A framework for integrating evidence to assess hazards and risk.

To accurately characterize human health hazards, human, animal, and mechanistic data must be integrated and the relevance to the research question of all three lines of evidence must be considered. Mechanistic data are often critical to the full integration of animal and human data and to characterizing relevance and uncertainty. This novel evidence integration framework (EIF) provides a method for synthesizing data from comprehensive, systematic, quality-based assessments of the epidemiological and toxicological literature, including in vivo and in vitro mechanistic studies. It organizes data according to both the observed human health effects and the mechanism of action of the chemical, providing a method to support evidence synthesis. The disease-based component uses the evidence of human health outcomes studied in the best quality epidemiological literature to organize the toxicological data according to authors' stated purpose, with the pathophysiology of the disease determining the potential relevance of the toxicological data. The mechanism-based component organizes the data based on the proposed mechanisms of effect and data supporting events leading to each endpoint, with the epidemiological data potentially providing corroborating information. The EIF includes a method to cross-classify and describe the concordance of the data, and to characterize its uncertainty. At times, the two methods of organizing the data may lead to different conclusions. This facilitates identification of knowledge gaps and shows the impact of uncertainties on the strength of causal inference.

Effects of hazardous chemicals on secondary sex ratio: A comprehensive review.

The secondary sex ratio (SSR), defined as the ratio of male to female offspring at birth, has garnered significant scientific interest due to its potential impact on population dynamics and evolution. In recent years, there has been a growing concern regarding the potential consequences of environmental chemicals on the SSR, given their widespread exposure and potential enduring ramifications on the reproductive system. While SSR serves as an indicator of health, ongoing research and scientific inquiry are being conducted to explore the potential relationship between chemicals and offspring ratio. Although some studies have suggested a possible correlation, others have yielded inconclusive results, indicating that the topic is intricate and still needs to be elucidated. The precise mechanism by which chemical agents exert their influence on the SSR remains ambiguous, with disruption of the endocrine system being a prominent justification. In light of the complex interplay between chemical exposure and SSR, the present review aims to comprehensively examine and synthesize existing scientific literature to gain a deeper understanding of how specific chemical exposures may impact SSR. Insights into chemical hazards that shift SSR patterns or trends could guide prevention strategies, including legislative bans of certain chemicals, to minimize environmental and public health risks.

Assessing pollution degree and human health risks from hazardous element distribution in soils near gold mines in a Colombian Andean region: Correlation with phytotoxicity biomarkers.

The assessment of human health risk due to the presence of hazardous elements in the environment is now necessary for environmental management and legislative initiatives. This study aims to determine the contamination by As, Cd, Pb, and Cr in soils near gold mines in three municipalities located in the Andean region of Colombia. One of the main objectives of the study is to explore possible correlations between the Lifetime Cancer Risk (LCR) and phytotoxicity biomarkers using a simple and rapid-response plant model, radish (Raphanus sativus L.). In the municipality of Yalí, Puerto Berrío, and Buriticá, the hazardous elements concentrations ranged from 8.1 to 35.5, 1.7 to 892, and 5.8 to 49.8 for As, 0.1 to 4.6, 0.1 to 65.2, and 0.5 to 18.2 for Cd, 18.5 to 201.3, 13.0 to 1908, and 189 to 2345 for Pb, and 5.4 to 118.4, 65.4 to 301, and 5.4 to 102.3 for Cr, respectively. The results showed that the biomarkers intracellular H2O2 concentration, antioxidant activity, and radicle elongation exhibited significant (P < 0.05) variations associated with the concentration of hazardous elements in the soils. Significant correlations (P < 0.05, r > 0.58) were found between the biomarkers and the LCR for Cd, Pb, and Cr, but not for As. The results using biomarkers reveal that soil pH and organic matter content are important variables that control the bioavailability of these elements in the soil. The use of indicators like LCR alone has limitations and should be accompanied by the use of biomarkers that allow for a better understanding of the biological system's response to exposure to potentially toxic elements. The results obtained show the urgent need to implement public policies to minimize exposure to hazardous substances in areas near gold mining projects.

Modeling mixtures interactions in environmental toxicology.

In the environment, organisms are exposed to mixtures of different toxicants, which may interact in ways that are difficult to predict when only considering each component individually. Adapting and expanding tools from pharmacology, the toxicology field uses analytical, graphical, and computational methods to identify and quantify interactions in multi-component mixtures. The two general frameworks are concentration addition, where components have similar modes of action and their effects sum together, or independent action, where components have dissimilar modes of action and do not interact. Other interaction behaviors include synergism and antagonism, where the combined effects are more or less than the additive sum of individual effects. This review covers foundational theory, methods, an in-depth survey of original research from the past 20 years, current trends, and future directions. As humans and ecosystems are exposed to increasingly complex mixtures of environmental contaminants, analyzing mixtures interactions will continue to become a more critical aspect of toxicological research.

Towards a more effective REACH legislation in protecting human health.

There is growing evidence indicating the substantial contribution of man-made products to an increase in the risk of diseases of civilization. In this article, the Belgian Scientific Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) Committee gives a critical view on the working of REACH. The current regulatory framework needs to further evolve taking into account data generated using modern science and technology. There is a need for improved assessment process not only before but also after entering the market. Objectivity, transparency, and the follow-up after market access can be optimized. Additionally, no guidance documents exist for regulation of mixture effects. Further, the lengthiness before regulatory action is a big concern. Decision-making often takes several years leading to uncertainties for both producers and end users. A first proposed improvement is the implementation of independent toxicity testing, to assure objectivity, transparency, and check and improve compliance. A "no data, no market" principle could prevent access of hazardous chemicals to the market. Additionally, the introduction of novel testing could improve information on endpoints such as endocrine disrupting abilities, neurotoxicity, and immunotoxicity. An adapted regulatory framework that integrates data from different sources and comparing the outputs with estimates of exposure is required. Fast toxicology battery testing and toxicokinetic testing could improve speed of decision-making. Hereby, several improvements have been proposed that could improve the current REACH legislation.

Opportunities in Assessing and Regulating Organohalogen Flame Retardants (OFRs) as a Class in Consumer Products.

BACKGROUND: In 2015, the U.S. Consumer Product Safety Commission (CPSC) received and then, in 2017, granted a petition under the Federal Hazardous Substances Act to declare certain groups of consumer products as banned hazardous substances if they contain nonpolymeric, additive organohalogen flame retardants (OFRs). The petitioners asked the CPSC to regulate OFRs as a single chemical class with similar health effects. The CPSC later sponsored a National Academy of Sciences, Engineering, and Medicine (NASEM) report in 2019, which ultimately identified 161 OFRs and grouped them into 14 subclasses based on chemical structural similarity. In 2021, a follow-up discussion was held among a group of scientists from both inside and outside of the CPSC for current research on OFRs and to promote collaboration that could increase public awareness of CPSC work and support the class-based approach for the CPSC's required risk assessment of OFRs. OBJECTIVES: Given the extensive data collected to date, there is a need to synthesize what is known about OFR and how class-based regulations have previously managed this information. This commentary discusses both OFR exposure and OFR toxicity and fills some gaps for OFR exposure that were not within the scope of the NASEM report. The objective of this commentary is therefore to provide an overview of the OFR research presented at SOT 2021, explore opportunities and challenges associated with OFR risk assessment, and inform CPSC's work on an OFR class-based approach. DISCUSSION: A class-based approach for regulating OFRs can be successful. Expanding the use of read-across and the use of New Approach Methodologies (NAMs) in assessing and regulating existing chemicals was considered as a necessary part of the class-based process. Recommendations for OFR class-based risk assessment include the need to balance fire and chemical safety and to protect vulnerable populations, including children and pregnant women. The authors also suggest the CPSC should consider global, federal, and state OFR regulations. The lack of data or lack of concordance in toxicity data could present significant hurdles for some OFR subclasses. The potential for cumulative risks within or between subclasses, OFR mixtures, and metabolites common to more than one OFR all add extra complexity for class-based risk assessment. This commentary discusses scientific and regulatory challenges for a class-based approach suggested by NASEM. This commentary is offered as a resource for anyone performing class-based assessments and to provide potential collaboration opportunities for OFR stakeholders. https://doi.org/10.1289/EHP12725.

Can "Hazard-Cost-Effectiveness Analysis" improve the risk management of chemicals under REACH?

Cost-Effectiveness Analysis (CEA) is a decision-making framework to prioritize policy decisions for chemicals. Differences in hazard profiles among chemicals are not integrated in CEA under the EU REACH Regulation, which could limit its relevance. Another concern is that two different economic decision support methods (CEA for chemicals considered as PBTs or vPvBs from a regulatory perspective and Cost Benefit Analysis (CBA) for others) are used under REACH. To address this situation, we define "Hazard" CEA by integrating a hazard score, based on persistence, bioaccumulation and (eco)toxicity, in the effect indicator of CEA. We test different designs and parameterizations of Hazard-CEA on a set of past socio-economic assessments under REACH for PBT and non-PBT chemicals. Weighing and thresholds in hazard scores do not have a significant impact on the outcome of Hazard-CEA but the design of the hazard scoring method does. We suggest using an integrated and unweighted scoring method with a multiplicative formulation based on the notion of risk. Hazard-CEA could be used for both PBT and non-PBT chemicals, to use a single method in REACH and therefore improve consistency in policy decisions. Our work also suggests that using Hazard-CEA could help make decision easier.

Highly efficient molecular film for inhibiting volatilization of hazardous nitric acid.

Nitric acid, an important basic chemical raw material, plays an important role in promoting the development of national economy. However, such liquid hazardous chemicals are easy to cause accidental leakage during production, transportation, storage and use. The high concentration and corrosive toxic gas generated from decomposition shows tremendous harm to the surrounding environment and human life safety. Therefore, how to inhibit the volatilization of nitric acid and effectively control and block the generation of the toxic gas in the first time are the key to deal with the nitric acid leakage accident. Herein, a new method of molecular film obstruction is proposed to inhibit the nitric acid volatilization. The molecular film inhibitor spontaneously spread and form an insoluble molecular film on the gas-liquid interface, changing the state of nitric acid liquid surface and inhibiting the volatilization on the molecular scale. The inhibition rate up to 96% can be achieved below 45 °C within 400 min. Cluster structure simulation and energy barrier calculation is performed to elucidate the inhibition mechanism. Theoretical analysis of energy barrier shows that the specific resistance of the inhibitor significantly increased to 460 s·cm-1 at 45 °C, and the generated energy barrier is about 17,000 kJ·mol-1, which is much higher than the maximum energy required for nitric acid volatilization of 107.97 kJ·mol-1. The molecular film obstruction strategy can effectively inhibit the volatilization of nitric acid. This strategy paves the way for preventing the volatilization of liquid hazardous chemicals in accidental leakage treatment.

An integrated in vitro approach to identifying chemically induced oxidative stress and toxicity in mitochondria.

Growing concerns exist about increasing chemical usage and the potential health risks. Developing an efficient strategy to evaluate or predict the toxicity of chemicals is necessary. The mitochondria are essential organelles for cell maintenance and survival but also serve as one of the main targets of toxic chemicals. Mitochondria play an important role in the pathology of respiratory disease, and many environmental chemicals may induce impairment of the respiratory system through mitochondrial damage. This study aimed to develop integrated in vitro approaches to identify chemicals that could induce adverse health effects by increasing mitochondria-mediated oxidative stress using the H441 cells, which have a club-cell-like phenotype. Twenty-six environmental toxicants (biocides, phthalates, bisphenols, and particles) were tested, and each parameter was compared with eleven reference compounds. The inhibitory concentrations (IC20 and IC50) and benchmark doses (BMD) of the tested compounds were estimated from three in vitro assays, and the toxic concentration was determined. At the lowest IC20, the effects of compounds on mitochondrial reactive oxygen species (ROS) production and mitochondrial membrane potential (MMP) were compared. Principal component analysis and k-mean clustering were performed to cluster the chemicals that had comparable effects on the cells. Chemicals that induce mitochondrial damage at different concentrations were used for an in-depth high-tier assessment and classification as electron transport system (ETS) uncoupling or inhibiting agents. Additionally, using in vitro to in vivo extrapolation (IVIVE) tools, equivalent administration doses and maximum plasma concentrations of tested compounds in human were estimated. This study suggests an in vitro approach to identifying mitochondrial damage by integrating several in vitro toxicity tests and calculation modeling.

The importance of variations in in vitro dosimetry to support risk assessment of inhaled toxicants.

In vitro methods provide a key opportunity to model human-relevant exposure scenarios for hazard identification of inhaled toxicants. Compared to in vivo tests, in vitro methods have the advantage of assessing effects of inhaled toxicants caused by differences in dosimetry, e.g., variations in con­centration (exposure intensity), exposure duration, and exposure frequency, in an easier way. Variations in dosimetry can be used to obtain information on adverse effects in human-relevant exposure scenarios that can be used for risk assessment. Based on the published literature of exposure approaches using air-liquid interface models of the respiratory tract, supplemented with additional experimental data from the EU H2020 project "PATROLS" and research funded by the Dutch Ministry of Agriculture, Nature and Food Quality, the advantages and disadvantages of dif­ferent exposure methods and considerations to design an experimental setup are summarized and discussed. As the cell models used are models for the respiratory epithelium, our focus is on the local effects in the airways. In conclusion, in order to generate data from in vitro methods for risk assessment of inhaled toxicants it is recommended that (1) it is considered what information really is needed for hazard or risk assessment; (2) the exposure system that is most suitable for the chemical to be assessed is chosen; (3) a deposited dose that mimics deposition in the human respiratory tract is used, and (4) the post-exposure sampling methodology should be carefully considered and relevant to the testing strategy used.

The impact of airborne pollutants on human health is determined by what pollutant it is, how much we breathe in, for how long and how often. Testing in animals is cumbersome and results may not reflect human health impacts. Advanced cell models of the human lung allow prediction of the health impact of many different exposure scenarios. Here, we compare different models and exposure methods and provide criteria that may assist in designing experiments, interpreting the results, and thus assessing the risks posed by airborne pollutants. We recommend (1) determining what infor­mation is needed to plan the experiment, (2) choosing an exposure method that is suitable for the pollutant of interest, (3) determining the amount of pollutant that interacts with the human lung, to relate this to realistic deposition in the lung, and (4) considering the time between the exposure and measurement of the effect.

A deep-learning approach for identifying prospective chemical hazards.

With the aim of helping to set safe exposure limits for the general population, various techniques have been implemented to conduct risk assessments for chemicals and other environmental stressors; however, none of these tools facilitate the identification of completely new chemicals that are likely hazardous and elicit an adverse biological effect. Here, we detail a novel in silico, deep-learning framework that is designed to systematically generate structures for new chemical compounds that are predicted to be chemical hazards. To assess the utility of the framework, we applied the tool to four endpoints related to environmental toxicants and their impacts on human and animal health: (i) toxicity to honeybees, (ii) immunotoxicity, (iii) endocrine disruption via ER-α antagonism, and (iv) mutagenicity. In addition, we characterized the predicted potency of these compounds and examined their structural relationship to existing chemicals of concern. As part of the array of emerging new approach methodologies (NAMs), we anticipate that such a framework will be a significant asset to risk assessors and other environmental scientists when planning and forecasting. Though not in the scope of the present study, we expect that the methodology detailed here could also be useful in the de novo design of more environmentally-friendly industrial chemicals.

Toxic Tire Wear Compounds (6PPD-Q and 4-ADPA) Detected in Airborne Particulate Matter Along a Highway in Mississippi, USA.

Tire wear particles (TWPs) are a major category of microplastic pollution produced by friction between tires and road surfaces. This non-exhaust particulate matter (PM) containing leachable toxic compounds is transported through the air and with stormwater runoff, leading to environmental pollution and human health concerns. In the present study, we collected airborne PM at varying distances (5, 15 and 30 m) along US Highway 278 in Oxford, Mississippi, USA, for ten consecutive days using Sigma-2 passive samplers. Particles (~ 1-80 µm) were passively collected directly into small (60 mL) wide-mouth separatory funnels placed inside the samplers. Particles were subsequently subjected to solvent extraction, and extracts were analyzed for TWP compounds by high resolution orbitrap mass spectrometry. This pilot study was focused solely on qualitative analyses to determine whether TWP compounds were present in this fraction of airborne PM. The abundance of airborne TWPs increased with proximity to the road with deposition rates (TWPs cm-2 day-1) of 23, 47, and 63 at 30 m, 15 m, and 5 m from the highway, respectively. Two common TWP compounds (6PPD-Q and 4-ADPA) were detected in all samples, except the field blank, at levels above their limits of detection, estimated at 2.90 and 1.14 ng L-1, respectively. Overall, this work suggests airborne TWPs may be a potential inhalation hazard, particularly for individuals and wildlife who spend extended periods outdoors along busy roadways. Research on the bioavailability of TWP compounds from inhaled TWPs is needed to address exposure risk.

The application of acute oral toxicity computational models in dangerous goods classification.

Acute oral toxicity (AOT) data inform the acute toxicity potential of a compound and guides occupational safety and transportation practices. AOT data enable the categorization of a chemical into the appropriate AOT Globally Harmonized System (GHS) category based on the severity of the hazard. AOT data are also utilized to identify compounds that are Dangerous Goods (DGs) and subsequent transportation guidance for shipping of these hazardous materials. Proper identification of DGs is challenging for novel compounds that lack data. It is not feasible to err on the side of caution for all compounds lacking AOT data and to designate them as DGs, as shipping a compound as a DG has cost, resource, and time implications. With the wealth of available historical AOT data, AOT testing approaches are evolving, and in silico AOT models are emerging as tools that can be utilized with confidence to assess the acute toxicity potential of de novo molecules. Such approaches align with the 3R principles, offering a reduction or even replacement of traditional in vivo testing methods and can also be leveraged for product stewardship purposes. Utilizing proprietary historical in vivo AOT data for 210 pharmaceutical compounds (PCs), we evaluated the performance of two established in silico AOT programs: the Leadscope AOT Model Suite and the Collaborative Acute Toxicity Modeling Suite. These models accurately identified 94% and 97% compounds that were not DGs (GHS categories 4, 5, and not classified (NC)) suggesting that the models are fit-for-purpose in identifying PCs with low acute oral toxicity potential (LD50 >300 mg/kg). Utilization of these models to identify compounds that are not DGs can enable them to be de-prioritized for in vivo testing. This manuscript provides a detailed evaluation and assessment of the two models and recommends the most suitable applications of such models.

Exposure to Hazardous Substances and Their Health Effects Among Residents Living Near Three Industrial Waste Incinerators in Korea.

BACKGROUND: This study aimed to evaluate exposure to various hazardous substances emitted by incineration facilities and their likely effect on the health for residents of Bugi-myeon, Cheongju, Korea, which has three incineration facilities. METHODS: Heavy metals, polycyclic aromatic hydrocarbons (PAHs), and dioxin concentrations in the air and soil of exposed and control areas were measured. Moreover, the exposure levels to harmful substances and its effects on health were investigated in 1,124 exposed and 232 control adults. RESULTS: PAHs and dioxin concentrations in the air in the exposed area were significantly higher than in the control area. Urinary cadmium and PAHs metabolite concentrations were significantly higher in the exposed group than in the control group. The exposure group also had a higher prevalence of depression and self-reported allergic symptoms than the control group. CONCLUSION: The possibility of residents in Bugi-myeon being exposed to hazardous substances at incineration facilities cannot be ruled out. To prevent them from further exposure to hazardous substances, it is necessary to prohibit the expansion of additional incineration facilities in this area and to implement continuous monitoring projects for residents.

A vision for safer food contact materials: Public health concerns as drivers for improved testing.

Food contact materials (FCMs) and food contact articles are ubiquitous in today's globalized food system. Chemicals migrate from FCMs into foodstuffs, so called food contact chemicals (FCCs), but current regulatory requirements do not sufficiently protect public health from hazardous FCCs because only individual substances used to make FCMs are tested and mostly only for genotoxicity while endocrine disruption and other hazard properties are disregarded. Indeed, FCMs are a known source of a wide range of hazardous chemicals, and they likely contribute to highly prevalent non-communicable diseases. FCMs can also include non-intentionally added substances (NIAS), which often are unknown and therefore not subject to risk assessment. To address these important shortcomings, we outline how the safety of FCMs may be improved by (1) testing the overall migrate, including (unknown) NIAS, of finished food contact articles, and (2) expanding toxicological testing beyond genotoxicity to multiple endpoints associated with non-communicable diseases relevant to human health. To identify mechanistic endpoints for testing, we group chronic health outcomes associated with chemical exposure into Six Clusters of Disease (SCOD) and we propose that finished food contact articles should be tested for their impacts on these SCOD. Research should focus on developing robust, relevant, and sensitive in-vitro assays based on mechanistic information linked to the SCOD, e.g., through Adverse Outcome Pathways (AOPs) or Key Characteristics of Toxicants. Implementing this vision will improve prevention of chronic diseases that are associated with hazardous chemical exposures, including from FCMs.

Innovate beyond PFAS.

New proposed legislation on "forever" chemicals is under consideration in Europe and the United States, where per- and polyfluoroalkyl substances (PFAS) are a hot topic for regulators and lawmakers. On both sides of the Atlantic, regulation of widely used PFAS has been complex and evolving. Their presence in hundreds of different products-from nonstick cookware to food packaging to firefighting foam-and their persistence in food, drinking water, and the environment have resulted in a pollution problem of unprecedented scale. Recently, for example, it was reported that 45% of the tap water in the United States contains at least one type of PFAS. Because these compounds are so chemically stable that they do not degrade in the environment (including in the human body), PFAS seriously challenge long-established ideas of how chemicals can be used, assessed, and regulated, and it remains to be seen whether the new regulations will solve this problem.

Toxicity of cadmium on dynamic human gut microbiome cultures and the protective effect of cadmium-tolerant bacteria autochthonous to the gut.

Cadmium (Cd) is a heavy metal toxic to the gut microbiome. In this study, we cultivated two human gut microbiomes (A and B) in bioreactors with Cd at 0 and 20 ppm for 7 days to investigate effects of Cd on the gut microbiome and to isolate Cd-tolerant bacteria autochthonous to the gut. Cd showed profound toxicity, abolishing butyrate production, depleting microbes in microbiome B, and simplifying microbiome A to a small Cd-tolerant community after 2 d of incubation. When spiked into the Cd-sensitive microbiome B, the Cd-tolerant community from microbiome A and isolates from that community worked synergistically with microbiome B to enhance butyrate production and maintained this synergism at Cd concentrations up to 5 ppm. Bacteria isolated from this Cd-tolerant community included Enterococcus faecium, Enterobacter cloacae, Lactococcus lactis, and Lactobacillus taiwanensis species. This work demonstrates a straightforward method for identifying Cd-tolerant bacteria autochthonous to the human gut that synergize with the microbiome to protect against Cd-related loss of butyrate production.

Does REACH provide sufficient information to regulate substances toxic to reproduction?

Hazard classification and risk assessment of substances, is essential to protect workers and consumers from hazardous substances including reproductive toxicants. The ability to classify substances for reproductive toxicity under the current REACH information requirements has been assessed. For low tonnage substances (<10 ton per annum (tpa)) information for classification is insufficient. When only a reproductive screening study is available (10-100 tpa), substances are mostly not classified in Category 1B as developmental and non-potent fertility effects may be missed. The information requirements could be improved by automatic triggering of follow-up studies in case of a Category 2 classification based on a screening study. Additionally, a study could be added to the information requirements for substances produced at 1-10 tpa. Performing a risk assessment is often problematic due to the limited study requirements at low tonnage levels. Only for substances produced at more than 100 tpa, there is a high likelihood to detect reproductive effects and perform accurate risk assessment provided that the extended-one-generation-reproductive-toxicity-study and/or extra cohorts are triggered where required. Regardless of the tonnage level, no specific studies on lactation are required. With this paper we intend to contribute to the discussion on the information requirements for reproductive toxicity in view of the REACH revision.

Genetic association between the APOE ε4 allele, toxicant exposures and Gulf war illness diagnosis.

INTRODUCTION: Exposure to nerve agents, pyridostigmine bromide (PB), pesticides, and oil-well fires during the 1991 Gulf War (GW) are major contributors to the etiology of Gulf War Illness (GWI). Since the apolipoprotein E (APOE) ε4 allele is associated with the risk of cognitive decline with age, particularly in the presence of environmental exposures, and cognitive impairment is one of the most common symptoms experienced by veterans with GWI, we examined whether the ε4 allele was associated with GWI. METHODS: Using a case-control design, we obtained data on APOE genotypes, demographics, and self-reported GW exposures and symptoms that were deposited in the Boston Biorepository and Integrative Network (BBRAIN) for veterans diagnosed with GWI (n = 220) and healthy GW control veterans (n = 131). Diagnosis of GWI was performed using the Kansas and/or Center for Disease Control (CDC) criteria. RESULTS: Age- and sex-adjusted analyses showed a significantly higher odds ratio for meeting the GWI case criteria in the presence of the ε4 allele (Odds ratio [OR] = 1.84, 95% confidence interval [CI = 1.07-3.15], p ≤ 0.05) and with two copies of the ε4 allele (OR = 1.99, 95% CI [1.23-3.21], p ≤ 0.01). Combined exposure to pesticides and PB pills (OR = 4.10 [2.12-7.91], p ≤ 0.05) as well as chemical alarms and PB pills (OR = 3.30 [1.56-6.97] p ≤ 0.05) during the war were also associated with a higher odds ratio for meeting GWI case criteria. There was also an interaction between the ε4 allele and exposure to oil well fires (OR = 2.46, 95% CI [1.07-5.62], p ≤ 0.05) among those who met the GWI case criteria. CONCLUSION: These findings suggest that the presence of the ε4 allele was associated with meeting the GWI case criteria. Gulf War veterans who reported exposure to oil well fires and have an ε4 allele were more likely to meet GWI case criteria. Long-term surveillance of veterans with GWI, particularly those with oil well fire exposure, is required to better assess the future risk of cognitive decline among this vulnerable population.