Polyphenol contents of five of medicinal plants from Cameroon and effects of their extracts on antioxidant capacities of human breast cancer cells

The aim of the study was to investigate the antiproliferative properties of extracts of five plants on breast cancer cells (MDA-MB-231) and their effects on antioxidant activities. The total phenol contents of the extracts were determined to establish a correlation to their antiproliferative effects which were evaluated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assay. Enzymes involved in oxidative stress were also determined. Total polyphenol contents of the five extracts varied from 170 to 440 mg L^?1 of gallic acid per mg of extract with the highest value for the extract of Sida cordifolia. Cell growth inhibition was observed within 24 h, with inhibitory concentration values ranging from 7.3 to 25 mg L^?1 depending on plant extract. At concentrations between 100 and 200 mg L^?1, all the extracts exhibited significant reduction of cell proliferation in time-dependent and linear manner. The activities of superoxide dismutase and catalase of cells treated for 24 h with the extract of S. cordifolia were 3.5 U per mg protein and 8 mole H₂O₂ consumed per min per mg protein higher than those of cells treated with an extract of Viscum album.     

离子液体氯化1-辛基-3-甲基咪唑对洋葱的遗传毒性作用

为探讨咪唑类离子液体氯化1-辛基-3-甲基咪唑([C_8mim][C1])对洋葱的遗传毒性,用浓度分别为2.5、5、10和20mmol·L~(-1)的[C_8mim][C1]处理洋葱内表皮细胞和洋葱根尖8 h,用浓度为10 mmol·L~(-1)的[C_8mim][C1]处理洋葱内表皮细胞和洋葱根尖4 h、8 h、12 h和24 h,研究[C_8mim][C1]对洋葱内表皮细胞核的影响和对洋葱根尖有丝分裂指数和染色体畸变的影响。结果显示,与对照组相比,随着[C_8mim][Cl]处理浓度和时间的增加,洋葱内表皮细胞核偏移率明显升高,根尖细胞有丝分裂指数显著降低,染色体畸变现象明显,畸变率升高。研究表明,当浓度达到2.5 mmol·L~(-1)时,[C_8mim][Cl]对洋葱具有遗传毒性。     

A study on genotoxicity of textile dyeing industry effluents from Rajshahi,Bangladesh, by the Allium cepa test

Textile dye effluents are believed to be toxic as they might exert various harmful effects on living organisms including genotoxicity. These effluents are the main source of direct and continuous input of pollutants into the aquatic ecosystems. In this study, dye effluents from a local silk dyeing industry were analysed for their genotoxic potential by the Allium cepa genotoxicity test. The A. cepa test is characterised as a genotoxicity test where the roots of A. cepa are grown in different concentrations of the test material. The macroscopic results clearly showed that the toxicity of the dye effluents prompted A. cepa root growth inhibition, and this effect increased with higher concentrations of the effluents. At the cellular level, no dividing cells were found at higher concentrations such as 60%, 80% and 100% of the effluents. However, at a lower concentration of 20%, dividing cells were identified, although the mitotic index was much lower than that of the control. Microscopic analysis showed that the dye effluents induced chromosomal aberrations at significant levels. Taken together, these results revealed that the textile dyeing industry effluents are toxic to eukaryotic cells and these dyes have genotoxic properties that can potentially lead to cancer development and adverse health conditions.     

Induction of apoptosis and cytokine markers in colon cancer cells by magnesium oxide (MgO) nanoparticles

Magnesium oxide nanoparticles (MgONP) are predominantly utilized in industrial products. This study was undertaken to elucidate the mechanisms underlying toxic effect of MgONP in human colon cancer (HT 29) cells over 48 hr period. Cytotoxicity was evaluated by using MTT and neutral red uptake assays. Data demonstrated that MgONP reduced cell viability in concentration- and time-dependent manner. MgONP induced oxidative stress by decreasing glutathione (GSH) concentrations and elevation of reactive oxygen species (ROS) and lipid peroxidation levels. Increased caspase-3 enzyme activity and greater condensed, damaged chromosome was observed following MgONP exposure in HT 29 cells. The level of interleukin-4 (IL-4), tumor necrosis factor (TNF-α), and DNA fragmentation were significantly higher in MgONP incubated cells. The results showed that MgONP-induced toxicity in HT 29 cells may be mediated through oxidative stress.     

Genotoxic Effect of Phenol on the Cells of Onion (Allium cepa) Roots

The roots of onion (Allium cepa) stand out for having cells with large size and small number of chromosomes.These characteristics make them useful in bioassays for the measurement of a variety of cytogenetic and morphological parameters , in which they can be used as toxicity indicators of the induction and formation of micronuclei and chromosomal aberrations.Based on this background , the potential genotoxic effect of phenol concentration on cells of A.ceparoots was investigated either in terms of induced aberrations or micronuclei formation.The results demonstrated that the higher the concentration of phenol , the higher the incidence of abnormalities , thus confirming the genotoxicity of this pollutant.     

The search for indigenous dune stabilizers: Germination requirements of selected South African species

The coastline of South Africa is characterized by extensive dune fields which are threatened by development, and thus the demand of stabilization of drift sand will increase. The non-invasive alien grass speciesAmmophila arenaria is at present the dominant sand stabilizer. Due to its foreign origin and invasiveness in North America its use was criticized and indigenous sand-binding species would be preferred. The germination requirements of the nativeArctotheca populifolia, Ipomoea pes-caprae. Myrica cordifolia andScaevola plumieri were investigated. The results showed that for all four species the total germination time and germination phase could be shortened and germination success (number of germinated seeds) improved. The different scarification and stratification treatments broke the dormancy ofI. pes-caprae, M. cordifolia andS. plumieri, whereas the seeds ofA. populifolia were not dormant. Seeds ofI. pes-caprae andM. cordifolia had induced an innate dormancy, respectively, whereas the type of dormancy in seeds ofS. plumieri dormancy processes remained unknown.     

Genotoxicity biomarkers in the amphipod Gammarus elvirae exposed in vivo to mercury and lead,and basal levels of DNA damage in two cell types

The present work deals with the application of genotoxicity biomarkers by means of the Comet assay in haemocytes and spermatozoa of the crustacean Gammarus elvirae exposed in vivo to heavy metals. Furthermore, a basal levels (BLs) study of DNA damage in the two cell types considered for two different gammarids species, G. elvirae and Echinogammarus veneris, was carried out. It is important to identify factors that influence the outcome of the assay in order to obtain reliable and reproducible results usable for risk assessment purposes. Our results highlight that the Italian legal limits for Hg and Pb, respectively, 0.5 and 50?µg/L, are inadequate for establishing safety thresholds in the aquatic environment. Furthermore, the freshwater invertebrate G. elvirae, used for the first time to measure the effect of genotoxicants, is a good candidate for evaluating the genotoxicity damage induced by heavy metals. Our results concerning spermatozoa show excessively variable responses and high BLs.     

Oxidative stress and genotoxicity of 1-methyl-3-octylimidazolium chloride on loach (Misgurnus anguillicaudatus)

This study was a preliminary step to evaluate the acute toxicity of 1-methyl-3-octylimidazolium chloride ([C₈mim]Cl) on loach (Misgurnus anguillicaudatus) by determining the effects on hepatic antioxidant enzyme activities and by the comet assay. The results showed that [C₈mim]Cl had acute toxicity at concentrations above 20 mg L^?1, inducing oxidative stress and genotoxicity on fish liver cells. In respect to enzyme activities, [C₈mim]Cl induced changes in the activities of superoxide dismutase, catalase, and glutathione content the livers of fish exposed at 20–80 mg L^?1. [C₈mim]Cl at the same exposure level caused a remarkable increase in malondialdehyde level. The comet assay indicated that [C₈mim]Cl at 20–80 mg L^?1 induced genotoxicity in liver cells. With increased exposure concentration and time, the two comet parameters trailing rate and tail moment were significantly increased, with significant differences (P < 0.05) observed between control group and each treatment group. The present study shows that ionic liquids can be a threat to the health of aquatic organism when accidentally released to aquatic ecosystems.     

Protective effect of Kappaphycus alvarezii (Rhodophyta) extract against DNA damage induced by mercury chloride in marine fish

Marine organisms are continuously exposed to agents, both exogenous and endogenous, that damage DNA. Consequently, it is important to determine the ability of compounds to provide protection against damaging chemicals. The aim of this study was to evaluate the anti-genotoxic activity of crude aqueous extracts of Kappaphycus alvarezii (Rhodophyceae), collected from the Southeast coast of India. This study focused on possible anti-genotoxic potential of aqueous extract of K. alverazii to interfere with clastogenicity induced by mercury chloride (HgCl₂) in marine fish, Therapon jarbua as measured by cytogenetic endpoints such as cell viability and comet assay. In the first set of experiments, fish were exposed to a single treatment of Hg at 0.125, 0.25, 0.5, 1, or 2?ppm along with controls. Mercury exposure produced significant DNA damage in all comet classes, maximum as >79% (Class 4) at 0.5, 1, and 2?ppm exposure in a time dependent manner. Algal extract did not induce genotoxicity when given alone and prevented Hg-induced genotoxicity. The algal extract reduction in genotoxicity was significant but not time- and concentration-dependent. Results suggested that under present experimental conditions, K. alvarezii extract exhibit potent anti-genotoxicity effects in this fish model; and thus these extracts may be recommended as a supplement in fish meal and may benefit humans ingesting Hg-contaminated fish.     

Mitochondrial dysfunction,impaired oxidative-reduction activity,degeneration, and death in human neuronal and fetal cells induced by low-level exposure to thimerosal and other metal compounds

Thimerosal (ethylmercurithiosalicylic acid), an ethylmercury (EtHg)-releasing compound (49.55% mercury (Hg)), was used in a range of medical products for more than 70 years. Of particular recent concern, routine administering of Thimerosal-containing biologics/childhood vaccines have become significant sources of Hg exposure for some fetuses/infants. This study was undertaken to investigate cellular damage among in vitro human neuronal (SH-SY-5Y neuroblastoma and 1321N1 astrocytoma) and fetal (nontransformed) model systems using cell vitality assays and microscope-based digital image capture techniques to assess potential damage induced by Thimerosal and other metal compounds (aluminum (Al) sulfate, lead (Pb)(II) acetate, methylmercury (MeHg) hydroxide, and mercury (Hg)(II) chloride) where the cation was reported to exert adverse effects on developing cells. Thimerosal-associated cellular damage was also evaluated for similarity to pathophysiological findings observed in patients diagnosed with autistic disorders (ADs). Thimerosal-induced cellular damage as evidenced by concentration- and time-dependent mitochondrial damage, reduced oxidative–reduction activity, cellular degeneration, and cell death in the in vitro human neuronal and fetal model systems studied. Thimerosal at low nanomolar (nM) concentrations induced significant cellular toxicity in human neuronal and fetal cells. Thimerosal-induced cytoxicity is similar to that observed in AD pathophysiologic studies. Thimerosal was found to be significantly more toxic than the other metal compounds examined. Future studies need to be conducted to evaluate additional mechanisms underlying Thimerosal-induced cellular damage and assess potential co-exposures to other compounds that may increase or decrease Thimerosal-mediated toxicity.     

In vitro evaluation of human liver cancer cells and the potential cytotoxicity of Tecoma stans (Bignoniaceae) and Brickellia cavanillesi (Asteraceae) both single and in combination

Medicinal herbs are steadily increasing in complementary use for chronic and alterative therapies. The health risks associated with herbal supplements have therefore been identified as a top research priority. Tecoma stans (Tronadora), a Bignoniaceae plant, is a herbal drug traditionally consumed as tea in South America for the control of diabetes. It contains the alkaloids of tecomine and tecostanine which were shown as functional compounds responsible for hypoglycemic activity. However, the side effects of aqueous extracts of this herbal tea have not been reported. In this study, studies sought to evaluate the cytotoxicity of water extracts from T. stans in human hepatoblastoma (HepG2). Toxic effects of T. stans were concentration- and time-dependent in the presence and absence of fetal bovine serum (FBS). Cells were incubated for up to 72-h with varying concentrations of herbal extracts (60–100%), cytotoxicity was determined spectrophotometrically by MTT and reported in terms of % cell viability. For IC₅₀ assay (24 h exposure), cytotoxicity was found at concentration of 60–100%. Evaluation of the effects of T. stans and Brickellia cavanillesi (Asteraceae) in combination revealed that extracts of both herbal extracts' nontoxic levels of T. stans significantly reduced cell viability in a time-dependent manner. Further studies are needed regarding the identification, toxicity, and molecular mechanism of action of active compounds, as well as their cytotoxicity to other cell lines following exposure to both single and combined herbal extracts of these plants with anti-diabetic properties.     

Oxidative stress in the freshwater cyprinid crucian carp (Carassius carassius L.) upon chronic exposure to endosulfan

Oxidative stress and antioxidant responses of crucian carp, upon chronic exposure to endosulfan, were evaluated in vivo. The lethal concentration (LC_50–96?h) was 70 μg L^?1; on its basis, the fish were exposed to endosulfan at 20, 35, and 50 μg L^?1 and autopsy was done on days 1, 2, 3, 4, 7, 14, 21, 28, and 35. Lipid peroxidation was induced in a concentration-dependent manner, being highest at 50 μg L^?1 (3/4 LC_{50–96 h}, sub-lethal concentration-I, SL-I) on day 4 (720% versus control), followed in its extent (490%) at 30 μg L^?1 (1/2 LC_{50–96 h}, sub-lethal concentration-II, SL-II) on day 7 and lowest (260%) at 10 μg L^?1 (1/4 LC_{50–96 h}, sub-lethal concentration-III, SL-III) on day 14. Glutathione showed a concentration- and time-dependent elevation in the initial phase, with highest level on day 4 (180%) at SL-I, but showed significant reduction in all test concentrations from day 21 of post-exposure. Superoxide dismutase was decreased significantly throughout the study, with highest reduction (63%) on day 4 at SL-I; catalase increased in all test concentrations up to day 14 but showed a significant decrease from the day 28 of post-exposure. The potential role of these parameters as indicators of pesticide pollution in aquatic systems is discussed.     

Effect of supplemented soybean (Glycine max L) diet and extracts on aluminum sulfate-induced genotoxicity

Aim of this study is to evaluate the protective role of Glycine max L. (soybean) against aluminum sulfate-induced genotoxicity. A diet containing 30% soybean and extracts obtained with various organic solvents were administered orally to male white Swiss mice prior to intraperitoneal injection of Al₂(SO₄)₃. Light microscopy was used for the examination of genotoxicity involving somatic cells, germ cells, and sperms. The main secondary metabolites contained in each extract were isolated and identified by chromatographic and spectral analyses. Administration of soy diet and extracts to the examined mice induced significant anti-mutagenic activity against bone marrow aberrations and spermatocyte and sperm abnormalities. At the end of this study, maximum protection was achieved using the methanolic extract (83% and 79%) followed by those obtained with chloroform (71% and 49%) and n-hexane (51% and 39%) for bone marrow aberrations and spermatocyte abnormalities, respectively. The observed activity of soybean extracts might be related to the main constituents isolated and identified. These main constituents were the isoflavone glycosides genistin and daidzin in addition to saponins (methanol extract), lignan matairesinol (chloroform extract), and β-sitosterol, unsaturated fatty acids as well as phospholipids (n-hexane extract). The results illustrated that soy extracts can be considered as alternative anti-mutagenic nutraceuticals and promising source for developing anti-mutagenic drugs.     

In vivo studies on the effect of Ocimum sanctum L. leaf extract in modifying the genotoxicity induced by chromium and mercury in Allium root meristems

In vivo cytogenetic assay in Allium cepa root tip cells has been carried out to detect the modifying effect of Ocimum sanctum aqueous leaf extract against chromium (Cr) and mercury (Hg) induced genotoxicity. It was observed that the roots post-treated with the leaf extract showed highly significant (p < 0.001) recovery in mitotic index (MI) and chromosomal aberrations (CA) when compared to pre-treated (Cr/Hg) samples and the lower doses of the leaf extract were found to be more effective than higher doses. The present study reveals that the Ocimum sanctum leaf extract possesses the protective effect against Cr/Hg induced genetic damage.     

Assessment of the in vivo genotoxicity of a new formulation of Bacillus thuringiensis var israelensis SH-14

Biological control agents have become a useful alternative for the reduction of the use of chemical insecticides. LABIOFAM (Cuba) is developing a new formulation of a biolarvicide that possesses as active biological agent Bacillus thuringiensis var israelensis serotype H14. In order to evaluate the genotoxicity of this new formulation, an in vivo battery test was used: micronucleus (MN), chromosome aberrations (CAs), and sperm morphology (SM) assays. A dose of 6.45?×?10⁸ spores was administered per animal via oral administration. Bone marrow cells were collected 24?h after a two day treatment for the MN assay, and 24?h after a unique treatment for the CA assay, using cyclophosphamide as the positive control. Sperm cells were collected at 5 weeks from the first of five administrations for the SM test, using acrylamide as positive control. Bacillus thuringiensis var israelensis serotype H14 failed to show either a significative increase of micronucleated polychromatic erythrocytes, chromosomal aberrations, or sperm abnormalities. Acute oral administration of a high dose of Bacillus thuringiensis var israelensis serotype H14 did not produce mutagenic effects in bone marrow or sperm cells.     

氧化石墨烯遗传毒性的实验研究

研究氧化石墨烯(GO)的遗传毒性,考察其致突变作用,为GO在生物领域的安全应用提供依据。采用Ames试验、体外CHL细胞染色体畸变试验和小鼠体内染色体畸变试验,分别在细菌水平、细胞水平及整体动物水平研究GO的遗传毒性。GO各剂量组的Ames试验结果为阴性。CHL试验中,CHL细胞染色体畸变率随GO浓度的增加而升高,其中1.000 mg·mL~(-1)剂量组(+S_9)和0.500 mg·mL~(-1)剂量组(-S_9)畸变率显著升高(P 0.05)。小鼠骨髓细胞染色体畸变试验中,骨髓细胞染色体畸变率同样随GO浓度的增加而升高,1.000和0.500 mg·kg~(-1)剂量组的畸变率显著提高(P 0.05)。虽然Ames试验结果没有反映出GO的遗传毒性,但在体外及体内染色体畸变试验中,GO均表现出对哺乳动物细胞染色体有潜在的遗传毒性。     

Evaluation and quantification of genotoxicity of urban waters by using Vicia faba bioassays

The ecotoxic effect of pollutants in the water environment has become a hot topic of research. Because of the genotoxicity describing the composite deleterious action of the toxicants in water, the Vicia faba root-tip cell micronucleus (MN) test was applied for genotoxicity assessment of organic and inorganic substances in four rivers, two lakes and effluent flows from two wastewater treatment plants in Xi’an, China. Results showed that the lowest MN frequency was between 2.0‰ and 3.0‰. However, the highest MN frequency of the water samples was as high as 30.0–34.0‰. The results well reflected that the seriously polluted river with high chemical oxygen demand and total organic carbon showed high genotoxicity, and also indicated the genotoxic impact of urban discharge on the surface waters and the ability of urban water bodies to absorb the impact. In addition, there were significant correlations between the genotoxicity expressed by MN rate and Cr⁶⁺ (5–80?mg/L) and phenol concentrations (2–40?mg/L). The genotoxicity of the urban water could be quantified as equivalent Cr⁶⁺ or phenol concentrations and which might be used as a set of quantitative indicators of the water genotoxicity.     

Uptake and genotoxicity of micromolar concentrations of cobalt chloride in mammalian cells

Literature data concerning the genotoxicity of cobalt salts have been conflicting. To establish appropriate incubation conditions, we conducted a series of uptake studies, before genotoxicity was determined by DNA strand break induction in HeLa cells and mutagenicity in V79 Chinese hamster cells. Co(II) is taken up by HeLa cells in a concentration‐dependent manner and is accumulated inside the cell. The uptake is preceded by a fast association step to the outer membrane, with no saturation up to 24 h. DNA strand breaks as determined by nucleoid sedimentation are induced at concentrations as low as 50μMCoCl₂. The induction is time‐dependent, showing the highest number of breaks after 4h incubation with no further increase up to 24h. CoCl₂ is mutagenic at the HPRT‐locus, enhancing the spontaneous mutation frequency 4.2‐fold at 100μ?. Besides direct interactions with DNA, the mutagenicity of CoCl₂ could also be due to a decrease in the Fidelity of DNA polymerisation.     

Genotoxicity of chromium in vitro on yeast: Interaction with DNA†

The genotoxicity of chromium chloride was investigated in cells of D₇ strain of Saccharomyces cerevisiae harvested either from logarithmic or stationary growth phase.

A weak induction of mifotic gene conversion and point reverse mutation was obtained when the incubations were performed using phosphate buffer. No genetic effect was observed when the incubations were performed using Tris‐HCl buffer.

The experiments with ⁵¹Cr radiotracer demonstrated that Cr³⁺ ion enters the yeast cells and binds to DNA even if the incubation mixture was performed with Tris HCl buffer. This behaviour could be due to the highest concentration of CrCl₃ that cause some damages to cytoplasmatic membrane.     

Effects of nonylphenol on ovarian cancer cells PEO4 c-myc and p53 mRNA and protein expression

The molecular mechanisms underlying proliferation and apoptosis induced by p-nonylphenol (NP) through regulation of mRNA and protein expression of c-myc, p53 mRNA and the protein was investigated using ovarian cancer PEO4 cells. PEO4 cells were grown in the Dulbecco's modified Eagle medium (DMEM), 3(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay was employed to investigate cell proliferation. RT-PCR and immunohistochemistry was performed to determine mRNA and protein expression of PEO4 c-myc and p53. Within the range of approximately 8–9×10^?7 mol L^?1, NP stimulated proliferation in PEO4 cells in a concentration-and time-dependent manner. RT-PCR and immunohistochemistry analyses revealed that treatment with 32 × 10^?7 mol L^?1 NP for 72 hr resulted in increased mRNA and protein expression of PEO4 c-myc and p53. Evidence indicates that NP exerted estrogenic actions and stimulated proliferation in estrogen responsive ovarian cancer PEO4 cells by up-regulation of c-myc mRNA and protein expression.