Michael D. Waters

IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans

The purpose of these guidelines is to provide concise guidance on the planning, performing and interpretation of studies to monitor groups or individuals exposed to genotoxic agents. Most human carcinogens are genotoxic but not all genotoxic agents have been shown to be carcinogenic in humans. Although the main interest in these studies is due to the association of genotoxicity with carcinogenicity, there is also an inherent interest in monitoring human genotoxicity independently of cancer as an endpoint. The most often studied genotoxicity endpoints have been selected for inclusion in this document and they are structural and numerical chromosomal aberrations assessed using cytogenetic methods (classical chromosomal aberration analysis (CA), fluorescence in situ hybridisation (FISH), micronuclei (MN)); DNA damage (adducts, strand breaks, crosslinking, alkali-labile sites) assessed using bio-chemical/electrophoretic assays or sister chromatid exchanges (SCE); protein adducts; and hypoxanthine-guanine phosphoribosyltransferase (HPRT) mutations. The document does not consider germ cells or gene mutation assays other than HPRT or markers of oxidative stress, which have been applied on a more limited scale.

Metal toxicity for rabbit alveolar macrophages in vitro

Abstract A model system in vitro has been employed to assess the relative cytotoxic properties of soluble salts of metals that occur as environmental contaminants. Rabbit alveolar macrophages obtained by lung lavage were exposed in tissue culture for 20 hours to chlorides of Cd2+, Ni2+, Mn2+, and Cr3+ and to ammonium vanadate (VO3−). All metals except Cd2+ produced significant decreases in numbers of cells at concentrations that affected cell viability. Cd2+ was unique in decreasing cell viability without causing cell lysis. In comparing the relative cytotoxicity of the various metals, the number of viable cells remaining after 20 hours was expressed as a percent of the total number of cells in control cultures at 20 hours. Thus, the net number of viable cells was decreased to 50% of control at concentrations of 0.1 m m Cd2+ and VO3− or 4–5 m m Ni2+, Mn2+, and Cr3+. The specific activity of acid phosphatase, a lysosomal indicator enzyme, was also decreased at similar concentrations. Using scanning electron microscopy it was possible to correlate surface alterations with exposure concentrations and cell viabilities so as to suggest a mode and sequence of cell injury which may ultimately lead to cell death.

Evaluation of the genetic activity profiles of 65 pesticides

We have previously reported the qualitative results of a major study on 65 pesticides (Waters et al., 1982). Dose information from this investigation (either lowest effective or highest ineffective dose tested) has now been incorporated into a computerized data management system. This report focuses on the qualitative profiles of genetic activity produced by these pesticides and our efforts to classify them according to their genotoxic effects and chemical structures. Three main categories may be distinguished based on the qualitative results: Category 1 pesticides were active in most of the in vitro and in vivo assays employed. These 9 compounds include the structurally similar organophosphate insecticides, acephate, demeton, monocrotophos and trichlorfon; the phthalimide fungicide analogues, captan and folpet; and the thiocarbamate herbicide analogues, diallate, sulfallate and triallate. The 26 Category 2 compounds demonstrated fewer positive results and may be subdivided into two parts, one of which contains 12 halogenated aromatic or heterocyclic ring compounds, including the phenoxy herbicides, 2,4-D, 2,4-DB and 2,4,5-T. The remaining part of Category 2 (14 compounds) consists of structurally similar organophosphate insecticides, azinphos-methyl, crotoxyphos, disulfoton, methyl parathion; three similar ethylenebisdithiocarbamate fungicides, maneb, mancozeb, and zineb; three similar pyrethroid insecticides, allethrin, chrysanthemic acid, and ethyl chrysanthemate; and four structurally diverse compounds, cacodylic acid, dinoseb, sec.-butylamine and benomyl. The third category of 30 pesticides gave negative results in all tests and represents structurally diverse compounds. Using the computerized profile matching methodology, from 2080 possible pairwise chemical combinations of the 65 pesticides, 20 statistically significant pairs were selected, 6 groups of pesticides were identified which were substantially similar to groups of pesticides we had formed previously (Waters et al., 1982) based on genetic activity and chemical structure. The matches showed excellent qualitative and, in most cases, excellent quantitative agreement. Hence it appears that specific patterns of test results present in the genetic activity profiles are related directly to chemical structure. Conversely, the data suggests that certain groups of compounds may be recognized by a well defined series of concordant tests results. As additional data is added, comparison of test results for new chemicals with existing data for known genotoxicants should aid in the evaluation of potential genetic health hazards.

CEBS—Chemical Effects in Biological Systems: a public data repository integrating study design and toxicity data with microarray and proteomics data

Abstract CEBS (Chemical Effects in Biological Systems) is an integrated public repository for toxicogenomics data, including the study design and timeline, clinical chemistry and histopathology findings and microarray and proteomics data. CEBS contains data derived from studies of chemicals and of genetic alterations, and is compatible with clinical and environmental studies. CEBS is designed to permit the user to query the data using the study conditions, the subject responses and then, having identified an appropriate set of subjects, to move to the microarray module of CEBS to carry out gene signature and pathway analysis. Scope of CEBS: CEBS currently holds 22 studies of rats, four studies of mice and one study of Caenorhabditis elegans. CEBS can also accommodate data from studies of human subjects. Toxicogenomics studies currently in CEBS comprise over 4000 microarray hybridizations, and 75 2D gel images annotated with protein identification performed by MALDI and MS/MS. CEBS contains raw microarray data collected in accordance with MIAME guidelines and provides tools for data selection, pre-processing and analysis resulting in annotated lists of genes of interest. Additionally, clinical chemistry and histopathology findings from over 1500 animals are included in CEBS. CEBS/BID: The BID (Biomedical Investigation Database) is another component of the CEBS system. BID is a relational database used to load and curate study data prior to export to CEBS, in addition to capturing and displaying novel data types such as PCR data, or additional fields of interest, including those defined by the HESI Toxicogenomics Committee (in preparation). BID has been shared with Health Canada and the US Environmental Protection Agency. CEBS is available at http://cebs.niehs.nih.gov. BID can be accessed via the user interface from https://dir-apps.niehs.nih.gov/arc/. Requests for a copy of BID and for depositing data into CEBS or BID are available at http://www.niehs.nih.gov/cebs-df/.

A survey of EPA/OPP and open literature on selected pesticide chemicals: II. Mutagenicity and carcinogenicity of selected chloroacetanilides and related compounds

With this effort, we continue our examination of data on selected pesticide chemicals and their related analogues that have been presented to the U.S. Environmental Protection Agencys (USEPAs) Office of Pesticide Programs (OPP). This report focuses on a group of selected chloroacetanilides and a few related compounds. As part of the registration process for pesticidal chemicals, interested parties (registrants) must submit toxicity information to support the registration including both mutagenicity and carcinogenicity data. Although this information is available to the public via Freedom of Information (FOI) requests to the OPP, publication in the scientific literature allows greater dissemination and examination of the data. For this Special Issue, graphic profiles have been prepared of the mutagenicity and carcinogenicity data available in the submissions to OPP. Also, a discussion is presented about how toxicity data are used to help establish tolerances (limits of pesticide residues in foods). The mutagenicity results submitted by registrants are supplemented by data on these chemicals from the open literature to provide a full perspective of their genetic toxicology. The group of chloroacetanilides reviewed here display a consistent pattern of mutagenic activity, probably mediated via metabolites. This mutagenic activity is a mechanistically plausible factor in the development of tumors seen in experimental animals exposed to this class of chemicals.

Use of computerized data listings and activity profiles of genetic and related effects in the review of 195 compounds

Computer-generated listings of data from short-term tests for genetic and related effects (activity profile listings) were prepared for 195 compounds that included for each compound, the test system (identified by a three-letter code word), qualitative results and the lowest effective dose (LED) or highest ineffective dose (HID) tested. A corresponding bar or line graph (activity profile) was also generated, in which test systems are displayed along the x-axis and the LED or HID values along the y-axis. The listings were reviewed and the data summarized by an IARC Working Group. The methodology used to generate these listings and plots is described, and results are given for one compound, benzene. The entire data base contains approximately 7000 entries from 4000 references.

Application of short-term bioassays in the fractionation and analysis of complex environmental mixtures

Section 1: Short-Term Bioassay Systems-An Overview.- The Use of Microbial Assay Systems in the Detection of Environmental Mutagens in Complex Mixtures.- Mutagenesis of Mammalian Cells by Chemical Carcinogens After Metabolic Activation.- Oncogenic Transformation of Mammalian Cells by Chemicals and Viral-Chemical Interactions.- Higher Plant Systems as Monitors of Environmental Mutagens.- The Role of Drosophila in Chemical Mutagenesis Testing.- The Cellular Toxicity of Complex Environmental Mixtures.- Section 2: Collection and Chemical Analysis of Environmental Samples.- Atmospheric Genotoxicants-What Numbers Do We Collect?.- State-of-the-Art Analytical Techniques for Ambient Vapor Phase Organics and Volatile Organics in Aqueous Samples from Energy-Related Activities.- Strategy for Collection of Drinking Water Concentrates.- Section 3: Current Research.- Short-term Bioassay of Complex Organic Mixtures: Part I, Chemistry.- Short-term Bioassay of Complex Organic Mixtures: Part II, Mutagenicity Testing.- Quantitative Mammalian Cell Genetic Toxicology: Study of the Cytotoxicity and Mutagenicity of Seventy Individual Environmental Agents Related to Energy Technologies and Three Subfractions of a Crude Synthetic Oil in the CHO/HGPRT System.- Environmental Testing.- Integrating Microbiological and Chemical Testing into the Screening of Air Samples for Potential Mutagenicity.- Chemical and Microbiological Studies of Mutagenic Pollutants in Real and Simulated Atmospheres.- Application of Bioassay to the Characterization of Diesel Particle Emissions.- Measurement of Biological Activity of Ambient Air Mixtures Using a Mobile Laboratory for In Situ Exposures: Preliminary Results from the Tradescantia Plant Test System.- Physical and Biological Studies of Coal Fly Ash.- Mutagenicity of Shale Oil Components.- Mutagenic Analysis of Drinking Water.- In Vitro Activation of Cigarette Smoke Condensate Materials to Their Mutagenic Forms.- Mutagenic, Carcinogenic, and Toxic Effects of Residual Organics in Drinking Water.- Mutagenic Analysis of Complex Samples of Aqueous Effluents, Air Particulates, and Foods.

Study of Pesticide Genotoxicity

With a limited supply of arable land supporting an ever-increasing human population, the threat of crop loss to agricultural pests becomes continually more acute. Thus pesticides have become an essential component of modern agriculture. As competing organisms evolve resistance to commonly used agents, new and more effective poisons and repellants must constantly be developed. The fundamental problem in pesticide development is to produce chemicals that act specifically against certain organisms without adversely affecting others. Because of the similarities in the structural, metabolic and genetic components of all life forms, absolute species specificity is frequently difficult to attain. Furthermore, such toxic chemicals improperly used may engender biological effects beyond those for which they were originally manufactured.

An overview of short‐term tests for the mutagenic and carcinogenic potential of pesticides

In the last few years, marked progress has been made in the development of methods for evaluating the mutagenic and carcinogenic potential of pesticide chemicals. The correlation of genetic and related biological activity in short-term tests with carcinogenic activity in whole animals allows the utilization of short-term mutagenicity bioassays to prescreen chemicals for effects related to mutation induction and presumptive carcinogenicity. In addition, bioassays now available can measure directly the chemical transformation of normal cells in culture into cells capable of producing tumors when injected into animals. This paper will review briefly the major types of relevant short-term tests and will develop a rationale for a phased approach to the evaluation of the mutagenic and carcinogenic potential of environmental chemicals. This approach involves the sequential application of bioassays which are organized into a three-level matrix emphasizing first detection, then confirmation, and finally hazard assessment. Chemicals demonstrating positive results in the short-term detection systems and confirmatory bioassays are pursued in higher level whole animal define a negative result. The phased approach should facilitate a cost effective utilization of limited testing resources and provide protection for human health in proportion to the anticipated hazard. Results obtained in evaluating a series of thirty-eight pesticide chemicals according to the phased approach discussed in detail.

Activity profiles of antimutagens: in vitro and in vivo data

In this review, retinol, chlorophyllin, and N-acetylcysteine are examined and compared with regard to their antimutagenic activity against some promutagens and a group of direct-acting alkylating agents. The promutagens included aflatoxin B1, certain polycyclic aromatic hydrocarbons (e.g., benzo[a]pyrene), and certain heterocyclic amines (e.g., food pyrolysates). Results of antimutagenicity testing selected from data surveyed in the published literature are displayed graphically as activity profiles of antimutagens showing both the doses tested and the extent of inhibition or enhancement of mutagenic activity. All three antimutagens are discussed in terms of their putative mechanisms of action in vitro and in vivo with emphasis on the xenobiotic metabolizing enzyme systems.