Evaluation of antimutagenic and cytotoxic activity of skin secretion extract of Rhinella marina and Rhaebo guttatus (Anura, Bufonidae) / Avaliação da atividade antimutagênica e citotóxica da secreção cutânea de Rhinella marina e Rhaebo guttatus (Anura, Bufonidae)

The skin secretion from toads of the Bufonidae family has great potential in the search for new active compounds to be used as drug candidates in treating some diseases, among them cancer. In this context, this study aimed to evaluate the cytotoxic and antimutagenic activity of the parotoid gland secretion extracts of Rhinella marina and Rhaebo guttatus, as well as biochemically analyze transaminases and serum creatinine for liver and renal damage, respectively. Cytotoxicity was performed by the colorimetric method based on MTT (3- [4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide) with different concentrations of the extracts in Walker or splenic tumor cell cultures from rats and mice. The micronucleus test was performed with male Swiss mice treated orally with the extracts for 15 days, and then intraperitoneally with N-ethyl-N-nitrosurea (50 mg kg-1). Micronucleated polychromatic erythrocytes (MNPCE) were evaluated in bone marrow. The extracts showed cytotoxic activity in the evaluated cells. There was a significant reduction in the frequency of MNPCE (R. marina = 56% and R. guttatus = 75%, p < 0.001), indicating antimutagenic potential of the extracts. The groups treated only with extract showed an increase in MNPCE frequency, evidencing mutagenic potential. Biochemical analyzes showed no significant difference between treatments. Thus, under our experimental conditions, the extracts of R. marina and R. guttatus skin secretions presented chemopreventive potential for cancer. (AU)

A secreção cutânea de anuros da família Bufonidae tem grande potencial na busca de novos compostos ativos para utilização como fármacos candidatos no tratamento de algumas doenças, entre elas o câncer. Neste contexto, este estudo teve como objetivo avaliar a atividade citotóxica e antimutagênica dos extratos da secreção da glândula parótida de Rhinella marina e Rhaebo guttatus, bem como a análise bioquímica de transaminases e creatinina séricas, para avaliar dano hepático e renal, respectivamente. A avaliação de citotoxicidade foi realizada pelo método colorimétrico baseado no MTT (3-[4, 5-dimethyl-2-thiazolyl]-2, 5-diphenyl-2H-tetrazolium bromide), com diferentes concentrações dos extratos em culturas de células do Tumor de Walker ou células esplênicas de rato e camundongo. O teste do micronúcleo foi realizado com camundongos Swiss machos que receberam tratamento oral com os extratos durante 15 dias, seguido de tratamento intraperitoneal com N-etil-N-nitrosuréia (50 mg kg-1). A frequência de eritrócitos policromáticos micronucleados (PCEMN) foi determinada em medula óssea. Os extratos apresentaram ação citotóxica nas células avaliadas. Houve uma redução significativa na frequência de PCEMN (R. marina = 56% e R. guttatus = 75%, p < 0,001), observando-se um potencial antimutagênico dos extratos. Os grupos tratados somente com os extratos apresentaram um aumento na frequência de PCEMNs, evidenciando um potencial mutagênico. As análises bioquímicas não apresentaram diferença significativa entre os tratamentos. Assim, nas condições experimentais testadas, as secreções cutâneas de R. marina e R. guttatus apresentaram potencial quimiopreventivo para câncer.(AU)

Camu-camu (Myrciaria dubia) from commercial cultivation has higher levels of bioactive compounds than native cultivation (Amazon Forest) and presents antimutagenic effects in vivo.

BACKGROUND: Camu-camu (Myrciaria dubia) is a typical Amazonian fruit and has high antioxidant capacity due to its high levels of vitamin C and phenolic compounds. This study aimed to determine the phytochemicals, antioxidant capacity and antimutagenic effects of camu-camu fruits with different maturity stages grown in dry (commercial cultivation) or flooded environments (native cultivation, Amazon). RESULTS: Total polyphenols, ascorbic acid and in vitro antioxidant capacity levels were higher in ripe fruits grown in a commercial cultivation. The extracts from ripe camu-camu grown in a commercial cultivation exerted antioxidant effects and high percentage of protection against doxorubicin and 1,2-dimethylhydrazine in all tested systems (liver, bone marrow and gut), for three camu-camu extract concentrations (17, 85 and 170 mg kg-1 body weight), as follows: bone marrow minocronucleus (37.91%, 41.75%, 43.95%); micronucleus gut test (61.01%, 64.40%, 50.28%); apoptosis index (60.26%, 62.44%, 58.22%); comet assay through the tail moment (71.64%, 72.31%, 70.70%), percent DNA in the tail (64.54%, 68.75%, 76.79%) and tail intensity (76.43%, 81.02%, 68.33%). CONCLUSION: The results of this study contribute to increasing the production of camu-camu fruits grown in dry environments and their use as a health-promoting food. © 2018 Society of Chemical Industry.

Nutraceutical potential of hemp (Cannabis sativa L.) seeds and sprouts.

In this study the antioxidant effect of Cannabis sativa L. seeds and sprouts (3 and 5 days of germination) was evaluated. Total polyphenols, flavonoids and flavonols content, when expressed on dry weight basis, were highest in sprouts; ORAC and DPPH (in vitro assays), CAA-RBC (cellular antioxidant activity in red blood cells) and hemolysis test (ex vivo assays) evidenced a good antioxidant activity higher in sprouts than in seeds. Untargeted analysis by high resolution mass spectrometry in negative ion mode allowed the identification of main polyphenols (caffeoyltyramine, cannabisin A, B, C) in seeds and of ω-6 (linoleic acid) in sprouts. Antimutagenic effect of seeds and sprouts extracts evidenced a significant decrease of mutagenesis induced by hydrogen peroxide in Saccharomyces cerevisiae D7 strain. In conclusion our results show that C. sativa seeds and sprouts exert beneficial effects on yeast and human cells and should be further investigated as a potential functional food.

Isolation and characterization of the compounds responsible for the antimutagenic activity of Combretum microphyllum (Combretaceae) leaf extracts.

BACKGROUND: Mutations play a major role in the pathogenesis and development of several chronic degenerative diseases including cancer. It follows, therefore that antimutagenic compound may inhibit the pathological process resulting from exposure to mutagens. Investigation of the antimutagenic potential of traditional medicinal plants and compounds isolated from plant extracts provides one of the tools that can be used to identify compounds with potential cancer chemopreventive properties. The aim of this study was to isolate and characterise the compounds responsible for the antimutagenic activity of Combretum microphyllum. METHODS: The methanol leaf extract of C. microphyllum was evaluated for antimutagenicity in the Ames/microsome assay using Salmonella typhimurium TA98. TA100 and TA102. Solvent-solvent fractionation was used to partition the extracts and by using bioassay-guided fractionation, three compounds were isolated. The antimutagenic activity of the three compounds were determined in the Ames test using Salmonella typhimurium TA98, TA100 and TA102. The antioxidant activity of the three compounds were determined by the quantitative 2,2-diphenyl-1-picrylhydrazyl (DPPH)-free radical scavenging method. The cytotoxicity was determined in the MTT assay using human hepatocytes. RESULTS: A bioassay-guided fractionation of the crude extracts for antimutagenic activity led to the isolation of three compounds; n-tetracosanol, eicosanoic acid and arjunolic acid. Arjunolic acid was the most active in all three tested strains with a antimutagenicity of 42 ± 9.6%, 36 ± 1.5% and 44 ± 0.18% in S. typhimurium TA98, TA100 and TA102 respectively at the highest concentration (500 µg/ml) tested, followed by eicosanoic acid and n-tetracosanol. The antioxidant activity of the compounds were determined using the quantitative 2,2 diphenyl-1-picryhydrazyl (DPPH)-free radical scavenging method. Only arjunolic acid had pronounced antioxidant activity (measured as DPPH-free scavenging activity) with an EC50 value of 0.51 µg/ml. The cytotoxicity of the isolated compounds were determined in the MTT assay using human hepatocytes. The compounds had low cytotoxicity at the highest concentration tested with LC50 values >200 µg/ml for n-tetracosanol and eicosanoic acid and 106.39 µg/ml for arjunolic acid. CONCLUSIONS: Based on findings from this study, compounds in leaf extracts of C. microphyllum protected against 4-NQO and MMC induced mutations as evident in the Ames test. The antimutagenic activity of arjunolic acid may, at least in part, be attributed to its antioxidant activity resulting in the detoxification of reactive oxygen species produced during mutagenesis.

The SOS Chromotest applied for screening plant antigenotoxic agents against ultraviolet radiation.

In this work, we investigated the usefulness of the SOS Chromotest for screening plant antigenotoxic agents against ultraviolet radiation (UV). Fifty Colombian plant extracts obtained by supercritical fluid (CO2) extraction, twelve plant extract constituents (apigenin, carvacrol, ß-caryophyllene, 1,8-cineole, citral, p-cymene, geraniol, naringenin, pinocembrin, quercetin, squalene, and thymol) and five standard antioxidant and/or photoprotective agents (curcumin, epigallocatechin gallate, resveratrol, α-tocopherol, and Trolox®) were evaluated for their genotoxicity and antigenotoxicity against UV using the SOS Chromotest. None of the plant extracts, constituents or agents were genotoxic in the SOS Chromotest at tested concentrations. Based on the minimal extract concentration that significantly inhibited UV-genotoxicity (CIG), five plant extracts were antigenotoxic against UV as follows: Baccharis nítida (16 µg mL-1) = Solanum crotonifolium (16 µg mL-1) > Hyptis suaveolens (31 µg mL-1) = Persea caerulea (31 µg mL-1) > Lippia origanoides (62 µg mL-1). Based on CIG values, the flavonoid compounds showed the highest antigenotoxic potential as follows: apigenin (7 µM) > pinocembrin (15 µM) > quercetin (26 µM) > naringenin (38 µM) > epigallocatechin gallate (108 µM) > resveratrol (642 µM). UV-genotoxicity inhibition with epigallocatechin gallate, naringenin and resveratrol was related to its capability for inhibiting protein synthesis. A correlation analysis between compound antigenotoxicity estimates and antioxidant activity evaluated by the oxygen radical absorbance capacity (ORAC) assay showed that these activities were not related. The usefulness of the SOS Chromotest for bioprospecting of plant antigenotoxic agents against UV was discussed.

Black tea: Phytochemicals, cancer chemoprevention, and clinical studies.

Tea (Camellia sinensis L.) is the most popular, flavored, functional, and therapeutic non-alcoholic drink consumed by two-thirds of the world's population. Black tea leaves are reported to contain thousands of bioactive constituents such as polyphenols, amino acids, volatile compounds, and alkaloids that exhibit a range of promising pharmacological properties. Due to strong antioxidant property, black tea inhibits the development of various cancers by regulating oxidative damage of biomolecules, endogenous antioxidants, and pathways of mutagen and transcription of antioxidant gene pool. Regular drinking of phytochemicals-rich black tea is linked to regulate several molecular targets, including COX-2, 5-LOX, AP-1, JNK, STAT, EGFR, AKT, Bcl2, NF-κB, Bcl-xL, caspases, p53, FOXO1, TNFα, PARP, and MAPK, which may be the basis of how dose of black tea prevents and cures cancer. In vitro and preclinical studies support the anti-cancer activity of black tea; however, its effect in human trails is uncertain, although more clinical experiments are needed at molecular levels to understand its anti-cancer property. This review discusses the current knowledge on phytochemistry, chemopreventive activity, and clinical applications of black tea to reveal its anti-cancer effect.

Assessment of multifunctional activity of bioactive peptides derived from fermented milk by specific Lactobacillus plantarum strains.

Milk-derived bioactive peptides with a single activity (e.g., antioxidant, immunomodulatory, or antimicrobial) have been previously well documented; however, few studies describe multifunctional bioactive peptides, which may be preferred over single-activity peptides, as they can simultaneously trigger, modulate, or inhibit multiple physiological pathways. Hence, the aim of this study was to assess the anti-inflammatory, antihemolytic, antioxidant, antimutagenic, and antimicrobial activities of crude extracts (CE) and peptide fractions (<3 and 3-10 kDa) obtained from fermented milks with specific Lactobacillus plantarum strains. Overall, CE showed higher activity than both peptide fractions (<3 and 3-10 kDa) in most of the activities assessed. Furthermore, activity of <3 kDa was generally higher, or at least equal, to the 3 to 10 kDa peptide fractions. In particular, L. plantarum 55 crude extract or their fractions showed the higher anti-inflammatory (723.68-1,759.43µg/mL of diclofenac sodium equivalents), antihemolytic (36.65-74.45% of inhibition), and antioxidant activity [282.8-362.3µmol of Trolox (Sigma-Aldrich, St. Louis, MO) equivalents]. These results provide valuable evidence of multifunctional role of peptides derived of fermented milk by the action of specific L. plantarum strains. Thus, they may be considered for the development of biotechnological products to be used to reduce the risk of disease or to enhance a certain physiological function.

Biological Activity of Conventional and Organic Pomegranate Juices: Antioxidant and Antimutagenic Potential.

None of the health claims about pomegranate juices has been approved yet by European Food Safety Authority (EFSA). There is a general perception among consumers that organic foods are healthier, tastier, and more nutritive than the conventional products. The aim of this research was to study the differences in the biological activity between ready-for-consumption juices obtained from pomegranates fruits grown under conventional and organic agricultural practices. Antioxidant activity has been evaluated by three methods (DPPH•, ABTS+, and FRAP), together with the total contents of phenolics and punicalagin (HPLC-DAD); besides, the Ames test was used to evaluate the antimutagenic potential of the juices. Pomegranate juice, either from conventionally or organically grown fruits, was antimutagenic (mean of 51 and 90 % for Salmonella typhimurium TA100 and TA98, respectively) and it was capable of protecting DNA from both, base-pair or frame-shift type of mutations. In fact, the antimutagenicity of conventional pomegranate juice was higher than that achieved by the organic sample; this finding was linked to a higher punicalagin content (201 and 104 mg L-1 for conventional and organic juices, respectively).

Antioxidant and genoprotective activity of selected cucurbitaceae seed extracts and LC-ESIMS/MS identification of phenolic components.

Cucurbitaceae are one of most widely used plant species for human food but lesser known members have not been examined for bioactive components. The purpose of this study was to evaluate the antioxidant and genoprotective activities from three cucurbitaceae seeds extracts and to identify phenolic components by LC-ESIMS/MS analysis. From the results, the yield of seeds extract was 20-41% (w/w) and samples had 16-40% total phenols as gallic acid equivalents (GAE). Compared with methanol solvent, using acidified methanol led to increased extraction yield by 1.4 to 10-fold, higher phenolic content (149.5 ± 1.2 to 396.4 ± 1.9 mg GAE/g), higher DPPH radical quenching and enhanced genoprotective activity using the pBR322 plasmid assay. LC-ESI-MS/MS analysis led to identification of 14-17 components, based on authentic standards and comparison with literature reports, as mainly phenolic acids and esters, flavonol glycosides. This may be the first mass spectrometric profiling of polyphenol components from cucurbitaceae seeds.

Morphoanatomic characterization of the stem and the leaf of Tabernaemontana catharinensis A. DC (Apocynaceae) and antimutagenic activity of its leaves / Caracterização morfoanatômica do caule e da folha de Tabernaemontana catharinensis A. DC (Apocynaceae) e atividade antimutagênica de suas folhas

ABSTRACT Tabernaemontana catharinensis A. DC (Apocynaceae) is used as a medicinal plant by the population. In order to contribute to the safe use of the plant as herbal medicine, this study aimed to morphoanatomically characterize the aereal vegetative organs of T. catharinensis and to evaluate the leaves’ mutagenic and antimutagenic activities. Histological blades of leaves and stem of T. catharinensis were performed; the methionine system (methG1) and Aspergillusnidulans conidia germination analysis were employed for mutagenic and antimutagenic evaluation. The morphoanatomic analysis did not show trichomes in the stem, petiole and leaf. Besides, it was observed both the presence of bi-collateral bundles - except in the foliar apex where the bundles were from the collateral type - as well as anamphistomatic leaf with paracyte stomata and sub-epidermal layer in the region of the leaf edges. The mutagenicity/antimutagenicity trial indicated a significant decrease of mutation frequency in comparison with the control group and showed that the T. catharinensis had antimutagenic activity within the type, time and form of treatment. Since the germination test showed that the conidia germination was accelerated from the bud phase, activities at the cell cycle level and polarized growth proved to be possible. The morphoanatomic analysis of the leaf and stem associated with the mutagenic and antimutagenic analyses contributes to the safe use of the plant by humans and also for the quality control of a possible phytotherapeutic drug.

RESUMO Tabernaemontana catharinensis A. DC (Apocynaceae) é utilizada como planta medicinal pela população. A fim de contribuir para o uso seguro da planta como medicinal, este trabalho teve como objetivo caracterizar morfoanatomicamente os órgãos vegetativos aéreos de T. catharinensis e avaliar a atividade mutagênica e antimutagênica de suas folhas. Foram realizados cortes histológicos da folha e do caule de T. catharinensis e, para a avaliação mutagênica e antimutagênica, foi utilizado o sistema metionina (methG1) e análise da germinação de conídios em Aspergillus nidulans. A análise morfoanatômica evidenciou a ausência de tricomas no caule, pecíolo e folha; presença de feixes bicolaterais, com exceção no ápice foliar cujos feixes são do tipo colateral; folha anfiestomática com estômatos paracíticos e camada subepidérmica na região do bordo foliar. O ensaio de mutagenicidade/antimutagenicidade mostrou uma diminuição significativa da frequência de mutação em relação ao controle, indicando que nesse tipo, tempo e forma de tratamento, T. catharinensis apresentou atividade antimutagênica. O ensaio de germinação evidenciou que houve aceleração da germinação dos conídios, a partir da fase de botão, indicando uma possível atuação em nível de ativação de ciclo celular e crescimento polarizado. A análise morfoanatômica da folha e do caule associados à análise mutagênica e antimutagênica, contribuem para o uso seguro da planta pela população e para o controle de qualidade de um possível fitoterápico.

Mutagenicity and antimutagenicity of six Brazilian Byrsonima species assessed by the Ames test.

BACKGROUND: In various regions of Brazil, several species of the genus Byrsonima (Malpighiaceae) are widely used to treat gastrointestinal complications. This genus has about 150 species of shrubs and trees distributed over the entire Neotropical region. Various biological activities have been identified in these plants, especially antioxidant, antimicrobial and topical and systemic anti-inflammatory activities. The aim of this study was to investigate the mutagenicity and antimutagenicity of hydroalcoholic leaf extracts of six species of Byrsonima: B. verbascifolia, B. correifolia, B. coccolobifolia, B. ligustrifolia, B. fagifolia and B. intermedia by the Salmonella microsome assay (Ames test). METHODS: Mutagenic and antimutagenic activity was assessed by the Ames test, with the Salmonella typhimurium tester strains TA100, TA98, TA97a and TA102, with (+S9) and without (-S9) metabolization, by the preincubation method. RESULTS: Only B. coccolobifolia and B. ligustrifolia showed mutagenic activity. However, the extracts of B. verbascifolia, B. correifolia, B. fagifolia and B. intermedia were found to be strongly antimutagenic against at least one of the mutagens tested. CONCLUSIONS: These results contribute to valuable data on the safe use of medicinal plants and their potential chemopreventive effects. Considering the excellent antimutagenic activities extracted from B. verbascifolia, B. correifolia, B. fagifolia and B. intermedia, these extracts are good candidate sources of chemopreventive agents. However, B. coccolobifolia and B. ligustrifolia showed mutagenic activity, suggesting caution in their use.

The protective effect of dietary Arthrospira (Spirulina) maxima against mutagenicity induced by benzo[alpha]pyrene in mice.

Benzo[alpha]pyrene (B[α]P) was used to test the possible antimutagenic effects of Arthrospira (Spirulina) maxima (SP) on male and female mice. SP was orally administered at 0, 200, 400, or 800 mg/kg of body weight to animals of both sexes for 2 weeks before starting the B[α]P (intraperitoneal injection) at 125 mg/kg of body weight for 5 consecutive days. For the male dominant lethal test, each male was caged with two untreated females per week for 3 weeks. For the female dominant lethal test, each female was caged for 1 week with one untreated male. All the females were evaluated 13-15 days after mating for incidence of pregnancy, total corpora lutea, total implants and pre- and postimplant losses. SP protected from B[α]P-induced pre- and postimplant losses in the male dominant lethal test, and from B[α]P-induced postimplantation losses in treated females. Moreover, SP treatment significantly reduced the detrimental effect of B[α]P on the quality of mouse semen. Our results illustrate the protective effects of SP in relation to B[α]P-induced genetic damage to germ cells. We conclude that SP, owing mainly to the presence of phycocyanin, could be of potential clinical interest in cancer treatment or prevention of relapse.

Nutritional and health perspectives of beans (Phaseolus vulgaris L.): an overview.

Beans, the variants of Phaseolus vulagris, are nutritionally and economically important food crop in each part of the world. Besides providing nutrients such as multifaceted carbohydrates, elevated proteins, dietary fiber, minerals, and vitamins, these also contain rich variety of polyphenolic compounds with prospective health benefits. This review mainly focuses the important nutritional aspects of beans as well as their contribution in decreasing the risks of chronically degenerative diseases.

Avaliação fitoquímica e atividade antioxidante, antimutagênica e toxicológica do extrato aquoso das folhas de Ocimum gratissimum L / Phytochemical Assessment And Antioxidant, Antimutagenic And Toxicological Activity Of Ocimum gratissimum L. Leaf aqueous extract

A espécie Ocimum gratissimum L., popularmente conhecida como alfavaca, é uma planta muito usada na medicina tradicional brasileira, à qual são atribuídas diversas atividades terapêuticas quando usada na forma de infuso de suas folhas. Neste estudo foi realizada a caracterização fitoquímica, a avaliação da ação antioxidante e a investigação dos efeitos antimutagênico e antigenotóxico, além do efeito mutagênico e genotóxico potencial do extrato aquoso liofilizado a parir das folhas de O. gratissimum (EAOG). O conteúdo de polifenóis totais no extrato foi determinado pelo método Folin-Ciocalteu, sendo encontrado 11,3 µg EAG/mg de EAOG. A atividade antioxidante foi avaliada pelo teste do 1,1-difenil-2-picril hidrazil (DPPH•), apresentando IC50 de 83,0 µg/mL. A antimutagenicidade e mutagenicidade foram avaliadas em cepas de Salmonella typhimurium (TA98 e TA100) utilizando o teste Salmonella/microssoma (Salmonella typhimurium/microssomas) em diferentes concentrações. EAOG induziu a atividade antimutagênica para a cepa TA98. A mutagenicidade não foi observada para o extrato em ambas as linhagens. Adicionalmente, a ação antigenotoxica avaliada pelo teste de clivagem do DNA-plasmidial também foi observada para EAOG. Os resultados também demonstraram que o extrato não foi capaz de induzir a genotoxicidade pelo teste empregado. Este estudo relata, pela primeira vez, as propriedades antimutagênica e antigenotóxica do extrato aquoso de O. gratissimum.

The species Ocimum gratissimum L., popularly known as Clove Basil, is a plant widely used in traditional Brazilian medicine, and several therapeutic activities are attributed to it when used as infusion of its leaves. In this study, we carried out a phytochemical characterization and the assessment evaluation and investigation of the antioxidant action of the antimutagenic and antigenotoxic effects and the potential mutagenic and genotoxic effects of the freeze-dried aqueous extract of the O. gratissimum (EAOG) leaves. The total polyphenol content in the extract was determined by the Folin-Ciocalteu method, and we found 11.3 µg EAG/mg of EAOG. The antioxidant activity was assessed by the 1,1-diphenyl-2-picryl hidrazil (DPPH·), with IC50 of 83.0 µg/mL. Antimutagenicity and mutagenicity were assessed in Salmonella typhimurium (TA98 and TA100) strains using the Salmonella/microsome (Salmonella typhimurium/microsome) test in different concentrations. EAOG induced antimutagenic activity for strain TA98. Mutagenicity was not observed for the extract in both strains. Additionally, antigenotoxic action, assessed by cleavage of the DNA-damage, was also observed for EAOG. The results also show that the extract was not able to induce genotoxicity by the test used. This study reports for the first time the antimutagenic and antigenotoxic properties of the O. gratissimum aqueous extract.

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.

Identification of antimutagenic properties of anthocyanins and other polyphenols from rose (Rosa centifolia) petals and tea.

Petals from different rose (Rosa centifolia) cultivars ("passion," "pink noblesse," and "sphinx") were assessed for antimutagenicity using Escherichia coli RNA polymerase B (rpoB)-based Rif (S) →Rif (R) (rifampicin sensitive to resistant) forward mutation assay against ethyl methanesulfonate (EMS)-induced mutagenesis. The aqueous extracts of rose petals from different cultivars exhibited a wide variation in their antimutagenicity. Among these, cv. "passion" was found to display maximum antimutagenicity. Upon further fractionation, the anthocyanin extract of cv. "passion" displayed significantly higher antimutagenicity than its phenolic extract. During thin-layer chromatography (TLC) analysis, the anthocyanin extract got resolved into 3 spots: yellow (Rf : 0.14), blue (Rf : 0.30), and pink (Rf : 0.49). Among these spots, the blue one displayed significantly higher antimutagenicity than the other 2. Upon high-performance liquid chromatography analysis, this blue spot further got resolved into 2 peaks (Rt : 2.7 and 3.8 min). The 2nd peak (Rt : 3.8 min) displaying high antimutagenicity was identified by ESI-IT-MS/MS analysis as peonidin 3-glucoside, whereas less antimutagenic peak 1 (Rt : 2.7) was identified as cyanidin 3, 5-diglucoside. The other TLC bands were also characterized by ESI-IT-MS/MS analysis. The least antimutagenic pink band (Rf : 0.49) was identified as malvidin 3-acetylglucoside-4-vinylcatechol, whereas non-antimutagenic yellow band (Rf : 0.14) was identified as luteolinidin anthocyanin derivative. Interestingly, the anthocyanin extracted from rose tea of cv. "passion" exhibited a similar antimutagenicity as that of the raw rose petal indicating the thermal stability of the contributing bioactive(s). The findings thus indicated the health protective property of differently colored rose cultivars and the nature of their active bioingredients.

Croton lechleri Müll. Arg. (Euphorbiaceae) stem bark essential oil as possible mutagen-protective food ingredient against heterocyclic amines from cooked food.

The Amazonian Croton lechleri stem bark essential oil was tested for its anti-mutagenic potential by performing the Ames test against heterocyclic amines (HCAs), in continuing research on applicative functional profile of this phytocomplex as food ingredient (Rossi et al., 2011). Salmonella typhimurium strain TA98 was used with and without metabolic activation (S9 mix). The anti-mutagenic properties was assayed with the following HCAs: 2-amino-3-methylimidazo-[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo-[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo-[4,5-f]quinoxaline (MeIQx), the imidazoles 2-amino-6-methyldipyrido-[1,2-a:3',2'-d]imidazole (Glu-P-1) and 2-aminodipirydo-[1,2-a:3',2'-d]imidazole (Glu-P-2). All HCAs with S9 induced mutagenicity at 10(-10) mol/plate. Without S9, IQ and MeIQ showed mutagenicity at 10(-8) mol/plate, MeIQx and Glu-P-1 at 10(-5) mol/plate, while Glu-P-2 was inactive. In presence of HACs (10(-9) mol/plate), C. lechleri essential oil was tested for mutagen-protective properties (concentration range: 0.01-0.10 mg/plate) taking the Highest Uneffective Dose (HUD) as threshold reference. With S9 mix, C. lechleri essential oil displayed a significant reduction of revertants at 0.05 mg/plate, from 21% to 34%. The essential oil showed mutagen-protective efficacy against IQ and MeIQ tested as direct mutagens (10(-7) mol/plate), with a revertants percentage reduction of 39% and 40%, respectively. No anti-mutagen capacity was noted for MeIQx and Glu-P-1 (10(-5) mol/plate). Since HACs are known as possible colon and liver cancer inducers, C. lechleri essential oil was tested for its cytotoxicity and anti-proliferative capacity against LoVo and HepG2 cancer cell lines showing IC50 of 74.95±0.05 µg/ml (LoVo) and 82.28±0.03 µg/ml (HepG2), displaying a promising role of this essential oil as a functional food ingredient with interesting mutagen preventing properties.

Antioxidant capacity and antimutagenic activity of anthocyanin and carotenoid extracts from nixtamalized pigmented Creole maize races (Zea mays L.).

Nixtamalization process is the first step to obtain maize based products, like tortillas; however, in both the traditional and commercial processes, white grain is generally preferred. Creole maize races, mainly pigmented varieties, have increasingly attention since these are rich in anthocyanins and carotenoids. The aim of this investigation was to evaluate the antioxidant and antimutagenic activity of rich anthocyanins and carotenoids extracts from creole maize races before (grain) and after (masa and tortilla) the nixtamalization process. Most anthocyanins and carotenoids were lost during nixtamalization. Before nixtamalization, blue and red genotypes contained either higher antioxidant capacity and anthocyanin contents (963 ± 10.0 and 212.36 ± 0.36 mg of cyanidin-3-glucoside eq/100 g, respectively) than the white and yellow genotypes. However, the highest carotenoid levels were displayed by red grains (1.01 ± 0.07 to 1.14 ± 0.08 µg of ß-carotene eq/g extract). Anthocyanins losses were observed when the blue grains were processed into masa (83 %) and tortillas (64 %). Anthocyanins content correlated with antiradical activity (r = 0.57) and with 2-aminoanthracene -induced mutagenicity inhibition on TA98 and TA100 (r = -0.62 and r = -0.44, respectively). For white grains, nixtamalization also reduced carotenoids (53 to 56 %), but not antioxidant activity and 2-Aa-induced mutagenicity. Throughout the nixtamalization process steps, all the extracts showed antimutagenic activity against 2-aminoanthracene-induced mutagenicity (23 to 90 %), displaying higher potential to inhibit base changes mutations than frameshift mutations in the genome of the tasted microorganism (TA100 and TA98, respectively). The results suggest that even though there were pigment losses, creole maize pigments show antioxidant and antimutagenic activities after nixtamalization process.

The effect of royal sun agaricus, Agaricus brasiliensis S. Wasser et al., extract on methyl methanesulfonate caused genotoxicity in Drosophila melanogaster.

The effect of culinary-medicinal Royal Sun Agaricus (Agaricus brasiliensis) hot water extract on methyl methane sulfonate (MMS) induced mutagenicity/genotoxity in Drosophila melanogaster was studied using a quick and broadly applicable in vivo assay, i.e., the wing somatic mutation and recombination test. We used 2nd instar larvae, trans-heterozygous for the third chromosome recessive markers, i.e., multiple wing hairs (mvh) and flare-3 [flr (3)], and fed them for 24 h with the aqueous extract of A. brasiliensis. For antigenotoxicity studies a 24-h pretreatment with the extract was done, followed by a 48-h treatment of the then 3rd instar larvae with MMS. The frequency of mutations of the wing blade changes (i.e., of the number of wing spots of different sizes) induced in somatic cells was determined as a parameter of genetic changes of the wing imaginal discs. The results showed that A. brasiliensis extract did not cause any genotoxic or mutagenic effects. No antigenotoxic and/or protective effect against the induction of mutations by MMS was observed. Instead, a possible enhanced mitotic recombination frequency by MMS was seen after pretreatment of the larvae with A. brasiliensis extract. Possible mechanisms of action are discussed.

In vitro antioxidant properties, DNA damage protective activity, and xanthine oxidase inhibitory effect of cajaninstilbene acid, a stilbene compound derived from pigeon pea [Cajanus cajan (L.) Millsp.] leaves.

The antioxidant properties, DNA damage protective activities, and xanthine oxidase (XOD) inhibitory effect of cajaninstilbene acid (CSA) derived from pigeon pea leaves were studied in the present work. Compared with resveratrol, CSA showed stronger antioxidant properties, DNA damage protective activity, and XOD inhibition activity. The IC(50) values of CSA for superoxide radical scavenging, hydroxyl radical scavenging, nitric oxide scavenging, reducing power, lipid peroxidation, and XOD inhibition were 19.03, 6.36, 39.65, 20.41, 20.58, and 3.62 µM, respectively. CSA possessed good protective activity from oxidative DNA damage. Furthermore, molecular docking indicated that CSA was more potent than resveratrol or allopurinol to interact with the active site of XOD (calculated free binding energy: -229.71 kcal mol(-1)). On the basis of the results, we conclude that CSA represents a valuable natural antioxidant source and may potentially be applicable in health food industry.