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1.  Mutagenicity and antimutagenicity of (−)-hinokinin a trypanosomicidal compound measured by Salmonella microsome and comet assays 
The dibenzylbutyrolactone lignan (−)-hinokinin (HK) was derived by partial synthesis from (−)-cubebin, isolated from the dry seeds of the pepper, Piper cubeba. Considering the good trypanosomicidal activity of HK and recalling that natural products are promising starting points for the discovery of novel potentially therapeutic agents, the aim of the present study was to investigate the (anti) mutagenic∕ genotoxic activities of HK.
The mutagenic∕ genotoxic activities were evaluated by the Ames test on Salmonella typhimurium strains TA98, TA97a, TA100 and TA102, and the comet assay, so as to assess the safe use of HK in the treatment of Chagas’ disease. The antimutagenic ∕antigenotoxic potential of HK were also tested against the mutagenicity of a variety of direct and indirect acting mutagens, such as 4- nitro-o-phenylenediamine (NOPD), sodium azide (SA), mitomycin C (MMC), benzo[a]pyrene (B[a]P), aflatoxin B1 (AFB1), 2-aminoanthracene (2-AA) and 2-aminofluorene (2-AF), by the Ames test, and doxorubicin (DXR) by the comet assay.
The mutagenicity∕genotoxicity tests showed that HK did not induce any increase in the number of revertants or extent of DNA damage, demonstrating the absence of mutagenic and genotoxic activities. On the other hand, the results on the antimutagenic potential of HK showed a strong inhibitory effect against some direct and indirect-acting mutagens.
Regarding the use of HK as an antichagasic drug, the absence of mutagenic effects in animal cell and bacterial systems is encouraging. In addition, HK may be a new potential antigenotoxic ∕ antimutagenic agent from natural sources. However, the protective activity of HK is not general and varies with the type of DNA damage-inducing agent used.
PMCID: PMC3545969  PMID: 23114276
Hinokinin; Ames test; Comet assay; Mutagenicity; Antimutagenicity
2.  Antimutagenic and free radical scavenger effects of leaf extracts from Accacia salicina 
Three extracts were prepared from the leaves of Accacia salicina; ethyl acetate (EA), chloroform (Chl) and petroleum ether (PE) extracts and was designed to examine antimutagenic, antioxidant potenty and oxidative DNA damage protecting activity.
Antioxidant activity of A. salicina extracts was determined by the ability of each extract to protect against plasmid DNA strand scission induced by hydroxyl radicals. An assay for the ability of these extracts to prevent mutations induced by various oxidants in Salmonella typhimurium TA102 and TA 104 strains was conducted. In addition, nonenzymatic methods were employed to evaluate anti-oxidative effects of tested extracts.
These extracts from leaf parts of A. salicina showed no mutagenicity either with or without the metabolic enzyme preparation (S9). The highest protections against methylmethanesulfonate induced mutagenicity were observed with all extracts and especially chloroform extract. This extract exhibited the highest inhibitiory level of the Ames response induced by the indirect mutagen 2- aminoanthracene. All extracts exhibited the highest ability to protect plasmid DNA against hydroxyl radicals induced DNA damages. The ethyl acetate (EA) and chloroform (Chl) extracts showed with high TEAC values radical of 0.95 and 0.81 mM respectively, against the ABTS.+.
The present study revealed the antimutagenic and antioxidant potenty of plant extract from Accacia salicina leaves.
PMCID: PMC3267653  PMID: 22132863
3.  Antioxidant and antigenotoxic activities of Angelica keiskei, Oenanthe javanica and Brassica oleracea in the Salmonella mutagenicity assay and in HCT116 human colon cancer cells 
BioFactors (Oxford, England)  2006;26(4):231-244.
Epidemiological studies indicate that consumption of green-yellow vegetables rich in chlorophyll, vitamin C, vitamin E, and carotenoids reduce the risk of cancer. We sought to examine the antigenotoxic and antioxidant properties of chlorophyll-rich methanol extracts of Angelica keiskei, Oenanthe javanica, and Brassica oleracea (kale). In the Salmonella mutagenicity assay, A. keiskei caused dose-dependent inhibition against three heterocyclic amine mutagens in the presence of S9, O. javanica was antimutagenic only at the highest concentration in the assay (2 mg/plate), and B. oleracea showed no consistent inhibitory activity at non-toxic levels. None of the extracts were effective against three direct-acting mutagens in the absence of S9. Extracts of A. keiskei and, to a lesser extent O. javanica, inhibited two of the major enzymes that play a role in the metabolic activation of heterocyclic amines, based on ethoxyresorufin-O-deethylase and methoxyresorufin-O-demethylase assays in vitro. All three plant extracts were highly effective in assays which measured ferric reducing/antioxidant power, oxygen radical absorbance capacity, and Fe2+/H2O2-mediated DNA nicking. Finally, using the ‘comet’ assay, all three plant extracts protected against H2O2-induced genotoxic damage in human HCT116 colon cancer cells. These findings provide support for the antigenotoxic and antioxidant properties of chlorophyll-rich extracts of A. keiskei, O. javanica, and B. oleracea, through mechanisms that include inhibition of carcinogen activation and scavenging of reactive oxygen species.
PMCID: PMC2267880  PMID: 17119270
Antimutagen; antioxidant; heterocyclic amines; phytochemical; comet assay; D NA breaks
4.  Antimutagenic and antioxidant activity of novel 4-substituted phenyl-2,2′-bichalcophenes and aza-analogs 
Evaluation of the potential antimutagenic activities of new compounds by Ames assay has been of great interest for the development of novel therapeutics for many diseases including cancer. Ten novel bichalcophenes with in vitro and in vivo broad spectrum activities against various microbial strains were investigated throughout the present study for their cytotoxic, antioxidant, and antimutagenic potential in a Salmonella reverse mutation assay system against sodium azide (NaN3) and benzo[a]pyrene (B[a]P). At nontoxic concentrations, all bichalcophenes alone or in combination with NaN3 (1 μg/plate) or B[a]P (20 μM) with S9 mix were not mutagenic. The bichalcophenes significantly reduced NaN3- and B[a]P-induced mutagenicity under pre-exposure and co-exposure conditions in a concentration-independent manner. However, the antimutagenic activity of bichalcophenes against B[a]P varied depending on the exposure regimen, being more effective under pre-exposure conditions. The antimutagenic activity was correlated with a high antioxidant activity that could promote the DNA repair system. Bichalcophenes are least likely to interfere with the microsomal bioactivation of B[a]P. Monocationic bichalcophenes were superior to the corresponding mononitriles as antimutagenic agents against both mutagens investigated, possibly due to the higher nucleophilic centers they have which could bind and protect the bacterial DNA. Three monocationic compounds were shown to have a strong anticancer activity against the 58 cell line. Based on the results of the present investigation, monocationic compounds (1, 4, and 5B) will be selected for further time consuming and costly chemoprevention studies in animal models.
PMCID: PMC3573810  PMID: 23430305
bichalcophenes; Salmonella typhimurium; sodium azide; benzo[a]pyrene; antimutagenicity
Mutation research  2006;627(2):136-145.
Selenium can have cancer chemopreventive activity, although the mechanism of action has not been well defined. Selenazolidine-4-(R)-carboxylic acids (SCAs) were devised as prodrugs of L-selenocysteine, to provide selenium in a form and at a concentration commensurate with cancer chemopreventive activity. In the present study, a series of selenazolidines has been evaluated in the Salmonella typhimurium TA98 tester strain and all were found to possess antimutagenic activity. There was little difference between the seven selenazolidines in their effectiveness against either benzo[a]pyrene (B[a]P) or 3,6-bis(dimethylamino)acridine (acridine orange), agents which differ in their requirement for mammalian enzyme bioactivation for mutagenicity. Antimutagenic activity against acridine orange was dependent on selenazolidine concentration, and EC50 values were in the 5 –10 μM range. At 25 μM, the concentration tested in common for the two mutagens, the selenazolidines were more effective antimutagens against acridine orange than against B[a]P, with reductions in mutant frequency ranging from 54–71% for B[a]P and 79–93% for acridine orange. Efficacy against B[a]P was not enhanced when the concentration was increased to 50 μM. The similarity in efficacy among the selenazolidines against B[a]P mutagenicity, contrasted with inter-compound differences in their ability to inhibit S9 CYP1A activity. The CYP1A Ki values ranged from a low of 63 μM (2-[2'-hydroxyphenyl]SCA) to a high of 1.1 mM (2-cyclohexylSCA), but all were above the concentration required to inhibit mutagenicity by 50%. Thus, all the SCAs possess antimutagenic activity against both B[a]P and acridine orange, the efficacy varies little between the individual selenazolidines, and for B[a]P, the efficacy is not proportional to the inhibitory effect on the mutagen bioactivating enzyme.
PMCID: PMC1805714  PMID: 17166761
Salmonella typhimurium TA98; selenazolidine-4-(R)-carboxylic acids; selenocysteine prodrugs; antimutagenic; benzo[a]pyrene; acridine orange; CYP1A inhibition
6.  Potential antimutagenic activity of berberine, a constituent of Mahonia aquifolium 
As part of a study aimed at developing new pharmaceutical products from natural resources, the purpose of this research was twofold: (1) to fractionate crude extracts from the bark of Mahonia aquifolium and (2) to evaluate the strength of the antimutagenic activity of the separate components against one of the common direct-acting chemical mutagens.
The antimutagenic potency was evaluated against acridine orange (AO) by using Euglena gracilis as an eukaryotic test model, based on the ability of the test compound/fraction to prevent the mutagen-induced damage of chloroplast DNA.
It was found that the antimutagenicity of the crude Mahonia extract resides in both bis-benzylisoquinoline (BBI) and protoberberine alkaloid fractions but only the protoberberine derivatives, jatrorrhizine and berberine, showed significant concentration-dependent inhibitory effect against the AO-induced chloroplast mutagenesis of E. gracilis. Especially berberine elicited, at a very low dose, remarkable suppression of the AO-induced mutagenicity, its antimutagenic potency being almost three orders of magnitude higher when compared to its close analogue, jatrorrhizine. Possible mechanisms of the antimutagenic action are discussed in terms of recent literature data. While the potent antimutagenic activity of the protoberberines most likely results from the inhibition of DNA topoisomerase I, the actual mechanism(s) for the BBI alkaloids is hard to be identified.
Taken together, the results indicate that berberine possesses promising antimutagenic/anticarcinogenic potential that is worth to be investigated further.
PMCID: PMC101396  PMID: 11943071
7.  Bilirubin and Related Tetrapyrroles Inhibit Food-Borne Mutagenesis: A Mechanism for Antigenotoxic Action against a Model Epoxide 
Journal of Natural Products  2013;76(10):1958-1965.
Bilirubin exhibits antioxidant and antimutagenic effects in vitro. Additional tetrapyrroles that are naturally abundant were tested for antigenotoxicity in Salmonella. Un-/conjugated bilirubin (1 and 2), biliverdin (4), bilirubin and biliverdin dimethyl esters (3 and 5), stercobilin (6), urobilin (7), and protoporphyrin (8) were evaluated at physiological concentrations (0.01–2 μmol/plate; 3.5–714 μM) against the metabolically activated food-borne mutagens aflatoxin B1 (9) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (10). Compound 8 most effectively inhibited the mutagenic effects of 9 in strain TA102 and 10 in TA98. Compound 7 inhibited 9-induced mutagenesis in strain TA98 most effectively, while 1 and 4 were promutagenic in this strain. This is likely due to their competition with mutagens for phase-II detoxification. Mechanistic investigations into antimutagenesis demonstrate that tetrapyrroles react efficiently with a model epoxide of 9, styrene epoxide (11), to form covalent adducts. This reaction is significantly faster than that of 11 with guanine. Hence, the evaluated tetrapyrroles inhibited genotoxicity induced by poly-/heterocyclic amines found in foods, and novel evidence obtained in the present investigation suggests this may occur via chemical scavenging of genotoxic metabolites of the mutagens investigated. This may have important ramifications for maintaining health, especially with regard to cancer prevention.
PMCID: PMC3812704  PMID: 24156291
8.  Bioactive Lipidic Extracts from Octopus (Paraoctopus limaculatus): Antimutagenicity and Antiproliferative Studies 
Fractions from an organic extract from fresh octopus (Paraoctopus limaculatus) were studied for biological activities such as antimutagenic and antiproliferative properties using Salmonella tester strains TA98 and TA100 with metabolic activation (S9) and a cancer cell line (B-cell lymphoma), respectively. A chloroform extract obtained from octopus tentacles was sequentially fractionated using thin layer chromatography (TLC), and each fraction was tested for antimutagenic and antiproliferative activities. Organic extract reduced the number of revertants caused by aflatoxin B1 showing a dose-response type of relationship. Sequential TLC fractionation of the active extracts produced several antimutagenic and/or antiproliferative fractions. Based on the results obtained, the isolated fractions obtained from octopus contain compounds with chemoprotective properties that reduce the mutagenicity of AFB1 and proliferation of cancer cell lines.
PMCID: PMC3562661  PMID: 23401709
9.  Antimutagenicity and Antiproliferative Studies of Lipidic Extracts from White Shrimp (Litopenaeus vannamei) 
Marine Drugs  2010;8(11):2795-2809.
An organic extract from fresh shrimp (Litopenaeus vannamei) was studied for antimutagenic and antiproliferative properties using Salmonella typhimurium tester strains TA98 and TA100 with metabolic activation (S9) and a cancer cell line (B-cell lymphoma), respectively. Shrimp extract was sequentially fractionated by thin layer chromatography (TLC) and each fraction was tested for antimutagenic and antiproliferative activities. Crude organic extracts obtained from shrimp reduced the number of revertants caused by aflatoxina B1, showing a dose-response type of relationship. Sequential TLC fractionation of the active extracts produced several antimutagenic and/or antiproliferative fractions. These results suggested that the lipid fraction of the tested species contained compounds with chemoprotective properties that reduce the mutagenicity of AFB1 and proliferation of a cancer cell line.
PMCID: PMC2996177  PMID: 21139845
antimutagenicity; antiproliferation; cultured shrimp
10.  Determination of the antimutagenicity of an aqueous extract of Rhizophora mangle L. (Rhizophoraceae), using in vivo and in vitro test systems 
Genetics and Molecular Biology  2010;33(1):176-181.
An aqueous extract of Rhizophora mangle L. bark is used as raw material in pottery making in the State of Espirito Santo, Brazil. This extract presents large quantities of tannins, compounds possessing antioxidant properties. Tannin antioxidant activity, as a plant chemical defense mechanism in the process of stabilizing free radicals, has been an incentive to studies on anti-mutagenicity. The present work aimed to evaluate possible antimutagenic activity of a R. mangle aqueous extract, using the Allium cepa test-system and micronuclear (MN) assay with blockage of cytokinesis in Chinese hamster ovary cells (CHO-K1). The Allium cepa test-system indicated antimutagenic activity against the damage induced by the mutagenic agent methyl methanesulfonate. A reduction in both MN cell frequency and chromosome breaks occurred in both the pre and post-treatment protocols. The MN testing of CHO-K1 cells revealed anti-mutagenic activity of the R. mangle extract against methyl methanesulfonate and doxorubicin in pre, simultaneous and post-treatment protocols. These results suggest the presence of phyto-constituents in the extract presenting demutagenic and bio-antimutagenic activities. Since the chemical constitution of Rhizophora mangle species presents elevated tannin content, it is highly probable that these compounds are the antimutagenic promoters themselves.
PMCID: PMC3036080  PMID: 21637623
Rhizophora mangle; antimutagenicity; Allium cepa; CHO-K1
11.  Pharmacological Assessment of the Medicinal Potential of Acacia mearnsii De Wild.: Antimicrobial and Toxicity Activities 
Acacia mearnsii De Wild. (Fabaceae) is a medicinal plant used in the treatment of microbial infections in South Africa without scientific validation of its bioactivity and toxicity. The antimicrobial activity of the crude acetone extract was evaluated by both agar diffusion and macrobroth dilution methods while its cytotoxicity effect was assessed with brine shrimp lethality assay. The study showed that both bacterial and fungal isolates were highly inhibited by the crude extract. The MIC values for the gram-positive bacteria (78.1–312.5) μg/mL, gram-negative bacteria (39.1–625) μg/mL and fungal isolates (625–5000) μg/mL differ significantly. The bacteria were more susceptible than the fungal strains tested. The antibiosis determination showed that the extract was more (75%) bactericidal than bacteriostatic (25%) and more fungicidal (66.67%) than fungistatic (33.33%). The cytotoxic activity of the extract was observed between 31.25 μg/mL and 500 μg/mL and the LC50 value (112.36 μg/mL) indicates that the extract was nontoxic in the brine shrimp lethality assay (LC50 > 100 μg/mL). These results support the use of A. mearnsii in traditional medicine for treatment of microbial infections. The extract exhibiting significant broad spectrum antimicrobial activity and nontoxic effects has potential to yield active antimicrobial compounds.
PMCID: PMC3344212  PMID: 22605976
Acacia mearnsii; antimicrobial activity; bactericidal; cytotoxic effects; extract
12.  Microbial Phylotype Composition and Diversity Predicts Plant Productivity and Plant-Soil Feedbacks 
Ecology letters  2012;16(2):167-174.
The relationship between ecological variation and microbial genetic composition is critical to understanding microbial influence on community and ecosystem function. In glasshouse trials using nine native legume species and forty rhizobial strains, we find that bacterial rRNA phylotype accounts for 68% of inter-isolate variability in symbiotic effectiveness and 79% of host-specificity in growth response. We also find that rhizobial phylotype diversity and composition of soils collected from a geographic breadth of sites explains the growth responses of two acacia species. Positive soil microbial feedback between the two acacia hosts was largely driven by changes in diversity of rhizobia. Greater rhizobial diversity accumulated in association with the less responsive host species, Acacia salicina, and negatively affected the growth of the more responsive A. stenophylla. Together this work demonstrates correspondence of phylotype with microbial function, and demonstrates that the dynamics of rhizobia on host-species can feed back on plant population performance.
PMCID: PMC3547158  PMID: 23216788
Mutualism; Symbiosis; Rhizobia; Plant-microbe interactions; Plant ecology; Microbial ecology; Plant-soil feedback; Diversity-productivity; Molecular ecology; Specificity; Ecosystem function
13.  Antibacterial activity of Artemisia nilagirica leaf extracts against clinical and phytopathogenic bacteria 
The six organic solvent extracts of Artemisia nilagirica were screened for the potential antimicrobial activity against phytopathogens and clinically important standard reference bacterial strains.
The agar disk diffusion method was used to study the antibacterial activity of A. nilagirica extracts against 15 bacterial strains. The Minimum Inhibitory Concentration (MIC) of the plant extracts were tested using two fold agar dilution method at concentrations ranging from 32 to 512 μg/ml. The phytochemical screening of extracts was carried out for major phytochemical derivatives in A. nilagirica.
All the extracts showed inhibitory activity for gram-positive and gram-negative bacteria except for Klebsiella pneumoniae, Enterococcus faecalis and Staphylococcus aureus. The hexane extract was found to be effective against all phytopathogens with low MIC of 32 μg/ml and the methanol extract exhibited a higher inhibition activity against Escherichia coli, Yersinia enterocolitica, Salmonella typhi, Enterobacter aerogenes, Proteus vulgaris, Pseudomonas aeruginosa (32 μg/ml), Bacillus subtilis (64 μg/ml) and Shigella flaxneri (128 μg/ml). The phytochemical screening of extracts answered for the major derivative of alkaloids, amino acids, flavonoids, phenol, quinines, tannins and terpenoids.
All the extracts showed antibacterial activity against the tested strains. Of all, methanol and hexane extracts showed high inhibition against clinical and phytopathogens, respectively. The results also indicate the presence of major phytochemical derivatives in the A. nilagirica extracts. Hence, the isolation and purification of therapeutic potential compounds from A. nilagirica could be used as an effective source against bacterial diseases in human and plants.
PMCID: PMC2830175  PMID: 20109237
14.  In Vitro Antimicrobial Activity of Acacia catechu and Its Phytochemical Analysis 
Indian Journal of Microbiology  2011;50(4):369-374.
Acacia catechu, commonly known as catechu, cachou and black cutch is an important medicinal plant and an economically important forest tree. The methanolic extract of this plant was found to have antimicrobial activities against six species of pathogenic and non-pathogenic microorganisms: Bacillus subtilis, Staphylococcus aureus, Salmonella typhi, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The maximum zone of inhibition (20 mm) was found to be exhibited against S. aureus. For this organism the minimum bactericidal concentration (MBC) of the crude extract was 1,000 μg/ml. The extract was found to be equally effective against gram positive and gram negative bacteria. The antimicrobial activity of the extract was found to be decreased during purification. The chemical constituents of organic plant extracts were separated by thin layer chromatography (TLC) and the plant extracts were purified by column chromatography and were further identified by Gas chromatography–mass selection (GC–MS) analysis. The composition of A. catechu extract had shown major components of terpene i.e. camphor (76.40%) and phytol (27.56%) along with other terpenes in minor amounts which are related with their high antibacterial and antifungal properties.
Electronic supplementary material
The online version of this article (doi:10.1007/s12088-011-0061-1) contains supplementary material, which is available to authorized users.
PMCID: PMC3209846  PMID: 22282602
Acacia catechu; Minimum inhibitory concentration; Zone of inhibition; In vitro antimicrobial activity; TLC; GC/MS
15.  Assessment of genotoxicity and antigenotoxicity of an aqueous extract of Cleistocalyx nervosum var. paniala in in vitro and in vivo models 
Interdisciplinary Toxicology  2012;5(4):201-206.
Cleistocalyx nervosum var. paniala, an edible fruit found in Northern Thailand, contains high amounts of phenolic compounds with in vitro antioxidant activity. The aqueous extract of the ripe fruit was evaluated for its safety and beneficial effects using genotoxicity and toxicity tests. The C. nervosum extract was not only non-mutagenic in Salmonella typhimurium strains TA98 and TA100 in the presence and absence of metabolic activation, but exhibited also moderate antimutagenic effects against aflatoxin B1 and 2-amino-3,4-dimethylimidazo[4,5-f]quinoline-induced mutagenesis. Electrospray ionization-mass spectrometric analysis revealed the major anthocyanins, which included cyanidin-3,5-diglucoside, cyanidin-3-glucoside and cyanidin-5-glucoside. The administration of C. nervosum at concentration of 5,000 mg/kg bw did not induce acute toxicity in rats. A liver micronucleus test was performed to detect clastogenicity and anticlastogenicity. The extract in the dose of 1,000 mg/kg did not cause micronucleus formation in the liver of rats. Furthermore, in rats administered 100–1,000 mg/kg of the extract, no anticlastogenic effect against diethylnitrosamine-induced hepatic micronucleus formation was observed. These studies provide data concerning the safety and antimutagenic potency of an aqueous extract of C. nervosum fruit.
PMCID: PMC3600524  PMID: 23554564
Cleistocalyx nervosum var. paniala; acute toxicity; ames test; liver micronucleus assay
16.  Cancer Chemopreventive Mechanisms of Tea Against Heterocyclic Amine Mutagens from Cooked Meat (44373) 
Cooking meat and fish under normal conditions produces heterocyclic amine mutagens, several of which have been shown to induce colon tumors in experimental animals. In our search for natural dietary components that might protect against these mutagens, it was found that green tea and black tea inhibit the formation of heterocyclic amine-induced colonic aberrant crypt foci (ACF) in the rat. Since ACF are considered to be putative preneoplastic lesions, we examined the inhibitory mechanisms of tea against the heterocyclic amines. In the initial studies using the Salmonella mutagenicity assay, green tea and black tea inhibited according to the concentration of tea leaves during brewing and the time of brewing; a 2–3-min brew of 5% green tea (w/v) was sufficient for >90% antimutagenic activity. N-hydroxylated heterocyclic amines, which are direct-acting mutagens in Salmonella, were inhibited by complete tea beverage and by individual components of tea, such as epigallocatechin-3-gallate (EGCG). Inhibition did not involve enhanced mutagen degradation, and EGCG and other catechins complexed only weakly with the mutagens, suggesting electrophile scavenging as an alternative mechanism. Enzymes that contribute to the metabolic activation of heterocyclic amines, namely microsomal NADPH-cytochrome P450 reductase and N,O-acetyltransferase, were inhibited by tea in vitro. Studies in vivo established that tea also induces cytochromes P450 and Phase II enzymes in a manner consistent with the rapid metabolism and excretion of heterocyclic amines. Collectively, the results indicate that tea possesses anticarcinogenic activity in the colon, and this most likely involves multiple inhibitory mechanisms.
PMCID: PMC2268949  PMID: 10202396
17.  Evaluation of the genotoxic and antigenotoxic potential of Melissa officinalis in mice 
Genetics and Molecular Biology  2011;34(2):290-297.
Melissa officinalis (L.) (Lamiaceae), a plant known as the lemon balm, is native to the east Mediterranean region and west Asia. Also found in tropical countries, such as Brazil, where it is popularly known as “erva-cidreira” or “melissa”, it is widely used in aqueous- or alcoholic-extract form in the treatment of various disorders. The aim was to investigate in vivo its antigenotoxicity and antimutagenicity, as well as its genotoxic/mutagenic potential through comet and micronucleus assaying. CF-1 male mice were treated with ethanolic (Mo-EE) (250 or 500 mg/kg) or aqueous (Mo-AE) (100 mg/kg) solutions of an M. officinalis extract for 2 weeks, prior to treatment with saline or Methyl methanesulfonate (MMS) doses by intraperitoneal injection. Irrespective of the doses, no genotoxic or mutagenic effects were observed in blood and bone-marrow samples. Although Mo-EE exerted an antigenotoxic effect on the blood cells of mice treated with the alkylating agent (MMS) in all the doses, this was not so with Mo-AE. Micronucleus testing revealed the protector effect of Mo-EE, but only when administered at the highest dose. The implication that an ethanolic extract of M. officinalis has antigenotoxic/antimutagenic properties is an indication of its medicinal relevance.
PMCID: PMC3115325  PMID: 21734832
Melissa officinalis; comet assay; micronucleus test; genotoxicity; antigenotoxicity
18.  Minor lipid components of some Acacia species: potential dietary health benefits of the unexploited seeds 
Oilseed samples from four Acacia species ( A. cyclops, A. ligulata, A. salicina and A. cyanophylla) were analyzed in order to evaluate the potential nutritional value of their unexploited seeds.
Samples were collected from different Tunisian geographic locations. Seed oils were extracted and carotenoids, tocopherols and sterols were analyzed using chromatographic methods.
The studied Acacia seeds seem to be quite rich in lipids (from 6% to 12%). All Acacia species contain mainly the xanthophylls zeaxanthin and lutein compounds: from ca. 38 of total lipids (A. cyclops) to ca. 113 of total lipids (A. cyanophylla). Total tocopherols varied from ca. 221 of total lipids (A. cyclops) to ca. 808 of total lipids (A. ligulata). Sterols are highly present and their contents ranged between ca. 7 g. kg-1 of total lipids (A. salicina) and 11 g. kg-1 of total lipids (A. cyclops).
This study highlights that these unexploited seeds might have a potential nutritional value and encourages researchers to more explore and find developments for these plants for healthy purposes.
PMCID: PMC3439337  PMID: 22577797
Unexploited Acacia; Oilseeds; Carotenoids; Tocopherols; Sterols
19.  In vitro antibacterial and antifungal activities of Cassia fistula Linn. fruit pulp extracts 
Ayu  2012;33(1):123-129.
Aim of the study is to assess the antimicrobial activity Cassia fistula fruit pulp extracts on some bacterial and fungal strains. Hydro alcohol and chloroform extracts of Cassia fistula fruit pulp were evaluated for the potential antimicrobial activity. The antimicrobial activity was determined in both the extracts using the agar disc diffusion method. Extracts were effective on tested microorganisms. The antibacterial and antifungal activities of solvent extracts (5, 25, 50, 100, 250 μg/mL) of C. fistula were tested against two gram positive, two gram negative human pathogenic bacteria and three fungi, respectively. Crude extracts of C. fistula exhibited moderate to strong activity against most of the bacteria tested. The tested bacterial strains were Staphylococcus aureus, Streptococcus pyogenes, Escherichia coil, Pseudomonas aeruginosa, and fungal strains were Aspergillus. niger, Aspergillus. clavatus, Candida albicans. The antibacterial potential of the extracts were found to be dose dependent. The antibacterial activities of the C. fistula were due to the presence of various secondary metabolites. Hence, these plants can be used to discover bioactive natural products that may serve as leads in the development of new pharmaceuticals research activities.
PMCID: PMC3456850  PMID: 23049197
Antibacterial activity; Antifungal activity; Bacteria; Cassia fistula
20.  Antimutagenic and mutagenic potentials of Chinese radish. 
Environmental Health Perspectives  1993;101(Suppl 3):247-252.
The edible part of fresh Chinese radish was chopped into small pieces, lyophilized, and then extracted sequentially with hexane, chloroform, and methanol. The solvent in each fraction was removed by evaporation under reduced pressure at 50-55 degrees C, and the residue was dissolved in dimethylsufoxide just before being tested for antimutagenicity as well as mutagenicity using the Salmonella/mammalian microsome mutagenicity test. We found that none of the three fractions exhibited any mutagenicity toward S. typhimurium strains TA98 and TA100 when tested either in the presence or absence of S-9 mix. Interestingly, however, hexane and chloroform extracts could strongly inhibit the mutagenicities of both direct mutagens (e.g., 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide and sodium azide) and indirect mutagens (e.g., aflatoxin B1). In contrast, however, these two fractions did not inhibit the mutagenicity of benzo[a]pyrene, which is also an indirect mutagen. Both hexane and chloroform extracts could also markedly inhibit the activities of rat liver aniline hydroxylase and aminopyrine demethylase. The methanol fraction could inhibit neither the mutagenicities of direct or indirect mutagens tested nor the activities of those two rat liver enzymes. Results of the present study demonstrate that Chinese radish may not contain any mutagenic compound but does contain some nonpolar compounds with antimutagenic activity toward both direct and indirect mutagens. In addition, the antimutagenic activity toward aflatoxin B1 may be partly due to the inhibition of enzymes necessary for activation of this mutagen.
PMCID: PMC1521162  PMID: 8143625
21.  Chemopreventive properties of chlorophylls towards aflatoxin B1: a review of the antimutagenicity and anticarcinogenicity data in rainbow trout 
Carcinogenesis  1999;20(10):1919-1926.
The anticarcinogenic activity of chlorophyllin (CHL), a water-soluble derivative of chlorophyll, was first reported in rainbow trout. This review describes certain experiments which set the stage for long-term tumor bioassays, in trout and other species, using CHL and various food-borne carcinogens. Initial work with trout and rat liver enzymes in the Salmonella assay showed that CHL was a potent antimutagen towards heterocyclic amines, polycyclic aromatic hydrocarbons, aflatoxins and other classes of mutagen. Antimutagenic activity was further demonstrated using the corresponding direct-acting mutagens in the absence of an exogenous metabolizing system. Mutagen-inhibitor interaction (molecular complex formation) was identified in spectrophotometry studies, suggesting that CHL acts as an ‘interceptor molecule’. In vivo, CHL reduced hepatic AFB1-DNA adducts and hepatocarcinogenesis when the inhibitor and carcinogen were co-administered in the diet. Finally, co-injection of inhibitor and AFB1 into trout embryos established that CHL was more effective than chlorophyll a in reducing AFB1-DNA adducts 2 weeks after injection, and liver tumors after 1 year.
PMCID: PMC2386994  PMID: 10506105
Chlorophyllin; Aflatoxin B1; Heterocyclic amine; Trout; Salmonella assay; DNA adduct; Anticarcinogen
22.  Evaluation of comparative and combined antimutagenic potential of vitamin C and vitamin E using histidine mutant Salmonella typhimurium strains 
Chemoprevention represents a new intervention strategy to control some type of carcinogenesis especially in subjects at high risk for cancer development. Experimental and epidemiological data indicate that a variety of nutritional factors including vitamin C and E are effective to lower the risk of some types of cancer. However large prospective studies have failed to find such significant association. A comparative and combined in vitro antimutagenic potential of two antioxidant vitamins ascorbic acid (vitamin C) and α-tocopherol (vitamin E) were evaluated using Ame’s Salmonella typhimurium test assay. Directly acting mutagens such as sodium azide (NaN3) and 4-Nitro-o-phenylenediamine (NPDA), and N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) were used to induce mutation in salmonella strains TA 98 and TA 100. Vitamin C significantly (P < 0.01) and dose dependently inhibited the mutagenicity induced by all the three mutagens. The percent inhibitions of vitamin C at 15 mg/plate were 33.8% (NaN3), 52.5 % (MNNG) and 55.4 % (NPDA). Vitamin E (15 mg/plate) was effective to inhibit mutagenicity induced by NaN3 and MNNG but did not inhibit mutation induced by NPDA. Combination of vitamins (vitamin C plus vitamin E) produced only an additive antimutagenic activity when compared to their activity at 5 mg/plate. The results of the study concluded that vitamin C is a better antimutagenic agent than vitamin E and combination of vitamins did not produce any synergistic activity.
PMCID: PMC3453667  PMID: 23105714
Antimutagenicity; Antioxidant; Cancer chemoprevention; Vitamins
23.  Antimicrobial activity of selected South African medicinal plants 
Nearly 3,000 plant species are used as medicines in South Africa, with approximately 350 species forming the most commonly traded and used medicinal plants. In the present study, twelve South African medicinal plants were selected and tested for their antimicrobial activities against eight microbial species belonging to fungi, Mycobacteria, Gram-positive and Gram-negative bacteria.
The radiometric respiratory technique using the BACTEC 460 system was used for susceptibility testing against Mycobacterium tuberculosis, and the liquid micro-broth dilution was used for other antimicrobial assays.
The results of the minimal inhibitory concentration (MIC) determinations indicated that the methanol extracts from Acacia karoo, Erythrophleum lasianthum and Salvia africana were able to prevent the growth of all the tested microorganisms. All other samples showed selective activities. MIC values below 100 μg/ml were recorded with A. karoo, C. dentate, E. lasianthum, P. obligun and S. africana on at least one of the nine tested microorganisms. The best activity (MIC value of 39.06 μg/ml) was noted with S. africana against E. coli, S. aureus and M. audouinii, and Knowltonia vesitoria against M. tuberculosis.
The overall results of the present work provide baseline information for the possible use of the studied South African plant extracts in the treatment of microbial infections.
PMCID: PMC3479042  PMID: 22704594
24.  Antibacterial and antifungal activities from leaf extracts of Cassia fistula l.: An ethnomedicinal plant 
This study was carried out with an objective to investigate the antibacterial and antifungal potentials of leaves of Cassia fistula Linn. The aim of the study is to assess the antimicrobial activity and to determine the zone of inhibition of extracts on some bacterial and fungal strains. In the present study, the microbial activity of hydroalcohol extracts of leaves of Cassia fistula Linn. (an ethnomedicinal plant) was evaluated for potential antimicrobial activity against medically important bacterial and fungal strains. The antimicrobial activity was determined in the extracts using agar disc diffusion method. The antibacterial and antifungal activities of extracts (5, 25, 50, 100, 250 μg/ml) of Cassia fistula were tested against two Gram-positive—Staphylococcus aureus, Streptococcus pyogenes; two Gram-negative—Escherichia coli, Pseudomonas aeruginosa human pathogenic bacteria; and three fungal strains—Aspergillus niger, Aspergillus clavatus, Candida albicans. Zone of inhibition of extracts were compared with that of different standards like ampicillin, ciprofloxacin, norfloxacin, and chloramphenicol for antibacterial activity and nystatin and griseofulvin for antifungal activity. The results showed that the remarkable inhibition of the bacterial growth was shown against the tested organisms. The phytochemical analyses of the plants were carried out. The microbial activity of the Cassia fistula was due to the presence of various secondary metabolites. Hence, these plants can be used to discover bioactive natural products that may serve as leads in the development of new pharmaceuticals research activities.
PMCID: PMC3217694  PMID: 22171301
Cassia fistula; in vitro antibacterial activity; antifungal activity; secondary metabolites
25.  Antimutagenicity of Cinnamaldehyde and Vanillin in Human Cells: Global Gene Expression and Possible Role of DNA Damage and Repair 
Mutation research  2006;616(1-2):60-69.
Vanillin (VAN) and cinnamaldehyde (CIN) are dietary flavorings that exhibit antimutagenic activity against mutagen-induced and spontaneous mutations in bacteria. Although these compounds were antimutagenic against chromosomal mutations in mammalian cells, they have not been studied for antimutagenesis against spontaneous gene mutations in mammalian cells. Thus, we initiated studies with VAN and CIN in human mismatch repair-deficient (hMLH1−) HCT116 colon cancer cells, which exhibit high spontaneous mutation rates (mutations/cell/generation) at the HPRT locus, permitting analysis of antimutagenic effects of agents against spontaneous mutation. Long-term (1–3-week) treatments of HCT116 cells with VAN at minimally toxic concentrations (0.5–2.5 mM) reduced the spontaneous HPRT mutant fraction (MF, mutants/106 survivors) in a concentration-related manner by 19% to 73%. A similar treatment with CIN at 2.5–7.5 μM yielded a 13% to 56% reduction of the spontaneous MF. Short-term (4–h) treatments also reduced the spontaneous MF by 64% (VAN) and 31% (CIN). To investigate the mechanisms of antimutagenesis, we evaluated the ability of VAN and CIN to induce DNA damage (comet assay) and to alter global gene expression (Affymetrix GeneChip) after 4-h treatments. Both VAN and CIN induced DNA damage in both mismatch repair-proficient (HCT116 + chr3) and deficient (HCT116) cells at concentrations that were antimutagenic in HCT116 cells. There were 64 genes in common whose expression was changed similarly by both VAN and CIN; these included genes related to DNA damage, stress responses, oxidative damage, apoptosis, and cell growth. RT-PCR results paralleled the Affymetrix results for 4 selected genes (HMOX1, DDIT4, GCLM, and CLK4). Our results show for the first time that VAN and CIN are antimutagenic against spontaneous mutations in mammalian (human) cells. These and other data lead us to propose that VAN and CIN may induce DNA damage that elicits recombinational DNA repair and, consequently, reduces spontaneous mutation.
PMCID: PMC1955325  PMID: 17178418
Cinnamaldehyde; Vanillin; Spontaneous mutagenesis; Microarray; Comet assay; Human cells; HPRT

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