Potential Bioassays For The Detection Of The Effects Of Underground Mining

INTRODUCTION Cross, et al. (1974), produced a retrospective study of standard setting for underground miners. This report had two distinct components; i) criteria of importance for the protection of the miners, and ii) economic considerations for standard setting. The methods for setting radiation safety standards reviewed were: dose calculations and consensus methods; epidemiology; pathology; bioassay; animal experiments; sputum cytology; chromosome aberrations; and, the Mantel-Bryan Model. By looking back, the authors intended to enable officials to look ahead in making future decisions based on reasonable conclusions. Now it may be time to consider underground miners' protection from another perspective: are there miners who may be especially susceptible to toxic environments?; and if so, are there any biomedical assays that might be indicative of exceptional sensitivities to toxic substances? The human population is genetically very heterogeneous. The data of Saccomanno, et al. (1973), reveal great variability in individual response to radon daughter exposure and only a small portion of the miner population subject to toxic inhalants develop squamous cell metaplasia (Saccomanno, et al., 1970). The majority of the miner population is either not susceptible or is resistant to the toxic agents. This information suggests the existence of a small subpopulation with increased sensitivity or reduced resistance and underscores the need for indicators from biomedical assays that might prove of value for the detection of such individuals. The heightened awareness of the contribution of pollutants in the environment for the potential induction of mutations and carcinogenesis lead to a profusion of short-term bioassays to circumvent the high cost and time-consuming large toxicity animal studies. Over 100 bioassays across taxa from microbes to man are at various stages of use or development (Hollstein, et al., 1979). Less than a dozen tests currently offer early promise for application to[ in vivo] effect studies of man. Many are still in early development, lack the sensitivity needed for a retrospective or prospective study at current permissible exposures, are impractical to conduct in the field, or are not cost effective. The purpose of this paper is to review some of the bioassays that may now, or in the near term, prove applicable for the detection of individual underground miners with increased susceptibility to toxic agents. Throughout this statement, it is assumed that any single test may give false negatives or false positives and, therefore, a tier of tests should be investigated. The possible tests are in various stages of development; some tests better proven than others with a firmer data base and, therefore, with greater probability of usefulness. Some of the less proven assays are not ruled out if they have practical or theoretical promise as indicators. Table I summarizes the assays critiqued for their potential to monitor the effects of [in vivo] exposure to genotoxic substances. POTENTIAL INDICATORS OF HIGHLY SENSITIVE MINERS Assays of Body Fluids It is desirable to have data on the agent(s) to which subjects are exposed when humans are monitored by biomedical effects. Obviously, to varying intensity, the underground mining environments are monitored for radon daughters and it is recognized that the miners are also exposed to other pollutants, most notably, uranium ore dust and diesel fumes. Further testing for the metabolites of the pollutants can be done on body fluid, urine. [High Performance Liquid Chromatography (HPLC)]: This is a very sensitive method for the detection of mutagenic metabolites in urine. The urine is treated with the enzyme sulfatase and beta-glucuronidase to permit identification of substances that are made nonmutagenic by conjugation as glucuronides. The sample is then passed through an XAD-2 resin column and the absorbed organic molecules eluted with acetone. The sample is then split and evaporated to 1 ml and used for direct chemical analysis using HPLC. One drawback to the test is the inability to measure cumulative exposure, but multiple samples can be obtained and comparison to baseline (control) and exposure samples can reveal qualitative differences as a consequence of exposure to mutagens. [The Ames/Salmonella Microbiological Assay]: The Ames/Salmonella microbiological mutagen test is the most extensively used short-term bioassay, with over 2,600 chemicals having undergone testing by this method (Hollstein, et al., 1979). The method, thoroughly worked out and tested for 10 years, consists of taking the second split urine sample from the HPLC preparation, evaporating to dryness and dissolving in dimethylsulfoxide (DMSO). The sample is then applied directly to