David L. Berry

Skin tumor initiating ability of benzo(a)pyrene 4,5- 7,8- and 7,8-diol-9,10-epoxides and 7,8-diol

The skin tumor initiating abilities of both K-region and non-K-region epoxides of benzo(a)pyrene(BP) were determined in mice using a two-stage system of tumorigenesis. BP-4,5-epoxide and BP-7,8-dihydrodiol-9,10-epoxide (anti) were found to be weak tumor initiators whereas BP-7,8-epoxide had about a third of the activity as the parent hydrocarbon, BP. However, the 7,8-dihydrodiol of BP was found to be approximately as potent as BP suggesting that it may be a proximate carcinogen.

Studies with chlorinated dibenzo-p-dioxins, polybrominated biphenyls, and polychlorinated biphenyls in a two-stage system of mouse skin tumorigenesis: potent anticarcinogenic effects

The chlorinated dibenzo-p-dioxins and chlorinated dibenzo-furans are among the most potent teratogens and toxins that are found as environmental contaminants. The polychlorinated biphenyls and polybrominated biphenyls are not nearly as toxic as the dioxins, but they are environmentally more prevalent than the dioxins. Both classes of compounds are inducers of cytochrome P/sub 450/-mediated microsomal monooxygenases and of aminolevulinic acid synthetase. The dioxins induce aryl hydrocarbon hydroxylase, the enzyme system responsible for converting polycyclic aromatic hydrocarbons to active and inactive metabolites. PCBs and PBBs induce the monooxygenase system with properties similar to both phenobarbital and 3-MC. Timing is important in the application of PCB to the diets of laboratory animals. Treatment with PCBs prior to the administration of carcinogen results in reduction of tumor response whereas treatment subsequent to application of the carcinogen results in enhancement of the tumorogenic response. The two-stage system of mouse skin tumorigenesis allows one to evaluate critically the initiation and promotion phases of carcinogenesis individually. (DLS)

Benzo(a)pyrene metabolism in mouse epidermis. Analysis by high pressure liquid chromatography and DNA binding

Mouse epidermal homogenates contain an inducible aryl hydrocarbon hydroxylase (AHH) complex that catalyzes the formation of benzo(a)pyrene-7,8-dihydrodiol from benzo(a)pyrene (BP) as assessed by high pressure liquid chromatography (HPLC). 5,6-Benzoflavone (5,6-BF), 7,8-benzoflavone (7,8-BF) and 17-beta-estradiol decreased and butylated hydroxytoluene (BHT) enhanced oxidative metabolism of BP when added in vitro. Epoxide hydrase activity (hydration of benzo(a)pyrene-4,5-epoxide) (BP-4,5-epoxide) was enhanced by 17-beta-estradiol, 5,6-BF, and 7,8-BF. BHT exhibited no significant effect and 1,2-epoxy-3,3,3-trichloropropane (TCPO) inhibited hydrase activity. The capacity of epidermal homogenates to catalyze the covalent binding of BP to DNA indicated that addition of both 5,6-BF and 7,8-BF decreased binding. BHT and TCPO did not significantly affect DNA-binding.

2,3,7,8-tetrachlorodibenzo-p-dioxin: potent anticarcinogenic activity in CD-1 mice

Abstract Topical pretreatment with non-toxic doses of 2,3,7,8-tetrachlorodibenzo-p-dioxin, a contaminant formed in the commercial synthesis of the herbicide 2,4,5-trichlorophen-oxyacetic acid, strongly inhibited the initiation of skin tumors by 7,12-dimethylbenz(a)-anthracene and benzo(a)pyrene in female CD-1 mice. 2,3,7,8-tetrachlorodibenzo-p-dioxin also produced marked induction of epidermal monooxygenase enzymes functional in the conversion of 7,12-dimethylbenz(a)anthracene to a variety of hydroxylated products. The time course of anticarcinogenic effects resulting from pretreatment with the dioxin correlated with the magnitude of induction as well as with a singnificant reduction in the quantity of 7,12-dimethylbenz(a)anthracene metabolites covalently bound in vivo to epidermal DNA and RNA but not protein.

Qualitative and quantitative separation of a series of phorbol-ester tumor promoters by high-pressure liquid chromatography.

A high-pressure liquid chromatographic (HPLC) method using a micro-particulate silica column and gradient elution was developed that separated 12-O-tetradecanoylphorbol-13-acetate (TPA) from 20-oxo-TPA; 12-O-tetradecanoylphorbol (TP); 13-O-acetylphorbol (PA), and from the diterpene alcohol, phorbol (P). A series of other phorbol-ester tumor promoters were also separated via HPLC. Spectrophotometric determination at 232 nm allowed detection sensitivities of 0.05 microgram of TPA. When tritiated TPA was applied to mouse skin, the majority of the tritiated product recovered was TPA, indicating only minimal metabolism of TPA and no need for metabolic activation for tumor promotion.

The ability of enteric bacteria to catalyze the covalent binding of bile acids and cholesterol to DNA and their inability to metabolize benzo(a)pyrene to a binding product and to known metabolites

The capacity of enteric bacteria (E. coli, Salmonella, Pseudomonas, Shigella and Klebsiella) to catalyze the covalent binding of benzo(a)pyrene (BP), cholic acid, deoxycholic acid and cholesterol was investigated. In general, these bacteria were incapable of activating BP to a covalently bound product with calf thymus DNA. Metabolism studies of BP by fluorometric assay failed to indicate any accumulation of BP-3-hydroxy in the incubation medium. Detailed metabolic investigation with high-pressure liquid chromatography indicated that these bacteria did not produce any known metabolites which are formed by mammalian systems. However, radioactivity was detected in all fractions, suggesting that the bacteria were readily metabolizing BP into smaller molecules for energy and carbon sources. Although the enteric bacteria did not metabolize BP into known metabolites, some were capable of activating cholesterol, cholic acid and deoxycholic acid to covalently bound products with DNA. The binding data with cholesterol and bile acids also suggested that the binding process required NADPH as a cofactor because binding level was rather low without NADPH.