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The cytotoxic and genotoxic effects of commercial endodontic sealers (AH Plus, Sealer 26 and Endomethasone N) incorporated with nanostructured silver vanadate decorated with silver nanoparticles (AgVO3 - at concentrations 2.5, 5, and 10%) on human gingival fibroblast (HGF), and the silver (Ag+ ) and vanadium (V4+ /V5+ ) ions release were evaluated. Cytotoxicity, cell death, and genotoxicity tests were carried out with extract samples of 24-hr and 7-days. The release of Ag+ and V4+ /V5+ was evaluated. Cytotoxicity in HGF was caused by AH Plus (AP) with 5 and 10% of AgVO3 (83.84 and 67.49% cell viability, respectively) with 24-hr extract (p

Alzheimer's disease (AD) is the primary dementia-causing disease worldwide, involving a multifactorial combination of environmental, genetic, and epigenetic factors, with essential participation of age and sex. Biochemically, AD is characterized by the presence of abnormal deposition of beta amyloid peptide (Aβ(1-42)), which in the brain is strongly correlated with oxidative stress, inflammation, DNA damage, and cholinergic impairment. The multiple mechanisms involved in its etiology create significant difficulty in producing an effective treatment. Neuroprotective properties of genistein and galantamine have been widely demonstrated through different mechanisms; however, it is unknown a possible synergistic neuroprotective effect against Aβ(1-42). In order to understand how genistein and galantamine combinations regulate the mechanisms of neuroprotection, we conducted a set of bioassays in vitro to evaluate cell viability, clonogenic survival, cell death, and anti-genotoxicity. Through molecular docking and therapeutic viability assays, we analyzed the inhibitory activity exerted by genistein on three major protein targets (AChE, BChE, and NMDA) involved in AD. The results showed that genistein and galantamine afforded significant protection at higher concentrations; however, combinations of sub-effective concentrations of both compounds provided marked neuroprotection when they were combined. In silico approaches showed that genistein has higher scores than the positive controls and low toxicity levels; nevertheless, the therapeutic viability indicated that unlike galantamine, genistein cannot undergo the action by P glycoprotein (PGP) and probably may be unable to cross the blood-brain barrier. In conclusion, our results show that genistein and galantamine exert neuroprotective by decreasing genotoxicity and cell death. In silico analysis, suggest that genistein modulates positively the expression of AChE, BChE, and NMDA. In this context, a combination of two or more drugs could inspire an attractive therapeutic strategy.

Made available in DSpace on 2020-12-12T02:06:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-01 Type 2 diabetes mellitus (T2DM), dyslipidemia and periodontitis are frequently associated pathologies; however, there are no studies showing the peripheral blood transcript profile of these combined diseases. Here we identified the differentially expressed genes (DEGs) of circulating lymphocytes and monocytes to reveal potential biomarkers that may be used as molecular targets for future diagnosis of each combination of these pathologies (compared to healthy patients) and give insights into the underlying molecular mechanisms of these diseases. Study participants (n = 150) were divided into groups: (H) systemically and periodontal healthy (control group); (P) with periodontitis, but systemically healthy; (DL-P) with dyslipidemia and periodontitis; (T2DMwell-DL-P) well-controlled type 2 diabetes mellitus with dyslipidemia and periodontitis; and (T2DMpoorly-DL-P) poorly-controlled type 2 diabetes mellitus with dyslipidemia and periodontitis. We preprocessed the microarray data using the Robust Multichip Average (RMA) strategy, followed by the RankProd method to identify candidates for DEGs. Furthermore, we performed functional enrichment analysis using Ingenuity Pathway Analysis and Gene Set Enrichment Analysis. DEGs were submitted to pairwise comparisons, and selected DEGs were validated by quantitative polymerase chain reaction. Validated DEGs verified from T2DMpoorly-DL-P versus H were: TGFB1I1, VNN1, HLADRB4 and CXCL8; T2DMwell-DL-P versus H: FN1, BPTF and PDE3B; DL-P versus H: DAB2, CD47 and HLADRB4; P versus H: IGHDL-P, ITGB2 and HLADRB4. In conclusion, we identified that circulating lymphocytes and monocytes of individuals simultaneously affected by T2DM, dyslipidemia and periodontitis, showed an altered molecular profile mainly associated to inflammatory response, immune cell trafficking, and infectious disease pathways. Altogether, these results shed light on novel potential targets for future diagnosis, monitoring or development of targeted therapies for patients sharing these conditions. Department of Diagnosis and Surgery School of Dentistry at Araraquara UNESP- São Paulo State University Department of Morphology Genetics Orthodontics and Pediatric Dentistry School of Dentistry at Araraquara UNESP- São Paulo State University Molecular Genomics and Therapeutics Section Genetics of Development and Disease Branch National Institute of Diabetes and Digestive and Kidney Diseases National Institutes of Health 10 Center Drive Building 10 Room 9D11 Department of Surgery Uniformed Services University of the Health Sciences and Henry Jackson Foundation for the Advancement of Military Medicine Postgraduate Program in Sciences of the University of Franca Department of Genetics Faculty of Medicine of Ribeirão Preto USP – University of São Paulo Department of Biology Faculty of Philosophy Sciences and Letters of Ribeirão Preto USP –University of São Paulo Department of Medical Genetics and Medicine Genomics University of Campinas – UNICAMP Department of Statistics Institute of Mathematics Statistics and Scientific Computing University of Campinas Department of Periodontology University of North Carolina at Chapel Hill – UNC School of Dentistry Advanced Research Center in Medicine Union of the Colleges of the Great Lakes (UNILAGO) Department of Diagnosis and Surgery School of Dentistry at Araraquara UNESP- São Paulo State University Department of Morphology Genetics Orthodontics and Pediatric Dentistry School of Dentistry at Araraquara UNESP- São Paulo State University

The Buccal Micronucleus Cytome Assay (BMCyt) has become an important biomonitoring tool for assessing cytogenetic damage in many studied populations. Each laboratory applies protocols that vary according to the method of collecting and preparing samples. Besides, Brazil is a country of great territorial extensions that received immigrants from various parts of the world with different genetic backgrounds. Therefore, the present study aimed to evaluate the inter-laboratory variation in scoring the same set of slides using the more comprehensive scoring criteria, to standardize the BMCyt protocol, to observe the basal alterations in populations of different Brazilian regions and to compare it with other places around the world. Our results showed that a valuable number of laboratories participated, ten laboratories from different regions of the country, for the validation of the BMCyt in human biomonitoring studies, resulting in the 804 healthy individuals. This was possible because we observed: a range of measures needs to be considered, such as the baseline frequency of DNA damage and cell death in non-exposed individuals; age when grouped showed an influence on DNA damage, although when evaluated by group we did not see an influence; association between smoking habit and all endpoints of the BMCyt (except karyolytic cells) was evident; the basal MN frequency, in the majority of groups, follows those around the world; and the BMCyt was confirmed as a good health status biomarker. We emphasize the need for constant discussions on the parameters of cell death due to greater difficulty among the analyzers.

Alzheimer's disease (AD) is the primary dementia-causing disease worldwide, involving a multifactorial combination of environmental, genetic, and epigenetic factors, with essential participation of age and sex. Biochemically, AD is characterized by the presence of abnormal deposition of beta amyloid peptide (Aβ

The Asociación Latinoamericana de Mutagénesis, Carcinogénesis y Teratogénesis Ambiental (ALAMCTA) is the organizational structure encompassing the five national environmental mutagenesis societies of Latin America. It was founded in 1980 and has held 10 congresses and had 10 presidents, representing members from throughout Latin America. This brief review describes the founding of ALAMCTA and the key events associated with it, including the initiation in 1993 of the influential Alexander Hollaender Courses in Mexico City, and the hosting of the 11th International Conference on Environmental Mutagens (ICEM) in 2013 in Foz do Iguaçu, Brazil. The ALAMCTA has proven to be a central organizing structure for scientists throughout the Latin America, aiming to meet, collaborate, exchange ideas, and promote the science of genetic toxicology and environmental mutagenesis. It has served to integrate scientists from diverse cultures and two language groups on a vast continent to know each other and to work towards common goals of improving public health, supporting basic research, and identifying and trying to solve environmental problems. Given its long history of 37 years and solid foundation due to the dedicated efforts of so many scientists from throughout the region, ALAMCTA is poised to play a critical role in Latin American science long into the future.

68 p.-17 fig.-1 tab. Chierrito, Talita P. C. et al., The lack of an effective treatment for Alzheimer' disease (AD), an increasing prevalence and severe neurodegenerative pathology boost medicinal chemists to look for new drugs. Currently, only acethylcholinesterase (AChE) inhibitors and glutamate antagonist have been approved to the palliative treatment of AD. Although they have a short-term symptomatic benefits, their clinical use have revealed important non-cholinergic functions for AChE such its chaperone role in beta-amyloid toxicity. We propose here the design, synthesis and evaluation of non-toxic dual binding site AChEIs by hybridization of indanone and quinoline heterocyclic scaffolds. Unexpectely, we have found a potent allosteric modulator of AChE able to target cholinergic and non-cholinergic functions by fixing a specific AChE conformation, confirmed by STD-NMR and molecular modeling studies. Furthermore the promising biological data obtained on human neuroblastoma SH-SY5Y cell assays for the new allosteric hybrid 14, led us to propose it as a valuable pharmacological tool for the study of non-cholinergic functions of AChE, and as a new important lead for novel disease modifying agents against AD., Financial support of this research provided by the Fundação de Pesquisa e Amparo do Estado de São Paulo-FAPESP (Proc. 12/04054-5; 12/14114-5; 13/50788-3; 14-10414-0); Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), CSIC (grant no. i-LINK-0801) and MINECO (grants no. SAF2016-76693-R to A.M., and CTQ2015-64597-C2-1 to J.J.B. and F.J.C.) is acknowledged

Made available in DSpace on 2019-10-06T16:41:10Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-07-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Inflammatory diseases, as periodontal disease (PD), has been associated with disturbance of lipid and glycemic metabolisms, as demonstrated by the increasing of PD patients with type 2 diabetes mellitus (T2D) and/or dyslipidemia comorbidities. We aimed to investigate the expression of inflammation and lipid metabolism genes, and correlations among clinical and biochemical characteristics in normoglycemic or T2D patients with dyslipidemia and PD, in comparison with healthy individuals. Five groups of 30 individuals each (150 patients) were formed based upon T2D, dyslipidemic and periodontal status. Blood analyses of lipid and glycemic profiles were carried out, and the gene expression was assessed by RT-qPCR. The systemic expression of IL6, TNFA and LEP genes were significantly higher in T2D, dyslipidemia and PD patients, while the PECAM1 gene showed the opposite. Higher RETN levels were found in patients with T2D independently of their glycemic control status. There were positive correlations between: TNFA, LEP and RETN with worse periodontal parameters; IL6, TNFA, ADIPOR1, LEP and RETN with waist-to-hip ratio; glycemic parameters with RETN; total cholesterol and triglycerides with LEP expression. We conclude that pro-inflammatory cytokines were related with worse lipid, glycemic and periodontal parameters, reinforcing that a hyper-inflammatory status connects systemic and oral inflammatory diseases. Department of Morphology School of Dentistry at Araraquara UNESP - São Paulo State University Department of Diagnosis and Surgery School of Dentistry at Araraquara UNESP - São Paulo State University Postgraduate Program in Sciences of the University of Franca Department of Genetics Faculty of Medicine of Ribeirão Preto and Department of Biology FFCLRP USP - University of São Paulo, Ribeirão Preto Department of Morphology School of Dentistry at Araraquara UNESP - São Paulo State University Department of Diagnosis and Surgery School of Dentistry at Araraquara UNESP - São Paulo State University FAPESP: 2007/08362-8 FAPESP: 2009/16233-9 FAPESP: 2010/10882-2 FAPESP: 2014/19699-7

Obesity is a major public health concern nowadays, as the risk of developing cancer and other chronic diseases has increased considerably in this population. Using in vitro tests (Comet Assay and Micronucleus Test), we evaluated whether lymphocytes from obese individuals (BMI: 40-62 kg / m2) are sensitive to genotoxic and cytotoxic damage when exposed to anorexigenics sibutramine, femproporex and phytotherapics Cordia ecalyculata and Echinodorus grandiflorus. We found that both possessed weak genotoxic activity and did not detect cytotoxic activity or interference in the cell division mechanisms of lymphocytes among obese individuals. La obesidad es una de las principales preocupaciones de salud pública en la actualidad, ya que el riesgo de desarrollar cáncer y otras enfermedades crónicas ha aumentado considerablemente en esta población. Mediante pruebas in vitro (Comet Assay y Micronucleus Test), evaluamos si los linfocitos de individuos obesos (IMC: 40-62 kg/m2) son sensibles al daño genotóxico y citotóxico cuando se exponen a los anoréxicos sibutramina, fenproporex y los fitoterapéuticos Cordia ecalyculata y Echinodorus grandiflorus. Encontramos que ambos tienen una actividad genotóxica débil y no detectamos actividad citotóxica o interferencia con los mecanismos de división celular de los linfocitos de individuos obesos. L'obesità è oggi uno dei principali problemi di salute pubblica, poiché il rischio di sviluppare il cancro e altre malattie croniche è aumentato considerevolmente in questa popolazione. Utilizzando test in vitro (Comet Assay e Micronucleus Test), abbiamo valutato se i linfociti di soggetti obesi (BMI: 40-62 kg/m2) sono sensibili al danno genotossico e citotossico quando esposti alla sibutramina anoressica, al fenproporex e ai fitoterapici Cordia ecalyculata e Echinodorus grandiflorus. Abbiamo scoperto che entrambi hanno una debole attività genotossica e non abbiamo rilevato attività citotossica o interferenza con i meccanismi di divisione cellulare dei linfociti di individui obesi. A obesidade é uma das principais preocupações de saúde pública atualmente, pois o risco de desenvolver câncer e outras doenças crônicas aumentou consideravelmente nessa população. Utilizando testes in vitro (Ensaio Cometa e Teste de Micronúcleo), avaliamos se linfócitos de indiví­duos obesos (IMC: 40-62 kg / m2) são sensí­veis a danos genotóxicos e citotóxicos quando expostos aos anorexígenos sibutramina, femproporex e fitoterápicos Cordia ecalyculata e Echinodorus grandiflorus. Descobrimos que ambos possuem fraca atividade genotóxica e não detectamos atividade citotóxica ou interferência nos mecanismos de divisão celular de linfócitos de indiví­duos obesos. A obesidade é uma das principais preocupações de saúde pública atualmente, pois o risco de desenvolver câncer e outras doenças crônicas aumentou consideravelmente nessa população. Utilizando testes in vitro (Ensaio Cometa e Teste de Micronúcleo), avaliamos se linfócitos de indiví­duos obesos (IMC: 40-62 kg / m2) são sensí­veis a danos genotóxicos e citotóxicos quando expostos aos anorexígenos sibutramina, femproporex e fitoterápicos Cordia ecalyculata e Echinodorus grandiflorus. Descobrimos que ambos possuem fraca atividade genotóxica e não detectamos atividade citotóxica ou interferência nos mecanismos de divisão celular de linfócitos de indiví­duos obesos.

Made available in DSpace on 2018-12-11T17:14:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 A high percentage of type 2 diabetes mellitus (T2D) patients are also affected by dyslipidemia and chronic periodontitis (CP), but no studies have determined the gene expression in patients that are simultaneously affected by all three diseases. We investigated the systemic expression of immune-related genes in T2D, dyslipidemia, and CP patients. One hundred and fifty patients were separated into five groups containing 30 individuals each: (G1) poorly controlled T2D with dyslipidemia and CP; (G2) well-controlled T2D with dyslipidemia and CP; (G3) normoglycemic individuals with dyslipidemia and CP; (G4) healthy individuals with CP; (G5) systemic and periodontally healthy individuals. Blood analyses of lipid and glycemic profiles were carried out. The expression of genes, including IL10, JAK1, STAT3, SOCS3, IP10, ICAM1, IFNA, IFNG, STAT1, and IRF1, was investigated by RT-qPCR. Patients with dyslipidemia demonstrated statistically higher expression of the IL10 and IFNA genes, while IFNG, IP10, IRF1, JAK1, and STAT3 were lower in comparison with nondyslipidemic patients. Anti-inflammatory genes, such as IL10, positively correlated with parameters of glucose, lipid, and periodontal profiles, while proinflammatory genes, such as IFNG, were negatively correlated with these parameters. We conclude that dyslipidemia appears to be the primary disease that is associated with gene expression of immune-related genes, while parameters of T2D and CP were correlated with the expression of these important immune genes. Department of Morphology School of Dentistry at Araraquara São Paulo State University (UNESP) Department of Diagnosis and Surgery School of Dentistry at Araraquara São Paulo State University (UNESP) Postgraduate Program in Sciences of the University of Franca Department of Genetics Faculty of Medicine of Ribeirão Preto University of São Paulo (USP) Department of Biology Faculty of Philosophy Sciences and Letters at Ribeirão Preto University of São Paulo (USP) Department of Morphology School of Dentistry at Araraquara São Paulo State University (UNESP) Department of Diagnosis and Surgery School of Dentistry at Araraquara São Paulo State University (UNESP)

Austdiol is a mycotoxin mainly produced by Aspergillus ustus and Mycoleptodiscus indicus. These fungi are found in rye, oats, barley, corn and feed grains; thus, as a potential contaminant of human food and animal feed, this mycotoxin is of great concern. As such, the elucidation of the cytotoxicity and mutagenicity of austdiol is important. In this study, austdiol was purified from a rice-oat solid medium culture of M. indicus using chromatographic separation techniques. Chinese hamster ovary (CHO-K1) cells were then used to study the effect of austdiol on mammalian cell cycle, clonogenicity and DNA damage. Austdiol induced cell cycle arrest in G2/M phase, with a decreased S phase population and increased sub-G1 population. Austdiol also increased the polyploid population. These events resulted in cell death detected 7 days after treatment by clonogenic assay. DNA damage represents the main mechanism of action of austdiol, which induces DNA breaks and increases the frequency of micronuclei and nucleoplasmic bridges in binucleated cells in a CHO-K1 cell line. Moreover, cells exposed to austdiol and doxorubicin (DXR) combined treatments presented a reduced number of colonies and increased frequencies of micronuclei and nucleoplasmic bridges compared with negative control and cells treated with austdiol or DXR alone.

This study aimed to evaluate the genotoxic effects in the oral epithelial cells of patients undergoing fixed orthodontic treatment and to compare these to a control group without treatment. The null hypothesis to be tested is that corrective orthodontic treatment at different periods does not cause genotoxic effects in patients. An observational cross-sectional study including 74 patients enrolled in corrective orthodontic treatment and 21 control patients, between 11 and 35 years of age, of both genders, participated in the research. Patients undergoing treatment were divided into four treatment groups differentiated by treatment periods: G1, n = 21 (1 month to 12 months); G2, n = 21 (13 to 24 months); G3, n = 23 (25 to 48 months); and G4, n = 9 (over 48 months). Cells were collected by scraping the internal side of the cheek and subsequently placed in tubes containing 0.9% sodium chloride solution. The sample underwent evaluation for genotoxic effects by means of the micronucleus test (MNT). Bivariate analyses were performed using parametric tests (t test or ANOVA) and nonparametric tests (Chi-square test, Kruskal-Wallis test, Dunn post-test). The adopted level of significance was 5%. Statistically significant differences for any of the genotoxic abnormalities (binucleated, trinucleated, karyolysis, piknosis, nuclear buds) were not found except for karyolysis, which was higher in the control group than in G4 (p

Objectives: To evaluate the genotoxic and cytotoxic effects of Haas appliances through micronuclei test and cytogenetic damage analysis in buccal mucosa epithelial cells of patients undergoing orthodontic treatment. Materials and Methods: Twenty-eight patients, 6–12 years of age and of both genders, who required a Haas appliance for the correction of a posterior crossbite were included. Epithelial cells from the mucosa were collected by gently scraping the inside of both the right and left cheeks. The cells were collected before the insertion of the appliance (T0), 1 month after the device was installed (T1), and again 3 months after the appliance was immobilized (T2). The cells were processed to obtain slides. Feulgen/Fast Green was used as the staining method, and the number of normal, karyolytic, pyknotic, nuclear buds, bi/trinucleated, and micronucleus cells were counted under light microscopy. Cellular abnormalities were evaluated with parametric and nonparametric tests for comparison of the means by analysis of variance testing, Tukey posttest, or the Kruskal-Wallis test and then by Dunn's posttest. The significance level was 5%. Results: There were no statistically significant changes in the micronuclei in the evaluated periods (P > .05). Nuclear buds increased at T1 (P < .05), returning to baseline levels at T2. Other abnormalities (cariolytic, pyknotic, and bi/trinucleated cells) showed a significant increase at T1 and T2 (P < .0001). Conclusions: The Haas appliance did not cause an increase in micronuclei in cells of the buccal mucosa. However, statistically significant increases in cariolytic, pyknotic, and bi/trinucleated cells were observed during treatment, suggesting possible DNA damage.

Background Mammaglobin A (MGA), mainly expressed in the breast epithelium, is overexpressed in breast cancer, and has been established as a tumor and promissory marker for the early detection of metastasis. Aim The main aim of this study was to evaluate the association between the presence of the MGA transcript in the peripheral blood of Brazilian breast cancer patients and healthy women and the development of breast cancer and tumor progression. Material and methods The expression of the MGA transcript in peripheral blood of 102 breast cancer patients and 102 healthy women was assessed by RT-PCR. Results MGA mRNA was expressed in the peripheral blood of 39 breast cancer patients and in none of the women from the control group. The presence of MGA was significantly associated with presence of metastasis and age at onset after 60 years. The presence of MGA mRNA in peripheral blood displayed a sensitivity of 38.2%, specificity of 100.0%, positive predictive value (PPV) of 100.0%, and negative predictive value (NPV) of 61.8% as a breast cancer marker. Conclusion This study provides additional evidence of the presence of MGA in the peripheral blood of breast cancer patients, and its applicability as an efficient biomarker for breast cancer (High specificity and PPV). To our knowledge, this is the first study to assess the expression of MGA mRNA in peripheral blood obtained from the Brazilian population.

Made available in DSpace on 2018-12-11T17:32:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-10-01 Despite advances in the field of biomaterials for bone repair/regeneration, some challenges for developing an ideal bone substitute need to be overcome. Herein, this study synthesized and evaluated in vitro a nanocomposite based on bacterial cellulose (BC), collagen (COL), apatite (Ap) and osteogenic growth peptide (OGP) or its C-terminal pentapeptide [OGP(10-14)] for bone regeneration purposes. The BC-COL nanocomposites were successfully obtained by carbodiimide-mediated coupling as demonstrated by spectroscopy analysis. SEM, FTIR and 31P NMR analyses revealed that in situ synthesis to apatite was an effective route for obtaining of bone-like apatite. The OGP-containing (BC-COL)-Ap stimulated the early development of the osteoblastic phenotype. Additionally, the association among collagen, apatite, and OGP peptides enhanced cell growth compared with OGP-containing BC-Ap. Furthermore, none of the nanocomposites showed cytotoxic, genotoxic or mutagenic effects. These promising results suggest that the (BC-COL)-Ap associated with OGP peptides might be considered a potential candidate for bone tissue engineering applications. São Paulo State University UNESP Institute of Chemistry Cell Culture Laboratory Faculty of Dentistry of Ribeirão Preto University of São Paulo USP São Paulo State University UNESP School of Dentistry at Araraquara Department of Genetics Faculty of Medicine of Ribeirão Preto University of São Paulo USP São Paulo State University UNESP Institute of Chemistry São Paulo State University UNESP School of Dentistry at Araraquara

The over-production of reactive oxygen species (ROS) can cause oxidative damage to a large number of molecules, including DNA, and has been associated with the pathogenesis of several disorders, such as diabetes mellitus (DM), dyslipidemia and periodontitis (PD). We hypothesise that the presence of these diseases could proportionally increase the DNA damage. The aim of this study was to assess the micronucleus frequency (MNF), as a biomarker for DNA damage, in individuals with type 2 DM, dyslipidemia and PD. One hundred and fifty patients were divided into five groups based upon diabetic, dyslipidemic and periodontal status (Group 1 - poor controlled DM with dyslipidemia and PD; Group 2 - well-controlled DM with dyslipidemia and PD; Group 3 - without DM with dyslipidemia and PD; Group 4 - without DM, without dyslipidemia and with PD; and Group 5 - without DM, dyslipidemia and PD). Blood analyses were carried out for fasting plasma glucose, HbA1c and lipid profile. Periodontal examinations were performed, and venous blood was collected and processed for micronucleus (MN) assay. The frequency of micronuclei was evaluated by cell culture cytokinesis-block MN assay. The general characteristics of each group were described by the mean and standard deviation and the data were submitted to the Mann-Whitney, Kruskal-Wallis, Multiple Logistic Regression and Spearman tests. The Groups 1, 2 and 3 were similarly dyslipidemic presenting increased levels of total cholesterol, low density lipoprotein cholesterol and triglycerides. Periodontal tissue destruction and local inflammation were significantly more severe in diabetics, particularly in Group 1. Frequency of bi-nucleated cells with MN and MNF, as well as nucleoplasmic bridges, were significantly higher for poor controlled diabetics with dyslipidemia and PD in comparison with those systemically healthy, even after adjusting for age, and considering Bonferroni's correction. Elevated frequency of micronuclei was found in patients affected by type 2 diabetes, dyslipidemia and PD. This result suggests that these three pathologies occurring simultaneously promote an additional role to produce DNA impairment. In addition, the micronuclei assay was useful as a biomarker for DNA damage in individuals with chronic degenerative diseases.

The mushroom Agaricus brasiliensis (sun mushroom), native from the southeast of Brazil, is well known by its medicinal properties that include effects on diabetes, cholesterol levels, and osteoporosis. The antimutagenic effects of A. brasiliensis has been investigated recently and revealed some controversial results depending on the temperature by which the A. brasiliensis tea is obtained. In the present study, we evaluated the effect of the A. brasiliensis extract prepared in two different temperatures, 4°C and 25°C, on the doxorubicin-induced DNA strand breaks and chromosomal aberrations (CAs) in human lymphocytes. The results demonstrated that A. brasiliensis was able to reduce the DXR-induced DNA damage in both temperatures; however, the CA test was more sensitive to demonstrate a better reduction when the cells were treated with an extract obtained at 25°C. A. brasiliensis extract obtained in different temperatures exhibited antigenotoxic and anticlastogenic effects in human lymphocytes.

Eugenia dysenterica ex DC Mart. (Myrtaceae) is a Brazilian tree with pharmacological and biological properties. The aqueous leaf extract, rich in polyphenols, was tested in the human neuroblastoma cell line SH-SY5Y to evaluate its effect on cell viability. The extract and two isolated compounds were also assessed for the potential inhibitory activity on acetylcholinesterase, an enzyme related to Alzheimer's disease. A simple chromatographic method using Sephadex LH-20 was developed to separate catechin and quercetin from the aqueous leaf extract of E. dysenterica. Identification was carried out by spectroscopic techniques IR, UV, and 1H and 13C NMR. The IC50 values were obtained by constructing dose-response curves on a graph with percentage inhibition versus log of inhibitor concentration and compared with physostigmine, a well-known AChE inhibitor. The extract was toxic for SH-SY5Y cells at concentrations higher than 7.8 μg/ml given for 24 h. The decline in SH-SY5Y cell viability appears to be related to its antiproliferative activity. The extract also showed relatively moderate acetylcholinesterase inhibitory activity of 66.33% ± 0.52% at 1.0 mg/ml with an IC50 value of 155.20 ± 2.09 μg/ml. Physostigmine, quercetin, and catechin showed IC50 values of 18.69 ± 0.07, 46.59 ± 0.49, and 42.39 ± 0.67 μg/ml, respectively.

Made available in DSpace on 2018-12-11T17:33:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Despite increasing research in type 2 diabetes mellitus (T2D), there are few studies showing the impact of the poor glycemic control on biological processes occurring in T2D. In order to identify potential genes related to poorly/well-controlled patients with T2D, our strategy of investigation included a primary screen by microarray (Human Genome U133) in a small group of individuals followed by an independent validation in a greater group using RT-qPCR. Ninety patients were divided as follows: poorly controlled T2D (G1), well-controlled T2D (G2), and normoglycemic individuals (G3). After using affy package in R, differentially expressed genes (DEGs) were prospected as candidate genes potentially relevant for the glycemic control in T2D patients. After validation by RT-qPCR, the obtained DEGs were as follows - G1 + G2 versus G3: HLA-DQA1, SOS1, and BRCA2; G2 versus G1: ENO2, VAMP2, CCND3, CEBPD, LGALS12, AGBL5, MAP2K5, and PPAP2B; G2 versus G3: HLA-DQB1, MCM4, and SEC13; and G1 versus G3: PPIC. This demonstrated a systemic exacerbation of the gene expression related to immune response in T2D patients. Moreover, genes related to lipid metabolisms and DNA replication/repair were influenced by the glycemic control. In conclusion, this study pointed out candidate genes potentially associated with adequate glycemic control in T2D patients, contributing to the knowledge of how the glycemic control could systemically influence gene expression. Department of Diagnosis and Surgery School of Dentistry Universidade Estadual Paulista (UNESP) Department of Morphology School of Dentistry Universidade Estadual Paulista (UNESP) Postgraduate Program in Sciences of the University of Franca Department of Genetics Faculty of Medicine of Ribeirão Preto and Department of Biology FFCLRP University of São Paulo (USP) Department of Medical Genetics University of Campinas (UNICAMP) Department of Diagnosis and Surgery School of Dentistry Universidade Estadual Paulista (UNESP) Department of Morphology School of Dentistry Universidade Estadual Paulista (UNESP)

Alzheimer's disease (AD) is a progressive condition, where dementia symptoms gradually worsen. Biochemically the disease is characterized by the presence of neuritic plaques, neurofibrillary tangles, in addition to cholinergic dysfunction in the central nervous system. The role of the cholinergic neurotransmission in AD is the basis of the widely accepted cholinergic hypothesis. Some of the most relevant therapies for the treatment of the disease are based on the acetylcholinesterase (AChE) inhibitor activity; however, these therapies are not effective to stop the disease progression, but only can temporarily slow down the worsening of dementia symptoms, and improve quality of life of patients and their caregivers. In recent years, plant alkaloids extracted from Amaryllidaceae family have received great attention due to the well-known anti cholinergic activity. In this context, the purpose of this study was to apply the docking molecular in silico analysis aiming to examine the recombinant human AChE enzyme (rhAChE) inhibitory activity displayed by different alkaloids from Amaryllidaceae family. Overall, the present results support the idea that alkaloids reported in this research are capable of interacting with rhAChE-binding sites.

The aim of this study was to characterize the physicochemical properties of bacterial cellulose (BC) membranes functionalized with osteogenic growth peptide (OGP) and its C-terminal pentapeptide OGP[10-14], and to evaluate in vitro osteoinductive potential in early osteogenesis, besides, to evaluate cytotoxic, genotoxic and/or mutagenic effects. Peptide incorporation into the BC membranes did not change the morphology of BC nanofibers and BC crystallinity pattern. The characterization was complemented by Raman scattering, swelling ratio and mechanical tests. In vitro assays demonstrated no cytotoxic, genotoxic or mutagenic effects for any of the studied BC membranes. Culture with osteogenic cells revealed no difference in cell morphology among all the membranes tested. Cell viability/proliferation, total protein content, alkaline phosphatase activity and mineralization assays indicated that BC-OGP membranes enabled the highest development of the osteoblastic phenotype in vitro. In conclusion, the negative results of cytotoxicity, genotoxicity and mutagenicity indicated that all the membranes can be employed for medical supplies, mainly in bone tissue engineering/regeneration, due to their osteoinductive properties.

ABH and Lewis antigen expression has been associated with cancer development and prognosis, tumor differentiation, and metastasis. Considering that invasive ductal breast carcinoma (IDC) presents multiple molecular alterations, the aim of the present study was to determine whether the polymorphism of ABO, Lewis, and Secretor genes, as well as ABO phenotyping, could be associated with tumor differentiation and lymph nodes metastasis. Seventy-six women with IDC and 78 healthy female blood donors were submitted to ABO phenotyping/genotyping and Lewis and Secretor genotyping. Phenotyping was performed by hemagglutination and genotyping by the polymerase chain reaction with sequence-specific primers. ABO, Lewis, and Secretor genes were classified by individual single nucleotide polymorphism at sites 59, 1067, 202, and 314 of the Lewis gene, 428 of the Secretor gene, and 261 (O1 allele), 526 (O2 and B allele), and 703 (B allele). No association was found between breast cancer and ABO antigen expression (P = 0.9323) or genotype (P = 0.9356). Lewis-negative genotype was associated with IDC (P = 0.0126) but not with anatomoclinical parameters. Nonsecretor genotype was associated with axillary lymph node metastasis (P = 0.0149). In conclusion, Lewis and Secretor genotyping could be useful to predict respectively breast cancer susceptibility and axillary lymph nodes metastasis.

Cordia ecalyculata Vell. and Echinodorus grandiflorus (Cham.Schltdl.) Micheli are extensively used in Brazil as therapeutic preparations for indigenous groups and the general population. These plants have been used in the folk medicine as: tonic, diuretic, anti-inflammatory, appetite suppressants, for the treatment of snake bites, and weight loss.In this study, it was verified the possible cytotoxic and genotoxic effects of the crude extracts of. Cordia ecalyculata and Echinodorus grandiflorus, as well as their effectiveness in treating obesity.The Micronucleus Test was used for the evaluation of possible clastogenic and aneugenic effects, and the Comet Assay was used for the evaluation of single-strand and double-strand DNA breaks. The cytotoxic effects of the crude extracts were verified by PCE/NCE ratio. Swiss mice (Mus musculus) were used as the experimental model.It was observed a significant (P0.05) increase, dose-independent, in the average frequency of micronucleated erythrocytes in peripheral blood in mice treated with either the Cordia ecalyculata or Echinodorus grandiflorus extracts, in comparison with the negative control. There were no significant differences (P0.05) in the frequency of micronucleated polychromatic erythrocytes for both extract treatment. We observed that treatment with the Cordia ecalyculata extract at concentrations of 1000 and 2000 mg/kg bw resulted in a PCE/NCE ratio that was larger (P0.05) than the negative control. After 15 days of daily treatment, a dose of 2000 mg/kg bw of either phytotherapeutic did not reduce body mass gain or the amount of food consumed by Swiss mice when compared with the negative control (P0.05).The results of this study allowed us to infer that the crude extracts of Cordia ecalyculata and Echinodorus grandiflorus do not display cytotoxic or genotoxic activities. However, they do possess weak clastogenic activity (without significance) on peripheral blood cells. Contrary to commonly held beliefs it was also found in this study that the extracts are not effective for obesity treatments.

A very appropriate method for antigenotoxicity evaluation of antioxidants is the comet assay, since this analytical method detects initial DNA lesions that are still subject to repair; in other words, lesions that are very associated to damages resulting from the generation and subsequent action of reactive species. However, a solid evaluation should be developed in order to avoid inexact interpretations. In our study, besides the association of curcumin with cisplatin, curcumin and cisplatin agents were also tested separately. Classical genotoxic compounds, when tested by the comet assay, present an increase in the nucleoid tail; however, the cisplatin treatment has resulted in a decrease of DNA migration. This was an expected effect, as the cross-links between cisplatin and DNA decrease the DNA electrophoretic mobility. A similar effect was observed with the curcumin treatment, which decreased the nucleoid tail. Such effect was not expected and reinforced the necessity of including in the study, separate treatment groups with potentially antigenotoxic substances. The comet assay results have been analyzed using specific software for image analysis, as well as the classical visual analysis, and we have observed that the effect of decrease in DNA electrophoretic mobility was more easily observed when the data were analyzed by the software.

Vitamin B(6) has shown to be a potentially effective antioxidant agent, and dietary antioxidants are also frequently valuable inhibitors of clastogenesis and carcinogenesis. The purpose of the present work was to study the clastogenicity of different doses of vitamin B(6) and to examine the possible modulating effect of this vitamin on chromosomal damage induced by the antitumor agent doxorubicin in Wistar rats. Experimental groups were set up for pre- and simultaneous treatment with vitamin B(6) alone or in combination with DXR. The data obtained from administering different doses of vitamin B(6) (12.5-100 mg/kg b.w.) showed no significant increase in total chromosomal aberrations when compared with the negative control. The administration of two doses of 25 mg/kg b.w. or one dose of 50 mg/kg b.w. of vitamin B(6) before doxorubicin injection seemed equally effective in protecting cells against doxorubicin clastogenicity. The anticlastogenic effect of vitamin B(6) on DXR-induced chromosomal damage could be ascribed to its antioxidant properties. Vitamin B(6) was not clastogenic or cytotoxic in rat bone marrow cells and it plays a role in inhibiting the clastogenicity induced by DXR.

Physalis angulata L (Solanaceae) is a medicinal plant from North of Brazil, whose different extracts and infusions are commonly used in the popular medicine for the treatment of malaria, asthma, hepatitis, dermatitis and rheumatism. However, the genotoxic effects of P. angulata on human cells is not well known. The main purpose of the present study was to evaluate the in vitro genotoxic effects of aqueous extract of P. angulata using the comet assay and the micronucleus assay in human lymphocytes provided from 6 healthy donors. Treatments with P. angulata extracts were performed in vitro in order to access the extent of DNA damage. The comet assay has shown that treatments with P. angulata at 0.5, 1.0, 2.0, 3.0 and 6.0 microg/mL in culture medium were genotoxic. Lymphocytes treated with P. angulata at the concentrations of 3.0 and 6.0 microg/mL in culture medium showed a statistically significant increase in the frequency of micronucleus (p

Nanoparticles (NPs) have been used in a range of products due to their unique properties. Nevertheless, these NPs can cause adverse biological effects and because of that, there is a great concern about the health and environmental risks related to their use. Recently, silver nanoparticles (Ag NPs) have been used in a variety of cytotoxicity and genotoxicity studies, but there are still controversies regarding the association between the size and the toxicity of these particles. Therefore, in this study, we aimed to evaluate the cytotoxicity and genotoxicity of Ag NPs (10 and 100 nm) in two different cell lines, CHO-K1 and CHO-XRS5, by performing cell viability assay (XTT), clonogenic assay, micronucleus test, comet assay, as well as by investigating the cell cycle kinetics using the flow cytometry. Cell cultures were exposed to different concentrations of AgNPs (0.025-5.0 μg/ml) for 24 h. Our results indicated that cytotoxicity and genotoxicity induced by the 100 nm-Ag NPs were greater than those induced by the 10 nm-Ag NPs for both cell lines, which suggests that the exposure to greater size particles (100 nm) can cause more adverse biological effects than the exposure to the smaller ones (10 nm).

Biochemically, Alzheimeŕs disease (AD) is characterized by the presence of abnormal deposition of beta amyloid peptide (Aβ(1-42)), which is generated by proteolytic processing from its precursor, the amyloid precursor protein (APP) in a non-physiological pathway. The presence of Aβ(1-42) in the brain is strongly correlated with cognitive impairment, cholinergic deficiency, bioenergetics disruption, cell death and DNA damage. Galanthamine is an acetylcholinesterase inhibitor (AChEI) used to symptomatic treatment of Alzheimeŕs disease (AD). Several studies have showed that galanthamine has antioxidant properties, anti-apoptotic action and also promotes neurogenesis; however, it is unknown whether galanthamine may present protection mechanisms against Aβ(1-42)-induced genomic instability. To understand the mechanisms of this neuroprotection, we studied the effects of galanthamine on the cell toxicity and DNA strand breaks induced by Aβ(1-42) using a set of biomarkers such as clonogenic assay, cytokinesis block micronucleus cytome (CBNM-cyt) and comet assay. The results showed that galanthamine treatments were capable to significantly reduce the Aβ(1-42)-induced cytotoxicity and genotoxicity. In conclusion, this study demonstrated that in addition to inhibition of acetylcholinesterase (AChE), galanthamine exerts antigenotoxic properties. This relevant property of galanthamine is worthwhile exploring further which may improve the development of new diseases-modifying agents.

The importance of selenium in human nutrition is well recognized. It is an important micronutrient involved in antioxidant defense and is of fundamental importance in the maintenance of genomic stability. However, questions relating to the appropriate chemical form of selenium used for supplementation and its beneficial dose is still being debated. Therefore, the present study investigated the ability of 2 selenium compounds-sodium selenite (SS) and selenomethionine (SM)-to protect DNA against the damage induced by doxorubicin (DXR). Wistar rats were supplemented orally for 10 consecutive days with SS or SM (1 and 2 mg/kg bw); 24 hours before euthanasia a single dose of DXR (90 mg/kg bw) was injected intraperitoneally. DNA lesions were assessed by the comet assay and micronucleus test; the concentrations of thiobarbituric acid reactive substances, vitamin E, and reduced glutathione were determined by using liver homogenates. The results obtained showed that SS and SM prevented the induction of DNA damage by DXR. It was also observed that these 2 selenium compounds increased the hepatic concentration of glutathione, maintained the thiobarbituric acid reactive substances, and allowed for the maintenance of hepatic concentrations of vitamin E even after DXR treatment, confirming the antioxidant properties of selenium compounds. Sodium selenite and SM supplementation exhibited very similar patterns of protection. In conclusion, SS and SM supplementation was effective in protecting DNA against DXR-induced DNA damage in Wistar rats, probably because of their antioxidant properties.

Insufficient intakes of many micronutrients found in fruits and vegetables, such as folic acid, vitamins C and B6 may lead to DNA damage, cancer, and degenerative disease. The investigation of dietary antioxidants is a field of great interest for elucidating mechanisms of mutagenesis/carcinogenesis. The present study was undertaken to investigate the effects of vitamin B6 on the induction of chromosomal aberrations in cultured human lymphocytes and to examine the possible anticlastogenic effect of this vitamin on chromosomal damage induced by the antitumor drug doxorubicin. The results showed that when the cultures treated with vitamin B6 were compared with the untreated control in terms of total chromosomal damage and abnormal metaphases, pre- and simultaneous treatment with this vitamin showed no significant differences. In the post-treatment, average and above average concentrations of vitamin B6 alone showed a clastogenic effect. In the simultaneous protocol, this vitamin (15, 90 and 120 microg/mL) was effective in inhibiting chromosomal aberrations induced by doxorubicin (p0.05), with a reduction of 33.1% with the highest concentration tested. However, in the post-treatment, the associations of vitamin B6 and doxorubicin exerted a more evident clastogenic effect than that observed in the cultures exposed only to the antitumor drug. In the present investigation, the inability of vitamin B6 to decrease chromosomal damage induced by doxorubicin in the pre- and post-treatments could be justified by the instability of this vitamin as a free radical scavenger. In conclusion, the results from this study confirmed that vitamin B6 is protective against chromosomal damage induced by doxorubicin in cultured human lymphocytes, but that the effects depend on concentration and form of treatment.

Metallic nanoparticles such as silver (Ag), cerium dioxide (CeO2) and titanium dioxide (TiO2) are produced at a large scale and included in many consumer products. It is well known that most metallic NPs are toxic to humans which raise concerns about these engineered particles. Various studies have already been published on the subject, however, almost all of these studies have been conducted in cancer or transformed cell lines. In this work we performed a comparative evaluation of these metallic NPs on normal untransformed human fibroblasts (GM07492) detecting cyto- and geno-toxic responses after exposure to these NPs. Our results showed that all three metallic NPs were able to cross the plasma membrane and were mainly found in endocytic vesicles. The Ag and TiO2 NPs affected mitochondrial enzymatic activity (XTT), increased DNA fragmentation, oxidative damage (Comet assay) and induced cell death mainly by the apoptotic pathway. Ag NPs increased GADD45α transcript levels and the phosphorylation of proteins γH2AX. Transient genotoxicity was also observed from exposure to CeO2 NPs while TiO2 NPs showed no increase in DNA damage at sub-cytotoxic concentrations. In comparison, Ag NPs were found to be the most cyto-genotoxic NPs to fibroblasts. Thus, these results support the use of normal fibroblast as a more informative tool to detect the mechanisms of action induced by metallic NPs. ispartof: Toxicology in Vitro vol:29 issue:7 pages:1319-31 ispartof: location:England status: published

Excessive alcohol consumption may cause the development of pathologies in the liver and pancreas and various digestive tract cancers. The enzymes GSTM1, GSTT1, GSTP1, CYP1A1 and CYP2E1 are involved in the bioactivation and detoxification of a variety of xenobiotics present in food, organic solvents, tobacco smoke, drugs, pesticides, environmental pollutants and alcoholic drinks. Polymorphisms in the genes coding for these enzymes have been associated with susceptibility to different diseases, including ethanol-related diseases. To investigate whether these polymorphisms represent risk-modifying factors for ethanol-related diseases, a study was conducted involving 120 Brazilian alcoholics and 221 controls with similar ethnic backgrounds. The distribution of alcoholics groups was as follows: 65 with liver cirrhosis, 14 with chronic pancreatitis and 41 without cirrhosis or pancreatitis. The data revealed that carriers of the rare GSTP1 Val allele were at higher risk of liver cirrhosis and pancreatitis, since we found higher frequencies of the Val/Val genotype in alcoholics with liver cirrhosis (15.4%) and pancreatitis (28.6%) in comparison with alcoholics without disease (7.3%). No differences were found in the prevalences of the GSTM1 and GSTT1 null genotypes between alcoholics and the controls and no association was found between the rare CYP2E1 c2 allele and liver cirrhosis and pancreatitis. However, when the mutant CYP1A1 allele was compared between alcoholics and controls, the m2/m2 genotype was more prevalent in the liver cirrhosis alcoholics (7.7%) than in the controls (1.4%) and this difference was statistically significant (P = 0.03, OR = 5.33). In conclusion, our data indicate an association between occurrence of the Val/Val GSTP1 genotype and chronic pancreatitis and an association between the m2/m2 CYP1A1 genotype and alcoholic liver cirrhosis. This could indicate that persons with these genotypes are genetically more prone to the development of alcoholic pancreatitis and alcoholic cirrhosis, respectively.

Acute lymphoblastic leukemia (ALL) is the most common form of pediatric cancer. Although exposure to environmental agents appears to predispose individuals to this disease, little attention has been paid to the role of genetic susceptibility to environmental exposures in the etiology of childhood ALL. The enzymes GSTM1, GSTT1, GSTP1, CYP1A1, and CYP2E1 are involved in the bioactivation and detoxification of a variety of xenobiotics present in food, organic solvents, tobacco smoke, drugs, alcoholic drinks, pesticides, and environmental pollutants. Polymorphisms in the genes coding for these enzymes have been associated with increased susceptibility to different cancers, including hematologic malignancies. To investigate whether these polymorphisms represent risk-modifying factors for childhood ALL, a study was conducted involving 113 Brazilian patients of childhood ALL and 221 controls with similar ethnic backgrounds. The data revealed that carriers of the rare GSTP1 Val allele were at higher risk of ALL (odds ratio [OR] = 2.7; 95% confidence interval [CI] = 1.1–6.8; P = 0.04). No difference was found in the prevalence of the GSTM1 and GSTT1 null genotypes between ALL patients and the controls, and no association was found between CYP1A1*2 and CYP2E1*3 variants and ALL. However, when the mutant CYP1A1 and CYP2E1 alleles were considered together with the GSTM1 and GSTP1 risk-elevating genotypes, the risk of ALL was increased further (OR = 10.3; 95% CI = 1.0–111.8; P = 0.05), suggesting a combined effect. These results imply that genetic variants of xenobiotic metabolizing genes influence the risk of developing childhood ALL. Environ. Mol. Mutagen. 43:100–109, 2004. © 2004 Wiley-Liss, Inc.

Objective . To investigate whether formocresol, in Buckley's original formulation, is mutagenic in vivo to lymphocyte cultures obtained from the peripheral blood of children aged from 5 to 10 years old. These children were recruited from those attending the dental clinics of Recife City Council and the University of Pernambuco School of Dentistry, Brazil. Method . The sample comprised 20 children who had primary teeth with cariously exposed vital pulps. Two venous blood samples were collected (6–8 ml) from each child, the first prior to vital pulpotomy (control group) and the second 24 h after pulpotomy (treated group). This research is a case-control study. The peripheral lymphocytes were grown in a complete culture medium consisting of 78% RPMI 1640 medium (a), supplemented with streptomycin (0.01 mg/ml), penicillin (0.005 ml −1 ), 20% fetal bovine serum (b) and 2% phytohemagglutinin (c). The lymphocytes were assessed for chromosomal aberrations via a previously published method which was modified. The cytogenetic analysis was performed in a blind test, where the slides were codified by an annotator and the scorers did not know which group they were analyzing. For each sample, this envolved the analysis of 200 metaphases. The level of significance adopted in the statistical test was 5.0% ( p Results . There was no statistically significant difference in clinical doses between the control and treated groups, using Wilcoxon's Signed Ranks test, for the chromosomal aberrations ( P =0.251) and for the total chromosomal breaks ( P =0.149). Although there were no statistically significant differences between the control and treated groups, Buckley's formocresol was mutagenic for one patient, raising doubt about the desirability of its use for pulpotomies in children. Conclusion . The results revealed that, from a statistical standpoint, formocresol is not mutagenic. However, further investigations are required, preferably with a larger sample, in patients needing more than one pulpotomy in order to observe whether an increase in the quantity of the drug would increase the quantity of chromosome aberrations and also to verify individual susceptibility to chromosome alterations with the use of formocresol.

In Pimelodidae fish the available chromosome numbers range from 2n=46 to 2n=63, the value 2n=56 being the most frequent one. In the present paper 6 species belonging to the Rhamdiinae subfamily were analyzed: Rhamdia hilarii, R. quelen, Cetopsorhamdia sp., C. iheringhi, Imparfinis cf. piperatus and Imparfinis aff. schubarti. Chromosome preparations were obtained by direct and short term culture methods from kidney cells and analysed under standard Giemsa staining, C-banding and NOR silver staining. Rhamdia species present a basic karyotype composed by 58 chromosomes and the NORs are located on the short arm of a subtelocentric chromosome pair. Cetopsorhamdia sp., C. iheringhi and Imparfinis aff. schubarti. also shows 2n=58, while I. cf. piperatus is the only analyzed species presenting 2n=56. However, these 4 species show an interstitial NOR location. These chromosomal data agree with the occurrence of subgroups in the Rhamdiinae subfamily, proposed on the basis of their morphological traits.

Diethylpropion has been available in the market for treating obesity for over 50 years. Refined studies are lacking to fully elucidate its action spectrum. The aim of our study was to evaluate possible toxic effects of anorectic diethylpropion in Chinese hamster ovary (CHO) cells. Comet assay (detects breaks in the DNA strand), micronucleus test (detects clastogenic/aneugenic damage), and cell survival test (detects cytotoxic damage) were used to evaluate the toxic effects. In comet assay, we found that the damage scores with diethylpropion treatments at the concentrations of 20 and 40 μg/mL were more significant ( p < 0.05) than that of the negative control. When assessing the possible aneugenic and/or clastogenic damage caused by the drug in CHO cells, we found no difference ( p > 0.05) in the values of micronucleated cells when comparing different diethylpropion treatments and the negative control. Regarding the cell viability, for all the diethylpropion concentrations tested, higher values ( p < 0.05) of apoptosis were found compared with those of the negative control. In relation to the number of necrotic cells, no difference ( p > 0.05) was noted between the means of the three concentrations of diethylpropion evaluated and the negative control. In the experimental conditions, we conclude that diethylpropion has weak genotoxic and cytotoxic activities.

The frequencies of chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) were determined in peripheral blood lymphocyte cultures from women with breast cancer treated by chemotherapy (CT) with FEC (5-fluorouracil, epirubicin, and cyclophosphamide) or CMF (cyclophosphamide, methotrexate, and 5-fluorouracil) cocktail in six CT cycles. The number of patients in each CT cycle were from 1 to 3 for SCE and 2 to 5 for CA. Samples were collected before and 48 h after CT. Although the size of the sample was limited and interindividual variability was wide, it appears that a 21-day interval between CT sessions is sufficient for cell recovery. This fact was demonstrated by the reduction in CA and SCE frequencies between cycles in parallel with the unchanged mitotic index and proliferative index values. Teratogenesis Carcinog. Mutagen. 22:257-269, 2002.

Natural dietary antioxidants are extensively studied for their ability to protect cells from damage to DNA, protein, and lipids induced by antitumor agents or radiation that leads to the generation of free radical in normal cells in vivo and in vitro. Curcumin is a natural antioxidant known to possess therapeutic properties and has been reported to scavenge free radicals and to inhibit clastogenesis in mammalian cells. However, curcumin has been reported to induce a significant increase in the frequency of chromosomal aberrations in Chinese hamster ovary (CHO) cells. To investigate whether the clastogenic activity of curcumin in CHO cells in culture can be ascribed to a pro-oxidant behavior, mediated by free radical generation, experiments were carried out with the combination of curcumin (15 microg/ml) and thiourea (10, 20, or 40 microg/ml), a potent hydroxyl radical scavenger. The results showed that the clastogenic action of curcumin was statistically decreased in a dose-dependent manner in the presence of thiourea. These data have shown that curcumin-induced chromosomal damage in CHO cells can be mediated by hydroxyl radical generation in the present experimental conditions. Teratogenesis Carcinog. Mutagen. 21:175-180, 2001.

Ethanol was supplied in the drinking water of Wistar rats at a concentration of 20% v/ v for up to 30 days. The animals treated with ethanol demonstrated a nonsignificant increase in chromosomal aberration frequency when compared with control animals. The mitotic index values obtained indicated no significant differences between ethanol treatment and control groups. The final weights of control rats were significantly greater than those of the ethanol-treated group. Chronic administration of ethanol showed no clastogenic or cytotoxic effect. After chronic ethanol consumption, the cytochromes P450 activity increases, thus possibly preventing the ethanol that has entered the circulation from reaching excessive levels, leading to metabolic adaptation and/or tolerance.

Different cell treatment protocols with the hypomethylating agent 5 azacytidine (5-aza C) were used in exponentially growing Chinese hamster ovary (CHO) cells in order to test its influence on the induction of chromosomal aberrations (CAs) induced by topoisomerase II inhibitors, ellipticine (EPC) and teniposide (VM-26). Cells pre-treated with 1 microg/ml 5-aza C for 1 h during the S-phase and post-treated in the last 2 h of incubation with 0.6 microg/ml EPC or 0.04 microg/ml VM-26 showed a reduction of 48% and 45%, respectively, in the frequencies of CAs as compared to the sum value of the frequencies obtained for each drug alone. 5-aza C added to the cultures for the last 2 h before cell fixation after a 30-min pulse treatment with EPC or VM-26 caused a 38% and 28% reduction, respectively. Simultaneous treatments with 5-aza C plus EPC, or 5-aza C plus VM-26 during the last 2 h of incubation (G2-phase), showed a significant effect of CA reduction (24%) only for the combination of 5-aza C + EPC. Preliminary assays with 5-aza C alone added to the cultures at different times demonstrated its effectiveness in inducing chromosome damage during the S-phase. Since S-phase-treated CHO cells showed a higher degree of reduction in the frequencies of CAs induced by EPC and VM-26, we suggest that 5-aza C incorporation into DNA may change the topo II cleavage sites, protecting the DNA from the induction of damage, or that the hypomethylation induced by incorporation of 5-aza C into DNA may change the chromatin structure facilitating the access to DNA repair enzymes. An alternative possibility is that 5-azaC can reactivate methylated genes involved in the repair of DNA double-strand breaks induced by topo II inhibitors.

Paepalantine is an isocoumarin isolated from Paepalanthus vellozioides which showed antimicrobial activity in in vitro experiments. In the present study, paepalantine was tested for possible clastogenic and cytotoxic action. Cultures from different individuals were treated with paepalantine at concentrations of 20, 40 and 80 microg/ml. The effect of isocoumarin was also tested in an in vivo assay using Wistar rat bone marrow cells. Paepalantine was administered intraperitoneally at concentrations of 6.25, 12.5 and 25 mg/kg body weight. Under these conditions paepalantine did not have a clastogenic effect, but was significantly cytotoxic in the in vitro and in vivo mammalian cell systems tested in the present work.

There is considerable interest in identifying dietary compounds which have the capacity to protect against chromosomal aberrations induced by antitumor agents. Fatty acids and their constituents are able to act as free radical scavengers. Doxorubicin (DXR) is an important chemotherapeutic agent, that also induces chromosome aberrations. Rat bone marrow cells treated simultaneously with olive oil (10 ml/kg body weight) and DXR (90 mg/kg body weight) developed significantly fewer chromosomal aberrations and abnormal metaphases than those treated with DXR alone. Existe considerável interesse na identificação de componentes da dieta que têm a capacidade de proteger contra as aberrações cromossômicas induzidas pelos agentes antitumorais. Os ácidos graxos e seus constituintes são capazes de atuar como seqüestradores de radicais livres. Na presente investigação, as células da medula óssea dos animais tratados simultaneamente com o óleo de oliva (10 ml/kg de peso corporal) e doxorubicina (DXR; 90 mg/kg de peso corporal) apresentaram uma significativa redução no total de aberrações cromossômicas e metáfases alteradas induzidas pela DXR.