RESUMO
Statins are inhibitors of cholesterol synthesis, but other biological properties, such as antimicrobial effects, have also been assigned to them, leading to their designation as pleiotropic agents. Our goal was to investigate the activity and selectivity of atorvastatin (AVA) against Trypanosoma cruzi by using in vitro models, aiming for more effective and safer therapeutic options through drug repurposing proposals for monotherapy and therapy in combination with benznidazole (BZ). Phenotypic screening was performed with different strains (Tulahuen [discrete typing unit {DTU} VI] and Y [DTU II]) and forms (intracellular forms, bloodstream trypomastigotes, and tissue-derived trypomastigotes) of the parasite. On assay of the Tulahuen strain, AVA was more active against intracellular amastigotes (selectivity index [SI] = 3). Also, against a parasite of another DTU (Y strain), this statin was more active (2.1-fold) and selective (2.4-fold) against bloodstream trypomastigotes (SI = 51) than against the intracellular forms (SI = 20). A cytomorphological approach using phalloidin-rhodamine permitted us to verify that AVA did not induced cell density reduction and that cardiac cells (CC) maintained their typical cytoarchitecture. Combinatory approaches using fixed-ratio methods showed that AVA and BZ gave synergistic interactions against both trypomastigotes and intracellular forms (mean sums of fractional inhibitory concentration indexes [∑FICIs] of 0.46 ± 0.12 and 0.48 ± 0.03, respectively). Thus, the repurposing strategy for AVA, especially in combination with BZ, which leads to a synergistic effect, is encouraging for future studies to identify novel therapeutic protocols for Chagas disease treatment.
Assuntos
Atorvastatina/farmacologia , Doença de Chagas/tratamento farmacológico , Nitroimidazóis/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Animais , Linhagem Celular , Doença de Chagas/parasitologia , Reposicionamento de Medicamentos/métodos , Sinergismo Farmacológico , Quimioterapia Combinada/métodos , Coração/parasitologia , Camundongos , Testes de Sensibilidade Parasitária/métodosRESUMO
Coffee is one of the most widely consumed beverages throughout the world. So far, many studies have shown the properties of coffee beverages, but little is known about its impacts on human and environmental health from its discard in the environment. So, the present work aims to investigate the mutagenic, genotoxic, cytotoxic and ecotoxic effects of leached (LE) and solubilized (SE) extracts from coffee waste, simulating the disposal of this residue in landfills and via sewage systems, respectively. Chemical analyses were also carried out. LE and SE induced mutagenicity in the TA98 Salmonella strain with and without exogenous metabolization (S9). In the TA100 only SE induced mutagenicity, what was observed without S9. An increase in the frequency of micronuclei was observed in HepG2 cell line after 3 and 24h of exposure to both extracts. No cytotoxic effects were observed in HepG2 cells by WST-1 assay. The EC50 values for the LE and SE were 1.5% and 11.26% for Daphnia similis, 0.12% and 1.39% for Ceriodaphnia dubia and 6.0% and 5.5% for Vibrio fischeri, respectively. Caffeine and several transition metals were found in both extracts. Coffee waste discarded in the environment may pose a risk to human and environmental health, since this compound can cause DNA damage and present toxicity to aquatic organisms.
Assuntos
Organismos Aquáticos/efeitos dos fármacos , Café/química , Mutagênicos/toxicidade , Resíduos/análise , Poluentes Químicos da Água/toxicidade , Aliivibrio fischeri/efeitos dos fármacos , Animais , Organismos Aquáticos/genética , Bioensaio , Sobrevivência Celular/efeitos dos fármacos , Dano ao DNA , Daphnia/efeitos dos fármacos , Saúde Ambiental , Células Hep G2 , Humanos , Testes de Mutagenicidade , Salmonella/efeitos dos fármacos , Esgotos/química , Testes de Toxicidade/métodosRESUMO
LLL-3, an anthracene derived compound, has been shown to be a promising therapeutic agent for the treatment of some kinds of cancer such as chronic myeloid leukemia and glioblastoma. However, no data regarding the toxic properties of this compound have yet been described in the literature. The present work aimed to investigate the mutagenic and genotoxic activities of LLL-3 using the TA97, TA98, TA100, TA102 and TA104 Salmonella/microsome strains for the Ames test and the micronucleus assay with the mouse macrophage cell line RAW 264.7. The findings showed that LLL-3, at doses of 0.001, 0.01, 0.1, 1.0 and 10.0 µg/plate, did not induce mutagenic activity in the Salmonella strains used under the conditions tested, and nor did it present genotoxicity in RAW 264.7 cells, at 10.0, 100.0 and 1000.0 µg/mL doses. Moreover, it is important to point out that the mitotic index of the cells decreased after exposure to LLL-3 under the same conditions tested, which may suggest some cytostatic effect, since this compound acts by inhibiting STAT3. Since most drugs used in the treatment of cancer present mutagenic activity as an adverse effect, these results suggest that LLL-3 is a promising drug for cancer therapy.
Assuntos
Antraquinonas/toxicidade , Antineoplásicos/toxicidade , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Fator de Transcrição STAT3/antagonistas & inibidores , Salmonella typhimurium/efeitos dos fármacos , Animais , Antraquinonas/farmacologia , Antineoplásicos/farmacologia , Técnicas de Cultura de Células , Linhagem Celular , Relação Dose-Resposta a Droga , Macrófagos/efeitos dos fármacos , Macrófagos/patologia , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Salmonella typhimurium/genéticaRESUMO
The use of strains of Salmonella enterica serovar Typhimurium with different metabolic capacities can indicate the class or classes of compounds present in an environmental sample and enable the diagnosis of the mutagenic activity of these pollutants adsorbed on particulate matter (PM) in the air. In the present study, the sensitivity of Salmonella strains TA98NR, TA98/1,8-DNP6, YG1021, and YG1024 to detect nitro compounds adsorbed on samples of PM 2.5 was compared from three sites in Rio de Janeiro city. Samples were collected using a high-volume sampler at three sites: one with light traffic and two with heavy traffic. The assays were performed in the presence of 10-50 µg/ plate organic extracts with and without exogenous metabolization. The YG1021 and YG1024 strains showed the highest rev/m(3) values, confirming their enhanced sensitivity. As YG1024 also demonstrated sensitivity to nitro and amino compounds, we suggest its use in research into environmental contamination.
Assuntos
Poluentes Atmosféricos/toxicidade , Mutagênicos/toxicidade , Nitrocompostos/toxicidade , Salmonella typhi/efeitos dos fármacos , Cidades , Humanos , Testes de Mutagenicidade , Salmonella typhi/genéticaRESUMO
Risk assessment can provide a comprehensive estimate of potential effects of contaminants under specific, well-defined, and well-described circumstances, providing quantitative relationships between exposure and effects to identify and to define areas of concern. We investigated the mutagenic activity of particulate matter in air samples collected from three sites in Rio de Janeiro city. Samples were collected using a high-volume sampler at Avenida Brasil, at Campus of Universidade do Estado do Rio de Janeiro, and at Rebouças Tunnel. Six polycyclic aromatic hydrocarbons were quantified by gas chromatography/mass spectrometry. Salmonella typhimurium TA98 and the derivative strains TA98/1.8-DNP(6), YG1021, and YG1024, commonly used in mutagenicity assays, were treated (10-50 µg/plate), with and without exogenous metabolization. The highest values for the polycyclic aromatic hydrocarbons were detected at Rebouças Tunnel. For chrysene, as an example, the concentration was nearly 200 times higher than that established by the US Environmental Protection Agency. Frequent traffic jams can place bus drivers who go through the Rebouças Tunnel at risk of exposure to up to 0.69 ng/m(3) benzo(a) pyrene. Independent of exogenous metabolization, mutagenicity was detected in strains YG1021 and YG1024 at all the sites, suggesting nitro and amino derivatives of polycyclic aromatic hydrocarbons. Rebouças Tunnel air samples gave the highest values for rev/µg and rev/m(3). This could be due to the fact that the long, enclosed passageway through a mountain restricts ventilation. The cancer risk estimate in this study was 10(-3) for the benzo(a)pyrene, at the two sites, indicating a high risk.
Assuntos
Poluentes Atmosféricos/análise , Cidades , Exposição Ambiental/efeitos adversos , Hidrocarbonetos Policíclicos Aromáticos/análise , Poluentes Atmosféricos/toxicidade , Brasil , Crisenos/análise , Crisenos/toxicidade , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Testes de Mutagenicidade , Material Particulado/análise , Material Particulado/toxicidade , Hidrocarbonetos Policíclicos Aromáticos/toxicidade , Medição de Risco , Fatores de Risco , Salmonella typhimurium/efeitos dos fármacosRESUMO
Coral reefs are diverse ecosystems that have a high density of biodiversity leading to intense competition among species. These species may produce unknown substances, many with pharmacological value. Chromonephthea braziliensis is an invasive soft coral from the Indo-Pacific Ocean that is possibly transported by oil platforms and whose presence can be a threat to a region's biodiversity. This species produces secondary metabolites that are responsible for inducing damage to the local ecosystem. In the present study, extracts were prepared from dried colonies of C. braziliensis (solvents: hexane, dichloromethane, ethyl acetate, and methanol). We evaluated their mutagenicity using the Salmonella reverse mutation assay (TA97, TA98, TA100, and TA102 strains), their genotoxicity using the DNA breakage analysis and micronucleus assay, and scavenging activity using the 1,1-diphenyl-2-picrylhydrazyl-free radical assay. Cytotoxicity and mutagenicity were not observed for any of the extracts. Genotoxicity was observed for the dichloromethane, ethyl acetate, and methanol extracts at high concentrations, but no DNA damage was observed in the micronucleus assay. Scavenging activity was not detected.
Assuntos
Antozoários/química , Dano ao DNA/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Animais , Linhagem Celular , Macrófagos/efeitos dos fármacos , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Salmonella , SolventesRESUMO
In rats, N-nitrosodiethylamine (NDEA) induces tumors mainly in the liver. This could be because various enzymes are responsible for the metabolic activation of NDEA, besides the hepatic NDEA metabolizing enzyme, CYP2E1. We examined NDEA genotoxicity and cytotoxicity in primary cultures of female rat hepatocytes; we also looked at how it affected CYP mRNA expression. Single incubation with 0.9% NaCl resulted in a mean of 0.2% apoptotic cells, which doubled with 105 µg NDEA/mL. The frequency of necrosis with NDEA treatment was also doubled. Besides the cytotoxic effects, there was also a 4-fold decrease in mitotic index and a 3-fold decrease in the percentage of cells with micronuclei. A significant increase in micronucleus cells when hepatocytes were incubated with 2.1 µg NDEA/mL suggests that DNA repair was inactive. The chromosomal aberration evaluation revealed a discrete dose-response curve. Treatment with NDEA induced increases in CYP mRNA: CYP2B2 (1.8 times) and CYP2E1 (1.6 times) with non-cytotoxic NDEA concentrations (0.21-21 µg/mL). CYP2B1 mRNA levels decreased at 0.21 µg NDEA/mL (2.5-fold), while CYP4A3 mRNA decreased 1.3-fold. NDEA treatment at 2.1 µg/ mL induced a 1.9-fold increase in CYP3A1 mRNA. Understanding the cumulative effects in target cells during precarcinogenesis is crucial to understanding the mode of action of potential carcinogens and in order to develop comprehensive chemical toxicity profiles.
Assuntos
Alquilantes/farmacologia , Hidrocarboneto de Aril Hidroxilases/biossíntese , Citocromo P-450 CYP2E1/biossíntese , Dano ao DNA/efeitos dos fármacos , Dietilnitrosamina/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Hepatócitos/enzimologia , Esteroide Hidroxilases/biossíntese , Animais , Apoptose/efeitos dos fármacos , Células Cultivadas , Relação Dose-Resposta a Droga , Feminino , Hepatócitos/patologia , Neoplasias Hepáticas/induzido quimicamente , Neoplasias Hepáticas/enzimologia , Neoplasias Hepáticas/patologia , Necrose/enzimologia , Necrose/patologia , RNA Mensageiro , Ratos , Ratos Endogâmicos F344RESUMO
The marine environment is a rich source of biological active compounds and the sponges can be considered the most productive one. This diversity gives rise to unique chemical compounds with potential pharmacological properties. Our study is focused on the genotoxic and antigenotoxic evaluation of two crude extracts obtained from the Brazilian endemic marine sponge Arenosclera brasiliensis. Salmonella typhimurium reverse mutation test with TA97, TA98, TA100 and TA102 strains were performed. For antimutagenic analysis, a pre-, co-, and post-treatment to evaluate, respectively, intracellular and extracellular reactions and possible modulation on DNA repair. Additionally, in order to verify the influence of the crude extracts on DNA damage induction, a plasmid-DNA treatment was assayed. No mutagenicity was observed in Salmonella reverse mutation test, neither DNA strand induced damage. Antimutagenic activity was observed in pre-, co-, and post-treatment. A significant antigenotoxic effect was observed in the crude extract, which suggests that A. brasiliensis extract has the potential to protect DNA from the action of 4NQO, 2-aminofluorene, sodium azide and mitomycin C.
Assuntos
Antimutagênicos/toxicidade , Poríferos/química , Salmonella typhimurium/efeitos dos fármacos , Animais , Antimutagênicos/isolamento & purificação , Brasil , Dano ao DNA/efeitos dos fármacos , Reparo do DNA/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Mutagênicos/isolamento & purificação , Mutagênicos/toxicidade , Plasmídeos/metabolismo , Salmonella typhimurium/genética , Extratos de TecidosRESUMO
The marine environment is a rich source of biologically active compounds with pharmacological properties. Marine organisms often produce secondary metabolites with structural features different from those produced by terrestrial ones, and the Phylum Porifera seems to be one of the most productive in this sense. This study was undertaken to provide data on mutagenic and antimutagenic activities from an acetone (Areac) and an ethanol (Areet) extract obtained from Arenosclera brasiliensis, an endemic Brazilian sponge. A qualitative Salmonella reverse mutation test was performed with the TA97, TA98, TA100, and TA102 strains by incubating cells with Areac and Areet in the presence and absence of a known mutagen. A cytotoxic evaluation of the extracts was also performed. A. brasiliensis did not display any mutagenic activity, but Areac showed significant toxicity against test strains. In the antimutagenic assay, a reduction in the number of his+ revertants was observed for the TA97, TA100 and TA102 strains treated with Areac when compared to the positive controls. Areet treatment showed protective activity against DNA lesions only for the TA100. These results are in agreement with those obtained previously with other A. brasiliensis extracts, suggesting an antimutagenic activity.
Assuntos
Antimutagênicos/farmacologia , Citotoxinas/farmacologia , Extratos Vegetais/farmacologia , Poríferos/química , Salmonella typhimurium/efeitos dos fármacos , Acetona/química , Animais , Etanol/química , Viabilidade Microbiana/efeitos dos fármacos , Testes de Mutagenicidade , Salmonella typhimurium/genética , Salmonella typhimurium/crescimento & desenvolvimentoRESUMO
Amazon mosses, such as Holomitriopsis laevifolia and Leucobryum sp. are naturally exposed to high levels of solar ultraviolet (UV) radiation. Theoretically, under environmental stress conditions these mosses have developed protective chemical and metabolic strategies against UV damage, by way of biosynthesis of secondary metabolites, such as flavonoids. The present paper aimed to evaluate the free-radical scavenging activity, and the photoprotective, mutagenic and photomutagenic potencies of the methanolic (ME), aqueous (AE), hydroalcoholic (HE), ethanolic (EE) extracts of H. laevifolia and Leucobryum sp. The phenolic contents were evaluated by spectrophotometry and by High-Performance Liquid Chromatography (HPLC). The present findings showed that the AE and HE of H. laevifolia and the AE of Leucobryum sp. presented the highest phenolic contents. The HPLC analysis indicated the presence mainly of phenolic and cinnamic acids, flavonols, flavones and flavanones. The AE and EE of H. laevifolia and the AE and HE of Leucobryum sp. efficiently scavenged the 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radical. All extracts showed significant values of in vitro Sun Protection Factor alone, and HE of Leucobryum sp. showed a synergistic effect in association with benzophenone-3. None of the extracts induced mutagenicity in the auxotrophic strains for histidine of Salmonella typhimurium, and photomutagenicity of the TA102 and TA104 strains was not detected after exposure to UV-A radiation. Besides, all extracts showed photoprotective activity against UV-A radiation for the TA104 strain, including synergistic protection in association with BP-3. Thus, the constituents in H. Laevifolia and Leucobryum sp. could be good candidates for cosmetic and dermatological applications, particularly in association with synthetic UV filters, since the concentration of the filters in the final product could be reduced.
Assuntos
Briófitas/química , Dano ao DNA/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/química , Raios Ultravioleta , Briófitas/metabolismo , Cromatografia Líquida de Alta Pressão , Dano ao DNA/efeitos da radiação , Flavonoides/análise , Flavonoides/química , Sequestradores de Radicais Livres/química , Testes de Mutagenicidade , Estresse Oxidativo/efeitos da radiação , Fenóis/análise , Fenóis/química , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/genética , Espectrofotometria , Fator de Proteção SolarRESUMO
In the present work, three commercial acid (pH 3.5-4) pyrogallol-containing hair gels, SunSet Alizador Negro (two formulations) and Embelleze Henê Gel, were tested for mutagenicity using two well-established assays. In the Salmonella mutagenicity assay using 648-5000 microg/plate of cosmetic samples, none of the samples reached a 2-fold increase in revertants relative to the controls. Both in the absence and in the presence of S9, the dose-response relation in strains TA98, TA100, TA102, TA1535, and TA1537 was not significant (p>0.01). In the mouse bone marrow micronucleus assay, 10 Swiss male mice were orally administered 2000 mg/kg of sample per body weight/day. The ratio between polychromatic and normochromatic erythrocytes as well as the presence of micronuclei in bone marrow cells were determined. Equal numbers of micronucleated polychromatic erythrocytes were detected between the cells of each treated group and the negative control, using ANOVA and chi-square analyses. Thus, none of the products induced mutagenesis in either assay. Previous studies have shown pyrogallol is mutagenic in various test systems, including Salmonella. However studies have also shown that acidic conditions may repress the reactive-oxygen species (ROS) produced by pyrogallol, and ROS is considered the primary mechanism for the mutagenicity of pyrogallol. Consistent with this are our results, which show that acidic, commercially available pyrogallol-containing hair gels are neither mutagenic in Salmonella nor induce micronuclei in mouse bone marrow in vivo.
Assuntos
Mutagênicos/toxicidade , Pirogalol/toxicidade , Animais , Géis , Cabelo , Concentração de Íons de Hidrogênio , Masculino , Camundongos , Testes para Micronúcleos , Testes de Mutagenicidade , Espécies Reativas de Oxigênio , Salmonella/efeitos dos fármacos , Salmonella/genéticaRESUMO
Antarctica moss Sanionia uncinata (Hedw.) Loeske is exposed in situ to damaging levels of ultraviolet (UV) radiation. This moss has the ability to respond to UV radiation exposure producing secondary metabolites such as flavonoids, and has been recommended as a potential source of photoprotective compounds and antioxidants. The aim of the present paper was to investigate the free-radical scavenging activity and mutagenic and photomutagenic properties of methanolic (ME), hydroethanolic (HE) and ethanolic (EE) extracts of S. uncinata. The phenolic contents were evaluated by high-performance liquid chromatography (HPLC) and spectrophotometry. The findings showed that ME and EE presented the highest phenolic contents and inhibited free radical-scavenging activity against 2,2'-diphenyl-1 picrylhydrazyl (DPPH) and the HPLC analysis indicated several classes of phenolic acids and flavonoids. The sun protection factors (SPF) were determined by an in vitro method and the results showed significant values. The SPF values of BZ-3 at 50µg/mL increased significantly in association with ME, HE and EE. The extracts did not induce mutagenicity in auxotrophic Salmonella typhimurium histidine and photomutagenicity was not detected in the TA102 and TA104 strains after exposure to UV-A at doses of up to 6.5J/cm2 for the TA102 strain and up to 0.24J/cm2 for the TA104 strain. In addition, with the exception of ME, all the extracts induced photoprotective effects in the presence of the TA104 strain at 0.04J/cm2. The present results suggest that S. uncinata extracts did not induce photomutation and showed promise for photoprotection against the photobiological and ROS-inducing effects of the UV-A radiation.
Assuntos
Briófitas , Extratos Vegetais/efeitos da radiação , Protetores Solares/efeitos da radiação , Raios Ultravioleta/efeitos adversos , Animais , Regiões Antárticas , Relação Dose-Resposta a Droga , Sequestradores de Radicais Livres/isolamento & purificação , Sequestradores de Radicais Livres/efeitos da radiação , Sequestradores de Radicais Livres/toxicidade , Testes de Mutagenicidade/métodos , Mutagênicos/toxicidade , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/toxicidade , Ratos , Salmonella typhimurium , Protetores Solares/isolamento & purificação , Protetores Solares/toxicidadeRESUMO
Ilex paraguariensis (yerba mate) has a beneficial effect in the management of obesity. Here, we studied the effects of yerba mate on hypothalamic changes in leptin and insulin signalling, oxidative stress and liver morphology and metabolism in postnatal early overfeeding (EO) Wistar rats. To induce EO, the litter size was reduced to three pups per dam, and litters with 10 pups per dam were used as a control (10 litters each). On postnatal day (PN) 150, EO offspring were subdivided into EO and EO+mate groups (10 animals each), which were treated with water or mate tea [1 g/kg body weight (BW)/day, by gavage], respectively, for 30 days. The C offspring received water. On PN180, yerba mate treatment prevented BW gain and reduced total body fat, visceral fat and food intake in comparison with the EO group. Leptin and insulin signalling in the hypothalamus measured by Western blotting was reduced only in the EO group. Yerba mate treatment had a greater impact on insulin signalling normalization. In the liver, yerba mate treatment normalized antioxidant enzyme activities and, consequently, decreased lipid peroxidation, determined by malondialdehyde content. In addition, the steatosis level and the liver triglyceride content were also restored. Thus, for the first time, yerba mate was demonstrated to increase antioxidant defences and improve liver metabolism in adult rats that were overfed during lactation, possibly through improvements in the hypothalamic action of insulin. These findings may be important for the treatment of obesity-related disorders.
Assuntos
Hipotálamo/metabolismo , Ilex paraguariensis/química , Insulina/metabolismo , Lactação , Leptina/metabolismo , Hepatopatias/metabolismo , Extratos Vegetais/farmacologia , Animais , Peso Corporal , Aleitamento Materno , Ingestão de Alimentos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hipotálamo/efeitos dos fármacos , Hepatopatias/tratamento farmacológico , Hepatopatias/etiologia , Masculino , Hipernutrição/complicações , Ratos , Ratos Wistar , Transdução de Sinais/efeitos dos fármacosRESUMO
The survival of different DNA repair mutant strains of Escherichia coli treated with H2O2 was evaluated in the presence or absence of an iron chelator (dipyridyl). Our results suggest that Fpg and UvrA proteins participate in vivo in the repair of DNA lesions produced by higher H2O2 concentrations in the presence of an iron chelator while UvrB and UvrC proteins seem to be ineffective in the repair of these lesions.
Assuntos
Adenosina Trifosfatases/farmacologia , Proteínas de Bactérias/farmacologia , Proteínas de Bactérias/fisiologia , Reparo do DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/farmacologia , Proteínas de Escherichia coli , Escherichia coli/genética , Peróxido de Hidrogênio/farmacologia , Quelantes de Ferro/farmacologia , N-Glicosil Hidrolases/farmacologia , 2,2'-Dipiridil/farmacologia , Proteínas de Bactérias/efeitos dos fármacos , Proteínas de Bactérias/genética , Morte Celular/efeitos dos fármacos , DNA-Formamidopirimidina Glicosilase , Relação Dose-Resposta a Droga , Escherichia coli/efeitos dos fármacos , Ferro/análise , MutaçãoRESUMO
Escherichia coli K-12 cells incubated in buffer can repair most of their X-ray-induced DNA single-strand breaks, but additional single-strand breaks are repaired when the cells are incubated in growth medium. While the radC102 mutant was proficient at repairing DNA single-strand breaks in buffer (polA-dependent repair), it was partially deficient in repairing the additional single-strand breaks (or alkali-labile lesions) that the wild-type strain can repair in growth medium (recA-dependent repair), and this repair deficiency correlated with the X-ray survival deficiency of the radC strain. In studies using neutral sucrose gradients, the radC strain consistently showed a small deficiency in rejoining X-ray-induced DNA double-strand breaks, and it was deficient in restoring the normal sedimentation characteristics of the repaired DNA.
Assuntos
Reparo do DNA , DNA Bacteriano/efeitos da radiação , Escherichia coli/efeitos da radiação , Mutação/efeitos da radiação , Meios de Cultura , Escherichia coli/genética , Cinética , Raios XRESUMO
A new radiation-sensitive mutant, radC , has been isolated. The radC gene is located at 81.0 min on the Escherichia coli K-12 linkage map. The radC mutation sensitized cells to uv radiation, but unlike most DNA repair mutations, sensitization to X rays was observed only for rich medium-grown cells. For cells grown in rich medium, the radC mutant was normal for gamma-radiation mutagenesis, but showed less uv-radiation mutagenesis than the wild-type strain; it showed normal amounts of X- and uv-radiation-induced DNA degradation, and it was approximately 60% deficient in recombination ability. The radC strain was normal for host cell reactivation of gamma-and uv-irradiated bacteriophage lambda; the radC mutation did not sensitize a recA strain, but did sensitize a radA and a polA strain to X and uv radiation and a uvrA strain to uv radiation. Therefore, we suggest that the radC gene product plays a role in the growth medium-dependent, recA gene-dependent repair of DNA single-strand breaks after X irradiation, and in postreplication repair after uv irradiation.
Assuntos
Escherichia coli/genética , Genes Bacterianos , Mutação , Radiogenética , Tolerância a Radiação , Sobrevivência Celular/efeitos da radiação , Mapeamento Cromossômico , Relação Dose-Resposta à Radiação , Raios gama , Raios UltravioletaRESUMO
We analyzed DNA lesions produced by H2O2 under low iron conditions, the cross adaptive response and the synergistic lethal effect produced by iron chelator-o-phenanthroline, using different Escherichia coli mutants deficient in DNA repair mechanisms. At normal iron levels the lesions produced by H2O2 are repaired mainly by the exonuclease III protein. Under low iron conditions we observed that the Fpg and UvrA proteins as well as SOS and OxyR systems participate in the repair of these lesions. The lethal effect of H2O2 is strengthened by o-phenanthroline if both compounds are added simultaneously to the culture medium. This phenomenon was observed in the wild type cells and in the xthA mutant (hypersensitive to H2O2). E. coli cells treated with low concentrations of H2O2 (micromolar) acquire resistance to different DNA damaging agents. Our results indicate also that pretreatment with high (millimolar) H2O2 concentrations protects cells against killing, by UV and this phenomenon is independent of the SOS system, but dependent on RecA and UvrA proteins. H2O2 induces protection against lethal and mutagenic effects of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). H2O2 also protects the cells against killing by cumene hydroperoxide, possibly with the participation of Ahp protein.
Assuntos
Proteínas de Escherichia coli , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Adenosina Trifosfatases/metabolismo , Proteínas de Bactérias/metabolismo , Dano ao DNA , Reparo do DNA , DNA Bacteriano/efeitos dos fármacos , Proteínas de Ligação a DNA/metabolismo , DNA-Formamidopirimidina Glicosilase , Sinergismo Farmacológico , Escherichia coli/genética , Escherichia coli/metabolismo , Ferro/metabolismo , Quelantes de Ferro/farmacologia , Metilnitronitrosoguanidina/farmacologia , N-Glicosil Hidrolases/metabolismo , Fenantrolinas/farmacologia , Proteínas Repressoras/metabolismo , Resposta SOS em Genética , Fatores de Transcrição/metabolismo , Raios UltravioletaRESUMO
In an attempt to determine whether electromagnetic field (EMF) exposure might lead to DNA damage, we exposed SnCl2-treated pBR322 plasmids to EMF and analysed the resulting conformational changes using agarose gel electrophoresis. An EMF-dependent potentiation of DNA scission (i.e. the appearance of relaxed plasmids) was observed. In confirmation of this, plasmids pre-exposed to EMF also were less capable of transforming Escherichia coli. The results indicate that EMF, in the presence of a transition metal, is capable of causing DNA damage. These observations support the idea that EMF, probably through secondary generation of reactive oxygen species, can be clastogenic and provide a possible explanation for the observed correlation between EMF exposure and the frequency of certain types of cancers in humans.
Assuntos
Dano ao DNA , Campos Eletromagnéticos/efeitos adversos , Eletroforese em Gel de Ágar , Escherichia coli/genética , Escherichia coli/metabolismo , Conformação de Ácido Nucleico , Plasmídeos , Compostos de Estanho/farmacologia , Transformação BacterianaRESUMO
The oil of Pterodon pubescens seeds (PpSO) is known for its cercaricidal and anti-inflammatory effects. Its anti-rheumatic activity was recently reported using mice with collagen II-induced arthritis treated with a hydroalcoholic extract of PpSO, mimicking the wine infusion used in popular medicine. In the present study, PpSO was tested for acute toxicity, mutagenic activity and cytotoxicity for human peripheral blood mononuclear cells (PBMNC). PpSO was obtained after seed extraction with 100% ethanol and evaporation. Cytotoxicity was estimated using the tetrazolium salt reduction test (MTT assay) by PBMNC (2.5 x 10(5) cells/ml) after exposure to 0.07, 0.7 and 7 microg PpSO/ml for 24 and 48 h. In the mutagenesis assay, the Salmonella/mammalian microsome assay was employed with or without metabolization. Acute toxicity was studied on 30 (n = 10/group) male DBA1/J mice (20 +/- 2 g) after a single oral dose of 2, 4, and 8 g PpSO/kg b.w. The animals were observed for 24 h, anesthetized, sacrificed and autopsied. The organs were processed for histopathology by staining with hematoxylin-eosin. The IC50 of PpSO to PBMNC in RPMI 1640 supplemented with 5% fetal calf serum (FCS) was 2 and 1 microg PpSO/ml after 24 and 48 h, respectively. The mutagenic test performed with or without metabolic activation of PpSO did not show mutagenic activity for the concentrations tested (7 and 70 microg/ml). Mouse mortality or significant signs of acute toxicity (ocular, cardiovascular, gastrointestinal, motor or respiratory signs) for the PpSO doses tested was not observed. The organs did not show any macroscopic alterations. Histopathologic analysis of the tissues also did not demonstrate any lesions. The present study provides data to classify PpSO as non-cytotoxic to PBMNC, non-mutagenic, and non-toxic after acute administration since the PpSO doses tested were extremely higher than those used by the population.