Evaluation of toxic effects of a diet containing fish contaminated with methylmercury in rats mimicking the exposure in the Amazon riverside population

This study was designed to evaluate the effects of a diet rich in fish contaminated with MeHg, mimicking the typical diet of the Amazon riverside population, in rats. Animals were randomly assigned to one of three groups with eight rats in each group: Group I—control, received commercial ration; Group II—received a diet rich in uncontaminated fish; Group III—received a diet rich in fish contaminated with MeHg. Treatment time was 12 weeks. Oxidative stress markers were evaluated, as well as the effects of this diet on DNA stability, systolic blood pressure (SBP), nitric oxide (NO) levels and histological damage in different tissues. There was a significant increase in SBP values in rats fed with MeHg-contaminated fish diet after the 10th week of the treatment. As far as oxidative stress biomarkers are concerned, no differences were observed in reduced glutathione and protein carbonyl levels, glutathione peroxidase, catalase, superoxide dismutase or δ-aminolevulinate dehydratase activities between the groups of animals receiving contaminated and uncontaminated fish diets. On the other hand, malondialdehyde levels increased significantly in rats fed with contaminated fish. NO levels were similar in all groups. DNA migration showed augmented in rats exposed to contaminated fish and histopathological analyses showed weak but significant leukocyte infiltration. Thus, we conclude that the MeHg-contaminated fish diet induced a slight lipid peroxidation and genotoxicity. However, these effects seem to be much less pronounced than when rats are exposed to aqueous solution containing CH₃HgCl.Our findings support the contention that the chemical form of MeHg in fish or fish nutrients such as polyunsaturated fatty acids, Se or vitamin E could minimize the toxic effects of MeHg exposure in fish-eating communities.     

Assessment in vitro of cytogenetic and genotoxic effects of propolis on human lymphocytes

We evaluated the genetic damage by ethanolic extract of propolis (EEP) induced to human lymphocytes which were exposed to increasing concentrations (0-2000μgml(-1)). The results indicated that EEP reduced significantly the mitotic index (MI) and proliferation index (PI) when high concentrations of EEP were used. Sister chromatid exchange (SCE) rates indicated that EEP could have genotoxic effects at high concentrations. Exposure of the cells to the amount of ethanol used as solvent did not alter either the MI and cell proliferation kinetics (CPK), or the rate of SCE. The results showed: (a) statistical increase in the percentage the cells with CAs and in the frequency of SCE at the highest concentrations, (b) a decrease in MI and in the CPK values was observed, (c) no effect was noticed in negative controls. In conclusion, it can be assumed that high concentrations of EEP have a cyto and genotoxic effect, in vitro, for human peripheral lymphocytes.     

Reactive oxygen species mediated DNA damage in human lung alveolar epithelial (A549) cells from exposure to non-cytotoxic MFI-type zeolite nanoparticles

Increasing utilization of engineered nanoparticles in the field of electronics and biomedical applications demands an assessment of risk associated with deliberate or accidental exposure. Metal based nanoparticles are potentially most important of all the nanoparticles in terms of health risks. Microporous alumino-silicates and pure silicates named as zeolites and zeo-type materials with variety of structures, chemical compositions, particle sizes and morphologies have a significant number of industrial uses such as in catalysis, sorption and ion-exchange processes. In particular, the nanosized particles due to their unique properties are used in hybrid organic-inorganic materials for photography, photonics, electronics, labeling, imaging, and sensing. The aim of the current study is to investigate pure silica MFI-type zeolites nanoparticles with sizes of 50 nm and 100 nm (samples MFI-50 and MFI-100) under suspended conditions and their toxicological effects on human lung alveolar (A549) cells under in vitro conditions.     

Aquilegia vulgaris extract protects against the oxidative stress and the mutagenic effects of cadmium in Balb/c mice

Cadmium (Cd) is a non-essential element and is a widespread environmental pollutant. Exposure to cadmium can result in cytotoxic, carcinogenic and mutagenic effects. The aim of the current work was to evaluate the protective effect of Aquilegia vulgaris extract against the oxidative stress and the genotoxicity induced by Cd using the chromosomal aberrations in somatic and germ cells assay and random amplified polymorphism DNA (RAPD-PCR) analysis. Forty male Balb/c mice were divided into four groups including the control group, Cd-treated group and the groups treated with the extract alone or plus Cd. The results indicated that Cd increased serum ALT, AST, urea, LDH, CK, lipid peroxidation in liver tissue accompanied with a significant decrease in GPX and SOD. Cd also increased the number of chromosomal aberrations in bone marrow and spermatocytes including structural and numerical aberrations. Animals treated with the extract alone were comparable to the control regarding all the tested parameters. The extract succeeded in preventing or diminishing the oxidative stress and the clastogenic effects of Cd. It could be concluded that Aquilegia vulgaris extract is a promising protective agent against oxidative stress and genotoxicity during the exposure to Cd.     

Comparative potency approach based on H2AX assay for estimating the genotoxicity of polycyclic aromatic hydrocarbons

Polycyclic Aromatic Hydrocarbons (PAHs) constitute a family of over one hundred compounds and can generally be found in complex mixtures. PAHs metabolites cause DNA damage which can lead to the development of carcinogenesis. Toxicity assessment of PAH complex mixtures is currently expressed in terms of toxic equivalents, based on Toxicity Equivalent Factors (TEFs). However, the definition of new TEFs for a large number of PAH could overcome some limitations of the current method and improve cancer risk assessment. The current investigation aimed at deriving the relative potency factors of PAHs, based on their genotoxic effect measured in vitro and analyzed with mathematical models. For this purpose, we used a new genotoxic assay (γH2AX) with two human cell lines (HepG2 and LS-174T) to analyze the genotoxic properties of 13 selected PAHs at low doses after 24 h treatment. The dose-response for genotoxic effects was modeled with a Hill model; equivalency between PAHs at low dose was assessed by applying constraints to the model parameters. In the two cell lines tested, we observed a clear dose-response for genotoxic effects for 11 tested compounds. LS-174T was on average ten times more sensitive than HepG2 towards PAHs regarding genotoxicity. We developed new TEFs, which we named Genotoxic Equivalent Factor (GEF). Calculated GEF for the tested PAHs were generally higher than the TEF usually used. Our study proposed a new in vitro based method for the establishment of relevant TEFs for PAHs to improve cancer risk assessment.     

The use of ex vivo human skin tissue for genotoxicity testing

As a result of the chemical legislation concerning the registration, evaluation, authorization and restriction of chemicals (REACH), and the Seventh Amendment to the Cosmetics Directive, which prohibits animal testing in Europe for cosmetics, alternative methods for safety evaluation of chemicals are urgently needed. Current in vitro genotoxicity assays are not sufficiently predictive for the in vivo situation, resulting in an unacceptably high number of misleading positives. For many chemicals and ingredients of personal care products the skin is the first site of contact, but there are no in vitro genotoxicity assays available in the skin for additional evaluation of positive or equivocal responses observed in regulatory in vitro genotoxicity assays. In the present study ex vivo human skin tissue obtained from surgery was used for genotoxicity evaluation of chemicals by using the comet assay. Fresh ex vivo human skin tissue was cultured in an air-liquid interface and topically exposed to 20 chemicals, including true positive, misleading positive and true negative genotoxins. Based on the results obtained in the present study, the sensitivity, specificity and accuracy of the ex vivo skin comet assay to predict in vivo genotoxicity were 89%, 90% and 89%, respectively. Donor and experimental variability were mainly reflected in the magnitude of the response and not the difference between the presence and absence of a genotoxic response. The present study indicates that human skin obtained from surgery is a promising and robust model for safety evaluation of chemicals that are in direct contact with the skin.     

Evaluation of antigenotoxic effects of carotenoids from green algae Chlorococcum humicola using human lymphocytes

Objective

To identify the available phytochemicals and carotenoids in the selected green algae and evaluate the potential genotoxic/antigenotoxic effect using lymphocytes.

Methods

Organic solvent extracts of Chlorococcum humicola (C. humicola) were used for the phytochemical analysis. The available carotenoids were assessed by HPLC, and LC-MS analysis. The genotoxicity was induced by the benzo(a)pyrene in the lymphocyte culture, the genotoxic and antigenotoxic effects of algal carotenoids with and without genotoxic inducer were evaluated by chromosomal aberration (CA), sister chromatid exchange (SCE) and micronucleus assay (MN).

Results

The results of the analysis showed that the algae were rich in carotenoids and fatty acids. In the total carotenoids lutein, β-carotene and α-carotene were found to be present in higher concentration. The frequency of CA and SCE increased by benzo(a)pyrene were significantly decreased by the carotenoids (P<0.05 for CA, P<0.001 for SCE). The MN frequencies of the cells were significantly decreased by the treatment with carotenoids when compared with the positive controls (P<0.05).

Conclusions

The findings of the present study demonstrate that, the green algae C. humicola is a rich source of bioactive compounds especially carotenoids which effectively fight against environmental genotoxic agents, the carotenoids itself is not a genotoxic substance and should be further considered for its beneficial effects.     

The Maillard reaction--illicite (bio)chemistry in tissues and food

We present a review of our early work on the Maillard reaction, at the interface of food chemistry and tissue biochemistry, as well as the reinterpretation of our early findings in the light of recent advances in the chemistry of the involved reactions. These concern specifically the role of lower aldehydes, produced during the glycolytic pathways and especially acetaldehyde. We also review some of our recent findings on the cytotoxic and genotoxic aspect of these “illicit” organic reactions, taking place in tissues (and also in food products) besides the genetically “programmed” metabolic pathways. Some recent results in organic-pharmaceutical chemistry confirm the potential importance of the reviewed reactions both in food chemistry and in tissues as well as the pathological importance of reactions taking place in tissues.     

Pharmacological, genotoxic and phytochemical properties of selected South African medicinal plants used in treating stomach-related ailments

Ethnopharmacological relevance

The evaluated medicinal plants are used in South African traditional medicine in treating stomach-related ailments.

Aims of the study

The study aimed at evaluating the pharmacological, genotoxic and phytochemical properties of the seven selected medicinal plants used for treating stomach-related ailments.

Materials and methods

: Ethyl acetate (EtOAc), ethanol (EtOH) 70% and water extracts of the selected plant parts were evaluated for their antimicrobial and anthelmintic activities using microdilution assays. Gram-positive bacteria (Enterococcus faecalis and Staphylococcus aureus), Gram-negative bacterium (Escherichia coli) and Candida albicans were used for antimicrobial assays. Caenorhabditis elegans was used for the anthelmintic assay. Plant extracts were also assayed for their cyclooxygenase-inhibitory activity against cyclooxygenase-1 and -2 enzymes. The Ames test was used to evaluate the genotoxicity of the plant extracts. A spectrophotometric method was used to determine the total phenolics, gallotannins, flavonoids and saponins.

Results

Twelve extracts exhibited minimum inhibitory concentration (MIC) <1 mg/mL against the bacterial test strains, and five extracts exhibited MIC <1 mg/mL against Candida albicans. The EtOAc extract of Tetradenia riparia had the best minimum lethal concentration (MLC) value (0.004 mg/mL) against Caenorhabditis elegans. All the EtOAc extracts exhibited percentage inhibition in the range of 50.7-94.7% against COX-1 and -2 enzymes at 250 μg/mL. All the plant extracts were non-mutagenic towards Salmonella typhimurium tester strains TA98, TA100 and TA1537 without metabolic activation. Phytochemical analysis revealed relatively high amounts of total phenolics, gallotannins and flavonoids in the evaluated plant extracts.

Conclusions

The general pharmacological activities exhibited by some of the plant extracts in this study support the traditional uses of the selected plants in treating stomach-related ailments. The Ames test showed that all the plant extracts were non-mutagenic but cytotoxicity tests are needed to ascertain the safety for long-term consumption.     

Genotoxicity assessment of chromium(III) propionate complex in the rat model using the comet assay

The aim of the study was to assess genotoxicity of a chromium(III) propionate complex in rat’s peripheral blood lymphocytes by the comet assay. The study was carried out on 18 12-weeks old female Wistar rats that were divided into three equal groups (six animals each): control (0), control-Cr(VI) and Cr(III)-tested rat fed ad libitum a basal diet and the diet supplemented either with 10 mg Cr(VI)/kg diet (given as K₂Cr₂O₇, equivalent of 1 mg/kg body mass/day) or 1000 mg Cr(III)/kg diet (given as [Cr₃O(O₂CCH₂CH₃)6(H₂O)₃]NO₃), equivalent of 100 mg Cr/kg body mass/day) for 4 weeks. High doses of supplementary Cr(III) were found to not affect body mass gain, feeding efficiency ratio and internal organ masses. Treatment of rats with the Cr(III) propionate complex, in contrast to Cr(VI), did not affect significantly the comet assay results in lymphocytes, which suggests that the compound does not exert genotoxic effects in rats.