Dione Silva Corrêa

Evaluation of the genotoxic potential of soil contaminated with mineral coal tailings on snail Helix aspersa.

Coal remains an important source of energy, although the fuel is a greater environmental pollutant. Coal is a mixture of several chemicals, especially inorganic elements and polycyclic aromatic hydrocarbons (PAH). Many of these compounds have mutagenic and carcinogenic effects on organisms exposed to this mineral. In the town of Charqueadas (Brazil), the tailings from mining were used for landfill in the lower areas of the town, and the consequence is the formation of large deposits of this material. The purpose of this study was to evaluate the genotoxic potential of soil samples contaminated by coal waste in different sites at Charqueadas, using the land snail Helix aspersa as a biomonitor organism. Thirty terrestrial snails were exposed to different treatments: 20 were exposed to the soil from two different sites in Charqueadas (site 1 and 2; 10 in each group) and 10 non-exposed (control group). Hemolymph cells were collected after 24h, 5days and 7days of exposure and comet assay, micronucleus test, oxidative stress tests were performed. Furthermore, this study quantified the inorganic elements present in soil samples by the PIXE technique and polycyclic aromatic hydrocarbons (PAH) by HPLC. This evaluation shows that, in general, soils from sites in Charqueadas, demonstrated a genotoxic effect associated with increased oxidative stress, inorganic and PAH content. These results demonstrate that the coal pyrite tailings from Charqueadas are potentially genotoxic and that H. aspersa is confirmed to be a sensitive instrument for risk assessment of environmental pollution.

Corrective effects of acerola (Malpighia emarginata DC.) juice intake on biochemical and genotoxical parameters in mice fed on a high-fat diet

Acerola contains high levels of vitamin C and rutin and shows the corresponding antioxidant properties. Oxidative stress on the other hand is an important factor in the development of obesity. In this study, we investigated the biochemical and antigenotoxic effects of acerola juice in different stages of maturity (unripe, ripe and industrial) and its main pharmacologically active components vitamin C and rutin, when given as food supplements to obese mice. Initial HPLC analyses confirmed that all types of acerola juice contained high levels of vitamin C and rutin. DPPH tests quantified the antioxidant properties of these juices and revealed higher antioxidant potentials compared to pure vitamin C and rutin. In an animal test series, groups of male mice were fed on a standard (STA) or a cafeteria (CAF) diet for 13 weeks. The latter consisted of a variety of supermarket products, rich in sugar and fat. This CAF diet increased the feed efficiency, but also induced glucose intolerance and DNA damage, which was established by comet assays and micronucleus tests. Subsequently, CAF mice were given additional diet supplements (acerola juice, vitamin C or rutin) for one month and the effects on bone marrow, peripheral blood, liver, kidney, and brain were examined. The results indicated that food supplementation with ripe or industrial acerola juice led to a partial reversal of the diet-induced DNA damage in the blood, kidney, liver and bone marrow. For unripe acerola juice food supplementation, beneficial effects were observed in blood, kidney and bone marrow. Food supplementation with vitamin C led to decreased DNA damage in kidney and liver, whereas rutin supplementation led to decreased DNA damage in all tissue samples observed. These results suggest that acerola juice helps to reduce oxidative stress and may decrease genotoxicity under obesogenic conditions.

Evaluation of acute and subacute toxicity and mutagenic activity of the aqueous extract of pecan shells [Carya illinoinensis (Wangenh.) K. Koch]

The infusion of pecan shells has been used to prevent and control hypercholesterolemia, diabetes and toxicological diseases. The aim of the present study was to evaluate toxicity and mutagenic effects of pecan shells aqueous extract (PSAE). Wistar rats were treated with a single dose of 300 or 2000 mg/kg of PSAE in the acute toxicity test. For the subacute test, the animals received 10 or 100 mg/kg of PSAE for 28 days. The mutagenicity was evaluated using Salmonella/microsome assay in TA1535, TA1537, TA98, TA100 and TA102 S. typhimurium strains in the presence and absence of metabolic activation (S9 mix) and micronucleus test in bone marrow. HPLC analyses indicated the presence of tannins, flavonoids, gallic and ellagic acids. Except for triglycerides, all treated groups presented normal hematological and biochemical parameters. Lower levels of triglycerides and weight loss were observed in the 100 mg/kg group. Mutagenic activities were not detected in S. typhimurium strains and by the micronucleus test. Based on these results, PSAE was not able to induce chromosomal or point mutations, under the conditions tested. The 100mg/kg dose showed significant antihyperlipidemic action, with no severe toxic effects.

DNA damage induced by coal dust, fly and bottom ash from coal combustion evaluated using the micronucleus test and comet assay in vitro

Coal mining and combustion generating huge amounts of bottom and fly ash are major causes of environmental pollution and health hazards due to the release of polycyclic aromatic hydrocarbons (PAH) and heavy metals. The Candiota coalfield in Rio Grande do Sul, is one of the largest open-cast coal mines in Brazil. The aim of this study was to evaluate genotoxic and mutagenic effects of coal, bottom ash and fly ash samples from Candiota with the comet assay (alkaline and modified version) and micronucleus test using the lung fibroblast cell line (V79). Qualitative and quantitative analysis of PAH and inorganic elements was carried out by High Performance Liquid Chromatography (HPLC) and by Particle-Induced X-ray Emission (PIXE) techniques respectively. The samples demonstrated genotoxic and mutagenic effects. The comet assay modified using DNA-glicosilase formamidopirimidina (FPG) endonuclease showed damage related to oxidative stress mechanisms. The amount of PAHs was higher in fly ash followed by pulverized coal. The amount of inorganic elements was highest in fly ash, followed by bottom ash. It is concluded that the samples induce DNA damage by mechanisms that include oxidative stress, due to their complex composition, and that protective measures have to be taken regarding occupational and environmental hazards.

Cytotoxicity and genotoxicity induced by coal and coal fly ash particles samples in V79 cells

AbstractExposure to coal and coal ashes can cause harmful effects in in vitro and in vivo systems, mainly by the induction of oxidative damage. The aim of this work was to assess cytotoxic and genotoxic effects using the V79 cell line treated with coal and coal fly ash particles derived from a coal power plant located in Santa Catarina, Brazil. Two coal samples (COAL11 and COAL16) and two coal fly ash samples (CFA11 and CFA16) were included in this study. COAL16 was co-firing with a mixture of fuel oil and diesel oil. The comet assay data showed that exposure of V79 cells to coal and coal fly ash particles induced primary DNA lesions. Application of lesion-specific endonucleases (FPG and ENDO III) demonstrated increased DNA effects indicating the presence of high amounts of oxidative DNA lesions. The cytokinesis-block micronucleus cytome assay analysis showed that exposure of V79 cells to high concentrations of coal and coal fly ash particles induced cytotoxic effects (apoptosis and necrosis) and chromosomal instability (nucleoplasmic bridges, nuclear buds, and micronucleus (MN) formation). These results may be associated with compounds contained in the surface of the particles as hazardous elements, ultrafine/nanoparticles, and polycyclic aromatic hydrocarbons (PAHs) which were detected in the samples. Graphical abstractᅟ

Chemical and toxicological effects of medicinal Baccharis trimera extract from coal burning area

The entire process of power generation, extraction, processing and use of coal strongly impact water resources, soil, air quality and biota leads to changes in the fauna and flora. Pollutants generated by coal burning have been contaminating plants that grow in area impacted by airborne pollution with high metal contents. Baccharis trimera is popularly consumed as tea, and is widely developed in Candiota (Brazil), one of the most important coal burning regions of the Brazil. This study aims to investigate the phytochemical profile, in vivo genotoxic and mutagenic potential of extracts of B. trimera collected from an exposed region to pollutants generated by coal burning (Candiota City) and other unexposed region (Bagé City), using the Comet assay and micronucleus test in mice and the Salmonella/microsome short-term assay. The HPLC analyses indicated higher levels of flavonoids and phenolic acids for B. trimera aqueous extract from Bagé and absence of polycyclic aromatic hydrocarbons for both extracts. The presence of toxic elements such as cobalt, nickel and manganese was statistically superior in the extract from Candiota. For the Comet assay and micronucleus test, the mice were treated with Candiota and Bagé B. trimera aqueous extracts (500-2000 mg/kg). Significant genotoxicity was observed at higher doses treated with B. trimera aqueous extract from Candiota in liver and peripheral blood cells. Micronuclei were not observed but the results of the Salmonella/microsome short-term assay showed a significant increase in TA98 revertants for B. trimera aqueous extract from Candiota. The extract of B. trimera from Candiota bioacumulated higher levels of trace elements which were associated with the genotoxic effects detected in liver and peripheral blood cells.

Assessment of the genotoxic and mutagenic properties of Himatanthus articulatus bark extracts used as phytotherapeutic drug in the Amazon.

ETHNOPHARMACOLOGICAL RELEVANCE Himatanthus articulatus (Apocynaceae) is a plant native to the Amazon, popularly used to treat external ulcers, tumors, inflammations, cancer, syphilis and malaria. AIM OF THE STUDY To investigate the in vivo genotoxic and mutagenic potential of this plant, using the comet assay and the micronucleus test. MATERIAL AND METHODS Female and male adult mice were treated with 500 mg/kg, 1000 mg/kg or 2000 mg/kg of Himatanthus articulatus aqueous or ethanolic bark extracts by gavage for two consecutive days. In addition, blood slides were exposed to hydrogen peroxide (ex vivo) to evaluate the anticlastogenic effect using the comet assay. The HPLC analyses indicated plumieride as the main constituent of both extracts from Himatanthus articulatus barks. RESULTS No differences between genders were observed. Micronuclei were observed only in the group treated with the highest dose of both extracts. Conversely, lower doses of these extracts showed protective effects to DNA against damage induced by hydrogen peroxide, indicating an important antigenotoxic effect. CONCLUSIONS The toxicological evaluation indicated that the extracts are non-genotoxic and reduce the clastogenic damage induced by hydrogen peroxide. In part, this result can be atributted to the phytochemical profile of Himatanthus articulatus, which presents iridoids and phenolic compounds.

Evaluation of Safety of Arrabidaea chica Verlot (Bignoniaceae), a Plant with Healing Properties

Arrabidaea chica Verlot (Bignoniaceae) has been used as a medicinal herb to treat anemia, hemorrhage, inflammation, intestinal colic, hepatitis, and skin infections in the Brazilian Amazon region. Studies have demonstrated the healing properties of extracts obtained from A. chica leaves, which contain anthocyanins and flavonoids. However, few investigations have assessed the safe use of this plant species. In this study, mutagenic and genotoxic effects of a crude aqueous extract, a butanolic fraction, and aqueous waste from A. chica leaves were evaluated using the Salmonella/microsome assay in TA98, TA97a, TA100, TA102, and TA1535 strains and the alkaline comet assay in Chinese hamster ovary (CHO) cell culture with and without metabolic activation. The crude aqueous extract, butanolic fraction, and aqueous waste were not mutagenic in any of the Salmonella typhimurium strains tested, and showed negative responses for genotoxicity in CHO cells. High-performance liquid chromatography (HPLC) analysis indicated the presence of phenolic acids and flavonoids such as rutin and luteolin. The lack of mutagenic/genotoxic effects might be due to phytochemical composition with high concentrations of known anti-inflammatory compounds. Thus, the crude aqueous extract, butanolic fraction, and aqueous waste from A. chica leaves do not appear to pose short-term genotoxic risks.

Chemical characterization and cytotoxic, genotoxic, and mutagenic properties of Baccharis trinervis (Lam, Persoon) from Colombia and Brazil

PHARMACOLOGY RELEVANCE Baccharis trinervis (Lam, Persoon) leaves are used in the traditional medicine for the treatment of high fevers, edema, inflammation, sores and muscle cramps, snakebites and as antiseptic. AIM OF THE STUDY To investigate the cytotoxic, genotoxic, and mutagenic effects of extracts and fractions of B. trinervis from Brazil and Colombia in Chinese Hamster Ovary (CHO) cells, and to examine the mutagenic activity in Salmonella typhimurium. MATERIAL AND METHODS Aqueous extracts (AE) of aerial parts of B. trinervis from Brazil (B) and Colombia (C) were fractioned in ethyl acetate fraction (EAF), butanol extract (BF), and aqueous residue fraction (ARF). Qualitative chemical screening and determination of total flavonoid content were made. Identification of chemical constituents was performed by High Performance Liquid Chromatography (HPLC) and High Resolution Mass Spectrometry (HRMS). For the in vitro tests, CHO cells were treated for 3h with extracts and fractions. The cytotoxic activity was evaluated by clonal survival and 3-(4.5-dimethylthiazole-2-yl)-2.5-biphenyl tetrazolium bromide reduction assay (MTT). Genotoxic and mutagenic effects were evaluated by the alkaline comet assay and Cytokinesis-blockage micronucleus test (CBMN), respectively. Additionally, Salmonella/microsome assay was carried out to determinate the mutagenic effects in EAF from Brazil and Colombia. RESULTS Phytochemical analyses indicated the presence of saponins and flavonoids. AE and EAF were the samples with the highest quantity of total flavonoids. HPLC showed the presence of luteolin only in AEC, and caffeic acid, ellagic acid, rosmarinic acid, and rutin were identified in AEB and AEC (AEC>AEB). The HRMS in positive mode of EAFB and EAFC showed presence of two carboxylic acids, coumarin, and two terpenoids. In addition, were identified one terpenoid and two carboxylic acids in AE, BF and ARF of B. trinervis from both countries in negative mode. Dose-dependent cytotoxic effects were observed in CHO cells treated with B. trinervis extracts and fractions by using clonal survival and MTT at concentrations higher than 0.05mg/mL. All the extracts and fractions induced DNA strand breaks in CHO cells with dose-dependent response, mostly EAFB and EAFC. The EAF from Brazil and Colombia showed mutagenic effect at 0.5mg/mL, while the other fractions did not show a significant difference in relation to the control. No mutagenic effects were found in EAF from both countries by the Salmonella/microsome assay. CONCLUSIONS Cytotoxic and genotoxic effects were demonstrated in all extracts and fractions used, although only EAF showed mutagenic effects by CBMN, but not by Salmonella/microsome assay. Our results suggest that flavonoids, phenylpropanoids, coumarins, and diterpenes may be responsible for the cytotoxic, genotoxic and mutagenic effects observed.

A 28-day Sub-acute Genotoxic and Behavioural Assessment of Garcinielliptone FC

Garcinielliptone FC (GFC) is a polyisoprenylated benzophenone isolated from Platonia insignis Mart (Clusiaceae) with promising anticonvulsant properties. However, its safe use and other effects on the central nervous system require assessment. This study assessed the toxicological effects of GFC using the comet assay and the micronucleus test in mice treated for 28 days. A behavioural model was employed to detect possible injuries on the central nervous system. Mice treated with GFC (2, 10 and 20 mg/kg; i.p.) daily for 28 days were submitted to rotarod test, open‐field test and tail suspension test (TST). After the behaviour tasks, biological samples were assessed to evaluate genotoxic and mutagenic effects using the comet assay and the micronucleus test. Garcinielliptone FC did not impair the performance of the animals in the rotarod and open‐field tests, with no antidepressant‐like effect in TST. No genotoxic effects in blood and cerebral cortex were observable in the comet assay; however, there was a significant increase in index and frequency of damage in liver after treatment with GFC 20 mg/kg. Garcinielliptone FC did not increase micronucleus frequency in bone marrow. At the tested doses, GFC was not toxic to the CNS and did not induce genotoxic damage to blood or bone narrow cells. DNA damage to liver tissue was caused only by the highest dose, although no mutagenic potential was observed.