Absence Of S9 Mixture Research Articles

Co-exposure to deltamethrin and thiacloprid induces cytotoxicity and oxidative stress in human lung cells.

Deltamethrin (DEL) and thiacloprid (THIA) are commonly used synthetic insecticides in agriculture either separately or in combination. There is limited information in human cells for the effects of the mixture of DEL + THIA on oxidative stress. Therefore, the present study was designed to examine the effects of the mixture on cell proliferation and oxidative stress in human lung fibroblast cells. Human telomerase reverse transcriptase (hTERT)-expressing human lung fibroblasts, WTHBF-6 cells, were treated with 2.5 + 37.5, 5 + 75, 12.5 + 187.5, and 25 +375 µM concentrations of DEL + THIA for the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay and 5 + 75, 12.5 + 187.5, and 25 + 375 µM for lipid peroxidation and reduced glutathione (GSH) assays for 24, 48, and 72 h in the absence and presence of metabolizing fractions of the mammalian liver (S9 mixture). Both the mixture of DEL + THIA and their metabolites significantly reduced cell viability and induced cytotoxicity in WTHBF-6 cells, especially at higher concentrations. The mixture of DEL + THIA significantly decreased GSH levels at the highest concentration for all treatment times and at the highest two concentrations (12.5 + 187.5 and 25 + 375 µM) for 72 h in the presence of S9 mixture. The highest concentration of DEL + THIA mixture caused a significant increase in malondialdehyde (MDA) level at 72 h in the absence of S9 mixture. There were also significant increases in MDA levels at the highest concentration for 48-h and all concentrations of DEL + THIA for 72-h treatment in WTHBF-6 cell cultures with S9. These data showed that the mixture of DEL + THIA and their metabolites can induce cytotoxicity and oxidative stress in human lung fibroblasts.

Cytotoxicity and genotoxicity of Hevea brasiliensis latex C-serum DCS sub-fraction as anticancer agents

Dialysed latex C-serum supernatant (DCS) sub-fraction from the rubber tree (Hevea brasiliensis) was reported to exert significant anti-proliferative effect on specific cancer cell lines, especially in human triple negative breast cancer (MDA-MB-231) and human liver carcinoma (HepG2) cells. In the present study, two human non-cancer origin cell lines, MCF-10A (human breast epithelial cells) and HDFa (human adult dermal fibroblast cells), showed least susceptibility when treated with DCS sub-fraction at concentration range 0–100 µg/mL using 2,5-diphenyl tetrazolium bromide (MTT) assay. Genotoxicity of DCS sub-fraction was assessed through Ames test, in vitro mouse lymphoma assay, and in vitro micronucleus assay. All genotoxicity experiments were tested with the presence and absence of S9 mixture as metabolic activation system. Ames test results showed negative result of mutagenicity as no clear dose-dependent relationship was observed despite mutagenic potential was detected in frame shift-based TA98 and TA1535 Salmonella typhimurium strains. Mouse lymphoma assay produced non-linear dose-dependent effect in experiments without the presence of S9. Meanwhile, experiment with the presence of S9 resulted in an inverse effect of the dose-dependent relationship. In micronucleus assay, the dose-dependent cytotoxicity effect of DCS sub-fraction showed no correlation with the increase number of cells containing micronuclei but appeared to be time-dependent. In conclusion, DCS sub-fraction showed no significant positive results in genotoxicity assays. In vivo genotoxicity assays should be conducted to ensure the safe use of DCS sub-fraction in further exploration for biological applications.

Antimutagenic and Multi-Biological Activities of Smilax excelsa L. Fruit Extract

In this study, antimutagenic, antimicrobial and antioxidant activities of Smilax excelsa L. fruit extract were evaluated. The antimicrobial effect was investigated by disk diffusion method and the antimutagenic effect was investigated by Ames/Salmonella/microsom test. The antioxidant properties of S. excelsa fruit samples were determined by investigating the total phenolic, flavonoid contents and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. It was observed that 200 mg plate-1 fruit extract was found to have a mutagenicity inhibition as 81% in the absence of S9 mixture and 67% in the presence of S9 mixture against Salmonella typhimurium TA1535. S. excelsa fruit extract produced an inhibition zone in the range of 11-16 cm against the tested microorganisms. Flavonoid and phenolic contents were found to be 0.7985±0.0124 mgQE 100 mL-1 and 11.9847±0.0041 mgGAE 100 mL-1 at the 200 mg mL-1 extract concentration, respectively. The DPPH removal rate was determined to be 55% at the 200 mg L-1 dose. As a result, it was observed that the S. excelsa fruit tissues exhibited a highly antimutagenic activity and has been determined as a potential natural antimicrobial and antioxidant source.

Potential role of mitochondrial damage and S9 mixture including metabolic enzymes in ZnO nanoparticles-induced oxidative stress and genotoxicity in Chinese hamster lung (CHL/IU) cells

The present study was designed to examine genotoxicity induced by 10–40 nm zinc oxide (ZnO) nanoparticles using the in vitro system. The frequency of micronuclei was significantly increased in a dose-dependent manner when cultured Chinese hamster lung (CHL/IU) cells were exposed to ZnO nanoparticles for 24, 48 and 72 h in the continuous treatment method. The maximal frequency of micronuclei was observed in exposure of CHL/IU cells to ZnO nanoparticles at a concentration of 125 μM. The frequency of micronuclei was profoundly enhanced when CHL/IU cells were exposed to ZnO nanoparticles in the presence vs. absence of S9 mixture including metabolic enzymes in the short-term treatment method, demonstrating an increase in the formation of micronuclei by S9 mixture. The maximal frequency of micronuclei was seen in exposure of CHL/IU cells to ZnO nanoparticles at a concentration of 140 μM. Similar results were obtained in chromosome aberrations, particularly structural aberrations. Surprisingly, administration of the superoxide radical scavenger, tempol, completely abolished an increase in the frequency of micronuclei in the presence or absence of S9 mixture, indicating a central role of superoxide radical in the formation of micronuclei. Indeed, reactive oxygen species (ROS) generation was elevated by simultaneous incubation of S9 mixture and ZnO nanoparticles and by exposure of CHL/IU cells to ZnO nanoparticles in the presence or absence of S9 mixture. An electron microscopic examination revealed mitochondrial damage in CHL/IU cells exposed to ZnO nanoparticles, indicating the participation of mitochondrial dysfunction in ROS generation in this setting. These observations suggest that ZnO nanoparticles evoke genotoxicity through superoxide radical-induced oxidative stress derived from mitochondrial damage in CHL/IU cells. S9 mixture appears to contribute to a further increase in genotoxicity through the generation of superoxide radical by metabolic activation of ZnO nanoparticles.

Evaluation of the Mutagenic Properties of Two Lignans from Acanthopanax koreanum Nakai

Acanthopanax koreanum Nakai, a well known traditional herb grown in Jeju Island, South of Korea, has been used as a tonic and sedative agent, as well as in the treatment of diabetes and immune diseases. Mutagenicity of two lignans, syringaresinol and tortoside A isolated from A. koreanum, was assessed using Salmonella/microsome (Ames) test. Tester strains used were Salmonella typhimurium TA98, TA100, TA1535, and Escherichia coli WP2uvrA. The mutagenic activity was determined both in the absence or presence of S9 mixture. As a result, tortoside A did not cause any increase in the number of his+ revertants in S. typhimurium and E. coli WP2uvrA strains in the presence or absence of S9 mix, compared to the controls. Similarly, low concentrations of syringaresinol (750 and 1,500 μg/plate) did not show any mutagenic properties in all bacterial strains, in the presence or absence of S9 mixture. However, in the high concentration of syringaresinol (3,000 μg/plate), the number of revertants were increased in TA1535 strains, in the absence of S9 metabolic activation. Therefore, in vivo experiments such as comet assay are needed to further determine the genotoxic/carciogenic potential of syringaresinol isolated from A. koreanum.

The Antimutagenic Effect of Mistletoe Lectin (Viscum album L. var. coloratum agglutinin)

A galactose- and N-acetyl-D-galactosamine-specific lectin (Viscum album L. var. coloratum agglutinin, VCA), which is known for its anticancer activity, was isolated from mistletoe. In this study, we investigated the antimutagenic potentials of VCA by using the pre-incubation method of the Ames test (Salmonella typhimurium TA98 and TA100) in the presence or absence of S9 mixture. Viscum album L. var. coloratum agglutinin was assessed for its antimutagenic properties against the mutagens 2-aminoanthracene (2AA) and furylfuramide (AF-2) for strain TA98, and sodium azide (NaN(3) ) and 2-aminoanthracene (2AA) for strain TA100. The concentrations used for this test compound were 100, 200 and 400 µg per plate. Viscum album L. var. coloratum agglutinin showed moderate, but not negligible, protective effects regarding the antimutagenic properties against the direct-acting mutagens NaN(3) and AF-2. Furthermore, VCA was more effective in preventing the mutagenicity of the indirect-acting mutagen 2-AA (in the presence of S9) when tested with both TA98 and TA100. In conclusion, this report has shown broad ranging antimutagenic effects of VCA to numerous mutagens in TA98 and TA100 Salmonella typhimurium strains. Although the data presented here cannot be applied in vivo, they can support other antimutagenic and anticarcinogenic findings for VCA.

Antimutagenic potential of the Thai herb, Mucuna collettii Lace

Mucuna collettii Lace is a Thai herb with a long record of consumption among mature Thai males for the promotion of sexual potency. The mutagenic and antimutagenic potentials of Mucuna collettii extract were carried out by using the Ames test pre-incubation method in the presence and absence of S9 mixture. Salmonella typhimurium strains TA 98 and TA 100 were applied as the tester strains. Prior to mutagenic and antimutagenic tests, the survival of the tester strains was performed by treating with the plant extract. Results showed Mucuna collettii extract exhibited strong cytotoxic effects in a dose-dependent manner. Toxicity of the plant was confirmed in mice in which negative adverse effect was found in kidney, uterus, ovary, and testis. Mucuna collettii extract in the presence and absence of S9 mixture was negative for mutagenic Ames test. Mucuna collettii extract in the presence and absence of S9 mixture was positive for antimutagenic Ames test towards either one or both of the tested mutagens: 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2) and benzo(a)pyrene. The antimutagenic activity of the plant extract was confirmed in rec-assays. Micronucleus test demonstrated that Mucuna collettii extract at high dose and a long incubation time could induce micronucleus formation in tested animals, but less than the response of the positive control. The overall mutagenic and antimutagenic assays are further evidences for the antimutagenic potential of Mucuna collettii.

Rec effect of certain textile dyes in Bacillus subtilis

A large number of compounds are toxic, genotoxic, mutagenic, teratogenic and/or carcinogenic. The genotoxicity of four textile dyes commonly used in India namely Sulphur Red Brown 360 (SRB), Jade Green 2G (JG), Reactofix Turquoise Blue 5GFL (RTB) and Direct Scarlet 4BS (DS) was determined by Bacillus subtilis spore Rec assay, both in the presence and absence of metabolizing activation mixture (S9 mix). Each dye was toxic at higher dose levels. A dose-dependent increase in the depth of growth inhibition zones was observed for all dyes. Zones of inhibition were usually clearer at higher doses of the dyes and with Rec− bacteria, but were translucent with Rec+ bacteria. SRB and DS were toxic to Rec+ and Rec− bacteria. JG was less genotoxic in the absence of S9 mix, however, its genotoxic potential increased in the presence of S9 mix. Reactofix T blue was more genotoxic in the absence of S9 mixture.

Mutagenicity monitoring of airborne particulate matter (PM10) in the Czech Republic

The mutagenic activities associated with inhalable airborne particulate matter (PM10) collected over a year in four towns (Czech Republic) have been determined. The dichloromethane extracts were tested for mutagenicity using the Ames plate incorporation test and the Kado microsuspension test both with Salmonella typhimurium TA98 and its derivative YG1041 tester strains in the presence and absence of S9 mixture. The aim of this study was to assess the suitability of both bacterial mutagenicity tests and to choose the appropriate indicator strain for monitoring purposes. To elucidate the correlation between mutagenicity and polycyclic aromatic hydrocarbons (PAHs), the concentration of PAHs in the air samples were determined by GC/MS. In general, the significant mutagenicity was obtained in organic extracts of all samples, but differences according to the method and tester strain used were observed. In both mutagenicity tests, the extractable organic mass (EOM) exhibited higher mutagenicity in the YG1041 strain (up to 97 rev/μg in the plate incorporation and 568 rev/μg in the microsuspension tests) than those in TA98 (up to 2.2 rev/μg in the plate incorporation and 14.5 rev/μg in the microsuspension tests). In the plate incorporation test, the direct mutagenic activity in YG1041 was on average 60-fold higher and in microsuspension assay 45-fold higher with respect to strain TA98. In the presence of S9 mix, the mutagenic potency in YG1041 declined ( P<0.001) in summer, but increased in TA98 ( P<0.05) in samples collected during the winter season. The microsuspension assay provided higher mutagenic responses in both tester strains, but in both strains a significant decrease of mutagenic potency was observed in the presence of S9 mix ( P<0.001 for YG1041, P<0.05 for TA98 in winter). The mutagenic potencies detected with both indicator strains correlated well ( r=0.54 to 0.87) within each mutagenicity test used but not (for TA98) or moderately ( r=0.44 to 0.66 for YG1041) between both of the tests. The mutagenic activity (in rev/m 3) likewise the concentration of benzo[ a]pyrene and sum of carcinogenic PAHs showed seasonal variation with distinctly higher values during winter season. A correlation between the PAH concentrations and the mutagenicity results for the plate incorporation, but not for the microsuspension tests was found. In samples from higher industrial areas, the higher mutagenicity values were obtained in plate incorporation test with TA98 and in both tests with YG1041 in summer season ( P<0.05). According to our results, plate incorporation test seems to be more informative than microsuspension assay. For routine ambient air mutagenicity monitoring, the use of YG1041 tester strain without metabolic activation and the plate incorporation test are to be recommended.

IN VITRO CHROMOSOME ABERRATION ASSAY USING HUMAN BRONCHIAL EPITHELIAL CELLS

The determination of chromosome aberrations (CA) in Chinese hamster lung fibroblast (CHL) cells was compared with that in normal human bronchial epithelial (BEAS-2B) cells, which upon adenovirus infection were reported to possess carcinogen metabolizing capacities similar to polycyclic aromatic hydrocarbons. CHL and BEAS-2B cells were treated with increasing concentrations of benzo\\[a]pyrene ( BaP) or N -nitrosodiethylamine (NDEA) . In BEAS-2B cells, BaP, at a concentration of 50 mug/ml, produced a significant increase in the CA frequency, while NDEA did not markedly alter the number of aberrations in the absence of S9 mixture. The CHL cells exposed to BaP and NDEA in the presence of S9 mixture responded as anticipated with a 30% and 14% frequency of CA observed in the BaP (50 mug/ ml) and NDEA (1000 mug/ ml) treated cells, respectively. The results of this study show that the CA assay using human cell line with intrinsic metabolic activation system, such as BEAS-2B cells, may be a useful model for predicting human clastogens and carcinogens.

Mixed disulfides from disulfiram inhibit the benzo[ a]pyrene induced mutagenesis

Some mixed disulfides derived from disulfiram and endogenous thiol compounds have been synthesized, biochemically characterized and their potential antigenotoxic effects have been proposed. The present study evaluated the mutagenic and antimutagenic specificities of two mixed disulfides using S. typhimurium reversion assay, namely S-( N, N-diethyldithiocarbamoly)- N acetylcysteine (AC-DDTC) and - L-glutathione (GS-DDTC). The two mixed disulfides were not mutagenic to Salmonella strains TA98 and TA100 in the presence or absence of S9 mixture. The increased number of revertants by benzo[ a]pyrene (B(a)P) has been reduced to the control level by the preincubation with AC-DDTC or GS-DDTC. It was not due to the killing effect of B(a)P, mixed disulfides or B(a)P-disulfide mixture. The antimutagenic effect of AC-DDTC was more potent than that of GS-DDTC. These results indicate that AC-DDTC and GS-DDTC may have a role to play in reducing the risk of mutagenic effects of B(a)P.

Suppressive effects of retinoids, carotenoids and antioxidant vitamins on heterocyclic amine-induced umu C gene expression in Salmonella typhimurium (TA 1535/pSK 1002)

Effects of retinoids. carotenoids and antioxidant vitamins were studied by mutagen-induced umu C gene expression system in Salmonella typhimurium (TA 1535/pSK 1002). Retinol (vitamin A), retinol acetate and retinoic acid showed remarkable inhibitory activities, whereas retinol palmitate exhibited significant but weak activity for umu C gene expression in tester bacteria induced by 3-amino-3.4-dimethyl-5 H-pyrido[4.3- b]indol (Trp-P-1) in the presence of hepatic metabolizing enzymes (S9 mixture). Carotenoids having provitamin A activity ( β-carotene and canthaxanthin) exhibited moderate suppressive effects on the same experimental system. The ranks of suppressive activities were retinol > retinol acetate > retinoic acid > canthaxanthin > β-carotene > retinol palmitate and their doses for inhibition by 50% (ID 50) were estimated to be 1.2 × 10 −7, 3.0 × 10 −7, 5.4 × 10 −7, 1.5 × 10 −6, 4.0 × 10 −5 and 6.0 × 10 −5 M, respectively. However, they did not cause significant inhibition on umu C gene expression induced by direct-acting mutagen (adriamycin or mitomycin C) in the absence of S9 mixture. Inhibition of umu gene expression appears to be due to inhibition of P450-mediated metabolic activation of the heterocyclic amine Trp-P-1. Ascorbic acid (vitamin C) showed weak but significant suppressive activity at high-dose concentrations (3 × 10 −6–10 −4M). However, α-tocopherol did not exhibit significant suppression at all dose concentrations. The significance of the experimental results is discussed from the viewpoint of the chemoprevention against genotoxicity associated with carcinogenesis.

Comparison of Antimutagenic Effect of Various Tea Extracts (Green, Oolong, Pouchong, and Black Tea)

The antimutagenic effects of various tea extracts prepared from nonfermented tea (green tea), semifermented tea (oolong tea and pouchong tea), and fermented tea (black tea) were investigated by Salmonella/microsome assay. No mutagenicity or toxicity in Salmonella typhimurium TA98 and TA100 was observed with any tea extract. The tea extracts markedly inhibited the mutagenicity of 2-amino-3-methylimidazo(4,5-f)quinoline, 3-amino-1,4-dimethyl-5H-pyridol(4,3-b)indole,2-amino-6-methyldipyrido(l,2-a:3′,2′-d)imidazole, benzo[a]pyrene, and aflatoxin B1 toward S. typhimurium TA98 and TA100 in the presence of S9 mixture, especially those of oolong and pouchong teas inhibited over 90% mutagenicity of these five mutagens at the dosage of 1 mg per plate. Among four tea extracts, black tea exhibited the weakest inhibitory effect on mutagenicity of these five mutagens. The mutagenicity of 4-nitroquinoline-N-oxide, a direct mutagen, was not inhibited by black and oolong tea extracts to S. typhimurium TA98 in the absence of S9 mixture but was increased by the tea extracts at the dose of 1 mg per plate to S. typhimurium TA100. As the antimutagenic effect of semifermented tea was stronger than nonfermented and fermented teas, some antimutagenic substances might be formed during manufacturing processes of tea.

Mutagenicity evaluation of riot control agent o-chlorobenzylidene malononitrile (CS) in the Ames Salmonella/microsome test.

o-Chlorobenzylidene malononitrile (CS), a riot control agent, was evaluated for its possible mutagenic activity in the Ames Salmonella/mammalian microsome mutagenicity test. Five histidine-deficient (His-) mutant tester strains of Salmonella typhimurium--TA97a, TA98, TA100, TA102 and TA104--were used. The liquid preincubation procedure was used with metabolic activation (presence of S9 mixture) and without metabolic activation (absence of S9 mixture). For the experiments with metabolic activation, three different concentrations of S9 fraction (supernatant of Aroclor 1254-induced rat liver homogenate at 9000 g)--5%, 15% and 30% in S9 mixture--were used. Along with mutagenic activity, CS was also evaluated for cytotoxic activity in all the five tester strains of Salmonella typhimurium, both in the presence and absence of S9 mixture. The mutagenic and cytotoxic activities of CS were assessed by counting the His+ revertant colonies and by counting the microcolonies (His-, auxotrophs in the background lawn), respectively, and the respective mean values were compared with the relative negative (solvent) control. A dose range of 12.5-800 micrograms plate-1 for CS did not induce a mutagenic response either in the presence or absence of S9 mix. No change in the negative mutagenic response of CS has been observed even in the presence of an elevated level of S9 fraction in the S9 mix. A dose of 200 micrograms plate-1 for CS was found to be cytotoxic by decreasing the surviving cells as well as His+ revertant colonies; however, the effect was reduced in the presence of an elevated level of S9 fraction in the S9 mix.

Effects of supercypermethrin, a synthetic developmental pyrethroid, on four biological test systems

The genotoxic potential of the insecticide supercypermethrin, a second-generation pyrethroid, was studied on four different test systems. It was non-mutagenic to Salmonella typhimurium strains TA1535, TA100, TA1538, TA98 and TA97 in the presence and absence of S9 mixture. It induced gene conversion at the tryptophan locus and induced point mutations at the isoleucine locus in Saccharomyces cerevisiae cells. A slight increase in the frequency of aberrant anaphases and telophases in root tips of Hordeum vulgare and Vicia faba was observed, but no genotoxic effects were detected in Drosophila melanogaster.

Mutagenicity of N, N'-dimethylurea and methylamine hydrochloride in the Ames Salmonella/microsome test: Absence of mutagenic response

Methyl isocyanate (MIC) in aqueous solution forms methylamine (MA) and N, N'-dimethylurea (DMU). MA in buffered system further converts into its salt form, methylamine hydrochloride (MAH). Therefore, MAH and DMU were evaluated for their mutagenic activity in the in vitro Ames Salmonella/ microsome mutagenicity test. The liquid preincubation protocol was followed, using tester strains TA98, TA100 and TA104 of Salmonella typhimurium, in the presence of 0, 5, 15 and 30% Aroclor 1254-induced rat liver S9 mixture. DMU and MAH did not induce a mutagenic response in any of the tester strains, both in the presence and in the absence of S9 mixture. The results therefore confirm that MIC in its native form or as its unknown metabolites is responsible for the mutagenic activity reported earlier by us in the his − tester strains TA100 and TA104 of Salmonella typhimurium (Mutation Res., 204 (1988) 123–129) and not due to its hydrolysis products, MA or DMU.

The evaluation of genotoxic activities of disinfectants and their metabolites by umu test

The genotoxic potential of 6 disinfectants and their 9 metabolites was investigated by umu test. In the tested disinfectants, glutaraldehyde showed positive genotoxicity independent of metabolic activation system and acrinol was positive only in the presence of S9 mixture. Alkylaminoethylglycine, benzalkonium chloride, chlorhexidine digluconate and methylrosaniline chloride were negative in the presence or absence of S9 mixture. In some metabolites of benzalkonium chloride, chlorhexidine digluconate or glutaraldehyde, only pyrogallol showed positive genotoxicity in the absence of S9 mixture and the activity was not affected by the metabolic activation system. Aniline, p-chloroacetoanilide, p-chloroaniline, p-chlorophenol, decabutyldimethylamine, glutaric acid, phenol and pyrocatechol did not induce umu gene expression independently of the presence of S9 mixture. The results in the umu test of these compounds were compared with their findings in the liquid rec-assay and Ames test. The umu test is a more useful and simplified method for the detection of genotoxicity of the compounds with killing effects on tester bacteria.

DNA repair synthesis in primary culture of bovine bile duct epithelial cells induced by chemical agents in relation to bile duct cancer

Unscheduled DNA synthesis (UDS) was measured autoradiographically in a primary culture of extrahepatic bile duct epithelial cells of Holstein cows following exposure to chemicals known to be capable of developing bile duct cancers in experimental animals, i.e., N-methyl- N′-nitro- N-nitrosoguanidine (MNNG, CAS No. 70-25-7), N-ethyl- N′-nitro- N-nitrosoguanidine (ENNG, CAS No. 4255-77-6), 20-methylcholanthrene (MCA, CAS No. 56-49-5), N-nitrosodimethylamine (DMN, CAS No. 62-75-9) and aflatoxin B 1 (AFB 1, CAS No. 1162-65-8). MNNG and ENNG induced UDS without addition of S9 mixture. MCA elicited UDS only if S9 mixture was added. Regardless of the presence or absence of S9 mixture, DMN failed to induce UDS. DNA repair by AFB 1 was enhanced by the presence of S9 mixture. Therefore, for MNNG, ENNG and high doses of AFB 1 activation by the liver is not necessary to exert genotoxic effects and they seem to be capable of direct action on bile duct epithelial cells in the presence of a bilioenteric fistula or anastomosis.

Induction of sister chromatid exchange by various selenium compounds in Chinese hamster cells in the presence and absence of S9 mixture

This study was designed to investigate the effect of 3 selenium compounds, namely, sodium selenite, sodium selenide and sodium selenate on induction of sister chromatid exchange (SCE) in the Chinese hamster V79 cell line in the presence and absence of S9 mixture. The results indicated that the most potent SCE inducer in the presence of S9 mixture was sodium selenite and this was followed by sodium selenide, while in the absence of S9 mixture the most effective SCE inducer was sodium selenide which was then followed by sodium selenite. For sodium selenate, the data indicated no increase in SCE rate as compared to the control values both in the presence and absence of S9 mixture. In addition, it was observed that growth inhibition as measured by no sister chromatid differentiation or no metaphases was produced by certain doses of the compounds tested. Based on this, the compound that induced the most growth inhibition both in the presence and absence of S9 mixture was sodium selenide and this was followed by sodium selenite. For sodium selenate no growth inhibition was observed. The different capabilities of the selenium compounds tested to induce SCE were clearly demonstrated in the system employed in this study. It is felt that this effect is an important cytogenetic characteristic of these compounds, yet how this activity relates to the antimutagenic and anticarcinogenic properties of these agents is difficult to discern.

Correlation of mutagenicity and tumorigenicity of betel quid and its ingredients.

The mutagenic activity of betel quid and its ingredients was determined using Salmonella typhimurium tester strains TA 100, TA 1535, TA 98, and TA 1538, both in the presence and absence of S9 mixture. Aqueous extracts of betel quid (BQ), betel quid with tobacco (BQT), and betel nut (BN) were mutagenic in strain TA 100. Aqueous extract of betel leaf (BL) was not mutagenic in any of the four strains. Arecoline and arecaidine, which are major alkaloids present in BN, were mutagenic in all four tester strains. Tumorigenicity studies in Swiss mice given the above constituents showed that BN and BQ induced lung tumors (47% and 26%, respectively). However, when BN was fed with BL, tumorigenicity was lowered to 38%. BL alone was not tumorigenic. Thus, the mutagenicity of betel quid and its ingredients is correlated with tumorigenicity.