Toxicology Of Chemical Mixtures Research Articles

Unravelling the Potential Cytotoxic Effects of Metal Oxide Nanoparticles and Metal(Loid) Mixtures on A549 Human Cell Line.

Humans are typically exposed to environmental contaminants’ mixtures that result in different toxicity than exposure to the individual counterparts. Yet, the toxicology of chemical mixtures has been overlooked. This work aims at assessing and comparing viability and cell cycle of A549 cells after exposure to single and binary mixtures of: titanium dioxide nanoparticles (TiO2NP) 0.75–75 mg/L; cerium oxide nanoparticles (CeO2NP) 0.75–10 μg/L; arsenic (As) 0.75–2.5 mg/L; and mercury (Hg) 5–100 mg/L. Viability was assessed through water-soluble tetrazolium (WST-1) and thiazolyl blue tetrazolium bromide (MTT) (24 h exposure) and clonogenic (seven-day exposure) assays. Cell cycle alterations were explored by flow cytometry. Viability was affected in a dose- and time-dependent manner. Prolonged exposure caused inhibition of cell proliferation even at low concentrations. Cell-cycle progression was affected by TiO2NP 75 mg/L, and As 0.75 and 2.5 μg/L, increasing the cell proportion at G0/G1 phase. Combined exposure of TiO2NP or CeO2NP mitigated As adverse effects, increasing the cell surviving factor, but cell cycle alterations were still observed. Only CeO2NP co-exposure reduced Hg toxicity, translated in a decrease of cells in Sub-G1. Toxicity was diminished for both NPs co-exposure compared to its toxicity alone, but a marked toxicity for the highest concentrations was observed for longer exposures. These findings prove that joint toxicity of contaminants must not be disregarded.

Basic principles of toxicological evaluation of mixtures

The vast majority of toxicity studies and risk assessments deals with single chemicals. In reality, however, humans are exposed simultaneously to a large number of chemicals, indicating that exposure assessment and safety evaluation should focus on mixtures rather than on single chemicals. This review on the toxicology of chemical mixtures covers widely varying issues, ranging from the basic concepts and terminology of joint action and interaction of chemicals to different designs for studying the toxicity of mixtures. Literature data indicate that exposure to mixtures which contain low levels of chemicals, levels lower than their no observed adverse effect levels (NOAELs), can exert toxic effects whether their components have similar modes of action or not. Different approaches and examples of studies on priority real-life mixtures, strategy and guidelines for mixture risk assessment as well as challenges ahead are also presented.

Effects of Developmental Exposure to Mixtures of Environmental Contaminants on the Hepatic Metabolism of Estradiol-17β in Immature Female Sprague Dawley Rats

Exposure to environmental contaminants induces the activation of cytochrome P450s (CYP) which lead to the hydroxylation of contaminants and endogenous hormones such as estrogens. The hydroxylation of estrogens forms catecholestrogens (CEs), one of them being the mutagenic 4-hydroxyestradiol-17β (4-OH-E2). Catecholestrogens are transformed by catechol-o-methyltransferases (COMTs) into nonreactive methoxyestrogens. To investigate the hepatic metabolism of estradiol-17β in female offspring at postnatal day (PND) 21, pregnant rats were dosed daily from gestation day 1 until PND 21 with 2 dose levels of organochlorine pesticides (OCPs; 0.019 or 1.9 mg/kg per d), methylmercury (MeHg; 0.02 or 2 mg/kg per d), polychlorinated biphenyls (PCBs; 0.011 or 1.1 mg/kg per d), or a mixture (M; 0.05 or 5 mg/kg per d) including all 3 groups of chemicals. Concentrations of organochlorines in the mixture M were based on their proportions in serum of the Canadian Arctic population. The messenger RNA (mRNA) expressions of CYP and COMT were analyzed by quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). High-performance thin layer chromatography and phosphor imaging were used to measure the transformation of (14)C substrates into estrogen metabolites. The low-dose treatments or the MeHg groups had no effect. The high-dose OCP, PCB, and M group increased the production of 2-OH-E2 and 6α-OH-E2, while only the PCB and M groups increased the 2-OH-CE/methoxyestrogen ratio. In all groups, the cytosolic COMT activity exceeded the microsomal production rate of 4-OH-E2. Although the M treatment included the PCB and OCP mixtures, it did not modify the estrogen metabolism more than did the PCB mixture alone. This endocrine disruption information contributes to our understanding of chemical interactions in the toxicology of chemical mixtures.

Introduction: Endocrine Disruptors—Exposure Assessment, Novel End Points, and Low-Dose and Mixture Effects

With the aim of discussing new research findings about chemicals able to interfere with the endocrine system, so-called endocrine disruptors, an international workshop was held in Prague, Czech Republic, 10–12 May 2005. The workshop was organized jointly by the EDEN project (Endocrine Disrupters: Exploring Novel Endpoints, Exposure, Low-Dose and Mixture-Effects in Humans, Aquatic Wildlife and Laboratory Animals; http://www.edenresearch.info) and the FIRE project (Risk Assessment of Brominated Flame Retardants as Suspected Endocrine Disrupters for Human and Wildlife Health; http://www.rivm.nl/fire), both large-scale consortia funded by the European Union (EU). The meeting was attended by more than 170 scientists from academia, industry, government agencies, and other organizations. EDEN and FIRE were part of the CREDO cluster (Cluster for Research on Endocrine Disruption), an umbrella organization of more than 60 EU-funded laboratories working on endocrine disruptors. Together with the COMPRENDO (Comparative Research on Endocrine Disruption) and the EURISKED projects (Multi-organic Risk Assessment of Selected Endocrine Disruptors; http://www.eurisked.org), EDEN and FIRE formed the core of the cluster. CREDO served as a platform for cooperation and exchange between its constituent projects. With the completion of almost all its member projects, CREDO has recently ceased to exist, but information about the cluster is still available on online (http://www.credocluster.info). The Prague workshop was the last in a series of meetings organized under the auspices of the CREDO cluster. The themes of previous workshops were risk assessment and the ecologic relevance of chemically induced endocrine disruption in wildlife (Jobling and Tyler 2006). This monograph contains papers presented at the Prague workshop and follows the session structure of the meeting, focusing on four main topics: “Indicators of Human and Wildlife Exposure to EDCs ” reports on new findings relating human exposure to endocrine disruptors in Europe to reproductive disorders, including cryptorchidism, hypospadias, and semen quality. New information about exposure trends for brominated flame retardants in freshwater and marine ecosystems is also given. “Novel models, end points, and biomarkers” summarizes research on the effects of endocrine disruptors on steroid-metabolizing enzymes and gives new data on alkylphenols as endocrine disruptors. Additional articles focus on the consequences of prolonged phytoestrogen exposure for reproductive organs and the outcomes of in utero exposure to endocrine disruptors for testicular dysgenesis, prostate development, and the thyroid hormone axis. “Low-dose effects of endocrine disruptors” gives a synthesis of recent observations in in vitro systems and highlights the importance of statistical power considerations in resolving the low-dose issue. “Mixture effects of endocrine disruptors and their assessment” contains reviews of combination effects, assesses the implications of low-level exposure to multiple chemicals and describes the joint effect of endocrine disruptors in fish and in a developmental toxicity rat model. Taken together, the articles presented in this monograph capture a great deal of the lively debates that took place during the Prague workshop and within the CREDO cluster as a whole. The workshop made considerable progress toward answering key questions that were unresolved a few years ago. It has also stimulated a consensus statement among scientists actively engaged in research in this field, the “Prague Declaration on Endocrine Disruption.” The Prague Declaration, also published as part of this monograph (available as Supplemental Material online at http://www.ehponline.org/docs/2007/10517/suppl.pdf), summarizes issues upon which the majority of scientists can agree, outlines research priorities for the next decade, and highlights steps that can be taken today to prevent health risks to humans and wildlife. It has been signed by more than 200 scientists from all over the world. All contributors to the workshop are thanked cordially. Without their enthusiasm and hard work the meeting and this monograph would not have happened.

A Unifying Concept for Assessing Toxicological Interactions: Changes in Slope

Robust statistical methods are important to the evaluation of toxicological interactions (i.e., departures from additivity) among chemicals in a mixture. However, different concepts of joint toxic action as applied to the statistical analysis of chemical mixture toxicology data or as used in environmental risk assessment often appear to conflict with one another. A unifying approach for application of statistical methodology in chemical mixture toxicology research is based on consideration of change(s) in slope. If the slope of the dose-response curve of one chemical does not change in the presence of other chemicals, then there is no interaction between the first chemical and the others. Conversely, if the rate of change in the response with respect to dose of the first chemical changes in the presence of the other chemicals, then an interaction is said to exist. This concept of zero interaction is equivalent to the usual approach taken in additivity models in the statistical literature. In these additivity models, the rate of change in the response as a function of the i(th) chemical does not change in the presence of other chemicals in a mixture. It is important to note that Berenbaum's (1985, J. Theor. Biol. 114, 413-431) general and fundamental definition of additivity does not require the chemicals in the mixture to have a common toxic mode of action nor to have similarly shaped dose response curves. We show an algebraic equivalence between these statistical additivity models and the definition of additivity given by Berenbaum.

Toxicology of chemical mixtures: a challenging quest along empirical sciences

This paper describes the “quest” of our institute trying to assess the toxicology of chemical mixtures. In this overview, we will discuss some critical developments in hazard identification and risk assessment of chemical mixtures during these past 15 years. We will stand still at empirical and mechanistic modeling. “Empirical” means that only information on doses or concentrations and effects is available in addition to an often empirically selected quantitative dose–response relationship. Empirical models have played a dominant role in the last decade to identify health and safety characteristics of chemical mixtures. Many of these models are based on the work of pioneers in mixture toxicology who defined three basic types of action for combinations of chemicals: simple similar action, simple dissimilar action and interaction. Nowadays, empirical models are mainly based on response-surface analysis and make use of advanced statistical designs. However, possible interactions between components in a mixture can also be given in terms of mechanistic models. In terms of “mechanistic” (or biological) understanding, interactions between compounds may occur in the kinetic phase (processes of uptake, distribution, metabolism and excretion) or in the dynamic phase (effects of chemicals on the receptor, cellular target or organ). A biological phenomenon such as competitive agonism as described for mixtures of drugs (biotransformation enzymes) or sensory irritants (nerve receptors) can accurately predict the effect of any of these mixtures. Thus, far mechanistic and empirical analyses of interactions are usually unrelated. It is one of the future challenges for mixtures research to combine information from both approaches. Also, our current biology-based models have their limitations, since they cannot integrate every relevant biological mechanism. In this respect, mechanistic modeling of mixtures may benefit from the developments coming from the arena of molecular biology (toxicogenomics) which offers an in-depth analysis of several involved enzymatic pathways in parallel through the use of a systems biology approach. This was illustrated with mixtures of food additives known to affect the liver. Key to further maturation of mixture toxicology is collaboration of experimental toxicologists, biomathematicians, biologists, pharmacologists, model developers, molecular biologists and bioinformaticians to ensure parallel and coordinated research in this challenging area of toxicology. For this reason, the next sequel will be even more challenging and exciting to that first 15 years of empirical testing.

Chemical mixture toxicology: from descriptive to mechanistic, and going on to in silico toxicology

Because of the pioneering vision of certain leaders in the biomedical field, the last two decades witnessed rapid advances in the area of chemical mixture toxicology. Earlier studies utilized conventional toxicology protocol and methods, and they were mainly descriptive in nature. Two good examples might be the parallel series of studies conducted by the U.S. National Toxicology Program and TNO in The Netherlands, respectively. As a natural course of progression, more and more sophistication was incorporated into the toxicology studies of chemical mixtures. Thus, at least the following seven areas of scientific achievements in chemical mixture toxicology are evident in the literature: (a) the application of better and more robust statistical methods; (b) the exploration and incorporation of mechanistic bases for toxicological interactions; (c) the application of physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) modeling; (d) the studies on more complex chemical mixtures; (e) the use of science-based risk assessment approaches; (f) the utilization of functional genomics; and (g) the application of technology. Examples are given for the discussion of each of these areas. Two important concepts emerged from these studies and they are: (1) dose-dependent toxicologic interactions; and (2) “interaction thresholds”. Looking into the future, one of the most challenging areas in chemical mixture research is finding the answer to the question “when one tries to characterize the health effects of chemical mixtures, how does one deal with the infinite number of combination of chemicals, and other possible stressors?” Undoubtedly, there will be many answers from different groups of researchers. Our answer, however, is first to focus on the finite (biological processes) rather than the infinite (combinations of chemical mixtures and multiple stressors). The idea is that once we know a normal biological process(es), all stimuli and insults from external stressors are merely perturbations of the normal biological process(es). The next step is to “capture” the biological process(es) by integrating the recent advances in computational technology and modern biology. Here, the computer-assisted Reaction Network Modeling, linked with PBPK modeling, offers a ray of hope to dealing with the complex biological systems.

Commentary: some critical issues and concerns related to research advances on the toxicology of chemical mixtures

Commentary: some critical issues and concerns related to research advances on the toxicology of chemical mixtures

Commentary: Some Critical Issues and Concerns Related to Research Advances on the Toxicology of Chemical Mixtures

This paper addresses some of the issues and concerns on research advances on the toxicology of chemical mixtures. Emphases will be selectively given to the following questions and answers: Can mechanistic studies be conducted on chemical mixtures? The fact that any studies, including mechanistic studies, of single chemicals are really the study of the parent chemical plus its metabolites underscores the relevance of mechanistic studies on chemical mixtures. Can predictions be made on the health effects of chemical mixtures? Some successes are already evident in the literature on simpler chemical mixtures. For more complex mixtures, it is possible and we propose an approach here. What can we learn from other disciplines (the importance of interdisciplinary collaboration)? Two aspects, the knowledge and methodologies available in clinical pharmacology and the latest advances in structure-oriented lumping in chemical engineering, are discussed in detail. Unrepeatable results: The possibility of magnification of biologic variability because of low-level exposures to chemical mixtures is suggested with special reference to some known examples, including the controversial study on synergistic interactions of endocrine disruptors. Is the driving force for scientific investigations on chemical mixtures the legislative and regulatory atmosphere? Two laws with chemical mixtures specifically in the language are quoted and discussed. Their implications regarding research funding and activities are described. What are the pitfalls of applying for research funding on investigating chemical mixtures? The dilemma at least one investigator faces in pursuing research funding is elaborated. The questions and issues listed above are not all inclusive, but they represent some of the aspects that need to be brought into the open in the scientific community for discussion and/or debate. Thus, the primary objective of this paper is to provide some momentum for the beginning of a fruitful and stimulating discussion. Environ Health Perspect 106(Suppl 4):1059-1063 (1998). http://ehpnet1.niehs.nih.gov/docs/1998/Suppl-4/1059-1063yang/abstract.html

Some critical issues and concerns related to research advances on toxicology of chemical mixtures.

This paper addresses some of the issues and concerns on research advances on the toxicology of chemical mixtures. Emphases will be selectively given to the following questions and answers: Can mechanistic studies be conducted on chemical mixtures? The fact that any studies, including mechanistic studies, of single chemicals are really the study of the parent chemical plus its metabolites underscores the relevance of mechanistic studies on chemical mixtures. Can predictions be made on the health effects of chemical mixtures? Some successes are already evident in the literature on simpler chemical mixtures. For more complex mixtures, it is possible and we propose an approach here. What can we learn from other disciplines (the importance of interdisciplinary collaboration)? Two aspects, the knowledge and methodologies available in clinical pharmacology and the latest advances in structure-oriented lumping in chemical engineering, are discussed in detail. Unrepeatable results: The possibility of magnification of biologic variability because of low-level exposures to chemical mixtures is suggested with special reference to some known examples, including the controversial study on synergistic interactions of endocrine disruptors. Is the driving force for scientific investigations on chemical mixtures the legislative and regulatory atmosphere? Two laws with chemical mixtures specifically in the language are quoted and discussed. Their implications regarding research funding and activities are described. What are the pitfalls of applying for research funding on investigating chemical mixtures? The dilemma at least one investigator faces in pursuing research funding is elaborated. The questions and issues listed above are not all inclusive, but they represent some of the aspects that need to be brought into the open in the scientific community for discussion and/or debate. Thus, the primary objective of this paper is to provide some momentum for the beginning of a fruitful and stimulating discussion.

Integrated Approaches for the Analysis of Toxicologic Interactions of Chemical Mixtures

Although an overwhelmingly large portion of the resources in toxicologic research is devoted to single chemical studies, the toxicology of chemical mixtures, not single chemicals, is the real issue regarding health effects of environmental and/or occupational exposure to chemicals. The relative lack of activities in the area of toxicology of chemical mixtures does not suggest ignorance of the importance of the issue by the toxicology community. Instead, it is a reflection of the difficulty, complexity, and controversy surrounding this area of research.Until recently, much of the literature on the toxicology of chemical mixtures has been either very focused on certain specific interaction studies or slanted toward broad-based, relatively vague theoretical deliberation. The typical interaction study involved binary mixtures at relatively high dose levels with acute toxicities as endpoints. Although the theoretical papers have been valuable contributions, little is available on actual, practical experimental approaches toward a systematic solution of this immensely complex area of research.We present here a broad discussion on the important issues of the toxicology of chemical mixtures. First, we provide some background information with respect to the problem and significance of toxicology of chemical mixtures in relation to some of the real life issues. Second, we review and compare the existing experimental approaches relevant to toxicologic interactions of chemical mixtures. Third, we propose three integrated approaches that involve the combination of physiologically based pharmacokinetic/pharmacodynamic (PBPK/PD) modeling with: (1) Monte Carlo simulation, (2) median effect principle (MEP), and (3) response surface methodology (RSM). These modeling approaches, coupled with very focused mechanistically based toxicology studies, could be the basis for solving the problems of toxicology and risk assessment of chemical mixtures.

Some current approaches for studying combination toxicology in chemical mixtures

Using conventional toxicology methodologies and at the mode and rate of studying chemicals in the last 25 yr, it is doubtful that society will ever have adequate toxicology information on the majority of the chemicals we used now or in the future. Considering further the issue of health effects of chemical mixture exposure (i.e. real-world issues), the problem of not being able to obtain adequate toxicology information is amplified. From a different perspective, concerns over animal rights have raised the consciousness of many biomedical researchers regarding animal experimentation. As many as 17–100 million animals are estimated to be killed for biomedical research in the US alone each year; therefore, minimizing animal usage judiciously in toxicological research should be in the mind of every responsible toxicologist. From these considerations, it is apparent that new, alternative, less animal-intensive, shorter-term and less expensive toxicology methods must be developed if there is to be a reasonable chance to deal with the hundreds of thousands of chemicals, as well as the near-infinite number of chemical mixtures, in the environment. Some of the recent advances indeed are heading towards that direction. In this article, a number of approaches for research work on combination toxicology of chemical mixtures are given; the examples are selected based on one or more of the following criteria: (1) minimizing animal usage; (2) shortening experimental durations; (3) studying environmentally realistic concentrations; (4) utilizing statistical/mathematical modelling; (5) advancing efficient experimental designs and (6) developing predictive toxicology.

Toxicology of chemical mixtures: challenges for today and the future

A major challenge for the toxicologist involved in safety evaluation of chemical mixtures is to test the hypothesis that as a rule exposure to mixtures of chemicals at (low) non-toxic doses of the individual chemicals is of no health concern. A series of repeated dose studies in rats with defined mixtures of chemicals with the same or different target organs revealed that exposure to a combination of chemicals compared with exposure to the individual compounds did not constitute an evidently increased hazard, provided each chemical was administered at a level similar to, or slightly lower than, its own 'No-Observed-Adverse-Effect-Lever. The results of subacute oral toxicity studies in rats with denned mixtures of nephrotoxicants with similar mode of action underlined the applicability of the additivity assumption for a mixture of chemicals with simple similar action. Safety evaluation of complex chemical mixtures is a challenge that can be tackled as follows: first, identify the (e.g. ten) most risky chemicals in the mixture, and, second, assess the hazard and the potential health risk of the mixture of the most risky chemicals, using procedures developed for defined mixtures. To identify interactions between individual compounds, a most promising testing strategy appeared to be a statistical approach using a fractional two-level factorial design. A challenge for today and the future is to gradually substitute mixture-oriented (real life-oriented) standard setting for (unrealistic) single chemical-oriented standard setting.

Use of tissue slices in chemical mixture toxicology and interspecies investigations

Precision-cut tissue slices have proven to be a useful in vitro system for biotransformation and toxicity studies. Since tissue slices can be readily prepared from a variety of tissues and species, they can easily be used for interspecies investigations and comparisons. Furthermore, slices can be readily prepared from human tissue, thus comparisons (extrapolation) can be made between laboratory animals and humans. Slices can also be used to examine the toxic interactions of chemicals in vitro. It is important to use the correct experimental design to demonstrate toxic interactions and to assure that the tissue slices are properly exposed to the chemicals. Overall, tissue slices offer a valid in vitro system for performing species comparisons and chemical-chemical interaction studies.

The use of physiologically-based pharmacokinetic/ pharmacodynamic dosimetry models for chemical mixtures

Human exposure to chemicals is rarely, if ever, limited to a single chemical. Therefore, it is essential that we consider multiple chemical effects and interactions in our risk assessment process. However, with the almost infinitely large number of chemical mixtures in the environment, systematic studies of the toxicology of these chemical mixtures with conventional methodologies and approaches are impossible because of the immense resources and unrealistically long durations required. Thus, the development of ‘Predictive and Alternative Toxicology’ is imperative. At Colorado State University (CSU), our research effort is entirely devoted to this challenge. In order to have a reasonable chance to deal with the complex issue of toxicology of chemical mixtures, we believe that the following concepts must be considered: (1) the utilization of computer; (2) the exploitation of mathematical/statistical methodologies; (3) developing very focused, mechanistically based, and short-term toxicology studies; (4) coupling computer/mathematical modeling with mechanistically-based toxicology. Our strategy is therefore the utilization of physiologically-based pharmacokinetic/pharmacodynamic (PBPK/PD) modeling, coupled with very focused, modeldirected toxicology experiments as well as other statistical/mathematical methodologies such as Monte Carlo simulation, isobolographic analysis, and response surface methodology. We believe that ‘Predictive and Alternative Toxicology’ in terms of tissue dosimetry at the pharmacokinetic and pharmacodynamic levels is achievable with simple and complex but chemically defined mixtures. In this presentation, we describe two ongoing research projects as an illustration of our ‘Bottom-Up’ and ‘Top-Down’ approaches for handling the chemical mixtures: (1) PBPK/PD modeling of toxicologie interactions between Kepone and carbon tetrachloride (CC1 4) and the coupling of Monte Carlo simulation for the prediction of acute toxicity; (2) the conceptual development of PBPK/PD modeling for a more complex chemical mixture of seven groundwater contaminants from hazardous waste sites and the consideration of subfractionation of this chemical mixture.

Physiologically based pharmacokinetic/pharmacodynamic modeling of chemical mixtures and possible applications in risk assessment

Human exposure to chemicals, be it environmental or occupational, is rarely, if ever, limited to a single chemical. Therefore, it is essential that we consider multiple chemical effects and interactions in our risk assessment process. However, with the almost infinitely large number of chemical mixtures in the environment, systematic studies of the toxicology of these chemical mixtures with conventional methodologies and approaches are impossible because of the immense resources and unrealistically long durations required. Thus, the development of predictive and alternative toxicology method is imperative. In order to have a reasonable chance to deal with the complex issue of toxicology of chemical mixtures, we believe that the following concepts must be considered: (1) the exploitation of recent advances in computational technology; (2) the utilization of mathematical/statistical modeling; (3) coupling computer modeling with very focused, mechanistically based, and short-term toxicology studies. Our approach is. therefore, the utilization of physiologically based pharmacokinetic/pharmacodynamic (PB-PK/PD) modeling, coupled with very focused, model-directed toxicology experiments as well as other statistical/mathematical modeling such as isobolo- graphic analysis and response surface methodology. Tissue dosimetry at the pharmacokinetic and pharmacodynamic levels is achievable with simple and complex, but chemically defined, mixtures. Our long-term goal is to formulate innovative risk assessment methodologies for chemical mixtures. In this presentation, one of our specific research projects is described: PB-PK/PD modeling of toxicologic interactions between Kepone and carbon tetrachloride (CCl 4) and the coupling of Monte Carlo simulation for the prediction of acute toxicity.

Book Reviews : Toxicology of Chemical Mixtures Edited by Raymond S.H. Yang Academic Press, London, 1994, pp. 720, £115.00

Book Reviews : Toxicology of Chemical Mixtures Edited by Raymond S.H. Yang Academic Press, London, 1994, pp. 720, £115.00

Enhancement of myelotoxicity induced by repeated irradiation in mice exposed to a mixture of groundwater contaminants.

As part of a program on the toxicology of chemical mixtures at the National Institute of Environmental Health Sciences/National Toxicology Program (NIEHS/NTP), hematopoietic functions were studied in female B6C3F1 mice treated with 0, 1%, and 5% of a chemical mixture stock of 25 groundwater contaminants in drinking water for 31.5 weeks. The toxicologic interaction between continuous exposure to groundwater contaminants and stress induced by multiple irradiation on hematopoiesis was investigated. For those mice receiving both the chemical mixture and irradiation, the exposure to the former was continuous throughout the 31.5-week experimental period, whereas whole body irradiations (4 times at 200 rads/each) were carried out at 7-week intervals with the first one at 3.5 weeks. Myelotoxicity assessment was made by determining the number of granulocyte-macrophage progenitor cells (CFU-GM) 1 week after each irradiation and also at 6 weeks following irradiation as a measure of recovery from stress. Non-irradiated mice treated with 5% chemical mixture solution showed suppression of CFU-GM after 15.5 weeks and became progressively more affected (only 70% of controls by 31.5 weeks of treatment). The population of CFU-GM in mice treated with 5% chemical mixture for 4.5 weeks plus irradiation (1 week after first irradiation) was only 22% of the non-irradiated vehicle control group. This combined (i.e., chemical mixture plus irradiation) suppression of CFU-GM intensified after repeated irradiation until the number of CFU-GM was only 10.7% following the fourth irradiation at 25.5 weeks. Thus, irradiation caused a significant reduction in CFU-GMs in all mice but the effects were more pronounced in mice treated with a chemical mixture.(ABSTRACT TRUNCATED AT 250 WORDS)

Strategy for studying health effects of pesticides/fertilizer mixtures in groundwater.

This paper is a "historical account" of the entire process leading to the final implementation of a toxicology research program on the health-effect studies of pesticide and fertilizer contaminants in groundwater conducted at the National Institute of Environmental Health Sciences/National Toxicology Program (NIEHS/NTP). Starting with the perception of a potential problem area, the program evolved through several stages, including planning and information-gathering, deliberation on experimental designs, formulation of a draft protocol, peer-review and public comments on the protocol, finalization of the protocol, announcement of open bidding to the NTP master agreement laboratories (i.e., a group of laboratories that has been previously evaluated and approved to conduct NTP toxicology contract work), implementation of chemistry developmental work, selection of toxicology laboratories and awarding of contracts, final implementation of toxicology studies, and cultivation of special collaborative efforts among NIEHS in-house scientists as well as colleagues at the neighbouring laboratories in Research Triangle Park. To put things in perspective, the author's present approach, a coupling of physiologically based pharmacokinetics/pharmacodynamics and computer modeling with the toxicology of chemical mixtures at Colorado State University, is used to address the issues of toxicology and risk assessment of chemical mixtures with respect to rationale and reality.

Toxicology of chemical mixtures: experimental approaches, underlying concepts, and some results

The toxicology of chemical mixtures will be the toxicology of the 1990s and beyond. While this branch of toxicology most closely reflects the actual human exposure situation, there is yet no standard protocol or consensus methodology for investigating the toxicology of mixtures. Thus, in this emerging science, experimentation is required just to develop a broadly applicable evaluation system. Several examples are discussed to illustrate the different experimental designs and the concepts behind each. These include the health effects studies of Love Canal soil samples, the Lake Ontario Coho salmon, the water samples repurified from secondary sewage in the city of Denver Potable Water Reuse Demonstration Plant, and the National Toxicology Program (NTP) effort on a mixture of 25 frequently detected groundwater contaminants derived from hazardous waste disposal sites. In the last instance, an extensive research program has been ongoing for the last 2 years at the NTP, encompassing general toxicology, immunotoxicology, developmental and reproductive toxicology, biochemical toxicology, myelotoxicology, genetic toxicology, neurobehavioral toxicology, and hepato- and renal toxicology. kwToxicology; Chemical mixtures; Experimental approach; Hazardous waste sites; Contaminated Great Lake salmon; Water reuse; Water recycle; Groundwater contaminants