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The naturally occurring flavouring agents trans -anethole and eugenol were evaluated for antigenotoxic effects in mice. The test doses of trans -anethole (40–400 mg/kg body weight) and eugenol (50–500 mg/kg weight) were administered by gavage 2 and 20 h before the genotoxins were injected intraperitoneally. Anti-genotoxic effects were assessed in the mouse bone marrow micronucleus test. Pretreatment with trans -anethole and eugenol led to significant antigenotoxic effects against cyclophosphamide (CPH), procarbazine (PCB), N -methyl- N ′-nitro- N -nitrosoguanidine (MNNG) and urethane (URE). In addition, trans -anethole inhibited the genotoxicity of ethyl methane sulfonate (EMS). Both trans -anethole and eugenol exerted dose-related antigenotoxic effects against PCB and URE. There was no significant increase in genotoxicity when trans -anethole (40–400 mg/kg body weight) and eugenol (50–500 mg/kg body weight) were administered alone.

The interaction between coffee (100 mg freeze-dried home brew/kg body weight) and dietary constituents was assessed for anti-genotoxic effects against cyclophosphamide, N-methyl-N-nitro-N- nitrosoguanidine (MNNG), N-nitroso-N-ethylurea, mitomycin C and urethane (URE) in the mouse bone marrow micronucleus test. Combinations of dietary constituents consisting of (1) chlorogenic acid, caffeic acid, ellagic acid and ferulic acid, (2) beta-carotene, curcumin and alpha-tocopherol, (3) chlorogenic acid, curcumin, alpha-tocopherol, anethole and eugenol, and (4) beta-carotene, curcumin, ellagic acid and chlorogenic acid were used in this study. Before the genotoxin was injected i.p., identical groups of mice were orally administered either vehicle control, coffee, dietary constituents, or coffee plus dietary constituents. Co-administration of coffee with the dietary constituents enhanced the anti-genotoxic effect compared with that of either coffee or the dietary constituents alone. Two-factor analysis of variance of the data suggests that there is a significant synergistic interaction between coffee and the dietary constituents for anti-genotoxic effects against MNNG (combination 1 and 2) and URE (combination 4).

In vivo anti-genotoxic effects of caffeinated and decaffeinated instant coffee were compared in mice after pretreatment either by gavage for 10 consecutive days or in the drinking water for 2 weeks. Changes in hepatic sulfhydryl (-SH) content and glutathione S-transferase (GST) activity were evaluated in pretreated animals. Both caffeinated and decaffeinated instant coffee induced a moderate increase in -SH content and GST activity following pretreatment (with 70, 140 and 280 mg/kg body weight) by gavage for 10 days. This enhancement was not always dose dependent. The maximum effect on GST activity was observed at a dose of 140 mg/kg body weight/day. However, such an effect was not observed after administration of drinking water containing 2% caffeinated/decaffeinated instant coffee for 2 weeks. Results of the bone marrow micronucleus test for evaluating genotoxic effects revealed that both caffeinated and decaffeinated instant coffee (140 mg/kg body weight/day) could exert significant anti-genotoxic effects against ip injected benzo[a]pyrene (BP), cyclophosphamide (CPH), 7,12-dimethylbenz[a]anthracene (DMBA), mitomycin C (MMC) and procarbazine (PCB) in animals pretreated by gavage. Anti-genotoxic effects against BP, DMBA and urethane (URE) were evaluated in animals that received drinking water containing 2% caffeinated/decaffeinated instant coffee for 2 weeks. With the exception of the anti-genotoxic effect of decaffeinated coffee against DMBA, there was no significant change in genotoxicity after the above pretreatment. From this work, there is no evidence for any significant difference in the in vivo anti-genotoxicity of caffeinated and decaffeinated instant coffee.

The protective effects of coffee against somatic mutation and mitotic recombination induced by cyclophosphamide (CPH), mitomycin C (MMC) and urethane (URE) were evaluated in the standard (ST) and high bioactivation (HB) crosses of the wing spot test in Drosophila melanogaster. These two crosses are characterized by different constitutive levels of cytochrome )-450-dependent enzyme activities. 3-day old larvae transheterozygous for the wing cell markers mwh (multiple wing hairs) and flr3 (flare3) were fed until pupation on medium containing a genotoxin alone or its combination with different concentrations of instant coffee. subsequently, the wings of the resulting adult flies were analysed for detecting single spots (mwh or flr3) originating from mutational or recombinational events as well as twin spots (mwh and flr3) originating exclusively from recombination. The results showed high sensitivity of the HB cross to URE. Co-administration of instant coffee was effective in exerting significant dose-related inhibitory effects on the genotoxicity of URE in the ST and the genetically susceptible HB cross. Similarly, coffee showed significant dose-related inhibitory effects on the genotoxicity of MMC in both crosses. The same protective effect was also observed with one concentration of coffee in combination with CPH. Pretreatment of 2-day-old HB larvae with coffee for 24 hr followed by treatment with URE was also effective in significantly reducing the induction of mutation and recombination. The magnitude of the protective effects of coffee against these three genotoxins was independent of the genotype of the larvae used for treatment, that is it was independent of the bioactivation capacity of these larvae. The study demonstrates the suitability of this assay for obtaining qualitative and quantitative data on the result of interactions among a genotoxin, an inhibitor of genotoxicity and bioactivation capacity of the host.

The possible role of coffee in modulating the in vivo genotoxicity of the well established genotoxic chemicals, mitomycin C, cyclophosphamide, procarbazine and adriamycin, was evaluated. Coffee was administered orally to mice that received the genotoxic chemicals ip. Genotoxicity was assessed in the bone-marrow micronucleus test. Doses of coffee in the range 225 to 1125 mg (dry weight)/kg body weight caused significant reductions in the in vivo genotoxicity of mitomycin C, cyclophosphamide and procarbazine but not adriamycin. The inhibitory effect was significant when the coffee was given about 2 hr before the genotoxin; there was a lesser effect when coffee was given together with the genotoxin but there was no inhibition when coffee was given 2-4 hr after the genotoxin. An experiment with mitomycin C demonstrated that the reduction in genotoxicity was dependent on the coffee dose. The inhibition of genotoxicity by coffee was observed in bone-marrow cells sampled 24, 48 or 68 hr after injecting cyclophosphamide. Freshly brewed coffee extract, standard instant coffee, decaffeinated instant coffee and freeze-dried home-brew coffee all exerted inhibitory effects.