated WT of 15 days, results indicate that the current WT has well robustness and resilience in the environment of higher temperatures. The optimal Bayesian population PBPK model provides effective analysis for determining WTs under scenario-specific conditions. It is a new insight into the increasing body of literature on developing the Bayesian-PBPK model and has practical implications for improving the regulation of food safety.

Shorter antibiotic regimens impact the control efforts in high tuberculosis burden regions of Taiwan.

OBJECTIVES: To assess the potential epidemiological impact and cost-effectiveness of shorter antibiotic regimens in high tuberculosis (TB) burden regions of Taiwan. METHODS: This study combined the TB population dynamic model and cost-effectiveness analysis with local data to simulate the disease burdens, effectiveness and costs of hypothetical 4-month, 2-month and 7-day regimens compared with the standard regimen. RESULTS: The main outcomes were the potential of shorter regimens for averted incidence, mortality and disability-adjusted life years, incremental cost-effectiveness ratio and net monetary benefit. Shorter regimens would lower incidence rates and mortality cases in a high TB burden region by an average of 19-33% and 27-41%, respectively, with the potential for cost-effectiveness or cost-saving. The 2-month and 7-day regimens would be more cost-effective than the 4-month regimen. The threshold daily drug prices for achieving cost-effectiveness and cost-saving for 4-month, 2-month and 7-day regimens were $US1, $US2 and $US70, respectively. Such cost-effectiveness would remain, even if the willingness-to-pay threshold was less than one gross domestic product per capita. CONCLUSIONS: The findings support the inclusion of shorter regimens in global guidelines and regional-scale TB control strategies, which would improve disease control, particularly in settings with high rates of incidence and poor treatment outcomes.

Risks of consuming cadmium-contaminated shellfish under seawater acidification scenario: Estimates of PBPK and benchmark dose.

We aim to assess the risks of renal dysfunction and osteoporosis that is attributed to the seawater acidification caused cadmium (Cd) level increase in human consumed shellfish. A physiology-based pharmacokinetic model was used to estimate Cd concentrations in urine and blood among shellfish-only consumers and among the general population. We used the benchmark dose (BMD) method to determine the threshold limits of Cd in urine for renal dysfunction and in blood for osteoporosis for assessing the human health risk. Our results revealed that seawater acidification could increase the Cd accumulation in shellfish by 10-13% compared to the situations under current pH levels. Under the lower seawater pH level, the daily intake of Cd could increase by 21%-67% among shellfish-only consumers, and by 13%-17% among the general population. Our findings indicated that seawater acidification would lead to a marginal increase in Cd intake among humans in shellfish-only consumers. The results of BMDs of urinary Cd showed that the threshold limits for renal dysfunction at 5% were 3.00 µg g-1 in males and 12.35 µg g-1 in females. For osteoporosis, the estimated BMDs of blood Cd were 7.95 µg L-1 in males and 1.23 µg L-1 in females. These results of the risk of Cd intake showed that the consumption of Cd-contaminated shellfish in the general population is largely unaffected by changes in seawater pH levels. Notably, the potential impact of seawater acidification on renal dysfunction for males in shellfish-only consumers face a 14% increase of risk.

Bayesian inference of nonylphenol exposure for assessing human dietary risk.

Nonylphenols (NPs) are endocrine-disrupting compounds commonly found in the environment and a number of food products. In this study, we constructed a probabilistic risk framework incorporating a Bayesian inference of exposure level in foodstuffs in conjunction with effect analysis of reproduction and renal disease. Our objective was to contrast the risk of dietary exposure to NPs among individuals in various age groups, with a particular focus on fertile females. In this study, seafood presented relatively high NP concentrations; however, seafood accounts for only a small proportion of the total food intake of most individuals. Rice was shown to make the largest contribution to NP daily intake among males and females in most age groups. Chicken made the largest contribution in the 12-16 and 16-18 year age groups. The mean average daily dose of NPs tended to decrease with age, regardless of gender. The estimated distribution of hazard quotients of <1 in all groups means that the risk of reproductive or renal abnormalities due to dietary exposure to NPs is negligible within most of the Taiwanese population. Nonetheless, preschoolers (3-6-year-olds) appear to be more vulnerable to NPs than do individuals in other age groups. There has been growing concern among researchers concerning the neurotoxic effects of NPs on offspring via maternal exposure. We recommend conducting a comprehensive assessment of exposure to NPs via multiple exposure routes, particularly among fertile women and preschoolers.

Impact of long-term parental exposure to Tamiflu metabolites on the development medaka offspring (Oryzias latipes).

Despite the widespread use of the antiviral drug, Tamiflu®, little is known about the long-term toxic effects of drug or its metabolites in an aquatic ecosystem. This study integrated epidemiological and ecotoxicological methods to determine environmentally relevant concentrations of Tamiflu. A model based on the species medaka (Oryzias latipes) was then used to determine the health status and reproductivity of adults exposed to the drug as well as the embryonic development of offspring. The proposed ecotoxicological model was also used to quantitatively and qualitatively evaluate the toxicodynamic parameters related to egg production, hatchability, and development. Our results revealed that at an environmentally relevant exposure, Tamiflu and its metabolites had no adverse effects on growth, survival, or fecundity of adult medaka. Nonetheless, we observed a reduction in hatchability under exposure to 300 µg L-1 and a reduction in body length under exposure exceeding 90 µg L-1. Under exposure to 300 µg L-1, the estimated spawning time to reach 50% of the maximum percentage of cumulative egg production (ET50) far exceeded that of the control group (without exposure to Tamiflu). We also observed a ∼ 3-fold decrease in maximum egg hatching (Emax). Based on an integrated epidemiological and ecotoxicological model, predictions of environmental concentrations of Tamiflu and its metabolites revealed that the influenza subtypes associated with increases in environmental concentrations: A(H3N2) > A(H1N1) > type B (in order of their effects). We also determined that A(H3N2) posed a potential risk to hatchability and development. Note however, the environmental concentrations of Tamiflu and its metabolites in most countries are lower than the effect concentrations derived in this study, indicating no hazards for aquatic environments. We recommend the use of hatchability and embryonic development as indicators in assessing the effects of long-term parental exposure to Tamiflu metabolites.

Assessing dietary exposure risk to neonicotinoid residues among preschool children in regions of Taiwan.

Neonicotinoids (NEOs) are a class of pesticides widely used worldwide. This study analyzed post-cooking residues of NEO pesticides and assessed their potential health risks for preschool children (0-6 years old) by conducting a total diet study (TDS). It involved food sampling, preparation, analysis of pesticide residues, estimation of food consumption data, and assessment of food safety risks. Food sampling was conducted between March and June 2015. A total of 128 food samples were obtained from 4 parts of Taiwan. After the food had been prepared, the 128 samples were aggregated into 32 composite food items and the NEO residues analyzed. Acetamiprid had the highest detection rate of the NEO residues (59.4%), and the concentrations ranged from not detected to 80.5 µg/kg. The estimated daily intake (EDI) of NEO residues among preschool children was found to be lower than the adjusted acceptable daily intake (ADI) even for highly exposed groups. The results showed that NEO pesticides were primarily detected in preserved fruits, cherry tomato, rape, bell fruit, and baby bok choy. The main health risk posed by detected NEO residues at high consumption rates for preschool children was attributed to acetamiprid (34.20 %ADI) and imidacloprid (23.69 %ADI), respectively. Therefore, this research implicates that the present level of NEO residues in the diets for preschool children in Taiwan does not exceed 100 %ADI.

Association Between Ambient Air Pollution and Elevated Risk of Tuberculosis Development.

BACKGROUND: Broad-scale evidence has shown the significant association between ambient air pollutants and the development of tuberculosis (TB). However, the impact of air quality on the risk of TB in Taiwan is still poorly understood. OBJECTIVE: To develop a probabilistic integrated population-level risk assessment approach for evaluating the contribution of ambient air pollution exposure to the risk of TB development among different regions of Taiwan. MATERIALS AND METHODS: A Bayesian-based probabilistic risk assessment model was implemented to link exposure concentrations of various air pollutants quantified in a probabilistic manner with the population-based exposure-response models developed by using an epidemiological investigation. RESULTS: The increment of the risk of TB occurred in a region with a higher level of air pollution, indicating a strong relationship between ambient air pollution exposures and TB incidences. Carbon monoxide (CO) exposure showed the highest population attributable fraction (PAF), followed by nitrogen oxides (NOX) and nitrogen dioxide (NO2) exposures. In a region with higher ambient air pollution, it is most likely (80% risk probability) that the contributions of CO exposure to development of TB were 1.6-12.2% (range of median PAFs), whereas NOX and NO2 exposures contributed 1.2-9.8% to developing TB. CONCLUSION: Our findings provide strong empirical support for the hypothesis and observations from the literature that poor air quality is highly likely to link aetiologically to the risk of TB. Therefore, substantial reductions in CO, NOX, and NO2 exposures are predicted to have health benefits to susceptible and latently infected individuals that provide complementary mitigation efforts in reducing the burden of TB. Considering that people continue to be exposed to both TB bacilli and ambient air pollutants, our approach can be applied for different countries/regions to identify which air pollutants contribute to a higher risk of TB in order to develop potential mitigation programs.

Assessing the population transmission dynamics of tilapia lake virus in farmed tilapia.

A novel virus, tilapia lake virus (TiLV), has been identified as a key pathogen responsible for disease outbreak and mass mortality of farmed tilapia. We used a deterministic susceptible-infectious-mortality (SIM) model to derive key disease information appraised with published TiLV-induced cumulative mortality data. The relationship between tilapia mortality and TiLV exposure dosages was described by the Hill model. Furthermore, a disease control model was proposed to determine the status of controlled TiLV infection using a parsimonious control reproduction number (RC )-control line criterion. Results showed that the key disease determinants of transmission rate and basic reproduction number (R0 ) could be derived. The median R0 estimate was 2.59 in a cohabitation setting with 2.6 × 105  TCID50 fish-1 TiLV. The present RC -control model can be employed to determine whether TiLV containment is feasible in an outbreak farm by quantifying the current level of transmission. The SIM model can then be applied to predict what additional control is required to manage RC  < 1. We offer valuable tools for aquaculture engineers and public health scientists the mechanistic-based assessment that allows a more rigorous evaluation of different control strategies to reduce waterborne diseases in aquaculture farming systems.

Probabilistic risk assessment of the effect of acidified seawater on development stages of sea urchin (Strongylocentrotus droebachiensis).

Growing evidence indicates that ocean acidification has a significant impact on calcifying marine organisms. However, there is a lack of exposure risk assessments for aquatic organisms under future environmentally relevant ocean acidification scenarios. The objective of this study was to investigate the probabilistic effects of acidified seawater on the life-stage response dynamics of fertilization, larvae growth, and larvae mortality of the green sea urchin (Strongylocentrotus droebachiensis). We incorporated the regulation of primary body cavity (PBC) pH in response to seawater pH into the assessment by constructing an explicit model to assess effective life-stage response dynamics to seawater or PBC pH levels. The likelihood of exposure to ocean acidification was also evaluated by addressing the uncertainties of the risk characterization. For unsuccessful fertilization, the estimated 50% effect level of seawater acidification (EC50 SW ) was 0.55 ± 0.014 (mean ± SE) pH units. This life stage was more sensitive than growth inhibition and mortality, for which the EC50 values were 1.13 and 1.03 pH units, respectively. The estimated 50% effect levels of PBC pH (EC50 PBC ) were 0.99 ± 0.05 and 0.88 ± 0.006 pH units for growth inhibition and mortality, respectively. We also predicted the probability distributions for seawater and PBC pH levels in 2100. The level of unsuccessful fertilization had 50 and 90% probability risks of 5.07-24.51 (95% CI) and 0-6.95%, respectively. We conclude that this probabilistic risk analysis model is parsimonious enough to quantify the multiple vulnerabilities of the green sea urchin while addressing the systemic effects of ocean acidification. This study found a high potential risk of acidification affecting the fertilization of the green sea urchin, whereas there was no evidence for adverse effects on growth and mortality resulting from exposure to the predicted acidified environment.