Assessment of DNA damage and molecular responses in Labeo rohita (Hamilton, 1822) following short-term exposure to silver nanoparticles

The increasing application of silver nanoparticles (Ag-NPs) both in industries and in agricultural fields has led to its accumulation in the aquatic ecosystem through water run-off. In the present study, the effects of Ag-NPs in the liver of Labeo rohita, were investigated at genomic and cellular level for seven days at the concentrations of 100, 200, 400 and 800 μg l⁻¹ by using 18 and 29 nm sizes of Ag-NPs. The Ag-NPs sizes of 18 and 29 nm were synthesized by a chemical method using atomic force microscopy with the zeta potential of −55 mV and−31.4 mV respectively. They were found to be spherical with smooth surfaces. Assessment of genotoxic effects of the particles in the fish using single-cell gel electrophoresis showed DNA damage on exposure to concentrations of 400 and 800 μg l⁻¹. Histopathological examination of the liver revealed vacuolar degeneration, hepatocytes have undergone total degeneration and high accumulation of Ag-NPs that depicted both time and dose-dependent relationships. Furthermore, the expression study of stress-related genes showed down-regulation, due to the production of free radicals and reactive oxygen species. Ag-NPs can cause both DNA damage and affect the cellular responses of L. rohita.     

Mercuric dichloride induces DNA damage in human salivary gland tissue cells and lymphocytes

Amalgam is still one of the most frequently used dental filling materials. However, the possible adverse effects especially that of the mercuric component have led to continued controversy. Considering that mercury may be released from amalgam fillings into the oral cavity and also reach the circulating blood after absorption and resorption, it eventually may contribute to tumorigenesis in a variety of target cells. The present investigation focuses on genotoxic effects below a cytotoxic dose level of mercuric dichloride (HgCl₂) in human samples of salivary glands and lymphocytes to elucidate a possible role in tumor initiation. DNA migration due to single strand breaks, alkali labile sites and incomplete excision repair was quantified with the aid of the single cell microgel electrophoresis (Comet) assay. The concepts of Olive Tail Moment, percentage of DNA in the Tail and Tail Length were used as measures of DNA damage. To control for cytotoxic effects, the trypan blue exclusion test was applied. Human samples of the parotid salivary gland and lymphocytes of ten donors were exposed to HgCl₂ concentrations from 1 to 50 μM. N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and dimethyl sulfoxide (DMSO) served as controls. Increasing dose-dependent DNA migration could be demonstrated after exposure to HgCl₂ in cells of the salivary glands and lymphocytes. In both cell types a significant increase in DNA migration could be shown starting from HgCl₂ concentrations of 5 μM in comparison to the negative control. The viability of the cell systems was not affected except at the highest concentration (50 μM) tested. These data indicate genotoxic effects of mercuric dichloride in human salivary glands and lymphocytes at concentrations not leading to cytotoxic effects or cell death. Consequently, a contributory role in oral salivary gland tumor initiation warrants further investigation. The authors declare that there is no conflict of interest.     

Variability in DNA damage of chub (Squalius cephalus L.) blood,gill and liver cells during the annual cycle

In this work the genotoxic potential of water in three localities in Serbia, which differ by the nature and degree of pollution, was determined in tissues of European chub (Squalius cephalus L.) on monthly basis over the 2011/2012 year season using the alkaline comet assay. Specimen samples of chub were taken from Special Nature Reserve “Uvac”, as control site, and Pestan and Beljanica Rivers, as polluted sites at Kolubara basin, surrounded with coal mines. Three tissues, blood, gills and liver were used for assessing the level of DNA damage. Analysis was done by software (Comet Assay IV). The control site at Reserve “Uvac” showed the lowest DNA damage values for all three tissues compared to Pestan and Beljanica. Blood has the lowest level of DNA damage in comparison with liver and gills. Decreased damage for all three tissues was observed at summer, while during the spring and autumn damage increased.     

In vitro genotoxic effects of the insecticide deltamethrin in human peripheral blood leukocytes: DNA damage (‘comet’ assay) in relation to the induction of sister-chromatid exchanges and micronuclei

Deltamethrin, a synthetic dibromo-pyrethroid insecticide, is extensively used in agriculture, forestry and in household products because of its high activity against a broad spectrum of insect pests (both adults and larvae), its low animal toxicity and its lack of persistence in the environment. Data on the genotoxicity and carcinogenicity of deltamethrin are rather controversial, depending on the genetic system or the assay used. The aim of this study was to further evaluate the potential genotoxic activity of deltamethrin. The in vitro genotoxicity of deltamethrin has been evaluated by assessing the ability of the insecticide to damage DNA (as evaluated using the single-cell microgel-electrophoresis or ‘comet’ assay) or induce sister-chromatid exchanges (SCE) and micronuclei (MN) in human peripheral blood leukocytes. All treatments were conducted with and without the presence of an external bioactivation source (±S9mix). The results indicate that deltamethrin, in the presence of metabolic activation (+S9mix), is able to induce DNA damage (double- and single-strand breaks, alkali-labile sites and open excision repair sites) as revealed by the increasing tail moment values observed with increasing doses. The frequency of SCE and MN were not statistically increased in deltamethrin-treated cells as compared to controls, both with and without S9mix. However, lower deltamethrin doses were tested, as compared to ‘comet’ assay, because of cytotoxicity.     

Genotoxicity evaluation of fullerene C60 nanoparticles in a comet assay using lung cells of intratracheally instilled rats

The genotoxicity of fullerene C₆₀ nanoparticles was evaluated in vivo with comet assays using the lung cells of rats given C₆₀ nanoparticles. The C₆₀ nanoparticles were intratracheally instilled as a single dose at 0.5 or 2.5 mg/kg or repeated dose at 0.1 or 0.5 mg/kg, once a week for 5 weeks, to male rats. The lungs were obtained 3 or 24 h after a single instillation and 3 h after repeated instillation. Inflammatory responses were observed in the lungs obtained 24 h after a single instillation at 2.5 mg/kg and repeated instillation at 0.5 mg/kg. Histopathological examinations revealed that C₆₀ nanoparticles caused slight changes including hemorrhages in alveoli and the cellular infiltration of macrophages and neutrophils in alveoli. In comet assays using rat lung cells, no increase in % Tail DNA was found in any group given C₆₀ nanoparticles. These findings indicate that C₆₀ nanoparticles had no potential for DNA damage in comet assays using the lungs cells of rats given C₆₀ even at doses causing inflammation.     

Genotoxic effects of produced waters in mosquito fish (Gambusia affinis)

The aim of this study was to assess the potential genotoxic effects of produced water (PW) from an Italian on-shore oil plant. Produced water is a complex mixture containing residual hydrocarbons, trace elements, naturally occurring radioactive material and potentially toxic treatment chemicals such as biocides, dispersants, detergents and scale inhibitors used in oil production. The test organism, mosquito fish (Gambusia affinis), was divided into male and female groups and exposed for 8 days in the laboratory to 50% concentrations of different produced waters: PW before treatment and after settling treatment. The fish were also exposed to lower concentrations (10%) of the same PW for 30 days. DNA damage was evaluated in erythrocytes by single cell gel electrophoresis (Comet assay) and micronucleus test, while an oxidative stress biomarker, was assessed. Polycyclic aromatic hydrocarbons (PAH) metabolites in bile were also evaluated. A higher sensitivity in biomarker responses was found in females in comparison to males. An increase in DNA strand breaks was observed in both genders after 30 days exposure and a statistically significant increase of micronucleated cells was found in females after 8 days exposure. A positive correlation between presence of micronucleated cells and PAH metabolites in bile was also observed.     

Tissue-specific genotoxicity and antioxidant imbalance of titanium dioxide nanoparticles (NPTiO2) and inorganic lead (PbII) in a neotropical fish species

The aquatic environment is the major recipient of wastes containing nanoparticles and other contaminants. Titanium dioxide nanoparticles (NPTiO₂) are one of the most produced and used nanoparticle worldwide. This study investigated the toxicity of NPTiO₂, as well as the toxicity interaction between NPTiO₂ and lead (Pb), in response to genetic and biochemical biomarkers using freshwater fish Rhamdia quelen, as an animal model. The results showed genotoxicity in blood and kidney tissues. No effect of NPTiO₂ alone or in co-exposure with Pb on liver genotoxicity were observed. Alterations in the antioxidant hepatic enzymes activities, as well as alterations in glutathione levels indicated that NPTiO₂ alone or in co-exposure with Pb can cause antioxidant imbalance. The lipid peroxidation was also raised after exposure to NPTiO₂. In general, the results of this study indicated that both NPTiO₂ alone and their co-exposure with Pb are capable of producing significant toxic effects in short-term exposure.     

Genotoxic and cytotoxic potential of titanium dioxide (TiO2) nanoparticles on fish cells in vitro

Intrinsic genotoxic and cytotoxic potential of titanium dioxide (TiO2) engineered nanoparticles (ENPs) were evaluated in a metabolically competent, established fish cell line derived from rainbow trout (Oncorhyncus mykiss) gonadal tissue (i.e. RTG-2 cells). Prior to evaluation of the toxic potential, mean size of the ENPs was determined using transmission electron microscopy (TEM). As a prerequisite, an extensive characterisation of the ENPs was carried out following sonication which enabled the synthesis of an efficient dosing strategy for the cells in which exposure in phosphate buffered saline (PBS) gave an optimal agglomeration effects compared to distilled water (H2O) and minimal essential media (MEM). Interaction of the ENPs with cells under scanning electron microscope (SEM) was also studied. The genotoxic and cytotoxic potential of the ENPs were determined either alone or in combination with ultraviolet radiation (i.e. UVA). Whilst genotoxic potential was determined by evaluating DNA strand breaks using single cell gel electrophoresis (SCGE) or the comet assay and induction of cytogenetic damage using cytokinesis-blocked micronucleus (MN) assay, cytotoxicity was determined by measuring the retention of supra vital stain, neutral red, by the lysosomes using the neutral red retention (NRR) assay. In addition, while performing the comet assay, lesion specific bacterial endonuclease, formamidopyrimidine DNA glycosylase (Fpg), which recognises oxidised purine bases, was used to determine oxidative DNA damage. The results suggested that the highest concentration of the ENPs (i.e. 50 microg ml(-1)) did not produce elevations in DNA damage over 4 h (comet assay), 24 h (modified comet assay) or 48 h (MN assay) exposures in the absence of UVA irradiation, although there was a significant reduction in lysosomal integrity over 24 h exposure (NRR assay). The induction of MN did not show any enhanced levels as a function of ENP concentration. A significantly increased level of strand breaks was observed in combination with UVA (3 kJ m(-2)). In general, the NRR assay suggested elevated levels of cytotoxicity when the UVA exposure was carried out with MEM compared to PBS, although both showed an increase when in combination with the highest concentration of ENPs (i.e. 50 microg ml(-1)). Overall, the study emphasises the need for adoption of an holistic approach while evaluating the potential toxic effects of ENPs in which appropriate measures should be taken to avoid agglomeration or aggregation to facilitate efficient cellular uptake to evaluate potential biological responses.     

In vitro study on the genotoxicity of dichloromethane extracts of valerian (DEV) in human endothelial ECV304 cells and the effect of vitamins E and C in attenuating the DEV-induced DNA damages

To study the genotoxicity of valepotriates in vitro, the degree of DNA damage in human endothelial cell line ECV304 treated with 5-60 microg/mL of dichloromethane extracts of valerian (DEV) was analyzed by the Comet assay. No DNA damage was observed in ECV304 cells after culture for 48 h in the presence of 5,10, and 20 microg/mL of DEV. But a moderate degree of DNA damage was observed in the cells treated with 40 or 60 microg/mL of DEV. Quantitative analyses of DNA damage in the presence of antioxidants vitamin E (VE) and vitamin C (VC) were also carried out. The study revealed that both VE and VC exhibited a biphasic effect, reducing DEV-induced DNA damages at low concentrations but increasing them at high concentrations. We concluded that (1). the observed DNA damage in ECV304 cells induced by high concentrations of DEV was mainly through epigenetic mechanisms, i.e., reactive oxygen species mediated oxidative DNA damage (2). at the low doses, DEV did not appear to have any significant genotoxicity in ECV304 cells, and (3). VE and VC, at proper concentrations, can reduce or eliminate the adverse effects derived from high doses of DEV. This study should serve as scientific guidance for clinical therapy of valerian preparation.     

Long-term exposure of A549 cells to titanium dioxide nanoparticles induces DNA damage and sensitizes cells towards genotoxic agents

Titanium dioxide nanoparticles (TiO₂-NPs) are one of the most produced NPs in the world. Their toxicity has been studied for a decade using acute exposure scenarios, i.e. high exposure concentrations and short exposure times. In the present study, we evaluated their genotoxic impact using long-term and low concentration exposure conditions. A549 alveolar epithelial cells were continuously exposed to 1–50?μg/mL TiO₂-NPs, 86% anatase/14% rutile, 24?±?6?nm average primary diameter, for up to two months. Their cytotoxicity, oxidative potential and intracellular accumulation were evaluated using MTT assay and reactive oxygen species measurement, transmission electron microscopy observation, micro-particle-induced X-ray emission and inductively-coupled plasma mass spectroscopy. Genotoxic impact was assessed using alkaline and Fpg-modified comet assay, immunostaining of 53BP1 foci and the cytokinesis-blocked micronucleus assay. Finally, we evaluated the impact of a subsequent exposure of these cells to the alkylating agent methyl methanesulfonate. We demonstrate that long-term exposure to TiO₂-NPs does not affect cell viability but causes DNA damage, particularly oxidative damage to DNA and increased 53BP1 foci counts, correlated with increased intracellular accumulation of NPs. In addition, exposure over 2 months causes cellular responses suggestive of adaptation, characterized by decreased proliferation rate and stabilization of TiO₂-NP intracellular accumulation, as well as sensitization to MMS. Taken together, these data underline the genotoxic impact and sensitization effect of long-term exposure of lung alveolar epithelial cells to low levels of TiO₂-NPs.     

Toxic effects of imidacloprid on adult loach (Misgurnus anguillicaudatus)

The present investigation was aimed to assess the effects of imidacloprid on the survival, genetic materials, hepatic transaminase activity and histopathology of loach (Misgurnus anguillicaudatus). The values of LC₅₀ (24, 48, 72 and 96 h) of imidacloprid were 167.7, 158.6, 147.9 and 145.8 mg/L, respectively, and the safety concentration was 42.55 mg/L. The erythrocyte micronuclei assays and the comet assay results showed that imidacloprid had genetic toxic effect on the loach erythrocytes. To assess the physiological and biochemical damage caused by imidacloprid, the activities of hepatic glutamic-pyruvic transaminase (GPT) and glutamic-oxalacetic transaminase (GOT) were measured and their values declined in treatment groups. Histological examination of testis revealed that imidacloprid treatment resulted in disorganized lobules and cysts structures. In the present work, we also investigated the joint toxicity of pesticides commonly used in paddy fields (imidacloprid and lambda-cyhalothrin) on M. anguillicaudatus, and confirmed that a synergistic effect existing in the binary mixtures. The results of our study provide relevant and comparable toxicity information that are useful for safety application of pesticides.     

Soy isoflavones have antimutagenic activity on DNA damage induced by the antileishmanial Glucantime (meglumine antimoniate)

Isoflavones are phytoestrogens reported to be potent antioxidant agents. In contrast, the antileishmanial meglumine antimoniate has mutagenic activities. This study evaluated the ability of soy isoflavones to reduce DNA damage induced by meglumine antimoniate. Antimutagenic effects (by micronucleus test) were tested using Swiss mice divided into seven groups treated with meglumine antimoniate (425?mg/kg bw pentavalent antimony); cyclophosphamide (50?mg/kg bw); water (negative control); single isoflavones dose (1.6?mg/kg bw), and three groups received one dose of isoflavones via gavage (0.4?mg/kg bw, 0.8?mg/kg bw or 1.6?mg/kg bw) plus meglumine antimoniate via intraperitoneal, simultaneously. To evaluate antigenotoxicity (by Comet assay), each group with 10 animals received the above-mentioned control doses; single dose of isoflavones 0.8?mg/kg bw, and three groups received isoflavones (0.8?mg/kg bw) by gavage along with intraperitoneal meglumine antimoniate, which were treated with isoflavones 24?h before or after receiving meglumine antimoniate (pre-treatment and post-treatment, respectively) or simultaneously. Cells were harvested 24?h after the treatment, and the data were evaluated by ANOVA followed by Tukey’s test (p?相似文献   

Black bean (Phaseolus vulgaris L.) as a protective agent against DNA damage in mice.

This study was designed to evaluate the toxicogenetic or protective effect of cooked and dehydrated black beans (Phaseolus vulgaris L.) in bone marrow and peripheral blood cells of exposed mice. The frequency of micronuclei detected using the bone marrow erythrocyte micronucleus test and level of DNA lesions detected by the comet assay were chosen as end-points reflecting mutagenic and genotoxic damage, respectively. Initially, Swiss male mice were fed with a 20% black bean diet in order to detect mutagenic and genotoxic activity. However, no increase in the frequency of bone marrow micronucleated polychromatic erythrocytes (MN PCEs) or DNA lesion in leukocytes was observed. In contrast, received diets containing 1, 10 or 20% of black beans, a clear, but not dose-dependent reduction in the frequency of MN PCEs were observed in animals simultaneously treated with cyclophosphamide, an indirect acting mutagen. Similar results were observed in leukocytes by the comet assay. Commercial anthocyanin was also tested in an attempt to identify the bean components responsible for this protective effect. However, instead of being protective, the flavonoid, at the highest dose administered (50 mg/kg bw), induced primary DNA lesion, as detected by the comet assay. These data indicate the importance of food components in preventing genetic damage induced by chemical mutagens, and also reinforce the role of toxicogenetic techniques in protecting human health.     

Effects of pesticides on human peripheral lymphocytes in vitro: induction of DNA damage

Because of the widespread use of pesticides for domestic and industrial applications the evaluation of their genotoxic effects is of major concern to public health. Although various experimental data have provided evidence that pesticides can possess genotoxic properties in animals and in in vitro test systems after acute and chronic exposure, the information on the genotoxic effects of some of pesticides is limited and inconsistent. In the present study, the genotoxic potential of commonly used pesticides (i.e., dimethoate and methyl parathion from the organophosphate class, propoxur and pirimicarb from carbamates, and cypermethrin and permethrin from pyrethroids) have been evaluated. The genotoxic effects of these substances were examined using the single cell gel electrophoresis (comet) assay in freshly isolated human peripheral lymphocytes. The cells were incubated with 10, 50, 100 and 200 µg/ml concentrations of the test substances for 0.5 h at 37°C and DNA damage was compared with that obtained in lymphocytes from the same donor not treated with substances. Hydrogen peroxide, 100 µM, was used as a positive control. Within the concentration ranges studied, no significant cytotoxic effects were observed. Dimethoate and methyl parathion at 100 and 200 µg/ml; propoxur at 50, 100 and 200 µg/ml, and pirimicarb, cypermethrin and permethrin at 200 µg/ml significantly increased DNA damage in human lymphocytes.     

Pathological effects in freshwater fish Poecilia reticulata (guppy) and Oryzias latipes (medaka) following methyl bromide and sodium bromide exposure

To study the effects of the fumigant methyl bromide (CH₃Br) and its anorganic residue, bromide (Br⁻), young guppies and medakas were exposed to these chemicals for 1 and 3 months. The concentration ranges were 10–32 000 mg/l (guppy) and 180–5 600 mg/l (medaka) for NaBr and 0.032–3.2 mg/l for CH₃Br. Besides monitoring of general toxicological parameters, histopathology was performed on total body sections. In addition, a short-term study with the two highest concentrations CH₃Br was conducted for the evaluation of (lethal) effects, in which scanning electron microscopy of the gills was included.In the guppy, a NaBr concentration of 100 mg/l and higher caused paresis and paralysis. Histopathology of these animals, exposed to concentrations of 3 200 mg/l, showed muscle changes indicative of a functional neuromuscular disturbance. In the thyroid, the epithelium appeared to be stimulated (at 32 and 100 mg/l and higher after 1 and 3 months' exposure, respectively), and the thyroid stimulating hormone producing cells in the pituitary were also activated. The development of gonads in both sexes was more advanced than in the controls after the 3-month exposure period, and there was inflammation in and around the larger blood vessels in the abdomen at concentrations of 3 200 mg/l and higher after 3 months. Medakas showed only activation of the thyroid epithelium even at the lowest concentration tested (180 mg/l).In the CH₃Br study, no specific effects were observed in the long-term study of either species. In the short-term study, however, major degenerative and regenerative changes were found in the superficial epithelia, especially of the gills and oral mucosa. These were apparently caused by the local irritating action of this compound. Moreover, at these lethal concentrations, necrotic alterations were seen in the thymic cortex and the testis which were considered stress-related. In this context, the importance of gill lesions is discussed.It is concluded that Br⁻ is goitrogenic in both species at low concentrations, and in guppies, at higher concentrations, it also affects skeletal muscle, blood vessels and gonads. Methyl bromide has no specific action but is irritating for superficial epithelia at relatively high concentrations.     

Genotoxic effect of Catha edulis (khat) crude extract after sub-chronic administration in rats

The aim of this study was to evaluate the genotoxic effects of a crude extract of khat (Catha edulis, Forsk) leaves in rats. Two groups were fed khat crude extract, 1000 and 2000 mg/kg body weight, for 90 days and were compared with a control group. The alkaline (pH > 13) version of comet assay was used in this study. However, no previous published work has been undertaken and showed the effect of khat on DNA migration in the comet assay. To compare the comet assay results with another genetic endpoint, blood samples were analyzed for chromosomal aberrations. These results showed no DNA damage detected using comet assay in both the khat treated groups, while the results of chromosomal aberrations assay showed a significant increase (P < 0.05) in the 2000 mg/kg body weight treated group compared to the control group.     

The utility of the guppy (Poecilia reticulata) and medaka (Oryzias latipes) in evaluation of chemicals for carcinogenicity.

There has been considerable interest in the use of small fish models for detecting potential environmental carcinogens. In this study, both guppies (Poecilia reticulata) and medaka (Oryzias latipes) were exposed in the aquaria water to three known rodent carcinogens for up to 16 months. Nitromethane, which caused mammary gland tumors by inhalation exposure in female rats, harderian gland and lung tumors in male and female mice, and liver tumors in female mice by inhalation, failed to increase tumors in either guppies or medaka. Propanediol, which when given in the feed was a multisite carcinogen in both sexes of rats and mice, caused increased liver tumors in male guppies and male medaka. There was reduced survival in female guppies and no increased tumors in female medaka. 1,2,3-Trichloropropane, which when administered by oral gavage was a multisite carcinogen in both sexes of rats and mice, caused an increased incidence of tumors in the liver of both male and female guppies and medaka and in the gallbladder of male and female medaka. The results of this study demonstrate that for these three chemicals, under these specific exposure conditions, the fish appear less sensitive and have a narrower spectrum of tissues affected than rodents. These results suggest that fish models are of limited utility in screening unknown chemicals for potential carcinogenicity.     

Ellagic acid, a natural polyphenol protects rat peripheral blood lymphocytes against nicotine-induced cellular and DNA damage in vitro: with the comparison of N-acetylcysteine

The present work is aimed at evaluating the protective effect of ellagic acid (EA), a natural polyphenolic compound that is widely distributed in fruits and nuts against nicotine-induced toxicity in rat peripheral blood lymphocytes. The effect of EA against nicotine toxicity was compared with N-acetylcysteine (NAC), a well-known antioxidant. Lymphocytes were exposed to nicotine at the doses of 0.125, 0.25, 0.5, 1, 2, 3 and 4 mM for 1h in culture media. Thiobarbituric acid reactive substances (TBARS), a lipid peroxidative marker and reduced glutathione (GSH), as indicative of endogenous antioxidant status were analyzed to fix the optimum dose. The lowest concentration eliciting significant damage was 1 mM nicotine and maximum damage was observed with 3 mM concentration, as evidenced by increased levels of TBARS and decreased levels of GSH. Hence, the test concentration was fixed at 3 mM nicotine. To establish most effective protective support we used five different concentrations of EA (10, 50, 100, 150 and 300 microM) against 3 mM nicotine. A dose-dependent inhibitory effect was observed with all doses of EA. Maximum protection was observed at the dose of 100 microM EA. So, 100 microM dose was used for further studies. We have tested five different concentrations of NAC-0.25, 0.5, 1, 2 and 4 mM to elucidate the optimum protective dose against nicotine toxicity. One millimolar NAC showed a significant protection against nicotine toxicity. Protective effect of EA against nicotine toxicity was elucidated by analyzing the lipid peroxidative index, viz., TBARS, hydroperoxides (HP) and endogenous antioxidant status, viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), Vitamins A, E and C. DNA damage and repair were assessed by using alkaline single-cell microgel electrophoresis (Comet assay) and micronucleus assay. There was a significant increase in the levels of lipid peroxidative index, severity in DNA damage and micronuclei number in nicotine-treated group, which was positively modulated by EA treatment. Antioxidant status was significantly depleted in nicotine-treated group, which was effectively restored by EA treatment. The protection of EA against nicotine toxicity was equally effective to that of NAC. EA and NAC treatment alone did not produce any damage to the normal lymphocytes at their effective doses. These findings suggest the potential use and benefit of EA as a modifier of nicotine-induced genotoxicity.     

Low levels of methylmercury induce DNA damage in rats: protective effects of selenium

In this study we examined the possible antigenotoxic effect of selenium (Se) in rats chronically exposed to low levels of methylmercury (MeHg) and the association between glutathione peroxidase (GSH-Px) activity and DNA lesions (via comet assay) in the same exposed animals. Rats were divided into six groups as follows: (Group I) received water; (Group II) received MeHg (100 μg/day); (Group III) received Se (2 mg/L drinking water); (Group IV) received Se (6 mg/L drinking water); (Group V) received MeHg (100 μg/day) and Se (2 mg/L drinking water); (Group VI) received MeHg (100 μg/day) and Se (6 mg/L drinking water). Total treatment time was 100 days. GSH-Px activity was determined spectrophotometrically and DNA damage was determined by comet assay. Mean GSH-Px activity in groups I, II, III, IV, V and VI were, respectively: 40.19 ± 17.21; 23.63 ± 6.04; 42.64 ± 5.70; 38.50 ± 7.15; 34.54 ± 6.18 and 41.39 ± 11.67 nmolNADPH/min/gHb. DNA damage was represented by a mean score from 0 to 300; the results for groups I, II, III, IV, V and VI were, respectively: 6.87 ± 3.27; 124.12 ± 13.74; 10.62 ± 3.81; 13.25 ± 1.76; 86.87 ± 11.95 and 76.25 ± 7.48. There was a significant inhibition of GSH-Px activity in group II compared with group I (P < 0.05). Groups V and VI did not show a difference in enzyme activity compared with groups III and IV, showing the possible protective action of Se. Comet assay presented a significant difference in DNA migration between group II and group I (P < 0.0001). Groups V and VI showed a significant reduction in MeHg-induced genotoxicity (P < 0.001) when compared with group II. A negative correlation (r = −0.559, P < 0.05) was found between GSH-Px activity and DNA lesion, showing that the greater the DNA damage, the lower the GSH-Px activity. Our findings demonstrated the oxidative and genotoxic properties of MeHg, even at low doses. Moreover, Se co-administration reestablished GSH-Px activity and reduced DNA damage.     

Cyto and genotoxicity of gold nanoparticles in human hepatocellular carcinoma and peripheral blood mononuclear cells

Engineered nanomaterials have been extensively applied as active materials for technological applications. Since the impact of these nanomaterials on health and environment remains undefined, research on their possible toxic effects has attracted considerable attention. It is known that in humans, for example, the primary site of gold nanoparticles (AuNps) accumulation is the liver. The latter has motivated research regarding the use of AuNps for cancer therapy, since specific organs can be target upon appropriate functionalization of specific nanoparticles. In this study, we investigate the geno and cytotoxicity of two types of AuNps against human hepatocellular carcinoma cells (HepG2) and peripheral blood mononuclear cells (PBMC) from healthy human volunteers. The cells were incubated in the presence of different concentrations of AuNps capped with either sodium citrate or polyamidoamine dendrimers (PAMAM). Our results suggest that both types of AuNps interact with HepG2 cells and PBMC and may exhibit in vitro geno and cytotoxicity even at very low concentrations. In addition, the PBMC were less sensitive to DNA damage toxicity effects than cancer HepG2 cells upon exposure to AuNps.