RESUMO
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
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
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
We previously demonstrated that reactive oxygen species (ROS) could be involved in ultraviolet-C (UVC)-induced DNA damage in Escherichia coli cells. In the present study, we evaluated the involvement of the GO system proteins in the repair of the 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxoG, GO) lesion, which is ROS-induced oxidative damage. We first found that the mutant strain Δfur, which produces an accumulation of iron, and the cells treated with 2,2'-dipyridyl, a iron chelator, were both as resistant to UVC-induced lethality as the wild strain. The 8-oxoG could be mediated by singlet oxygen ((1)O(2)). The Fpg protein repaired this lesion when it was linked to C (cytosine), whereas the MutY protein repaired 8-oxoG when it was linked to A (adenine). The survival assay showed that the Fpg protein, but not the MutY protein, was important to UVC-induced lethality and interacted with the UvrA protein, a nucleotide excision repair (NER) protein involved in UVC repair. The GC-TA reversion assay in the mutant strains from the '8-oxoG-repair' GO system showed that UVC-induced mutagenesis in the fpg mutants, but not in the MutY strain. The transformation assay demonstrated that the Fpg protein is important in UVC repair. These results suggest that UVC could also cause indirect ROS-mediated DNA damage and the Fpg protein plays a predominant role in repairing this indirect damage.
Assuntos
Quebras de DNA , Reparo do DNA , DNA-Formamidopirimidina Glicosilase/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/efeitos da radiação , Raios Ultravioleta/efeitos adversos , 8-Hidroxi-2'-Desoxiguanosina , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/metabolismo , DNA Glicosilases/genética , DNA Glicosilases/metabolismo , DNA Bacteriano/efeitos da radiação , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , DNA-Formamidopirimidina Glicosilase/genética , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Escherichia coli/enzimologia , Escherichia coli/genética , Proteínas de Escherichia coli/genética , Regulação Bacteriana da Expressão Gênica , Mutagênese , Plasmídeos/genética , Plasmídeos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Oxigênio Singlete/metabolismo , Transformação BacterianaRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as "carqueja", have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism. AIM OF THE STUDY: To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions. MATERIALS AND METHODS: Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 µg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102. RESULTS: The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 µg/kg) with AFBt reduced the paw edema (3h) at lower levels (29.2-37.3%; P<0.01), than those observed for AEBt (44.7-54.2%; P<0.001), EFBt (49.3-58.2%; P<0.001) or indomethacin (64.6%, P<0.001, 10mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC(50) 182.6 µg/ml), EFBt (IC(50) 78.1 µg/ml) and AFBt (IC(50) 86.2 µg/ml), with lower effects on HTC hepatic cell (IC(50) 308.8 µg/ml, 396.5 µg/ml and 167.9 µg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC(50) 372.5 µg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC(50) 2.2 µg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC(50) of 0.42 µg/ml; the IC(50) for doxorubicin for HEK and HTC cells was 0.28 µg/ml and 14.4 µg/ml, respectively, at 96h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively. CONCLUSIONS: Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.
Assuntos
Anti-Inflamatórios/toxicidade , Baccharis/química , Extratos Vegetais/toxicidade , Animais , Anti-Inflamatórios/farmacologia , Células Cultivadas , Ensaio Cometa , Glutationa Transferase/metabolismo , Humanos , Técnicas In Vitro , Masculino , Camundongos , Extratos Vegetais/farmacologia , Ratos , Células Tumorais Cultivadas , ÁguaRESUMO
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 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
Animais , Antimutagênicos/farmacologia , Citotoxinas/farmacologia , Extratos Vegetais/farmacologia , Poríferos/química , Salmonella typhimurium , Acetona/química , Etanol/química , Testes de Mutagenicidade , Salmonella typhimurium/crescimento & desenvolvimento , Salmonella typhimurium/genética , Viabilidade MicrobianaRESUMO
Carotenoids are 40-carbon molecules with conjugated double bonds, making them particularly effective for quenching free radicals. They have always been believed to possess anticancer properties, which could be due to their antioxidant potential. Norbixin is an unusual dicarboxylic water-soluble carotenoid present as a component in the pericarp of the seeds of Bixa orellana L. (from the Bixaceae family), a tropical shrub commonly found in Brazil. The main carotenoids present in these seeds, bixin and norbixin, form a coloring material, known as annatto, which is mainly used in the food industry. As annatto is only used as a coloring material, most studies of annatto pigments have focused on the determination of annatto levels in food. However, little attention has been given to the biological properties of bixin and norbixin. We evaluated the effect of norbixin on the response of Escherichia coli cells to DNA damage induced by UV radiation, hydrogen peroxide (H2O2) and superoxide anions (O2*-)) and found that norbixin protects the cells against these agents. Norbixin enhanced survival at least 10 times. The SOS induction by UVC was inhibited 2.3 times more when cells were grown in the presence of norbixin. We also found that norbixin has antimutagenic properties, with a maximum inhibition of H2O2-induced mutagenic activity of 87%, based on the Salmonella mutagenicity test
Assuntos
Antimutagênicos/farmacologia , Carotenoides/farmacologia , Dano ao DNA/efeitos dos fármacos , Escherichia coli/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Bixaceae/química , DNA Bacteriano/efeitos dos fármacos , DNA Bacteriano/efeitos da radiação , Dano ao DNA/efeitos da radiação , Escherichia coli/citologia , Peróxido de Hidrogênio/toxicidade , Resposta SOS em Genética , Superóxidos/toxicidade , Testes de Mutagenicidade/métodos , Raios UltravioletaRESUMO
The SoxRS regulon is induced when bacterial cells are exposed to redox-cycling agents such as menadione or paraquat. In this paper it is shown that a physical agent, such as ultraviolet radiation with a wavelength of 312 nm (UVB) can induce soxS gene expression. The results indicate that this induction involves the RpoS protein. Moreover, an unexpected increase of soxS gene expression independent of a functional soxR gene in UVB-irradiated cells has been verified. This increase could be explained by transcription of soxS gene in a rpoS-dependent pathway.
Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/fisiologia , Escherichia coli/efeitos da radiação , Regulação Bacteriana da Expressão Gênica/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Fator sigma/metabolismo , Transativadores/metabolismo , Raios Ultravioleta , Sobrevivência Celular/efeitos da radiação , Relação Dose-Resposta à Radiação , Escherichia coli/citologia , Proteínas de Escherichia coli/genética , Estresse Oxidativo/fisiologia , Doses de Radiação , Transativadores/genéticaRESUMO
The chemical compound temephos (0,0,0',0'-tetrametyl-0,0'-thiodi-p-phenylene phosphorothioate) is an organophosphorous pesticide that has been used in Brazil since 1967 in control campaigns against the mosquito Aedes aegypti, the vector of dengue and yellow fever. We used single cell gel electrophoresis (SCGE), SOS/umu and Ames/Salmonella assays to test the toxicity and mutagenicity of temephos. Temephos was genotoxic in the SCGE assay, inducing severe DNA lesions (type IV lesions) at doses above 1.34 micro M. It was mutagenic, but not toxic, in the SOS/umu assay to Escherichia coli strain PQ37, but not to PQ35, at concentrations above 1.33 micro M, particularly when the S9 mixture was not used in the assay. Temephos was not mutagenic in the Ames assay with S. typhimurium strains TA97, TA98, TA100 and TA102, both with and without metabolic activation. However, temephos at concentrations above 3.33 micro M was mutagenic to TA98NR, YG7104 and YG7108, both with and without metabolic activation. In conclusion, temephos was genotoxic and mutagenic in all the three tests used, and in two of them at concentrations similar to those routinely used to combat Aedes aegypti
Assuntos
Animais , Masculino , Inseticidas/toxicidade , Mutagênicos/toxicidade , Temefós/toxicidade , Ensaio Cometa , Ratos , Ratos Wistar , Resposta SOS em Genética , Salmonella typhimurium/efeitos dos fármacos , Testes de Mutagenicidade/métodosRESUMO
In the present study, we evaluated the sensitivity of different Escherichia coli strains to Cumene hydroperoxide (CHP) treatment under distinct conditions of Fe2+ availability. Our results showed that the pretreatment with an iron chelator (dipyridyl) protects all the tested strains against CHP toxic effects, but it was not sufficient to abolish the CHP induced mutagenesis. On the other hand, simultaneous pretreatment with both dipyridyl and neocuproine (copper chelator) leads to a complete protection against CHP mutagenic effects. Our data suggest the participation of copper ion in the CHP mutagenesis induced in E. coli.
Assuntos
Derivados de Benzeno/farmacologia , Dano ao DNA , Reparo do DNA/efeitos dos fármacos , DNA Bacteriano/efeitos dos fármacos , Ferro/farmacologia , Mutagênicos/farmacologiaRESUMO
Naturally occurring antioxidants such as carotenoids are extensively studied for their potential in reducing the risk for cancer and other chronic diseases. In the present study, the radical-scavenger activity of the food additive norbixin, a water-soluble carotenoid extracted from Bixa orellana seeds and commercialized as annatto, was evaluated under conditions of DNA damage induced by reactive oxygen species, particularly by hydroxyl radicals. The cell-free scavenger activity of norbixin was evaluated using plasmid DNA as target molecule and Sn2+ or Fe2+ as oxidant. The addition of H2O2 enhanced DNA breakage induced by metal ions, particularly Fe2+. Under these conditions, norbixin started to protect plasmid DNA against single- and double-strand breakage at a metal:norbixin ratio of 1:1 (Sn2+) and 1:10 (Fe2+). However, at lower ratios to Sn2+, norbixin enhanced Sn2+-induced DNA breakage (P < 0.05). The ability of norbixin to protect genomic DNA against oxidative damage was assessed in murine fibroblasts submitted to H2O2-induced oxidative stress and the results were evaluated by the comet assay. Under low serum conditions (2 % fetal bovine serum (FBS)), a protective effect of norbixin against H2O2-induced DNA breakage was inversely related to its concentration, a protection ranging from 41 % (10 microm) to 21 % (50 microm). At higher concentrations of norbixin, however, oxidative DNA breakage was still enhanced, even in the presence of a high serum concentration (10 % FBS). Under normal conditions, norbixin per se has no detectable genotoxic or cytotoxic effects on murine fibroblasts. The antimutagenic potential of norbixin against oxidative mutagens was also evaluated by the Salmonella typhimurium assay, with a maximum inhibition of 87 % against the mutagenicity induced by H2O2. Although plasmid DNA and Ames data indicated that norbixin can protect DNA against oxidative damage, it seems to be a risky guardian of genomic DNA as it can also increase the extent of oxidative damage.
Assuntos
Carotenoides/farmacologia , Dano ao DNA , Aditivos Alimentares/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Animais , Antimutagênicos/farmacologia , Técnicas de Cultura de Células , Relação Dose-Resposta a Droga , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Peróxido de Hidrogênio/farmacologia , Metais/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Plasmídeos/genéticaRESUMO
Three tinctures samples from extracts of the popular medicinal plant Thuya occidentalis were tested in vitro through two short term tests for measuring the activity of genotoxic chemicals. Using the Salmonella/mammalian-microsome (Mutatest) assay and the SOS-chromotest (induction of beta-galactosidase in Escherichia coli), none of the extract was effective in inducing mutagenesis or beta-galactosidase synthesis (as an indicator of general and early sign of DNA damage), even with metabolization
Assuntos
Corantes/farmacologia , Escherichia coli/efeitos dos fármacos , Testes de Mutagenicidade/métodos , Extratos Vegetais/farmacologia , Salmonella typhimurium/efeitos dos fármacos , Resposta SOS em GenéticaRESUMO
Toxicity of an aqueous extract prepared from Echinodorus macrophyllus dried leaves, a plant used in folk medicine to treat inflammation and kidney malfunctions, was estimated by different bioassays. Mutagenicity of the aqueous extract was evaluated in the Salmonella/microsome assay (TA97a, TA98, TA100 and TA102 strains), with or without metabolic activation. No mutagenic activity (lyophilized extract tested up to 50 mg/plate) could be detected to any of the tester strain. Furthermore, no cytotoxic effect has been observed when a crude extract of E. macrophyllus (up to 7.5 mg/ml) was tested on the exponential growth of hepatoma and normal kidney epithelial cells in culture. Toxicity of E. macrophyllus was also evaluated in male Swiss mice after 6 weeks of continuous ingestion of the aqueous extract in drinking water. Average daily ingested doses were 3, 23 and 297 mg/kg for a lyophilized extract, and 2200 mg/kg for a crude extract, with dose two being equivalent to the daily dose recommended to humans. At the end of the treatment, all animals revealed a deficit in final body weight ranging from 5 to 47%. Biochemical analysis of the plasma revealed some minor alterations indicating subclinical hepatic toxicity. Genotoxic effect on liver, kidney and blood cells has been also evaluated by the comet assay, being negative to liver and blood cells. However, DNA analyses of the kidney cells detected some genotoxic activity for the highest dose tested of E. macrophyllus extract, either lyophilized or crude. On the other hand, exposure dose of 23 mg/kg, equivalent to the daily dose recommended to humans, did not revealed any genotoxic effect and hence this herb seems to be safe to human organism.
Assuntos
Plantas Medicinais , Animais , Peso Corporal/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , DNA/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Masculino , Camundongos , Testes de Mutagenicidade , Tamanho do Órgão/efeitos dos fármacos , Extratos Vegetais/toxicidade , Ratos , Células Tumorais CultivadasRESUMO
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.
Assuntos
Citotoxinas/toxicidade , Fabaceae/química , Mutagênicos/toxicidade , Óleos de Plantas/toxicidade , Plantas Medicinais/química , Animais , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Técnicas In Vitro , Leucócitos Mononucleares/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos DBA , Testes de Mutagenicidade , Sementes/químicaRESUMO
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
Cross-adaptive response is defined as the capacity of cells to become resistant to a lethal agent when pretreated with a different lethal substance. In the present paper, the cross-adaptive response between hydrogen peroxide and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) was studied in Escherichia coli repair mutants. Our results suggest that high doses of H2O2 induces protection against the lethal effects of MNNG in wild-type strain, ada, ogt, ada-ogt, aidB and alkA mutants. On the other hand, the MNNG induced mutagenesis is reduced by H2O2 pretreatment in wild-type and ogt mutant strains, but not in ada mutant. Furthermore, the protecting effect induced by H2O2 is time dependent: it decreases 15 min after the pretreatment and, after 30 min, is almost abolished. This reduction in the protecting effect is followed by an augmentation in the mutation frequency when MNNG is added 30 min after H2O2 pretreatment. This cross-adaptive response may be due to a modification of the MNNG alkylation pattern in the oxidized DNA.
Assuntos
Antimutagênicos/farmacologia , Proteínas de Escherichia coli , Escherichia coli/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Metilnitronitrosoguanidina/toxicidade , Metiltransferases , Mutagênicos/toxicidade , Oxidantes/farmacologia , Proteínas de Bactérias/genética , Escherichia coli/genética , NADH NADPH Oxirredutases/genética , O(6)-Metilguanina-DNA Metiltransferase , Fatores de Tempo , Fatores de TranscriçãoRESUMO
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.