Genistein at maximal physiologic serum levels induces G0/G1 arrest in MCF-7 and HB4a cells, but not apoptosis.

Several studies have demonstrated that a balanced diet can contribute to better human health. For this reason, soy-based food and pure isoflavones (pills) are one of the most consumed. The association of this consumption and lower risks of chronic diseases and cancer is well established for the Asian population and has been attracting the attention of people worldwide, especially women at menopause who seek to alleviate the symptoms associated with the lack of estrogen. Despite positive epidemiological data, concerns still exist because of conflicting results found in scientific literature with relation to the role of isoflavones in breast and hormone-related cancers. The aim of our study was to investigate the cytotoxicity, induction of apoptosis, and changes in apoptosis-related genes of maximal physiological serum levels of the isoflavone genistein (Gen) in MCF-7 tumoral cells and in HB4a non-tumoral cells. In addition, induction of cell cycle arrest was also investigated. Only supraphysiological levels of Gen (50 and 100 µM) were cytotoxic to these cell lines. Concentrations of 10 and 25 µM did not induce apoptosis and significant changes in expression of the studied genes. Positive results were found only in cell cycle analysis: G0/G1 delay of MCF-7 cells in both concentrations of Gen and at 25 µM in HB4a cells. It is the first study investigating effects of Gen in the HB4a cell line. Thus, despite the lack of apoptosis induction (generally found with high concentrations), Gen at physiologically relevant serum levels still exerts chemopreventive effects through the modulation of cell cycle.

Lentinula edodes (shiitake) modulates chemically induced mutagenesis by enhancing pitting.

This study was undertaken to understand how Lentinula edodes modulates in vivo mutagenesis induced by alkylating agents in bone marrow and peripheral blood as described in our previous article. Male Swiss mice were pretreated for 15 consecutive days with aqueous extracts prepared from L. edodes, after which, the number of circulating blood cells, normal erythroid bone marrow cell cycling, and phagocytosis of micronucleated reticulocyte (MNRET) and activation of spleen macrophages were assessed. The results indicate that the antimutagenicity seen in bone marrow and peripheral blood is exerted by distinct compounds with different actions. The antimutagenic effect in bone marrow is exerted by compounds subject to degradation at deep-freeze storage temperature of -20°C. On the other hand, compounds responsible for antimutagenicity in peripheral blood are not subject to degradation at -20°C. The results also indicate that the antimutagenic action in peripheral blood leading to the reduction of circulating MNRET occurs in the spleen primarily through a phagocytic activity due to higher macrophage numbers and probably not due to the enhanced activation state of individual cells.

In vitro evaluation of the genotoxic activity and apoptosis induction of the extracts of roots and leaves from the medicinal plant Coccoloba mollis (Polygonaceae).

Coccoloba mollis (Family Polygonaceae) is a medicinal plant popularly used in cases of memory loss, stress, insomnia, anemia, impaired vision, and sexual impotence, but the scientific literature, to date, lacks studies on the biological effects of this species, particularly with regard to cytotoxicity and induction of DNA damage. The aim of the present study was to assess in vitro (in hepatic HTC cells) ethanolic extracts of the roots and leaves of C. mollis for cytotoxicity, genotoxicity, and induction of apoptosis. For these evaluations the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity assay, comet assay, micronucleus test with cytokinesis block, and an in situ test for detection of apoptotic cells with acridine orange staining were used. The results showed that the extract obtained from the roots of C. mollis is more cytotoxic than that obtained from the leaves and that the reduction in cell viability observed in the MTT assay was a result, at least in part, from the induction of apoptosis. Both extracts induced DNA damage at a concentration of 20 microg/mL in the comet assay, but no genotoxicity was detected with any of the treatments carried out in the micronucleus test.

Protective effects of beta-glucan extracted from barley against benzo[a]pyrene-induced DNA damage in hepatic cell HepG2.

The aim of the present study was to assess the genotoxic and antigenotoxic effect of beta-glucan (BG) extracted from barley. The genotoxicity of BG was tested in the single-cell gel electrophoresis assays (SCGE)/HepG2 test system. Moreover, the protective effects of BG against the genotoxicity of B[a]P were studied to delineate its mechanism of antigenotoxicity using four different treatment protocols - pre-treatment, simultaneous simple, simultaneous with pre-incubation, and post-treatment. The results showed that the compound itself was devoid of mutagenic activity at the three lower concentrations studied (1, 5, and 25microg/mL); however, genotoxic and cytotoxic effects were seen at 100 and 200microg/mL, respectively. In combination experiments with B[a]P, pronounced inhibition of DNA migration in the SCGE assay was observed in the two simultaneous treatments, and a smaller reduction was observed in the two other treatments. Thus, the data suggest that BG acts through binding to the genotoxic compound or capturing free radicals produced during its activation. However, the protective effects observed with pre-treatment and post-treatment suggest that the BG may be modulating cell metabolism.

beta-Glucans in promoting health: prevention against mutation and cancer.

The polysaccharides beta-glucans occur as a principal component of the cellular walls. Some microorganisms, such as yeast and mushrooms, and also cereals such as oats and barley, are of economic interest because they contain large amounts of beta-glucans. These substances stimulate the immune system, modulating humoral and cellular immunity, and thereby have beneficial effect in fighting infections (bacterial, viral, fungal and parasitic). beta-Glucans also exhibit hypocholesterolemic and anticoagulant properties. Recently, they have been demonstrated to be anti-cytotoxic, antimutagenic and anti-tumorogenic, making them promising candidate as pharmacological promoters of health.

DNA damage and aberrant crypt foci as putative biomarkers to evaluate the chemopreventive effect of annatto (Bixa orellana L.) in rat colon carcinogenesis.

Chemoprevention opens new perspectives in the prevention of cancer and other degenerative diseases. Use of target-organ biological models at the histological and genetic levels can markedly facilitate the identification of such potential chemopreventive agents. Colon cancer is one of the highest incidence rates throughout the world and some evidences have indicated carotenoids as possible agents that decrease the risk of colorectal cancer. In the present study, we evaluate the activity of annatto (Bixa orellana L.), a natural food colorant rich in carotenoid, on the formation of aberrant crypt foci (ACF) induced by dimethylhydrazine (DMH) in rat colon. Further, we investigate, the effect of annatto on DMH-induced DNA damage, by the comet assay. Male Wistar rats were given s.c. injections of DMH (40 mg/kg body wt.) twice a week for 2 weeks to induce ACF. They also received experimental diets containing annatto at 20, 200 or 1000 ppm for five 5 weeks before (pre-treatment), or 10 weeks after (post-treatment) DMH treatment. In both protocols the rats were sacrificed on week 15th. For the comet assay, the animals were fed with the same experimental diets for 2 weeks. Four hours before the sacrifice, the animals received an s.c. injection of DMH (40 mg/kg body wt.). Under such conditions, dietary administration of 1000 ppm annatto neither induce DNA damage in blood and colon cells nor aberrant crypt foci in rat distal colon. Conversely, annatto was successful in inhibiting the number of crypts/colon (animal), but not in the incidence of DMH-induced ACF, mainly when administered after DMH. However, no antigenotoxic effect was observed in colon cells. These findings suggest possible chemopreventive effects of annatto through their modulation of the cryptal cell proliferation but not at the initiation stage of colon carcinogenesis.

Toxicological and toxicogenetic effects of plants used in popular medicine and in cattle food

Toxicological and toxicogenetic effects of aqueous (tea) and hexanica fruit extract of Indigofera suffruticosa Mill, and hydroalcoholic root extract od Solanum agrarium Stendt. Were evaluated in Balb C male mice intraperitoneally exposed. A hepatotoxic effect was observed just for animals treated with aqueous fruit extract of I. suffruticosa. In relation to the toxicogenetic effect, just the group trreated with 12.5% of toxic dose of aqueous fruit extract of I. suffruticosa showed a statistically significant increase in the frequency of cells with chromosome aberrations (cytogenetic effect), although a slight increase was also observed for the highest dose (25% of LF50_ of hydroalcoholic root extract of S. agrarium. The results obtanied show that before S. agrarium is used as medicine and before the wide use of I. suffruticosa in cattle food, careful evaluation must be done.

Toxicological and toxicogenetics evaluation of plant used in cattle food and in popular medicine: indigofera suffruticosa mill

A planta Indigofera suffruticosa Mill é uma leguminosa que ocorre em pastagens. É utilizada como forrageira pelo gado e como planta medicinal por populaçöes carentes. Neste trabalho, foi avaliada a capacidade dos extratos e hexânico de folhas na induçäo de aberraçöes cromossômicas em células de medula óssea de camundongos. Grupos de animais foram tratados intraperitonealmente com 3 diferentes concentraçöes do extrato aquoso, 0,312, 0,625 e 1,250 g/Kg p.c., e 3 diferentes concentraçöes do extrato hexânico, 0,625, 1,250 e 2,50 g/Kg p.c.. Os animais foram sacrificados 24 horas após o tratamento. Näo se observou diferença significativa na freqüência de aberraçöes cromossômicas entre os grupos tratados com o extrato aquoso (3,0; 3,33; 5,0%) e o grupo controle negativo (4,6%). Para o extrato hexânico, um discreto aumento (5,7%) foi observado apenas para a menor concentraçäo, quando comparado com o controle negativo (4,8%). Outros estudos säo necessários, ainda, para avaliaçäo de um possível efeito genotóxico desta planta