Maria do Carmo Pimentel Batitucci

Antimutagenic activity of ipriflavone against the DNA-damage induced by cyclophosphamide in mice.

In the present study we evaluated the potential of ipriflavone against the cytotoxic and mutagenic effects induced by cyclophosphamide chemotherapeutic agent in bone marrow cells of mice, using the micronucleus assay in vivo on cells of bone marrow. The study was performed following three protocols: pre-treatment, simultaneous treatment and post treatment. The results demonstrated that ipriflavone has a protective effect against mutagenicity induced by cyclophosphamide in the pre-treatment and post-treatment and against the cytotoxicity in all treatments. There was variation between the genders in some of the experimental groups. To evaluate their possible mechanisms of action, it was performed the DPPH assay, which showed no ability to donate hydrogens, suggesting that it acts through other mechanisms. Due to its ability to prevent chromosomal damage, ipriflavone is likely to open an interest field concerning its possible the use in clinical applications.

[Study of mutations causing hypertrophic cardiomyopathy in a group of patients from Espirito Santo, Brazil].

FUNDAMENTO: La cardiomiopatia hipertrofica (CH) es la enfermedad cardiaca hereditaria mas frecuente, causada por mutaciones en los genes codificadores para proteinas del sarcomero. Aunque se hayan identificado mas de 430 mutaciones en varios continentes y paises, no hay relato de que esto se haya estudiado en Brasil. OBJETIVO: Conducir un estudio genetico para identificar mutaciones geneticas que causan la CH en un grupo de pacientes en el estado de Espirito Santo, Brasil. METODOS: Usando la tecnica SSCP, se estudiaron 12 exones de los tres principales genes involucrados con la CH: exones 15, 20, 21, 22 y 23 del gen de la cadena pesada de la β-miosina (MYH7), exones 7, 16, 18, 22 y 24 del gen de la proteina C unida a la miosina (MYBPC3) y exones 8 y 9 del gen de la troponina T (TNNT2). RESULTADOS: Se encontraron 16 alteraciones, incluyendo dos mutaciones, una de ellas posiblemente patogenica en el gen MYBPC3 gen (p. Glu441Lys) y otra patogenica ya descrita en el gen TNNT2 (p. Arg92Trp); 8 variaciones de secuencia raras y 6 variaciones de secuencia con frecuencia alelica mayor que el 1% (polimorfismos). CONCLUSIONES: Con estos datos, es posible concluir que el genotipaje de los pacientes es factible en nuestro medio. Es posible que la variante p.Glu441Lys en el exon 16 del gen MYBPC3 sea patogenica, resultando en un fenotipo mas leve que el encontrado en asociacion con otras mutaciones. La variante p.Arg92Trp en el exon 9 del gen TNNT2 no resulta en un fenotipo tan homogeneo como el descrito anteriormente y puede llevar a hipertrofia grave.BACKGROUND Hypertrophic cardiomyopathy (HC) is the most frequent cardiac hereditary disease, caused by mutations in sarcomere protein coding genes. Although more than 430 mutations have been identified in several continents and countries, there have been no reports of mutations in Brazil. OBJECTIVE To carry out a genetic study to identify genetic mutations that cause HC in a group of patients in Espirito Santo, Brazil. METHODS Using the SSCP technique, 12 exons from the three main genes involved in HC were studied: exons 15, 20, 21, 22 and 23 of the beta-myosin heavy chain gene (MYH7), exons 7, 16, 18, 22 and 24 of the myosin binding protein C gene (MYBPC3) and exons 8 and 9 of troponin T gene (TNNT2). RESULTS 16 alterations were found, including two mutations, one of them possibly pathogenic in the MYBPC3 gene (p. Glu441Lys) and another pathogenic one, previously described in the TNNT2 gene (p.Arg92Trp), 8 rare sequence variations and 6 sequence variations with allelic frequency higher than 1% (polymorphisms). CONCLUSION These data allow the conclusion that the genotyping of patients is feasible in our country. It is possible that the isolated p.Glu441Lys variant identified in exon 16 of the MYBPC3 gene is pathogenic, promoting a milder phenotype than that found when in association with other mutations. The p.Arg92Trp variant in the exon 9 of TNNT2 gene does not promote such a homogeneous phenotype as previously described and it can lead to severe hypertrophy.

Cardiovascular effects induced by the aqueous fraction of the ethanol extract of the stem of Solanum stipulaceum in rats

The cardiovascular effects induced by the aqueous fraction of the ethanol extract of the stem (AFS) of Solanum stipulaceum Roem. & Schult were studied in rats. In non-anesthetized rats, AFS injections produced significant and dosedependent hypotension associated with increase in heart rate. In isolated rat superior mesenteric rings, AFS was able to antagonize the contractions induced by phenylephrine and KCl. The vasorelaxant activity of AFS was not inhibited by either removal of vascular endothelium, L-NAME, atropine or indomethacine. In isolated rat atrial preparations, AFS produced concentration-related negative inotropic and chronotropic responses. These results suggest that the hypotensive effect of AFS is due to a peripheral vasodilation, which can not be attributed to the participation of vascular endothelium. Finally, AFS acts directly on the heart decreasing contractility and heart rate.

Mutagenicity of ipriflavone in vivo and in vitro.

Ipriflavone (7-isopropoxy-isoflavone) is a semisynthetic isoflavone derivative from daidzein and prescribed to prevent and treat osteoporosis in postmenopausal women. In the present study, ipriflavone was investigated with regard to their cytotoxic and mutagenic effects using the micronucleus assay (MN) in vivo on cells of bone marrow and peripheral blood of Swiss albino mice and the micronucleus test with the cytokinesis-blocked micronucleus assay (CBMN assay) on human peripheral blood lymphocytes. The studies were performed in mice with three dosages of the drug, 1.71, 8.57 and 42.85 mg/kg bw in single oral exposure, and for two dosages, 5 and 10 μg/mL in the CBMN assay. Ipriflavone, in the dosages tested, did not differ from controls neither in the induction of MN nor induced cytotoxicity to cells in the in vivo test. However, in the CBMN assay, the concentration of 10 μg/mL induced a statistically significant increase in MN formation and decreased cell proliferation, demonstrating to be mutagenic and cytotoxic at this concentration.

Association of MTHFR and PICALM polymorphisms with Alzheimer’s disease

Alzheimer’s disease (AD) is a complex neurodegenerative disorder and the primary cause of dementia in the elderly and causes a decrease in cognition, functionality, and behaviour. Genetic risk factors play an important role in the pathogenesis of AD. In this case–control study, we aimed to investigate whether single nucleotide polymorphisms in MTHFR (rs1801133), PICALM (3851719), CLU (rs11136000), and CR1 (rs6701713) are associated with AD. Genotype frequencies were evaluated in 82 late-onset AD patients and 161 elderly healthy controls matched by age and gender. We detected a significant association of the MTHFR rs1801133 and PICALM rs3851179 polymorphisms with AD. The results of this study support the hypothesis that several genes are involved in the aetiology of AD.

Cystic fibrosis Δf508 mutation screening in Brazilian women with altered fertility

Cystic Fibrosis (CF) is an autosomal recessive disease, caused by mutations in the Cystic Fibrosis Transmembrane Regulator gene (CFTR). The most frequent mutation in CF is ΔF508. The disease is clinically characterized by elevated concentrations of sweat chlorides and abnormally thick mucus. It affects organs such as lung, pancreas, gastrointestinal and reproductive tract. Women with CF commonly present delayed puberty and amenorrhea due to malnutrition. Our objective was to screen the presence of ΔF508 mutation in 24 women with altered fertility. Nine of these women presented reduced fertility without a known cause, four showed polycystic ovaries and two had early menopause. One woman with early menopause was a carrier of the ΔF508 mutation. Our study demonstrates that it is possible that the frequency of CF mutations among patients with altered fertility may be higher than expected. Previous data showed that fibrocystic women can show reduced fertility, maternal mortality associated with pregnancy and increased incidence of spontaneous abortion. We therefore recommend that women with reduced fertility undertake genetic tests for a better evaluation of pregnancy risks and clinical monitoring.

Estudo de mutações causadoras de cardiomiopatia hipertrófica em um grupo de pacientes no Espírito Santo, Brasil

FUNDAMENTO: La cardiomiopatia hipertrofica (CH) es la enfermedad cardiaca hereditaria mas frecuente, causada por mutaciones en los genes codificadores para proteinas del sarcomero. Aunque se hayan identificado mas de 430 mutaciones en varios continentes y paises, no hay relato de que esto se haya estudiado en Brasil. OBJETIVO: Conducir un estudio genetico para identificar mutaciones geneticas que causan la CH en un grupo de pacientes en el estado de Espirito Santo, Brasil. METODOS: Usando la tecnica SSCP, se estudiaron 12 exones de los tres principales genes involucrados con la CH: exones 15, 20, 21, 22 y 23 del gen de la cadena pesada de la β-miosina (MYH7), exones 7, 16, 18, 22 y 24 del gen de la proteina C unida a la miosina (MYBPC3) y exones 8 y 9 del gen de la troponina T (TNNT2). RESULTADOS: Se encontraron 16 alteraciones, incluyendo dos mutaciones, una de ellas posiblemente patogenica en el gen MYBPC3 gen (p. Glu441Lys) y otra patogenica ya descrita en el gen TNNT2 (p. Arg92Trp); 8 variaciones de secuencia raras y 6 variaciones de secuencia con frecuencia alelica mayor que el 1% (polimorfismos). CONCLUSIONES: Con estos datos, es posible concluir que el genotipaje de los pacientes es factible en nuestro medio. Es posible que la variante p.Glu441Lys en el exon 16 del gen MYBPC3 sea patogenica, resultando en un fenotipo mas leve que el encontrado en asociacion con otras mutaciones. La variante p.Arg92Trp en el exon 9 del gen TNNT2 no resulta en un fenotipo tan homogeneo como el descrito anteriormente y puede llevar a hipertrofia grave.BACKGROUND Hypertrophic cardiomyopathy (HC) is the most frequent cardiac hereditary disease, caused by mutations in sarcomere protein coding genes. Although more than 430 mutations have been identified in several continents and countries, there have been no reports of mutations in Brazil. OBJECTIVE To carry out a genetic study to identify genetic mutations that cause HC in a group of patients in Espirito Santo, Brazil. METHODS Using the SSCP technique, 12 exons from the three main genes involved in HC were studied: exons 15, 20, 21, 22 and 23 of the beta-myosin heavy chain gene (MYH7), exons 7, 16, 18, 22 and 24 of the myosin binding protein C gene (MYBPC3) and exons 8 and 9 of troponin T gene (TNNT2). RESULTS 16 alterations were found, including two mutations, one of them possibly pathogenic in the MYBPC3 gene (p. Glu441Lys) and another pathogenic one, previously described in the TNNT2 gene (p.Arg92Trp), 8 rare sequence variations and 6 sequence variations with allelic frequency higher than 1% (polymorphisms). CONCLUSION These data allow the conclusion that the genotyping of patients is feasible in our country. It is possible that the isolated p.Glu441Lys variant identified in exon 16 of the MYBPC3 gene is pathogenic, promoting a milder phenotype than that found when in association with other mutations. The p.Arg92Trp variant in the exon 9 of TNNT2 gene does not promote such a homogeneous phenotype as previously described and it can lead to severe hypertrophy.

In vivo antimutagenic and antiatherogenic effects of the (1 → 3)(1 → 6)-β-d- glucan botryosphaeran

The antimutagenic effect of botryosphaeran, an exocellular (1 → 3)(1 → 6)-β-d-glucan, from the ascomyceteous and plant-borne endophytic fungus, Botryosphaeria rhodina MAMB-05, was evaluated in young (6-8 weeks) and elderly (18 months) Swiss albino mice of both genders. The hypolipidemic, hypoglycemic and antiatherogenic potential was also evaluated in 18-month old male LDL receptor knockout (LDLr-/-) mice. Administration of botryosphaeran by gavage (doses: 7.5, 15, 30 mg/kg b.w./day) in a 30-day pretreatment protocol (young mice), or 15-day protocol (older mice), did not cause genotoxicity as assessed by the micronucleus test in peripheral blood (PB) and bone marrow cells (BMCs). Furthermore, there was no cytotoxic effect of this β-d-glucan in the treatments. A lower frequency of micronuclei was observed in BMCs from young and old mice that received botryosphaeran, indicating its antimutagenic effect. Botryosphaeran (30 mg/kg b.w./day) promoted 102.22% (young) and 103.45% (elderly) reductions in cyclophosphamide-induced damage in male mice. Botryosphaeran also exerted chemoprotective effects in LDLr-/- and wild-type (C57BL/6) mice. Botryosphaeran treatment for 15 days at a dose of 30 mg/kg b.w./day improved the lipidic profile (reductions of 53.8-84.3%), and decreased aortic lipid deposition (32.8%) in the LDLr-/- atherosclerotic mice. The results indicate botryosphaeran has relevant biologic effects, making it a promising candidate for the development of new therapeutic agents.

Coriandrum sativum grown under organic or chemical fertilizer effectively prevents DNA damage: Preliminary phytochemical screening, flavonoid content, ESI (-) FTICR MS, in vitro antioxidant and in vivo (mice bone marrow) antimutagenic activity against cyclophosphamide

Objective: To evaluate the influence of fertilization and phenological stages on secondary metabolites production and chemoprotective effects of Coriandrum sativum (C. sativum) L. Methods: The plants were grown under organic or chemical fertilizer, collected at vegetative and flowering development stages and their hydroalcoholic extracts were analyzed by phytochemicals methods, mass spectrometry, antioxidant and antimutagenic assays. Results: All extracts exhibited metabolites such as coumarins, flavonoids and steroids, and mass spectrometry showed similar molecular peaks among the extracts evaluated, suggesting the presence of palmitic and α -linolenic acids. Vegetative C. sativum extract grown under chemical fertilizer showed better antioxidant activity, according to the DPPH assay. Vegetative C. sativum extracts grown under organic and chemical fertilizer were able to effectively reduce micronucleous frequency in the simultaneous and pre-treatment protocols, especially reaching 55.90% of damage reduction in the pre-treatment protocol. Conclusions: These findings suggest that chemical fertilization promotes an increase in the content of flavonoids in C. sativum and, consequently, leads to better antioxidant and antimutagenic activities, as well as reinforces the potential uses of this culinary plant in health promotion and disease prevention.

Mutagenic Effect of Three Invasive Species through Allium Cepa Bioassay

Invasive alien species are a global threat to biodiversity that affects protected areas around the world. The occupation of new environments by these plants is a problem to be solved and it is essential to investigate all the aspects that allow this successful to find solutions to this question, such as its mutagenic effects. Thus, this study aimed to evaluate the mutagenic effect of leaves extracts of Acacia mangium Willd, Artocarpus heterophyllus Lam and Eriobothrya japonica (Thunb.) Lindl through Allium cepa bioassay. For this, A. cepa seeds were submitted to continuous and discontinuous (acute and chronic) treatments in medium with water (negative control) or four concentration of each extract (1, 5, 10 and 50 mg/mL). The mitotic index was affected at all concentrations of three extracts tested in all treatments, continuous and discontinuous. Aneugenic effects were not related to any treatment tested. E. japonica extract induced clastogenic effects at 1, 5 and 10 mg/mL in continuous treatment, 5 and 10 mg/mL in acute discontinuous treatment and at 10 mg/mL in chronic discontinuous treatment. Clastogenic effect was also observed at 10 mg/mL of A. heterophyllus extract in continuous and acute discontinuous treatments.