The distribution of dermal tumorigens in coal liquids: relationship of tumorigenicity and microbial mutagenicity.

Polycyclic aromatic hydrocarbons and/or their pyrolle derivatives were found to be the primary contributors to the skin tumorigenicity of the neutral fractions of two coal oils. Mutagenicity of the neutral fraction in Salmonella test strains was found to be due primarily to polycyclic aromatics containing polar substituents. Thus, the chemical classes responsible for skin tumorigenicity differ from those responsible for mutagenicity.

Effects of genotype and age on mixed-function oxidase activities in adult Drosophila melanogaster.

The mixed-function oxidases that metabolize dimethylnitrosamine, aminopyrine, benzphetamine, 7-ethoxycoumarin and benzo[alpha]pyrene were measured in adults of the Canton-S, Oregon-R and Hikone-R strains of Drosophila melanogaster. The expression of these activities is both genotype and age dependent.

Studies on the relationship between dimethylnitrosamine-demethylase activity and dimethylnitrosamine-dependent mutagenesis in Drosophila melanogaster.

The relationship between dimethylnitrosamine (DMN) demethylase activity and DMN-induced mutagenesis was investigated in Drosophila melanogaster. The activity of DMN-demethylase was at least 10-fold greater in the Hikone-R strain than in three other Drosophila strains. However, the sex-linked recessive lethal (SLRL) mutations induced by DMN in the four strains differed by less than 2-fold. Several possibilities to explain the lack of correlation between DMN-demethylase activity and DMN-induced mutations were tested and eliminated. They include: (i) the presence of inhibitors of DMN-demethylase in extracts of low-activity strains, (ii) a sex bias in the Hikone-R strain in which the enzyme activity is confined to the females, (iii) the possibility that DMN treatment induces DMN-demethylase activity in the low-activity strains and (iv) the possibility that Hikone-R has a much more efficient DNA repair system than the other strains. The results are discussed in terms of what is known about the role of DMN-demethylase in the metabolic activation of DMN in other systems.

Preparation of oils for bacterial mutagenicity testing.

4 procedures used to prepare fossil-derived oils for bacterial mutagenicity testing have been examined. These are, (a) dewaxing by partitioning the oil between dimethyl sulfoxide (DMSO) and cyclohexane, (b) incorporating a surfactant to increase compatibility of the oil with the bioassay media, (c) directly slurrying the oil in DMSO, and (d) computing the mutagenicity of the oil by summing the contributions of individual chemical class fractions. DMSO slurries generally exhibit higher mutagenicities than computed by summing the contributions of chemical class fractions. Results of testing DMSO-slurries correlate (r = 0.87) well, however, with those obtained by summation. Mutagenicity results agree within a factor of two for the samples tested by 4 sample preparation procedures.

Enhancement of mutagenic activity in Salmonella by contraceptive steroids.

Two oral contraceptive steroids, mestranol and norethynodrel, were evaluated for mutagenicity in the Salmonella histidine reversion assay. The pure forms of the hormones were not mutagenic when tested with either missense (TA1535, TA100) or frameshift (TA98, TA1538, TA1537) strains. In vitro activation of the hormones with liver homogenates from rats induced either with phenobarbital or Aroclor did not influence these results. However, mestranol was capable of enhancing the mutation yield obtained by an ineffective subthreshold dose of 2-acetylaminofluorene. Dimethyl sulfoxide extracts of two contraceptive pills, Ovulen-21 (containing mestranol) or Enovid-E (containing mestranol or norethynodrel), also were nonmutagenic. But again, both these extracts were capable of enhancing the mutation yield induced with an ineffective dosage of 2-acetylaminofluorene and N-nitrosopiperidine. These studies point to the possible promotional effect and subsequent potential hazard to the female consumers who use these hormones as a means of pregnancy control.

Mutagenicity of 4,4'-methylenedianiline derivatives in the Salmonella histidine reversion assay.

4,4'-Methylenedianiline and its derivatives were assayed for mutagenicity in the Salmonella/microsomal mutagenicity assay develop by Ames. A specificity to revert strain TA98 suggests a mechanism of frameshift mutagenesis. Liver microsomal preparations (S-9) from rats induced with phenobarbital were most effective for metabolic activation. Alkyl substitution of 4,4'-methylenedianiline did not alter its mutagenic activity; however, substitution of both positions ortho to the amino group eliminated mutagenic activity. Substitution with alkoxy-carbonyl groups eliminated mutagenic activity, whereas halogen substitution (chlorine, fluorine) enhanced the mutagenic activity. The results presented here show the use of structure-activity studies as predictive tools for the assessment of genotoxic properties of industrial chemicals.

Structure activity studies with N-nitrosamines using Salmonella typhimurium and Escherichia coli.

Mutagenic activities of a large number of nitrosamines were determined using Salmonella histidine reversion and Escherichia coli arginine reversion assays. The cyclic nitrosamines exhibited a close correlation between their mutagenic and carcinogenic properties, while no such relationship was evident with the aliphatic nitrosamines. Substitution of cyclic nitrosamines with methyl, hydroxy and oxy groups did not alter the mutagenic activities. However, when positions alpha to the N-nitroso groups were substituted with methyl groups, the biological activity was eliminated. Substitution with halogens enhanced, whereas carboxyl substitution eliminated the biological activity. The E. coli assay not only substantiated the observations made with Salmonella, but also demonstrated mutagenic activity in the case of certain carcinogenic nitrosamines which were not mutagenic in the Salmonella assay.

Analytical and biological analysis of test materials from the synthetic fuel technologies.

Nitrogen-containing organic compounds from environmental sources are receiving increasing attention because of uniquely active mutagens which have been found in this class (Chrisp et al., 1978; Nagao and Sugimura, 1978: Guerin et al., 1980) Differences in mutagenic activities among the various organo-nitrogen compounds, i.e., pyrrole types, pyridine types and aniline types, have been noted consistently. Furthermore, differences among homologs of a particular compound type are often striking. Information in this paper engages the question of chemical structure/biological activity relationships. Activity data for several N-heterocyclic, nitro-, amino- (primary, secondary and tertiary), and amino-N-heterocyclic aromatic compounds are presented. The number of fused rings and the substituent type affect the mutagenic activities greatly. The trends observed are discussed generally with reference to molecular structural features.

Nitrosamine-induced mutagenesis in Escherichia coli K12 (343/113). 1. Mutagenic properties of certain aliphatic nitrosamines.

The Escherichia coli K12 (343/113) test system developed by G. Mohn was used to detect the mutagenic activity induced by a group of aliphatic nitrosamines. Metabolic activation was incorporated into the assay by the addition of liver homogenates induced in either Sprague-Dawley rats or C3H mice with the addition of 0.1% phenobarbital to the drinking water. Nitrosodiethylamine (NDEA) was mutagenic upon metabolic activation and exhibited a preference to revert the missense mutation at the arginine locus. NDEA was also capable of inducing the forward mutation, selected as an ability to utilize galactose. NDEA was converted effectively into a mutagen in a time period of 30 min to 2 h. Metabolic activation with the mouse and rat liver preparations did not result in quantitative differences. Aliphatic nitrosamines that gave unexpected results with the Salmonella assay [4-10] were examined in the E. coli system. Nitrosodipropylamine (NDPA) and nitrosodiallylamine (NDAA) were mutagenic in both E. coli and Salmonella. Nitrosomethylethylamine (NMEA) was not mutagenic in Salmonella but was mutagenic in E. coli, and a strong carcinogen, nitrosomethylneopentylamine (NMNA), was not mutagenic in either assay. These results indicate the use of multiple genetic assays for the detection of genotoxic chemicals in our environment.

Mutagenicity of N-nitrosopyrrolidine derivatives in Salmonella (Ames) and Escherichia coli K-12 (343/113) assays.

The mutagenicity of nitrosopyrrolidine (NPYR) and its derivatives was determined by use of the Ames Salmonella assay. A clear specificity to revert the missense stain of TA1535 and a requirement for the phenobarbital-induced rat-liver activation system (S9 mix) were noted. 3,4-Dichloronitrosopyrrolidine was more mutagenic than NPYR, whereas 3-hydroxynitrosopyrrolidine was weakly mutagenic. The carcinogenic nitroso-3-pyrrolidine was not mutagenic under the test conditions. The noncarcinogenic derivatives (2,5-dimethylnitrosopyrrolidine, nitrosoproline and 4-hydroxynitrosoproline) were not mutagenic. Liquid preincubation assays were not any more effective than the pour-plate assays. Selected derivatives of NPYR were tested in the Escherichia coli K-12 (343/113) assay A specificity to revert the missense mutation at the arg locus and a dependence on phenobarbital-induced rat-liver S9 mix were noted with NPYR and its derivatives. 3,4-Dibromonitrosopyrrolidine, which was not mutagenic in Salmonella, was effective in E. coli, and the weakly carcinogenic NPRL was a weak mutagen resulting in a 2-fold enhancement in the E. coli arginine reversion assay.

Microsomal activation of selected polycyclic aromatic hydrocarbons and aromatic amines in Drosophila melanogaster.

Benzo[a]pyrene, 7,12-dimethylbenz[a]anthracene, 2-acetylaminofluorene, 2-aminoanthracene, and 1-aminopyrene, when fed to adult Drosophila melanogaster males, gave a negative mutagenic response in the X-linked recessive lethal assay. Benzo[a]pyrene was also ineffective in inducing "Minutes". Aflatoxin B1, EMS and DMN gave a positive response which was dependent on the concentration of mutagen fed. Whole fly homogenates prepared from adult Drosophila were assayed for mixed-function oxidase activity in the Salmonella/microsome test. Crude Drosophila microsomes activated 2-acetylaminofluorene, 2-aminofluorene, 2,7-diaminofluorene, 2-aminoanthracene, 1-aminopyrene, and aflatoxin B1. Tests with benzo[a]pyrene, pyrene, 1,2,3,4-dibenz[a]anthracene, and 7-12-dimethylbenz[a]anthracene were negative.

Analytical and biological analyses of test materials from the synthetic fuel technologies. III. Use of sephadex LH-20 gel chromatography technique for the bioassay of crude synthetic fuels.

To determine the health effects associated with newly emerging energy technologies, we have subjected a group of synthetic fuels to mutagenicity evaluation, using the Ames Salmonella assay. Coupling of chemical fractionation to the mutagenicity assays was necessary. Fractions obtained by use of Sephadex LH-20 gel chromatography on crude-coal-derived oils and shale oil were tested for mutagenicity with strain TA98 (with Aroclor S9 mix). Mutagenicity results obtained with synthetic fuels were compared with those from a mixture of natural petroleum crude oils. Merits of the Sephadex LH-20 separation technique and precautions in interpreting experimental results are discussed.

Effects of methylation and ring size on mutagenicity of cyclic nitrosamines in Drosophila melanogaster.

The mutagenic activity of 7 nitrosopiperazines, 2 nitropyrrolidines, and 3 nitrosomorpholines was examined in the X-linked recessive-lethal assay of Drosophila melanogaster. Mutagenicity is also reported for a series of cyclic nitrosamines that differ in structure only in the number of carbon atoms in the ring. Of the 18 compounds tested, 6 (nitrosopiperazine; 2,3,5,6-tetramethyldinitrosopiperazine; nitrosoproline; 2,5-dimethylnitrosopyrrolidine; nitrosothiomorpholine; and nitrosooctamethyleneimine) were nonmutagenic. As we reported earlier in investigations with the nitrosopiperidines, substitutions with methyl groups at all of the alpha-carbon atoms reduce or eliminate the mutagenic activity of dinitrosopiperazine and nitrosopyrrolidine.

Mutagenicity of alkaline constituents of a coal-liquified crude oil in mammalian cells.

Employing the CHO/HGPRT system, we have shown that the acetone fraction of a crude coal--liquid crude oil is the major contributor to the mutagenicity of synthetic oil. The system appears to be useful for determination of mutagenicity of organic mixtures and for corroboration of results from other biological assays.