Acacia salicina extracts protect against DNA damage and mutagenesis in bacteria and human lymphoblast cell K562 cultures

Three extracts were prepared from the leaves of Acacia salicina: aqueous, methanol, and ethyl acetate extracts. The antigenotoxic properties of these extracts were investigated by assessing the inhibition of mutagenicity of the indirect-acting mutagen benzo[a]pyrene using the Ames assay and the genotoxicity of the direct-acting mutagen, hydrogen peroxide, using the “Comet assay.” Aqueous, methanol, and ethyl acetate extracts at doses of 500, 50, and 500 μg per plate reduced benzo[a]pyrene mutagenicity by 95%, 82%, and 40%, respectively, in Salmonella typhimurium TA98 strain and by 91%, 66% and 63%, respectively, at the same doses with a TA97 assay system. Human lymphoblast cells K562 were pretreated with 50% inhibition concentration of each extracts and then treated by H₂O₂, for the Comet assay. The Comet assay results showed that ethyl acetate and methanol extracts decreased the DNA damage caused by H₂O₂ by, respectively, 34.8% and 31.3%. We envisaged also the study of the antioxidant effect of these extracts by the enzymatic xanthine/xanthine oxidase assay. Results indicated that methanol and ethyl acetate extracts were potent inhibitors of xanthine oxidase and superoxide anion scavengers. We conclude that these integrated approaches to antigenotoxicity and antioxidant assessment may be useful to help compare the beneficial effects associated with using A salicina as medicinal and dietary plant.     

Evaluation of the mutagenicity and antimutagenicity of extracts from oat,buckwheat and wheat bran in the Salmonella/microsome assay

This study examined the mutagenic and antimutagenic activities of the DMSO extracts from the oat, buckwheat and wheat bran, which are good sources of polyphenols with antioxidant and anticarcinogenic properties. Extracts from buckwheat and wheat bran showed no mutagenic activity. Oat extract showed slight mutagenic effect in Salmonella typhimurium TA102. The antimutagenic activities against direct-acting (3-(5-nitro-2-furyl)acrylic acid, 2-nitrofluorene, hydrogen peroxide) and indirect-acting (aflatoxin B1) mutagens were also investigated using Ames test with S. typhimurium TA98, TA100 and TA102. Cereal extracts exhibit concentration-dependent protective antigenotoxic activity against all used mutagens. The total phenolic content in studied cereal extracts expressed as gallic acid equivalent increases in the order: buckwheat < wheat bran < oat. Total flavonoid content expressed as rutin equivalent increases in the order: oat < wheat bran < buckwheat.     

Protective effects of methanol extracts from Cladonia rangiformis and Umbilicaria vellea against known mutagens sodium azide and 9-aminoacridine

Lichens and their various extracts have been occasionally used in the treatment of many diseases. Cladonia rangiformis and Umbilicaria vellea are two important species of these lichens and they have several biological activities. In the present study, methanol extracts of these lichens, which are grown in the Eastern Anatolia Region of Turkey, were isolated, and their mutagenic and antimutagenic properties were investigated by using AMES-Salmonella and Zea mays Root Tip Mitotic Index mutagenicity and antimutagenicity assay systems. Known mutagens sodium azide (NaN(3)) and 9-Aminoacridine (9-AA) were used to determine antimutagenic properties of methanol extracts. The results showed that all methanol extracts, investigated in the present study, can be considered genotoxically safe because they do not have mutagenic activity at the tested concentrations. Besides, all of them have antimutagenic activity against 9-AA known as a model intercalator agent in the AMES-Salmonella test system. The inhibition rates obtained from the antimutagenicity assays ranged from 37.07% (C. rangiformis-5 μg/plate) to 54.39% (C. rangiformis-5 μg/plate). Furthermore, all the methanol extracts have significant antimutagenic activity against NaN(3) mutagenicity in Z. mays Root Tip Mitotic Index assay system. These activities are valuable towards an extension of the employ of these drugs as new phytotherapeutic or preservative ingredients.     

Mutagenicity and antimutagenicity testing of six chemicals associated with the pungent properties of specific spices as revealed by the Ames Salmonella/microsomal assay

Three compounds, capsaicin, thymol and borneol, were initially screened for mutagenic activity using Salmonella typhimurium strains TA97, TA98 and TA100, with and without S9 metabolic activation, and 20 min standard preincubation time. Three other compounds, allyl isothiocyanate, eugenol and cinnamaldehyde, were screened for mutagenic activity as above, but with a prolonged, nonstandard preincubation time of up to 120 minutes. All six test compounds used in the assays are associated with the pungent properties of some specific spices in which the test compounds can be found to exist naturally. The first objective of this study was to observe if mutagenic activity can be correlated to the pungent properties of these six test compounds. However, due to toxicity and the observation that only capsaicin was mutagenic, using strain TA100 in the presence of S9 metabolic activation, it was not possible to deduce any relationship between mutagenicity and the test chemials' pungent properties. Naturally occurring capsaicin, found in the spice Capsicum annum, was detected and quantified using thin layer and gas chromatographic techniques.The final objective was to detect the presence of antimutagenic factor(s) in C. annum that would suppress the mutagenicity of capsaicin. When the mutagenic capsaicin and 2-aminoanthracene were assayed in the presence of C. annum acetone extract, using strain TA100 with S9 metabolic activation, the mutagenic response of both the mutagens were reduced by approximately 50%. Assaying capsaicin and 2-aminoanthracene in the presence of chlorophyll, the mutagenic response of the two mutagens was reduced by less than 40%. From this observation it was inferred that chlorophyll can successfully suppress the mutagenicity activities of capsaicin and 2-aminoanthracene, together with other antimutagenic factors that were present in the acetone extract of C. annum.     

松茸提取物体外抑制染发剂致突变作用

目的　寻找安全有效的抗突变生物资源 ,为降低染发剂致突变危害提供实验依据。方法　采用修改的Ames试验 ,观察真菌植物松茸提取物对染发剂致突变作用的影响。结果　松茸提取物能显著抑制染发剂的致突变作用 (P <0 0 1) ,对测试菌株TA97、TA98和TA10 0 的诱发回变菌落形成的抑制率最高分别达到 96 9% ,99 6 %和 95 4 % ,其抑制作用间存在明显的剂量 反应关系。结论　松茸提取物对染发剂致突变具有显著的抑制和抗突变作用     

氯化粪水有机致突变组分的分离

目的：探讨实验室模拟条件下氯化粪水有机浓集物致突变组分的分离。方法：采用Ames试验与硅胶柱分离技术相结合，将采集的粪水经大孔树脂XAD-2／8混合柱吸附，用乙酸乙酯和甲醇进行洗脱，洗脱浓集物用丙酮溶解分别置于大孔硅胶层析柱上过柱，然后用石油醚、二氯甲烷、丙酮、甲醇和水依次对层析柱作径向淋洗展开。用Ames试验对各淋洗液浓集物进行致突变性检测。结果：粪水氯化后有机浓集物呈现强致突变性，乙酸乙酯洗脱浓集液(EA)残渣量和比活性高于甲醇洗脱浓缩液(M)。用该程序分离的EA和M的主要活性组分都集中于二氯甲烷和丙酮两个部分，其残渣回收率分别高达96％、100％，重量比活性分别提高了4．06、15.27倍(TA98菌株)和2．85、5．29倍(TA100菌株)。结论：该分离程序是有效的。     

Nonmutagenicity of curcumin and its antimutagenic action versus chili and capsaicin

Turmeric, which is one of the commonly used spices in Indian cooking, was tested for mutagenicity using the Ames test. The alcoholic extract of fresh or dried turmeric, its principal components, and pyrolyzed turmeric powder and curcumin were tested for mutagenicity in Salmonella typhimurium strains with and without metabolic activation. None of these were mutagenic in all the tester strains. Chilies (which are used with turmeric powder) and their principal alkaloid capsaicin were mutagenic in the TA 98 with S9 mixture. We tested curcumin, which is the principal component of turmeric, for its antimutagenic effect. It showed dose-dependent decreases in mutagenicity of chili extract and capsaicin. Also, we compared the antimutagenicity of curcumin with other known antioxidants, including BHA, vitamins E and C, and vegetable oils. These all showed dose-dependent decreases in mutagenicity of chili extract and capsaicin. These studies show that although there are few mutagenic principles in Indian food, there is still quite a large number of antimutagenic principles in the Indian diet that will modulate the activity of environmental mutagens.     

新疆红树莓果实不同有效部位提取及其抗氧化活性测定

目的测定树莓果实不同提取部位总酚含量并比较不同提取部位抗氧化活性。方法树莓果实95%乙醇提取物经石油醚脱脂后,依次用乙酸乙酯、正丁醇萃取,对得到的各提取部位浸膏,①以没食子酸为标准品进行总酚含量测定,同时用相对标准偏差(RSD)表示稳定性;②采用1,1-二苯-2-苦肼基和羟自由基清除能力实验测定抗氧化活性。结果①Folin-Ciocalteu比色法具有精密度高、稳定性和重现性好等优点;②各提取部位物总酚含量从GAE10.56mg/g到96.76mg/g,其中以乙酸乙酯部位总酚含量最高(GAE96.76mg/g);③抗氧化实验中,树莓不同提取物均显示了较好的抗氧化活性,其中以乙酸乙酯部位抗氧化活性最高。结论树莓是良好的天然抗氧化剂。     

Antimutagenic,Antigenotoxic, and Anticytotoxic Activities of Silybum Marianum [L.] Gaertn Assessed by the Salmonella Mutagenicity Assay (Ames Test) and the Micronucleus Test in Mice Bone Marrow

Silymarin (SM), a standardized extract from Silybum marianum (L.) Gaertn., is composed mainly of flavonolignans, and silibinin (SB) is its major active constituent. The present study aimed to evaluate the antimutagenic activities of SM and SB using the Ames mutagenicity test in Salmonella Typhimurium, as well as their anticytotoxic and antigenotoxic activities using the mouse bone marrow micronucleus test. To assess antimutagenicity, Salmonella Typhimurium strains were treated with different concentrations of SM or SB and the appropriate positive control for each strain. To assess antigenotoxicity and anticytotoxicity, Swiss mice were treated with different concentrations of SM or SB and mitomycin C (MMC). The results showed that SM was not significantly effective in reducing the number of frameshift mutations in strain TA98, while SB demonstrated significant protection at higher doses (P < 0.05). Regarding strain TA 100, SM and SB significantly decreased mutagenicity (point mutations) (P < 0.05). The results of the antigenotoxic evaluation demonstrated that SM and SB significantly reduced the frequency of micronucleated polychromatic erythrocytes (MNPCE) (P < 0.05). The results also indicated that SM and SB significantly attenuated MMC-induced cytotoxicity (P < 0.05). Based on these results, both SM and SB presented antimutagenic, antigenotoxic, and anticytotoxic actions.     

Nonmutagenicity of curcumin and its antimutagenic action versus chili and capsaicin

Turmeric, which is one of the commonly used spices in Indian cooking, was tested for mutagenicity using the Ames test. The alcoholic extract of fresh or dried turmeric, its principal components, and pyrolyzed turmeric powder and curcumin were tested for mutagenicity in Salmonella typhimurium strains with and without metabolic activation. None of these were mutagenic in all the tester strains. Chilies (which are used with turmeric powder) and their principal alkaloid capsaicin were mutagenic in the TA 98 with S9 mixture. We tested curcumin, which is the principal component of turmeric, for its antimutagenic effect. It showed dose‐dependent decreases in mutagenicity of chili extract and capsaicin. Also, we compared the antimutagenicity of curcumin with other known antioxidants, including BHA, vitamins E and C, and vegetable oils. These all showed dose‐dependent decreases in mutagenicity of chili extract and capsaicin. These studies show that although there are few mutagenic principles in Indian food, there is still quite a large number of antimutagenic principles in the Indian diet that will modulate the activity of environmental mutagens.     

Effect of different polarities leaves crude extracts of Omani juniperus excels on antioxidant,antimicrobial and cytotoxic activities and their biochemical screening

ObjectiveTo prepare different polarities leave crude extracts of Juniperus excels (J. excels) and to determine their phytochemical screening, antioxidant, antimicrobial and cytotoxic activities.MethodsThe phytochemical screening of different crude extracts was determined by well-established methods. The antioxidant activity was determined by free radical scavenging (2,2-diphenyl-1-picrylhydrazyl, DPPH) method. The antimicrobial activity was evaluated by agar disc diffusion method and cytotoxic activity was determined through brine shrimp lethality assay.ResultsThe phytochemical tests showed that alkaloids, flavonoids, saponins, steroide, triterpenoids and tannins are present in all leave crude extracts except chloroform crude extract do not contain tannins. The antioxidant results of crude extracts were found to be in the order of hydro alcoholic > chloroform> ethyl acetate >hexane. All leave crude extracts showed moderate antibacterial against gram positive and gram negative food borne pathogen bacteria. The cytotoxicity results revealed that hexane and chloroform extracts have killed all the shrimp larvae at the concentration of 500 μg/mL.ConclusionMore in-vivo and in-vitro studies along with detailed phytochemical investigations are needed in order to potentially use this plant in the prevention and therapies of some oxidative damage related diseases.     

Genotoxicity of urban air particulate matter: Correlations between mutagenicity data,airborne micropollutants,and meteorological parameters

The mutagenicity of airborne particulate matter was monitored at a site representative of the high traffic density of the city of Rome. Inhalable (less than 10 μm) particles were collected every other day with a high‐volume sampler from November 1990 to April 1991. Mutagenicity of particle extracts was evaluated by the microsuspension procedure with Salmonella typhimurium strain TA98. Mutagenic activities of particle extracts displayed a fourfold variation during the period of sampling, with the lowest values in April (about 600 induced revertants mg^‐1), and the highest in February (about 2500 revertants mg^‐1). Multivariate statistical analyses on the interrelationships between mutagenicity, micropollutants levels and meteorological parameters highlighted a close inverse relationship between air mutagenicity and ambient temperature. Lower temperatures determined both an increased content of organic matter on air particles, and an increased mutagenicity (on a weight basis) of particle extracts, which was associated with a relative enrichment of the more volatile PAHs.     

不同增/溶剂对Ames试验致突变性的影响

目的观察8种增/溶剂对Ames试验致突变性检出的影响,为合理选用增/溶剂提供参考。方法依照《化妆品卫生规范》(2007)中鼠伤寒沙门氏菌/回复突变试验方法,对95%乙醇、甲醇、丙酮、吐温-20、吐温-60、吐温-80、DMSO和乙酸乙酯8种增/溶剂进行测定和结果判定。结果吐温-20在不大于5.0mg/皿剂量下,其余7种增/溶剂在最大剂量100.0 mg/皿情况下对TA97、TA98、TA100和TA102菌株的回变菌落数对比判定均显阴性。结论 Ames试验中,95%乙醇、甲醇、丙酮、吐温-60、吐温-80、DMSO和乙酸乙酯最大使用剂量可为100.0 mg/皿;吐温-20最大使用剂量为5.0 mg/皿。     

Antigenotoxic properties of lactic acid bacteria in the S. typhimurium mutagenicity assay

A high percentage of human tumors is reported to be related to dietary habits. One way to improve the nutritional impact is to increase the intake of protective factors, such as inhibitors of DNA damage and other types of anticarcinogens. Specific strains of lactic acid bacteria used to ferment milk are promising candidates that may be antimutagenic and anticarcinogenic. We have studied the antimutagenicity of 10 isolated strains of beneficial lactic acid bacteria. Four types of fermented milk products were also studied for their protective properties. The effect of these bacteria on the yield of revertants induced by nitrosated beef extract was investigated in the Salmonella typhimurium mutagenicity assay. Eight of 10 isolated Lactobacillus strains reduced the yield of his+ revertants almost back to the levels of the untreated controls. Different fermented fresh yogurts containing viable bacteria (probably Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus or Lactobacillus acidophilus and Bifidobacteria) showed protective effects as well. The degree of suppressing revertants was independent of the yogurt's fat content. In contrast, yogurt products that had been heat treated were not inhibitory. The other fresh fermented milk products (e.g., buttermilk, kefir, and “Dickmilch”) were not antimutagenic in this study. The results imply that some bacteria used in milk processing have an antimutagenic potential and that this property is specific for the bacterial strain.     

Polyphenols,vitamin C and antioxidant activity in wines from Rosa canina L. and Rosa rugosa Thunb.

The purpose of this study was to determine the concentration of biologically active compounds (polyphenols and l-ascorbic acid) in Rosa canina L. and Rosa rugosa Thunb. wines. The antioxidant capacity and antimutagenicity of the wines were also investigated. Aged and young wines contained phenolics levels of 2786–3456 and 3389–3990 mg/L GAE, respectively. The final concentrations of ascorbic acid were 1200 for Rosa rugosa Thunb. and 600 mg/L for Rosa canina L. R. rugosa and R. canina wines revealed high antioxidant activity in different assays (with ABTS, DPPH, and DMPD radicals). Expressed in terms of Trolox equivalent antioxidant capacity (TEAC), the activity ranged from 8 to 13.5 mM. Significant differences were found between the tested wines terms of their reactivity against the ABTS and DMPD radicals. The wines inhibited in vitro N-methyl-N′-nitro-nitrosoguanidine (MNNG) and the number of induced His⁺ revertants increased in a dose-dependent manner by 16–48% in Salmonella Typhimurium TA98 and 12–52% in Salmonella Typhimurium TA100. Wines from dog rose (Rosa canina L.) showed a greater ability to reduce mutations.     

2,3,7,8 Tetrachlorodibenzodioxin in fish from the pigeon river of Eastern Tennessee,USA: Its toxicity and mutagenicity as revealed by the Ames Salmonella assay

Levels of 2,3,7,8 tetrachlorodibenzodioxin (TCDD) were determined in both striated muscle (fillets) and whole body extracts of fish specimens harvested during a two-year period (1987–1989) from the Pigeon River (between Hartford and Newport) of Eastern Tennessee (USA). Whole body (wet weight) fish extract levels as high as 117 g/kg body weight and composite fish fillet (wet weight) extract levels as high as 87 g/kg fillet weight were observed. Pure TCDD was found to be highly toxic to theSalmonella typhimurium strains TA97, TA98, and TA100 at dosages which exceeded 825 ng TCDD/ml in the top agar of the Ames Salmonella assay. An 825 ng/ml TCDD dosage was not mutagenic to any of the tested Salmonella strains, either with or without metabolic activation (S9 mix). However, when acidic fish extracts from the Pigeon River were tested for mutagenicity, most of the fish extracts were mutagenic to Salmonella strains TA97, TA98, and TA100. These mutagenic extracts also demonstrated mutagenic dose-response curves. Other chemicals within the extracts as well as synergistic effects may account for the mutagenicity.     

Influence of ethanol and methanol gasoline blends on the mutagenicity of particulate exhaust extracts

TheSalmonella mutagenicity test was used to evaluate the influence of alcohol fuel extenders on the genetic toxicity of particulate exhaust extracts. Four spark-ignition engine equipped vehicles were operated on gasoline alone, 10% blends of ethanol or methanol in gasoline, and a commercially available “gasohol.” The tests were conducted on a chassis dynomometer and the particulate exhaust was collected on high volume filters after dilution in a tunnel. The vehicles used were a 1980 Chevrolet Citation, a 1980 Mercury Monarch, a 1981 Ford Escort and a 1981 Oldsmobile Cutlass. Dichloromethane extracts of the exhaust particles from all tests were mutagenic inSalmonella typhimurium strains TA 100 and TA 98. The extracts were less mutagenic in the nitroreductase deficient strains TA 98NR and TA 98DNPR suggesting that nitro substituted polycyclic aromatic hydrocarbons may be responsible for part of the mutagenicity. In all the alcohol blended fuel tests, the mass of particle associated organics emitted from the exhaust was lower than that observed during the control tests using gasoline alone. Thus, in most cases, estimates of the emission of mutagenic combustion products from the exhaust were lower in the alcohol blend tests.     

Modulatory effect of Byrsonima basiloba extracts on the mutagenicity of certain direct and indirect-acting mutagens in Salmonella typhimurium assays

Byrsonima basiloba A. Juss. species is a native arboreal type from the Brazilian "cerrado" (tropical American savanna), and the local population uses it to treat diseases, such as diarrhea and gastric ulcer. It belongs to the Malpighiaceae family, and it is commonly known as "murici." Considering the popular use of B. basiloba derivatives and the lack of pharmacological potential studies regarding this vegetal species, the mutagenic and antimutagenic effect of methanol (MeOH) and chloroform extracts were evaluated by the Ames test, using strains TA97a, TA98, TA100, and TA102 of Salmonella typhimurium. No mutagenic activity was observed in any of the extracts. To evaluate the antimutagenic potential, direct and indirect mutagenic agents were used: 4 nitro-o-phenylenediamine, sodium azide, mitomycin C, aflatoxin B(1), benzo[a]pyrene, and hydrogen peroxide. Both the extracts evaluated showed antimutagenic activity, but the highest value of inhibition level (89%) was obtained with the MeOH extract and strain TA100 in the presence of aflatoxin B(1). Phytochemical analysis of the extracts revealed the presence of n-alkanes, lupeol, ursolic and oleanolic acid, (+)-catechin, quercetin-3-O-alpha-L-arabinopyranoside, gallic acid, methyl gallate, amentoflavone, quercetin, quercetin-3-O-(2"-O-galloyl)-beta-D-galactopyranoside, and quercetin-3-O-(2"-O-galloyl)-alpha-L-arabinopyranoside.     

Mutagenicity of urine from nurses handling cytostatic drugs,influence of smoking

Summary Mutagenicity towards Salmonella typhimurium TA 100 of urine from smoking nurses, who were occupationally involved in the treatment of patients with cytostatic drugs, was significantly increased in comparison with that of smoking control subjects. Mutagenicity towards Salmonella typhimurium TA 100 was not increased in exposed non-smokers when compared to control non-smokers. In smoking subjects urinary mutagenicity appeared increased towards Salmonella typhimurium TA 1538 in the presence of S-9 mix.Rats pretreated with Aroclor 1254 showed higher mutagenicity in their urine than untreated rats after cyclophosphamide administration. Therefore, the synergistic effect of smoking might be due in part to induction of enzymes involved in the mutagenic activation of cytostatic drugs. Further, the animal experiments showed that cyclophosphamide (the most frequently used mutagenic cytostatic drug) can be absorbed after oral or percutaneous administration. Therefore, it is not excluded that differences in working hygiene between smokers and non-smokers also play a role.Support by the General Directorate of Labour, Dutch Ministry of Social Affairs     

Anticarcinogenic Activity of Strawberry,Blueberry, and Raspberry Extracts to Breast and Cervical Cancer Cells

Freeze-dried fruits of two strawberry cultivars, Sweet Charlie and Carlsbad, and two blueberry cultivars, Tifblue and Premier were sequentially extracted with hexane, 50% hexane/ethyl acetate, ethyl acetate, ethanol, and 70% acetone/water at ambient temperature. Each extract was tested separately for in vitro anticancer activity on cervical and breast cancer cell lines. Ethanol extracts from all four fruits strongly inhibited CaSki and SiHa cervical cancer cell lines and MCF-7 and T47-D breast cancer cell lines. An unfractionated aqueous extract of raspberry and the ethanol extract of Premier blueberry significantly inhibited mutagenesis by both direct-acting and metabolically activated carcinogens.