Mechanism Of Antimutagenicity Research Articles

Interaction of heavy metals in Drosophila melanogaster larvae: Fourier transform infrared spectroscopy and single-cell electrophoresis study

The present study evaluates the Murraya Koenigii (CuLE) and Tinospora Crispa (TiSE) antimutagenic effect and the impact of industrial soil and solid waste leachate on Drosophila larvae. Larvae were exposed to leachate prepared at different pH (7, 4.93, 2.88) and treated with TiSE and CuLE at different concentration (4 g/L and 6 g/L) mixed with standard Drosophila medium. Emphasis was given to the binding interaction of heavy metals with proteins in Drosophila. The change in structure and molecular composition in Drosophila by leachate containing heavy metals induced toxicity has been studied by using Fourier transform infrared (FTIR) spectroscopy. Results from the study demonstrated that CuLE/TiSE administration restored the level of oxidative stress as evidenced by an enhanced antioxidant system and a decrease in lipid peroxidation and protein oxidation. The amide I and amide II bands spectral shifting revealed the binding interaction. The shift in the peak of PO2− asymmetric stretching might be due to compositional changes in nucleic acids. Single-cell electrophoresis was performed to detect the DNA damage which also proved to be ameliorated by administration of CuLE/TiSE. The result concludes that CuLE/TiSE may have great potential in the protection of Drosophila larvae from leachate induced oxidative stress through antioxidant and antimutagenic mechanisms this might help to cope with environmental toxicants. Communicated by Ramaswamy H. Sarma

Protective Effect ofNigella sativaandNigella damascenaFixed Oils Against Aflatoxin Induced Mutagenicity in the Classical and Modified Ames Test

The antioxidant and mutagenic/antimutagenic activities of the fixed oils from Nigella sativa (NSO) and Nigella damascena (NDO) seeds, obtained by cold press-extraction from the cultivar samples, were comparatively investigated for the first time. The antimutagenicity test was carried out using classical and modified Ames tests. The fatty acid composition of the fixed oils was characterized by gas chromatography-mass spectrometry (GC-MS) while the quantification of thymoquinone in the fixed oils was determined by UPC2 . The main components of the NSO and NDO were found to be linoleic acid, oleic acid, and palmitic acid. The results of the Ames test confirmed the safety of NSO and NDO from the viewpoint of mutagenicity. The results of the three antioxidant test methods were correlated with each other, indicating NDO as having a superior antioxidant activity, when compared to the NSO. Both NSO and NDO exhibited a significant protective effect against the mutagenicity induced by aflatoxin B1 in Salmonella typhimurium TA98 and TA100 strains. When microsomal metabolism was terminated after metabolic activation of the mycotoxin, a significant increase in antimutagenic activity was observed, suggesting that the degradation of aflatoxin B1 epoxides by these oils may be a possible antimutagenic mechanism. It is worthy to note that this is the first study to assess the mutagenicity of NSO and NDO according to the OECD 471 guideline and to investigate antimutagenicity of NDO in comparison to NSO against aflatoxin.

Molecular mechanism of antimutagenicity by an ethoxy-substituted phylloquinone (vitamin K1 derivative) from spinach (Spinacea oleracea L.)

In our previous study, an antimutagenic compound from spinach (Spinacea oleracea L.), ethoxy-substituted phylloquinone (ESP) was isolated and characterized. The current study deals with elucidation of the possible mechanism of antimutagenicity of ESP against ethyl methanesulfonate (EMS) deploying model systems such as human lymphoblast (TK+/- or TK6) cell line (thymidine kinase gene mutation assay) and Escherichia coli MG1655 (rifampicin resistance assay). Findings of the study ruled out the possibility of direct inactivation of EMS by ESP. DAPI competitive binding assay indicated the DNA minor groove binding activity of ESP. Interestingly, ESP did not display major groove binding or intercalating abilities. Further, proteomics study using 2-D gel electrophoresis in E. coli and subsequent studies involving single gene knockout strains revealed the possible role of tnaA (tryptophanase) and dgcP (diguanylate cyclase) genes in observed antimutagenicity. These genes have been reported to be involved in indole and cyclic-di-GMP biosynthesis, respectively, which eventually lead to cell division inhibition. In case of TK+/- cell line system, ADCY genes (adenylate cyclase), a functional analogue of dgcP gene, were found to be transcriptionally up-regulated. The generation/doubling time were significantly higher in E. coli or TK+/- cells treated with ESP than control cells. The findings indicated inhibition of cell proliferation by ESP through gene regulation as a possible mechanism of antimutagenicity across the biological system. Cell division inhibition actually provides additional time for the repair of damaged DNA leading to antimutagenicity.

Antimutagenic components in Spatholobus suberectus Dunn against N-methyl-N-nitrosourea

BackgroundAn extract from Spatholobus suberectus (S. suberectus) Dunn has been reported to show potent antimutagenic effects against N-alkyl-N-nitrosoureas in umu screening. The aim of this study was to identify the antimutagenic components from extracts of S. suberectus against N-methyl-N-nitrosourea (MNU) in the Ames assay with Salmonella typhimurium strain TA1535 and to elucidate the antimutagenic mechanism of the flavonoids.ResultsFrom the ethyl acetate fraction obtained from fractionation of the methanol extract of S. suberectus Dunn, medicarpin, formononetin and isoliquiritigenin were successfully isolated through a combination of normal- and reversed-phase chromatography. Genistein and naringenin, which were already reported to be contained in S. suberectus Dunn, were also tested for their antimutagenicity towards MNU, along with formononetin, isoliquiritigenin and medicarpin. Our results demonstrated that genistein, isoliquiritigenin, medicarpin and naringenin were antimutagenic against MNU without showing cytotoxicity. MNU is reported to cause not only DNA alkylation but also induce reactive oxygen species. The hydroxyl radical scavenging capacity of the flavonoids was correlated with the antimutagenic capacity, indicating that the hydroxyl radical scavenging activity was involved in their antimutagenicity towards MNU.ConclusionsIt is important to prevent DNA damage by N-nitrosamines for cancer chemoprevention. Genistein, isoliquiritigenin, medicarpin and naringenin were demonstrated to possess an antigenotoxic effects against carcinogenic MNU due to their radical scavenging activity.

Genotoxicity inhibition by Syzygium cumini (L.) seed fraction and rutin: understanding the underlying mechanism of DNA protection.

Considering the ethnopharmacological importance of Syzygium cumini's seed and the lack of information on the antimutagenic and DNA-protecting mechanisms, a fraction-based study was conducted. Four different (hexane, chloroform, ethyl acetate, and aqueous) fractions were obtained from the sequential extraction of the methanolic extract of the seed. The most active antioxidant fraction (ethyl acetate) contained significant amount of phenolics and flavonoids. LC-qTOF-MS analysis of the ethyl acetate fraction revealed the presence of rutin, myricetin, naringin, cuscohygrin, and epoxycarryophyllone as constituent phytocompounds. The ethyl acetate fraction (100 μg ml-1) and a selected compound (rutin, 40 μg ml-1) showed remarkable decrease in the revertants frequency range from 74-77% and 66-84%, respectively, against both the mutagens (sodium azide (NaN3) and methyl methane sulfonate (MMS)) in the Salmonella typhimurium tester strains. All the statistical analyses were at a significance level of 0.05 between the different treatment groups. Moreover, the underlying mechanism of antimutagenicity using different treatment regime for rutin was explored. MMS-mediated DNA fragmentation and oxidation in lymphocytes were also shown to be decreased significantly when treated with the ethyl acetate fraction and rutin. Oxidative damage to pBR322 plasmid DNA was also reduced when incubated with different concentration of the ethyl acetate fraction and rutin. Biophysical (UV, fluorescence, ITC, etc.) and computational methods were employed to obtain a closer look at the DNA-rutin interaction. The data obtained clearly revealed that the ethyl acetate fraction exhibited promising antimutagenic and DNA-protective activity and its flavonoid constituents, including rutin, contribute significantly to the observed activity.

Dietary prevention of Helicobacter pylori-associated gastric cancer with kimchi.

To prove whether dietary intervention can prevent Helicobacter pylori-induced atrophic gastritis and gastric cancer, we developed cancer preventive kimchi (cpKimchi) through special recipe and administered to chronic H. pylori-initiated, high salt diet-promoted, gastric tumorigenesis mice model. H. pylori-infected C57BL/6 mice were administered with cpKimchi mixed in drinking water up to 36 weeks. Gross and pathological gastric lesions were evaluated after 24 and 36 weeks, respectively and explored underlying molecular changes to explain efficacies. Cancer preventive actions of anti-inflammation and anti-mutagenesis were compared between standard recipe kimchi (sKimchi) and special recipe cpKimchi in in vitro H. pylori-infected cell model. The erythematous and nodular changes, mucosal ulcerative and erosive lesions in the stomach were noted at 24th weeks, but cpKimchi administration significantly ameliorated. After 36th weeks, scattered nodular masses, some ulcers, and thin nodular gastric mucosa were noted in H. pylori-infected mice, whereas these gross lesions were significantly attenuated in cpKimchi group. On molecular analysis, significant expressions of COX-2 and IL-6, activated NF-κB and STAT3, increased apoptosis, and marked oxidative stresses were noted in H. pylori-infected group relevant to tumorigenesis, but these were all significantly attenuated in cpKimchi group. cpKimchi extracts imparted significant selective induction of apoptosis only in cancer cells, led to inhibition of H. pylori-induced proliferation, while no cytotoxicity through significant HO-1 induction in non-transformed gastric cells. In conclusion, daily dietary intake of cpKimchi can be an effective way either to rejuvenate H. pylori-atrophic gastritis or to prevent tumorigenesis supported with the concerted actions of anti-oxidative, anti-inflammatory, and anti-mutagenic mechanisms.

Mutagenicity of cigarette butt waste in the bacterial reverse mutation assay: The protective effects of β-caryophyllene and β-caryophyllene oxide.

Cigarette filters pose a serious litter and toxic waste disposal problem, because of their not biodegradability and to the leaching of toxins in the environment. Therefore, cigarette butts need to be manipulated as special waste, with potential risks to human health and environment. In the present study, the genotoxic potential of a methanol extract from commonly discharged cigarette butts (CBE) was evaluated in the bacterial reverse mutation assay on Salmonella typhimurium TA98 and TA100 and Escherichia coli WP2uvrA strains, both in the absence and presence of the S9 exogenous metabolic activator. Furthermore, the ability of the natural sesquiterpenes β-caryophyllene (CRY) and β-caryophyllene oxide (CRYO) to inhibit the mutagenicity of CBE was studied as a possible preventive strategy. In order to identify the potential antimutagenic mechanisms, three different protocols (pretreatment, cotreatment, and posttreatment) were applied. CBE showed to increase the number of revertant colonies in all the strains tested in presence of S9, so resulting mutagenic. In the antimutagenicity assay, both CRY and CRYO significantly reduced the revertant colonies induced by CBE, although with different potency and specificity. For both sesquiterpenes, the antimutagenicity was strong in all experimental conditions, except for the cotreatment of CRY with CBE in WP2uvrA, which produced a moderate inhibition. Both desmutagenic and bioantimutagenic mechanisms seem to be involved in the antimutagenicity of the test substances. Taking into account the potential genotoxicity of cigarette butts, CRY and CRYO appear as possible further candidates as environmental decontaminants against this hazardous waste. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1319-1328, 2016.

Mutagenicity and antimutagenicity of hydrophilic and lipophilic extracts of Thai northern purple rice.

Purple rice (Oryza sativa L. var. indica) cv. Kum Doisaket is cultivated in northern Thailand. This study evaluated the mutagenic and antimutagenic properties of hydrophilic and lipophilic components of purple rice using the Ames test. The seed and hull of purple rice were extracted with hexane, methanol, ethanol, and water. The methanol extracts had the highest amounts of phenolic acids and flavonoids, while the hexane extracts contained large amount of tocols and γ-oryzanol. None of the extracts were mutagenic in Salmonella typhimurium strains TA98 and TA100. The hexane extract of rice hull and the methanol extract of rice seed were strongly effective against aflatoxin B1- and 2-amino-3, 4 dimethylimidazo (4, 5-f) quinoline-induced mutagenesis, while aqueous extracts showed weakly antimutagenic properties. All extracts with the exception of aqueous extracts enhanced the number of revertant colonies from benzo (a) pyrene induced-mutagenesis. None of the extracts inhibited mutagenesis induced by the direct mutagens 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide and sodium azide. The hull extracts showed more potent antimutagenicity than the seed extracts. Based on a chemical analysis, γ-oryzanol and γ-tocotrienol in the hull and cyanidin-3-glucoside and peonidin-3-glucoside in the seed are candidate antimutagens in purple rice. The antimutagenic mechanisms of purple rice might be related to either modulation of mutagen metabolizing enzymes or direct attack on electrophiles. These findings supported the use of Thai purple rice as a cancer chemopreventive agent.

Antimutagenic compounds and their possible mechanisms of action

Mutagenicity refers to the induction of permanent changes in the DNA sequence of an organism, which may result in a heritable change in the characteristics of living systems. Antimutagenic agents are able to counteract the effects of mutagens. This group of agents includes both natural and synthetic compounds. Based on their mechanism of action among antimutagens, several classes of compounds may be distinguished. These are compounds with antioxidant activity; compounds that inhibit the activation of mutagens; blocking agents; as well as compounds characterized with several modes of action. It was reported previously that several antitumor compounds act through the antimutagenic mechanism. Hence, searching for antimutagenic compounds represents a rapidly expanding field of cancer research. It may be observed that, in recent years, many publications were focused on the screening of both natural and synthetic compounds for their beneficial muta/antimutagenicity profile. Thus, the present review attempts to give a brief outline on substances presenting antimutagenic potency and their possible mechanism of action. Additionally, in the present paper, a screening strategy for mutagenicity testing was presented and the characteristics of the most widely used antimutagenicity assays were described.

Paradoxically Augmented Anti-Tumorigenic Action of Proton Pump Inhibitor and Gastrin in APCMin/+ Intestinal Polyposis Model1

Though long-term administration of proton pump inhibitor (PPI) imposed the risk of gastrointestinal track tumorigenesis by accompanied hypergastrinemia, no overt increases of colon cancer risk were witnessed after a long-term cohort study. Our recent investigation revealed that PPI prevented colitis-associated carcinogenesis through antiinflammatory, anti-oxidative, and anti-mutagenic mechanisms in spite of hypergastrinemia. Therefore, we hypothesized that PPI might either antagonize the trophic action of gastrin on gastrointestinal tumorigenesis or synergize to exert augmented anti-tumorigenic actions.We challenged APCMin/+ mice with gastrin, PPI, PPI and gastrin together for 10 weeks and counted intestinal polyposis accompanied with molecular changes. Gastrin significantly increased intestinal polyposis, but combination of PPI and gastrin markedly attenuated intestinal polyposis compared to gastrinpromoted APCMin/+ mice (P < .001), in which significant β-catenin phosphorylation and inhibition of β-catenin nuclear translocation were observed with PPI alone or combination of PPI and gastrin, whereas gastrin treatment significantly increased β-catenin nuclear translocation. Significant footprints of apoptosis, G0/G1 accumulation, inactivation of p38 and extracellular signal-regulated kinase, decreased expressions of CD31, and inhibition of tumor necrosis factor-α and cyclooxygenase-2 were noted in the combination group. In vitro investigations were similar to in vivo findings as shown that PPI treatment inhibited the binding of gastrin to its receptor, inactivated β-catenin-associated signaling including Tcf/Lef and glycogen synthase kinase β, and paradoxically inhibited β-catenin-associated proliferative activities. Our investigations explain why colon cancer risk has not increased despite long-term use of PPIs and provide a rationale for using PPI to achieve anti-tumorigenesis beyond acid suppression.

Suppression of SOS repair in E. coli: possible mechanism of antimutagenicity and protective effects of common vegetables

This study aims to understand variations in antimutagenic potential of vegetables, as observed in reduction of UV-induced mutation (RifS→RifR) in Escherichia coli cells. On further investigation, the juice of vegetables [eggplant (small-violet), pepper (hot Arbol), bean (French), and tomato] was found to suppress mutagenic SOS response as measured by cell filamentation, LexA degradation, and induction of defective prophage as per their antimutagenic potential. Cell filamentation which was observed in 25 and 60% of the UV- and gamma-induced cells, reduced to 2–8%, and 3–16%, respectively in the presence of the vegetable juice; moreover, LexA was also not significantly affected. Phage induction frequency reduced upto 76% compared to control UV-exposed cells. The antimutagenic effect was found to be partially dependent on recB, ruvB gene functions, and was independent of uvrA function. Phenolic compounds were found to be the major contributors to the observed antimutagenicity.

Comparative Evaluation of Antimutagenicity of Commonly Consumed Fruits and Activity-Guided Identification of Bioactive Principles from the Most Potent Fruit, Java Plum (Syzygium cumini)

Commonly consumed fruits showed remarkable variation in antimutagenicity when assayed by E. coli rifampicin resistance assay, and Java plum (Syzygium cumini) was found to be one of the most potent fruits. Its anthocyanins contributed maximally to the observed antimutagenicity and resolved into three distinct bands in HPTLC. Although these bands displayed similar antioxidant capacity, the band at R(f) 0.22 was the most antimutagenic and resolved into two peaks in HPLC. The second peak (t(R) 3.8 min) displayed a strong and broad spectrum antimutagenicity and was identified as petunidin-3,5-diglucoside by analysis of its molecular ion and fragmentation pattern by ESI-MS/MS. The presence of glucose moiety was confirmed by TLC analysis of acid-hydrolyzed products. This purified anthocyanin was found to suppress mutagenic SOS DNA repair process in E. coli and thus indicated suppression of the error-prone DNA repair pathway as one of the major mechanisms of antimutagenicity of this fruit.

The antimutagenic and antioxidant properties of Spirogyra neglecta (Hassall) Kützing

Spirogyra species is a freshwater filamentous green alga. Consumed as food in Northern Thailand, the algae have shown hypoglycemic ability in rats. The aims of this study were to investigate the chemical composition, antioxidant and antimutagenic activities in vitro of the aqueous extract of Spirogyra neglecta (Hassall) Kutzing. The antioxidant activity was evaluated by assessing the inhibition of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals. The IC50 value of S. neglecta extract was 48.67±3.06 µg/ml in the DPPH scavenging assay. The mutagenicity and antimutagenicity were determined via the Ames test with and without metabolic activation. The extract did not enhance the numbers of revertant colonies ofSalmonella typhimurium strains TA98 and TA100 in the absence and presence of metabolic activation; this indicates non-mutagenicity. The extract was strongly effective in reducing the mutagenicity induced by 2-aminoanthracene (2-AA), 2-aminofluorene (2-AF), aflatoxin B1 (AFB1) and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ) and weakly inhibited mutagenesis induced by 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). The algae extract did not reduce the numbers of revertant colonies induced by a direct mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)-acrylamide (AF-2) and NaN3. The water extract of S. neglectashowed antioxidant activity and protection from indirect mutagens in S. typhimurium. The possible antimutagenic mechanism of S. neglecta aqueous extract might be due to the inhibition of some metabolizing enzymes of mutagen metabolism Key words: Ames test, antimutagenicity, antioxidant, Spirogyra species.

Antimutagenicity mechanisms of the Rhoeo discolor ethanolic extract

Introduction Rhoeo discolor, a medical plant from Mexico, is known to be an antioxidant and chemoprotective antimutagen. Rhoeo discolor ethanolic extract (EERD) is a complex mixture, so in this study its antimutagenic mechanisms were further evaluated. Material and methods Employing Ames test, with uvrB− and uvrB+ strains, its antimutagenic activity against frameshift mutagens 2-amino-anthracene (AA), and 2 amino-fluorene (AF), alkylating: methyl-N′-nitro-N-nitrosoguanidine (MNNG) and ethyl-N′-nitro-N-nitrosoguanidine (ENNG) and mitomycin C were evaluated. Induction of ogt, alkyl-DNA-glycosylases were studied with Salmonella typhimurium strains deficient in ada and ogt genes (YG7100 ada−/ogt+ YG7104 ada+/ogt−, G7108 ada−/ogt−). Results EERD, was not antimutagenic against AA or AF on S. typhimurium TA98 neither in UTH8413 uvrB+ strains. It significantly reduced mutations induced by Mitomycin C on strain TA102. EERD was antimutagenic to mutations induced by alkylating compounds on S. typhimurium TA100 or UTH8414 uvrB+. This antimutagenic effect was not observed on strains lacking ogt gene. Conclusions EERD, did not affect CYP450 in vitro microsomal activation of AF or AA, on the Ames test, neither improved DNA uvrB excision repair system. EERD reduced oxidative damages on strain TA102, caused by Mitomycin C. This plant extract might be used to avoid DNA damage by alkylation, corrected mainly alkylguanine transferase protein encoded by ogt gene.

Evaluation of the genotoxic and anti-genotoxic activities of Silybin in human hepatoma cells (HepG2)

Silybin (SB), a constituent of the medicinal plant Silybum marianum, is reported to be a potent hepatoprotective agent, but little is currently known regarding its genotoxicity, mutagenicity and potential chemopreventive properties. In this study, we evaluated the ability of SB to induce DNA migration and micronuclei (MN) formation in human hepatoma cells (HepG2). Also, possible preventive effects of SB on MN formation induced by three different mutagens, bleomycin (BLEO), benzo[a]pyrene (B[a]P) and aflatoxin B(1) (AFB(1)), were studied. To clarify the possible mechanism of SB antimutagenicity, three treatment protocols were applied: pretreatment, in which SB was added before the application of the mutagens; simultaneous treatment, in which SB was added during treatment and post-treatment, in which SB was added after the application of the mutagens. At concentrations up to 100 microM, SB was non-genotoxic, while at a concentration of 200 microM, SB induced DNA migration, generated oxidized DNA bases, reduced cell viability, decreased the replicative index of the cells and induced oxidative stress. It is noteworthy that SB was able to reduce the genotoxic effect induced by B[a]P, BLEO and AFB(1) in pretreatment and simultaneous treatments but had no significant effect on DNA damage induction in post-treatment. Taken together, our findings indicate that SB presents anti-genotoxic activity in vitro, which suggests potential use as a chemopreventive agent.

Effect of annatto on micronuclei induction by direct and indirect mutagens in HepG2 cells

Annatto (AN), a natural food colorant rich in carotenoids, has been reported as being an effective antioxidant, but little is known about its potential chemopreventive properties. In this study, we evaluated the ability of AN to protect human hepatoma cells (HepG2) from micronucleus (MN) induction against three different mutagens: benzo(a)pyrene (B(a)P), doxorubicin (DXR), and methyl methanesulfonate (MMS). In an attempt to clarify the possible mechanism of antimutagenicity of AN, three protocols of treatment were applied (pretreatment; simultaneous treatment, and post-treatment with AN following treatment with the mutagens). Also, cells exposed only to AN were assayed for cytotoxicity and mutagenicity. A dosage up to 10 microg/ml of AN was devoid of mutagenic activity. Protective effects were seen on micronuclei induced by B(a)P and DXR using pre and simultaneous treatment, but AN had no significant effect on MN induction by MMS in any of the protocols. Our results also show that exposure of cells to concentrations of AN higher than 10 microg/ml decreased cell viability. Taken together, our findings indicate that AN presents antimutagenic activity in vitro, but its protective effect is dependent on the mutagen and on type of treatment suggesting its potential use as a chemopreventive agent.

Antimutagenic activity of Aspergillus awamori-fermented black soybean response to simulated digestive juice treatments and its antimutagenic mechanisms

Aspergillus awamori-fermented black soybeans, a potential healthy food ingredient, possess antimutagenicity against 4-nitroquinoline- N-oxide (4-NQO); a direct mutagen and benzo[ a]pyrene (B[ a]P; an indirect mutagen). Results of this study revealed that antimutagenic, desmutagenic, and blocking effects all contributed to the antimutagenicity of the methanol of fermented black soybean. However, the desmutagenic effect of the fermented black soybean extract on B[ a]P is not caused by the interaction of intact B[ a]P or S9 mixture with the extract. Instead, it is due to the interaction of B[ a]P metabolites with the antimutagenic factors in the extract. Exposure to simulated gastric juice (pH 1.2) for 30 min or intestinal juice for 2 h resulted in a reduced antimutagenicity of fermented black soybean extract. Nevertheless, the simulated gastric juice treated-fermented black soybean extracts and the simulated intestinal juice treated-fermented black soybean extracts still possessed a considerable antimutagenic effect against 4-NQO and B[ a]P.

Possible Mechanisms of Antimutagenicity in Fermented Soymilk Prepared with a Coculture of Streptococcus infantis and Bifidobacterium infantis

The possible mechanisms of antimutagenicity against 4-nitroquinoline-N-oxide (4-NQO; a direct mutagen) and 3,2′-dimethyl-4-amino-biphenyl (DMAB; an indirect mutagen) were examined in fermented soymilk prepared with a coculture of Streptococcus thermophilus and Bifidobacterium infantis. The antimutagenicity in the fermented soymilk was not due to the bioantimutagenic effect of modulation of DNA repair processes. The mutagenicity of DMAB decreased with increased pre-incubation of fermented soymilk and the DMAB metabolite but not with intact DMAB or an S9 mixture. Mutagenicity of 4-NQO was not affected by preincubation of fermented soymilk with this mutagen. Mutagenicity of both 4-NQO and DMAB was reduced when Salmonella Typhimurium TA 100 was pretreated with fermented soymilk, indicating that fermented soymilk affected the function of the bacterial cell, which might also lead to reduced mutagenicity of the tested mutagens. Desmutagenic and blocking effects were the main mechanisms of antimutagenicity in the fermented soymilk against DMBA. In contrast, the antimutagenic effect of the fermented soymilk on 4-NQO was primarily due to a blocking effect.

Anti-clastogenic effect of b-glucan extracted from barley towards chemically induced DNA damage in rodent cells

beta-Glucan (BG) was tested in vitro to determine its potential clastogenic and/or anti-clastogenic activity, and attempts were made to elucidate its possible mechanism of action by using combinations with an inhibitor of DNA polymerase. The study was carried out on cells deficient (CHO-k1) and cells proficient (HTC) in phases I and II enzymes, and the DNA damage was assessed by the chromosomal aberration assay. BG did not show a clastogenic effect, but was anti-clastogenic in both cell lines used, and at all concentrations tested (2.5, 5 and 10 microg/mL) in combination with damage inducing agents (methylmethane sulfonate in cell line CHO-k1, and methylmethane sulfonate or 2-aminoanthracene in cell line HTC). BG also showed a protective effect in the presence of a DNA polymerase beta inhibitor (cytosine arabinoside-3-phosphate, Ara-C), demonstrating that BG does not act through an anti-mutagenic mechanism of action involving DNA polymerase beta.

Usefulness of the SOS Chromotest in the study of medicinal plants as radioprotectors

Purpose: The aim of this work is to investigate the usefulness of a modified protocol of the SOS Chromotest to detect antigenotoxicity activities against γ-rays of plant extracts with proven antioxidant activity, and to elucidate the antigenotoxic mechanisms involved in radioprotection using this system.Materials and methods: The methodology developed was assayed with amifostine, the most studied radioprotector, and with Phyllanthus orbicularis HBK, Cymbopogon citratus (DC) Stapf and Pinus caribaea Morelet extracts, using pre- and post-treatment procedures.Results: The P. caribaea and C. citratus extracts were antigenotoxic against γ-rays when the cells were pre-treated with both extracts, suggesting a possible antigenotoxic action through a free radical scavenging mechanisms. Amifostine and the P. orbicularis extract were also antigenotoxic under pre- and post-treatment conditions, indicating that several antimutagenic components of this plant extract may also operate by some intracellular mechanism, unlike its antioxidant activity.Conclusions: The results have demonstrated the usefulness of the modified SOS Chromotest assay in the screening of phytochemical radioprotectors as well as in the study of their antimutagenic mechanisms.