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At least 19 recordsLinked to original sources

Mutagens in a river heavily polluted with paper recycling wastes: results of field and laboratory mutagen assays.

Paper recycling industries generate considerable quantities of waterborne wastes, and thus water pollution constitutes the greatest environmental problem associated with this industrial activity [Hunt and Franklin, 1973]. Generally the impact of this water pollution is considered in terms of aesthetic blight and deterioration of water quality. We present data that document another aspect of this pollution, the environmental contamination of an aquatic ecosystem with mutagenic materials. A natural population of the fern Osmunda regalis growing in a river heavily polluted with paper recycling wastes had a high incidence of chromosome mutations. This population was monitored for four years for the frequency of two-break chromosome mutations. These mutations were postzygotic in origin and suggested the presence of mutagens in the river water. The fern population is downstream from the outfalls of a paper recycling mill, which was discharging 13.3 X 10(6) liters of untreated paper recycling waste per day. In 1977 a waste-water-treatment facility was constructed to remove the solid waste previously discharged into the river. This facility generates 69,300 kg of solid waste daily, which is taken to a landfill. Periodic samples of this solid waste were collected from the waste-treatment facility in the summer of 1978, extracted with various solvents, and the extracts tested for mutagenic activity with the Salmonella/mammalian microsome mutagenicity test [Ames et al, 1975]. A majority of the solid waste samples contained mutagenic materials, but in all cases S-9 activation was required for mutagenic activity. The samples also were assayed with the soybean mitotic crossing-over assay [Vig, 1975]. Four out of six samples were positive. These results document the presence of mutagens in the solid waste generated by a paper recycling industry and the genetic impact of these mutagens on the local biota.

Chromosome Aberrations

The efficiency and extent of mutagenic activity of some new mutagens of base-analogue type.

N4-Hydroxycytidine, 5-methyl-N4-hydroxydeoxycytidine and 2-amino-N6-hydroxyadenine were tested for their mutagenic activity in S. typhimurium and E. coli cells. Reversion analysis of different markers was applied in a plate-test system, and 2-aminopurine was used as a reference mutagen. (i) 2-Amino-N6-hydroxyadenine was the most potent mutagen. In some cases it gave more than 1000 colonies of revertants per plate. (ii) N6-Hydroxycytidine was the least specific mutagen. Almost all the tested markers were inducible to revert by this analogue. (iii) The mutagenic specificity of 5-methyl-N4-hydroxydeoxycytidine seemed to be opposite to that of 2-aminopurine. This suggests that the former can induce transition of CG to TA. (iv) A comparison of the mutagenic actions of N4-hydroxycytidine and 5-methyl-N4-hydroxy-deoxycytidine showed that deoxyriboside analogues are not necessarily more efficient mutagens than ribonucleosides. (v) No purine or pyrimidine deficiency was needed for mutagenesis to occur for any of the mutagens investigated. (vi) The results on bacteria with different repair abilities suggest that base-analogue mutagenesis (except perhaps for BrdUrd) occurs mainly during replication of nucleic acids containing substituted nucleosides with bi-functional specificity.

2-Aminopurine

Microsomal activation to mutagens of antischistosomal methyl thioxanthenones and initial tests on a possibly non-mutagenic analogue.

Five methylthioxanthenone and methylbenzothiopyranoindazole analogues, including lucanthone (Miracil D), are non-mutagenic for Salmonella typhimurium but are activated to mutagens by a rat liver microsome preparation. Hydroxymethyl analogues, including hycathone (Etrenol), are mutagenic in the absence of microsomes. It seems reasonable to assume that the hydroxymethyl derivatives are the more proximal mutagens and that Salmonella is unable to carry out the hydroxylation necessary for mutagen activation. During the pase 24 years, several million patients with schistosomiasis have been treated with lucanthone, and in recent years about 700 000 persons with hycanthone. The possible long-term deleterious effects of these agents for man even now remain to be determined. Our studies indicate that particular modifications in the structure of thioxanthenones drastically alter their mutagenicity. One apparently non-mutagenic thioxanthenone has been found. A number of the less mutagenic compounds also exhibit decreased acute toxicity in the mouse while retaining appreciable antischistosomal activity, suggesting that genetic and schistosomicidal activities may be dissociated from each other.

Animals

The mutagenic action of nitroimidazoles. IV. A comparison of the mutagenic action of several nitroimidazoles and some imidazoles.

The mutagenic action of 51 imidazoles was investigated. The fluctuation test of Luria and Delbrück was used, with Klebsiella pneumoniae as test organism. 8 compounds, including 5 with a weak mutagenic action in the fluctuation test, were also investigated by the Ames test in which Salmonella typhimurium TA100 was used. Of the 51 imidazoles examined, 33 were nitroimidazoles. 31 of the latter appeared to be mutagenic, whereas out of the 18 other imidazoles without a nitro group only 2 were mutagenic. Several of the substances tested for mutagenicity showed an antimicrobial activity. No direct relationship between antimicrobial action, growth inhibition and mutagenicity was established. With methyl-nitroimidazoles a relationship was found between the chemical structure and mutagenic action. However, when the nitroimidazoles had a more complex chemical structure, a relationship between this structure and mutagenicity could not be established.

Drug Evaluation, Preclinical

Testing the environment for dispersed mutagens: use of plant bioconcentrators coupled with microbial mutagen assays.

Mutagens dispersed in ecosystems are usually in low concentration and episodic in occurrence. The possibility of detecting such dispersed mutagens by utilizing indigenous bioconcentrator organisms coupled with a microbial mutagen assay offer a useful screening protocol. There are numerous examples of plant and animal species which concentrate toxic substances from the environment. Body extracts of these bioconcentrators can be suitably fractioned and tested for mutagens with various microbial mutagen assays. The fractions may be tested with a broad range of microbial assays covering numerous genetic end points as well as both with and without mammalian microsomal activation. This kind of environmental screening has an advantage over physicochemical techniques, in that sampling techniques are simpler and a wider chemical spectrum can be screened. There are problems inherent with testing a complex biological extract, however. If a reversion assay is used, the metabolite necessary for growth may be present. Toxins may be introduced, either concentrated from the environment in the same way as the mutagen, or produced by the concentrator itself. Finally, the concentrator may also produce an endogenous mutagen which will give spuriously active extracts. Methods for minimizing some of these difficulties are discussed.

Bacteria

Metabolic and mutagenicity studies on DDT and 15 derivatives. Detection of 1,1-bis(p-chlorophenyl)-2,2-dichloroethane and 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethyl acetate (kelthane acetate) as mutagens in Salmonella typhimurium and of 1,1-bis(p-chlorophenyl) ethylene oxide, a likely metabolite, as an alkylating agent.

Using a novel in vitro technique, whereby microsomal enzymes were embedded in an agar layer to prolong their viability, 1,1-bis(p-chlorophenyl) ethylene(DDNU), a mammalian metabolite of 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), was converted by microsomal mono-oxygenases of mouse liver into 1,1-bis(p-chlorophenyl)-1,2-ethanediol (DDNU-diol). The putative epoxide intermediate, 1,1-bis(p-chlorophenyl)ethylene oxide (DDNU-oxide), a new compound, was synthesized; it showed weak alkylating activity with 4-(4-nitrobenzyl)pyridine but was not mutagenic in Salmonella typhimurium strains TA100 and TA98. DDT and 13 of its metabolites or putative synthetic derivatives, including 1,1-bis(p-chlorophenyl)-2,2-dichloroethylene (DDE), 1 1,1-bis(p-chlorophenyl)-2-chloroethylene (DDMU), 1,1-bis(p-chlorophenyl)-2-chloroethane (DDMS)-DDNU, 2,2-bis(p-chlorophenyl)ethanol (DDOH), bis(p-chlorophenyl)acetic acid (DDA) and 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethanol (Kethane), caused no mutagenic effects in S. typhimurium strains TA100 or TA98, either in the presence or absence of a mouse-liver microsomal fraction. 1,1-Bis(p-chlorophenyl)-2,2,2-trichloroethyl acetate (Kelthane acetate) was a direct-acting mutagen in strain TA100, whereas 1,1-bis(p-chlorophenyl)-2,2-dichloroethane (DDD) was mutagenic in TA98, only in the presence of a mouse-liver microsomal system. The results are discussed in relation to possible pathways whereby DDT is activated to mutagenic and/or carcinogenic metabolites.

Alkylation

Mutagenicity of diallate, sulfallate, and triallate and relationship between structure and mutagenic effects of carbamates used widely in agriculture.

In an investigation of the mutagenic properties of 20 carbamate herbicides and fungicides by use of the Salmonella/microsome mutagenicity test as developed by Ames et al. (Mutation Res., 31: 347-364, 1975), we have found that three thiocarbamate compounds, diallate, sulfallate and triallate, are mutagenic in the presence of a liver microsomal fraction on strains TA1535 and TA100. This indicates that the metabolic products of these thiocarbamates are causing base-pair substitutions. Since the 2-chloro-allyl group is common to the three mutagenic compounds but is not common to the 17 nonmutagenic compounds, a metabolic derivative of this group is probably responsible for the mutagenic activity.

Animals

Influence of subcellular fractions of mammalian testes on the mutagenic activity of nitrofurans toward Escherichia coli; presence of a co-mutagen-like factor.

The activation of nitrofurans to mutagenic intermediates by testicular tissue was investigated. AF-2 and nitrofurazone were tested in a microsomal suspension assay with strain E. coli K-12 343/113 as indicator and subcellular fractions from rabbit testes. Different mutation patterns were observed in the presence or absence of testicular homogenate, indicating the presence of different mutagenic intermediates. The frequency of arg+ reversion increased proportionally to the homogenate concentration suggesting that the nitrofurans were activated by testicular components to intermediates that induced base-pair substitutions. Other experiments showed that a component of low molecular weight, present in the soluble fraction of homogenates of testes from rabbits, rats and monkeys, was responsible for the increased mutation frequency. It is concluded that this "co-mutagen-like" factor either alters the metabolism of nitrofurans in E. coli and/or promotes the formation of base-pair substitution-type mutations. This direct interaction between a nonenzymic component of mammalian testes and the mutation induction/expression process in E. coli suggests the role of co-mutagen-like factors in the sensitivity of testes to nitrofurans.

Animals

Cytotoxicity and mutagenicity of alkylating agents in cultured mammalian cells (CHO/HGPRT system): mutagen treatment in the presence or absence of serum.

Cytotoxicity and mutation induction at the hypoxanthine-guanine phosphoribosyl transferase locus in Chinese hamster ovary cells (CHO/HGPRT system) were measured for a range of concentrations of 6 alkylating agents [methyl and ethyl methanesulfonate (MMS, EMS), N-methyl- and N-ethyl-N'-nitro-N-nitrosoguanidine (MNNG, ENNG), and methyl- and ethyl-nitrosourea (MNU, ENU)] to determine the effect of the presence or absence of serum during the time of mutagen treatment. Cultures were treated with the mutagens for 5 h, a time period which results in no growth inhibition in the absence of serum, to estimate the potential decrease in effective mutagen dose to the cells which might result from reactivity with the serum proteins. With all 6 agents, identical results were found for cytotoxicity and for mutagenicity regardless of the presence or absence of serum during treatment. This finding demonstrates that the use of serum in cell-culture medium does not present any problems in apparent dosimetry studies, at least with these alkylating agents.

Alkylating Agents

Mutagenicity of plant flavonoids: structural requirements for mutagenic activity in Salmonella typhimurium.

40 compounds structurally related to the plant flavonol quercetin were tested for mutagenic activity in Salmonella typhimurium strain TA98. 10 flavonols, quercetin, myricetin, rhamnetin, galangin, kaempferol, tamarixetin, morin, 3'-O-methylquercetin, 7,4'-di-O-methylquercetin and 5,7-di-O-methyl-quercetin, exhibited unequivocal mutagenic activity. 4 compounds, quercetin, myricetin, rhamnetin and 5,7-di-O-methylquercetin, were active without metabolic activation, although metabolic activation markedly enhanced their activity. All 4 have free hydroxyl groups at the 3' and 4' positions of the B ring. The other active compounds required an in vitro rat-liver metabolizing system for significant activity. Structural features which appear essential for mutagenic activity in this strain are a basic flavanoid ring structure with (1) a free hydroxyl group at the 3 position, (2) a double bond at the 2, 3 position, (3) a keto group at the 4 position, and (4) a structure which permits the proton of the 3-hydroxyl group to tautomerise to a 3-keto compound. The data are consistent with the requirement for a B ring structure that permits oxidation to quininoid intermediates. Free hydroxyl groups in the B ring are not essential for activity if a rat-liver metabolic activating system is employed. Data from 12 compounds which differ only at the essential sites described above indicate that the structural requirements for mutagenicity in strain TA100 are the same as those for activity in strain TA98. Based on the above structural requirements, a metabolic pathway for flavonol activation to DNA-reactive species is proposed.

Animals

Mutagenicity and mutagenic specificity of metronidazole and niridazole in Neurospora crassa.

Mutagenicity and mutagenic specificity of niridazole and metronidazole, two chemotherapeutic agents used in the treatment of human parasitic diseases, were studied with the ad-3 test system of Neurospora crassa. The results show that neither compound is mutagenic in resting conidia. In growing vegetative cells, however, both compounds are mutagenic in N. crassa. Genetic analysis of the mutants indicated that niridazole induces predominantly base-pair substitution mutations. None of the niridazole-induced mutants resulted from multilocus deletions. The spectra of genetic alterations induced by metronidazole are similar to those induced by the mono-functional alkylating agents ethyleneimine (EI), ethylmethanesulfonate (EMS), and ICR-177. It is therefore suggested that the mechanism of mutation induction by metronidazole in Neurospora is similar to that of monofunctional alkylating agents.

Genotype

Mutagenicity and mutagenic specificity of metronidazole and niridazole in Neurospora crassa.

Mutagenicity and mutagenic specificity of niridazole and metronidazole, two chemotherapeutic agents used in the treatment of human parasitic diseases, were studied with the ad-3 test system of Neurospora crassa. The results show that neither compound is mutagenic in resting conidia. In growing vegetative cells, however, both compounds are mutagenic in Neurospora crassa. Genetic analysis of the mutants indicate that niridazole induces predominantly base-pair substitution mutations. None of the niridazole-induced mutants resulted from multilocus deletions. The spectra of genetic alterations induced by metronidazole are similar to those induced by the monofunctional alkylating agents, EI, EMS and ICR-177. It is suggested, therefore, that the mechanism of mutation induction by metronidazole in Neurospora is similar to that of monofunctional alkylating agents.

Genetic Complementation Test

The mutagenicity of saccharin impurities. I. Detection of mutagenic activity.

Sodium saccharin, ortho-toluenesulfonamide and impurities extracted from commercially produced saccharin with water and organic solvents were tested for mutagenicity with strains of Salmonella typhimurium. The organic solvent soluble impurities exhibited strong mutagenic activity for TA98 and slight activity for TA100. Mutagenic activity for S. typhimurium TA98 was demonstrated in extracts of some but not all lots of sodium saccharin produced by both Maumee and Remsen-Fahlberg processes. The significance of the mutagenic impurity to the carcinogenicity of saccharin is discussed.

Drug Contamination

Mutagenicity tests on anthelmintics: microsomal activation of viprynium embonate to a mutagen.

Eight anthelmintic preparations readily available in Australia were tested for mutagenicity in the Salmonella typhimurium test system. A slightly modified version of the procedure recommended by Ames et al. [2] was adopted, in that the test samples were placed in "wells" cut out of the agar of a plate previously seeded with the appropriate tester strain. Addition of a mixture of rat liver microsomal enzymes and appropriate co-factors ("S-9 mix") to one of the two wells on a single plate allowed a possible requirement for metabolic activation to be recognised. Using this procedure, viprynium embonate was found to be non-mutagenic. It was however, activated by the rat liver microsome preparation to a mutagen capable of causing both base-pair substitution (detected with strain TA100) and frameshift (detected with strain TA98) mutations. The other seven compounds tested all gave negative results in this system.

Animals

Mutagenicities of styrene oxide derivatives on Salmonella typhimurium (TA 100): relationship between mutagenic potencies and chemical reactivity.

The lethal and mutagenic effects of p-methyl-, m-chloro-, p-chloro- and unsubstituted styrene oxide on Salmonella typhimurium (TA 100) were investigated. At equal concentrations, p-chlorostyrene oxide was more lethal than p-methyl-, m-chloro- or unsubstituted styrene oxide. When the survival fraction was 0.8 or more, the mutagenicities of these compounds increased in the order: m-chlorostyrene oxide = p-chlorostyrene oxide less than styrene oxide less than p-methyl-styrene oxide. The mutagenicities of these compounds depended only on the reactivity of their benzylic site; the reactivity at their primary site and their partition coefficients appeared to have no effect.

Dose-Response Relationship, Drug

Concentration of mutagens from urine by absorption with the nonpolar resin XAD-2: cigarette smokers have mutagenic urine.

A method is described for concentrating mutagens/carcinogens from human urine about 200-fold for subsequent assay in the Salmonella/mammalian microsome mutagenicity test. The method is also applicable for other aqueous liquids and for other in vitro tests for mutagens/carcinogens. The urine (up to 500 ml) is put through a column with a 1.5-cm3 bed volume of XAD-2 (styrene-divinylbenzene polymer) and the adsorbed material is then eluted with a few milliliters of acetone. The acetone is taken to dryness and the residue is dissolved in dimethyl sulfoxide. This is the urine concentrate that is assayed for mutagenicity. Various mutagens/carcinogens have been added to human urine and the recoveries have been measured after adsorption on XAD-2, XAD-4, and Tenax GC (diphenyl-p-phenylene oxide polymer). We propose that this method be used in monitoring the urine of human populations and of experimental animals in toxicological studies. It is shown with this procedure that cigarette smokers have mutagenic urine while nonsmokers do not.

Chromatography

[Salmonella typhimurium--test system for detecting the mutagenic activity of environmental pollutants. II. Mutagenic effect of heavy metal salts in an in vitro system with metabolic activation].

Mutagenic effect of zinc chloride on Salmonella typhimurium strain was detected using in vitro metabolic activation system. Cadmium chloride showed no significant mutagenic activity in the same system. It is recommended to use both in vitro and in vivo metabolic activation systems in mutagenicity testing of chemicals.

Animals

Mutagenicity studies with praziquantel, a new anthelmintic drug: tissue-, host-, and urine-mediated mutagenicity assays.

Praziquantel, a new anthelmintic drug with activity against all species of schistosomes pathogenic to man, and against a wide range of Cestodes, was tested for mutagenic potential. For the detection of both base substitutions and frameshift mutations, Salmonella typhimurium TA 100 and TA 98 were used as tester strains. Using the plate assay with and without added S-9, host-mediated assay and urine-mediated assay without and after incubation with beta-glucuronidase/arylsulfatase, no mutagenic activity could be detected.

Administration, Oral