Antimutagenic and antioxidant activity of the essential oils of Citrus sinensis and Citrus latifolia.

The essential oils of Citrus sinensis and Citrus latifolia showed antimycotic activity against Candida spp. isolated from the oral cavity; they are neither mutagenic on the Ames test nor cytotoxic. Their main components are R-(+)-limonene, ß-thujene, α-myrcene and γ-terpinene. The aim of this work was to evaluate their antimutagenic and antioxidant capacities. Antimutagenic properties were evaluated against MNNG and ENNG on S. typhimurium TA100; against 2AA on strain TA98 and in front of 4NQO and NOR on strain TA102. Both were antimutagenic against MNNG (p < 0.001) but only C. latifolia was antimutagenic against ENNG (p < 0.001). Both presented antimutagenic activity against 2AA (p < 0.001). They were antioxidant against the ROS-generating compound 4NQO (p < 0.001) and the antibiotic NOR (p < 0.001). In the antioxidant evaluation, the activity in DPPH assay was in a range of 6-23% for C. sinensis and of 22-71% for C. latifolia. Both were antioxidant compared with BHT in ß-carotene bleaching assay and were able to decreased apoptosis in HaCat cells stimulated with H2O2. The levels of intracellular superoxide ion were lower in the presence of both oils. In conclusion, the essential oils of C. sinensis and C. latifolia are antimutagenic against at least three types of mutagens and have antioxidants properties.

Antimutagenicity of Stevia pilosa and Stevia eupatoria evaluated with the Ames test.

Stevia pilosa and Stevia eupatoria are plants used for various purposes in traditional medicine. In this report we studied the antimutagenic effect of methanolic extracts obtained from leaves, root, and flowers of the two species using the Ames test with and without metabolic activation. We tested the effect of the extracts on the damage induced by three mutagens with the following results: 1 - we found an inhibitory effect of both species on the mutagenicity induced by 2-aminoanthracene in the strain TA98. The best antimutagenic effect was obtained with leaves of both species and the flowers of S. eupatoria (99%), 2 - the mutations induced with N-ethyl-N'-nitro-N-nitrosoguanidine in the strain TA100 was also reduced. The flowers of S. pilosa and the root of S. eupatoria showed about 93% of inhibition, 3 - finally, the mutations induced by mitomycin-C on the strain TA102 had a reduction of 87% with the leaves of S. eupatoria. Besides, we determined the radical scavenging potential of the extracts with the DPPH method, and found a potent effect produced by all extracts, with an efficacy of more than 90%. The present study showed both antimutagenic and antioxidant potential of the tested extracts, and suggest the pertinence to confirm these effects in other models, and to accurately determine their mechanism of action.

The amoebicidal aqueous extract from Castela texana possesses antigenotoxic and antimutagenic properties.

Due to long-term treatment toxicity and clinical resistance to drugs commonly used against E. histolytica, new drugs against amoebiasis are urgently needed. Castela texana ("chaparro amargo") is a shrub taken traditionally in teas and capsules of dry plant to treat intestinal amoebic infections. An aqueous extract was prepared and its mutagenic, genotoxic and cytotoxicity properties were evaluated in prokaryotic and eukaryotic systems. This extract was neither mutagenic when evaluated with the Ames test in Salmonella typhimurium strains TA98, TA100 and TA102, nor genotoxic in unscheduled DNA synthesis in hepatocyte cultures, even at the highest concentrations tested. In fact, C. texana extract showed antimutagenic activity on S. typhimurium strains TA98 and TA100 in the Ames test. Furthermore, it was capable of protecting liver cell cultures against unscheduled DNA synthesis induced by 2-acetylaminofluorene at a concentration of 6.77 microg/ml. A free-radical scavenging test was used in order to explore the antioxidant capacity of C. texana extract with S. typhimurium strain TA102 pretreated with norfloxacin, a free radical producer. This extract showed a free radical withdrawal effect. The effective chemoprotective activity of this extract could be due to the antioxidant capacity of the C. texana extract components. In this paper it is shown that the antiamoebic natural product, C. texana, is also antimutagenic and protects against induction of preneoplastic lesions in rat liver. These results justify further studies to extend it use to human beings.

Antigenotoxic, antimutagenic and ROS scavenging activities of a Rhoeo discolor ethanolic crude extract.

Rhoeo discolor is a legendary plant used for treatment of superficial mycoses in Mexican traditional medicine. Despite its extended use, it is not known whether it has side-effects. An ethanolic crude extract from Rhoeo discolor was prepared, its mutagenic capacity was investigated by the Ames test, and its genotoxic activity in primary liver cell cultures using the unscheduled DNA synthesis assay. This extract was not mutagenic when tested with Salmonella typhimurium strains TA97, TA98 and TA100, and it did not elicit unscheduled DNA synthesis in hepatocyte cultures. In addition, we explored the antimutagenic and antigenotoxic activities of the extract and its ROS scavenger behaviour. Our results show that Rhoeo extract is antimutagenic for S. typhimurium strain TA102 pretreated with ROS-generating mutagen norfloxacin in the Ames test, and protects liver cell cultures against diethylnitrosamine induction of unscheduled DNA synthesis even at 1.9 ng per dish, which was the lowest dose tested. A free radical scavenging test was used in order to explore the antioxidant capacity of Rhoeo extract, as compared with three commercial well-known antioxidants quercetin, ascorbic acid and tocopherol. Rhoeo extract showed less radical scavenging effect than quercetin, but similar to that of alpha-tocopherol and more than ascorbic acid. It is important to note that this extract was neither mutagenic in S. typhimurium nor genotoxic in liver cell culture, even at concentrations as high as four- and 166-fold of those needed for maximal antimutagenic or chemoprotective activities, respectively.

S9 induction by the combined treatment with cyclohexanol and albendazole.

Cyclohexanol (CH) is an industrial solvent capable of inducing cytochrome P450 (CYP) enzymes including the CYP2E and CYP2B subfamilies. S9 from CH treated rats is able to activate several N-nitrosamines that are poorly activated by Aroclor 1254, phenobarbital/beta-naphthoflavone (PB/NF) or 3-methylcholanthrene S9 fractions into mutagens detected by the Salmonella typhimurium Ames test. Additionally, albendazole (ABZ) is a widely used anthelmintic drug and a potent inducer of the CYP1A subfamily. Since CYP1A, -2B and -2E subfamilies are implicated in the activation of several environmental mutagens/carcinogens, we studied CYP induction in the rat liver by the combined effect of these two compounds, and used S9 derived from it in the Salmonella/microsome assay to compare with S9 obtained from Aroclor or PB/NF treated rats. Total CYP content in hepatic microsomes was induced by Aroclor, but not by any of the other chemical combinations. Western blot and enzymatic activity analysis revealed quantitative but not qualitative differences in the CYP subfamilies present in the different microsomal fractions; all of the chemicals used increased the levels of CYP1A1/2, CYP2B1/2 and CYP2E1 with respect to control microsomes. CYP3A was not modified by the different treatments. When tested in the Ames test, Aroclor S9 and PB/NF S9 were the most effective in the activation of benzo[a]pyrene and 3-methylcholanthrene which are metabolized mainly by CYP1A1; additionally, the highest mutagenic potency of 2-aminofluorene and N-nitrosodipropylamine, which are activated by CYP1A2 and CYP2B, respectively, were obtained with PB/NF S9. All these compounds were also activated when CH/ABZ S9 was used as the exogenous source of metabolism. Mutagens like N-nitrosopyrrolidine and N-nitrosodimethylamine, activated by CYP2E1, were detected only when CH/ABZ S9 was used, and the effectiveness of the different S9 fractions in activating cyclophosphamide decreased in the following order: Aroclor = PB/NF > CH/ABZ > control. From these experiments we can conclude that the individual CYP- inducing properties of ABZ and CH complement each other when the two compounds are administered in conjunction and that the resulting S9 fraction is able to activate several known mutagens in the Ames test.

[Frequency of colicin and hemolysins in Escherichia coli isolated from pregnant patients with urinary tract infection, symptomatic and asymptomatic]. / Frecuencia de colicina y hemolisinas en Escherichia coli aisladas de pacientes embarazadas con infección de vías urinarias, sintomática y asintomática.

Colicin production was studied among 137 Escherichia coli strains isolated from pregnant patients with symptomatic or asymptomatic, urinary tract infection (IVU). The observed colicinogeny frequency was 72.92% (n = 96), among the symptomatic patients and 29.26% (n = 41) among the asymptomatic group. The most frequently identified colicins from symptomatic patients were: V (23.9%), A (18.7%) and E1 (17.7%) and within the patients with asymptomatic IVU the most frequently observed colicins were A (9.7%), E1 (17.0%) y V (2.4%). The major frequency of colicin V among the E. coli strains isolated from the symptomatic group against the asymptomatic one, was statistical significant (p > or = 0.025). The results on the observed frequencies of colicins E1 and A were not statistical significant. Among the 137 studied E. coli strains, 37.2% were hemolytic. 83.3% of the colicin V producing strains (n = 24) were hemolytic, among the strains producing other colicin different than colicin V 34% (n = 58) were hemolytic and 12.0% of non colicinogenic strains were hemolytic. These results were statistical significant (p > 0.05). The present data, suggest that colicins production is an important pathogenicity factor among the E. coli strains, specially for those strains producing colicin V. The observed association among hemolysin and colicin V production may be an interesting pathogenicity factor which suggests an increasing ability of uropathogenic strains to produce symptomatic urinary tract infections.

Antimutagenesis of beta-carotene to mutations induced by quinolone on Salmonella typhimurium.

BACKGROUND: Quinolone-induced mutagenesis in the Salmonella typhimurium hisG48 strains suggests that these antibiotics are oxygen free radical generators. The use of beta-carotene as antioxidant was evaluated as an alternative to reduce oxidative cell damage in patients who need therapy with nalidixic acid, norfloxacin, or pipemidic acid. The studied beta-carotene (30%), used by pharmaceutical laboratories as dietary complements, was not toxic or mutagenic for the S. typhimurium TA102 strain at a dose equivalent to 1,500 I.U. At the studied concentrations, the evaluated antimutagen did not modify the minimum inhibitory concentration of nalidixic acid, norfloxacin, or pipemidic acid against uropathogenic Escherichia coli strains. METHODS: The mutagenic effect of nalidixic acid and norfloxacin against hisG48 strains was inhibited with 1500 I.U. of beta-carotene. The antimutagenic effect of beta-carotene against mutations induced by pipemidic acid was observed even with 150 I.U. of beta-carotene. The antimutagenic effect against mutations induced on S. typhimurium TA102 or TA104 strains was observed only when the aroclor 1254 rat-induced liver S9 mixture was used. RESULTS: This mutagenic effect was detected only when the strains were exposed to quinolones and the beta-carotene simultaneously with the S9 mixture, suggesting that quinolones induce oxygen free radicals that may be scavenged by beta-carotene. CONCLUSIONS: The antimutagenic effect of this vitamin A precursor is probably due to the active molecule of vitamin A, a desmutagen with the ability of radical capture. A diet rich in beta-carotene or vitamin A could be a good alternative to reduce genotoxic risk to patients being treated with quinolone.

Genotoxic evaluation of norfloxacin and pipemidic acid with the Escherichia coli Pol A-/Pol A+ and the ames test.

BACKGROUND: Genotoxicity of antibiotics has not been well evaluated, and there is not much information on the genetic risk of quinolone drugs, even though they are widely used as alternative choice drugs in urinary infections. METHODS: Pipemidic acid and norfloxacin were tested for their capacity to induce point mutations using the Ames test and DNA damage on Escherichia coli PolA-/PolA+. RESULTS: At non-toxic doses, all of the drugs studied were negative on the E. coli PolA-/PolA+ test with or without in vitro metabolic activation with induced arochlor 1254 rat liver (S9). They did not produce frameshift mutations in TA98, or base-pair substitutions in S. typhimurium hisG46 strains TA100, or UTH8414. Norfloxacin and its induced metabolites in vitro with S9 rat liver were mutagenic to hisG48 strains TA102 and TA104, both of which detect oxidative chemicals. Pipemidic acid induced mutations in S. typhimurium hisG48 strains only when they had an efficient DNA excision repair system. CONCLUSIONS: These results suggest that the risk of oxygen-free radical generation from quinolones should be considered.

[Assessment of genetic toxicity of the exposure to clomiphene citrate, with various bacterial test systems]. / Valoración de riesgo gentóxico de exposición a citrato de clomifeno, con diferentes sistemas de prueba bacterianos.

Clomiphene citrate (CC) induces framshift mutations on the Salmonella typhimurium Ames strains TA1538, TA97 and TA100 employing in vitro metabolic activation with S9 aerochlor 1254 induced rat livers, but not base pair substitution mutations with neither the standard or the preincubation method. CC induced genolethal DNA damages on the Escherichia coli PolA-/PolA+ with S9 on the preincubation method or without S9 on the disk diffusion one. The severe primary DNA damages produced b CC was verified by the SOS induction on the lysogenic lambda phage induction with De Marini (1988) method and the induction of colicin E1 plasmid on E. coli. These results are suggestive that CC may be an adduct forming compound which is able to inhibit replication if the cell lacks DNA polymerase, or it may produce framshift mutations after replications. CC induced damages could be large lesions conducing to unicatenary DNA strains, that are able to induce the lexA regulated genes. So, the use of this ovulation inductor is a risk of genotoxic damage and it is advisable to do a risk-benefit evaluation in any particular case before its prescription.

Mutagenicity of urine from mice exposed orally to nitrite and various aminated antiparasitic drugs.

Mutagenic N-nitroso compound formation from the in vitro reaction of amebicides and anthelmintic drugs, which are pyrimidine derivatives or contain secondary aliphatic amines or heterocyclic nitrogens, has been previously described (Arriaga Alba et al.: Environmental and Molecular Mutagenesis 12:65-73, 1988). Under similar conditions, antiparasitic drugs containing halogenated derivatives of tertiary amines or quaternary ammonium salts do not form mutagenic nitrosated compounds. In the present study the mutagenic activity of mouse urine was determined after oral administration of sodium nitrite and the two above-mentioned groups of drugs (pyrantel pamoate, chloroquine, piperazine and dehydroemetine/iodochlorhydroxyquin and bephenium hydroxynaphthoate, respectively). Results show that the simultaneous administration of piperazine or chloroquine with sodium nitrite produced urinary mutagens that appeared conjugated as glucuronides, whereas pyrantel pamoate and dehydroemetine in the presence of nitrite caused only slightly mutagenic urine. No mutagenic activity was detected in the urine of mice to which halogenated derivatives of tertiary amines (iodochlorhydroxyquin) or quaternary ammonium salts (bephenium hydroxynaphthoate) were administered together with nitrite.

Survey of drug and phage resistance and colicin and hemolysin production among coliforms isolated in the Ivory Coast.

Analysis of 178 strains isolated as total and fecal coliforms in the Ivory Coast revealed that (i) hemolytic activity was scarce (0.6%) among this bacterial population; (ii) the most prevalent colicins detected were, in decreasing order, E, I, A, and G; (iii) the frequency of coliphage and drug resistance was similar to that observed in other countries, except for those of drug-resistant strains in animal feces, which were lower than in countries where animals are antibiotic fed; and (iv) one of the drug resistance plasmids seemed to possess a restriction-modification system and another seemed to code for capsular material.