946 resultados para CNPQ::CIENCIAS DA SAUDE::MEDICINA
Resumo:
Epilepsies are neurological disorders characterized by recurrent and spontaneous seizures due to an abnormal electric activity in a brain network. The mesial temporal lobe epilepsy (MTLE) is the most prevalent type of epilepsy in adulthood, and it occurs frequently in association with hippocampal sclerosis. Unfortunately, not all patients benefit from pharmacological treatment (drug-resistant patients), and therefore become candidates for surgery, a procedure of high complexity and cost. Nowadays, the most common surgery is the anterior temporal lobectomy with selective amygdalohippocampectomy, a procedure standardized by anatomical markers. However, part of patients still present seizure after the procedure. Then, to increase the efficiency of this kind of procedure, it is fundamental to know the epileptic human brain in order to create new tools for auxiliary an individualized surgery procedure. The aim of this work was to identify and quantify the occurrence of epilepticform activity -such as interictal spikes (IS) and high frequency oscillations (HFO) - in electrocorticographic (ECoG) signals acutely recorded during the surgery procedure in drug-resistant patients with MTLE. The ECoG recording (32 channels at sample rate of 1 kHz) was performed in the surface of temporal lobe in three moments: without any cortical resection, after anterior temporal lobectomy and after amygdalohippocampectomy (mean duration of each record: 10 min; N = 17 patients; ethic approval #1038/03 in Research Ethic Committee of Federal University of São Paulo). The occurrence of IS and HFO was quantified automatically by MATLAB routines and validated manually. The events rate (number of events/channels) in each recording time was correlated with seizure control outcome. In 8 hours and 40 minutes of record, we identified 36,858 IS and 1.756 HFO. We observed that seizure-free outcome patients had more HFO rate before the resection than non-seizure free, however do not differentiate in relation of frequency, morphology and distribution of IS. The HFO rate in the first record was better than IS rate on prediction of seizure-free patients (IS: AUC = 57%, Sens = 70%, Spec = 71% vs HFO: AUC = 77%, Sens = 100%, Spec = 70%). We observed the same for the difference of the rate of pre and post-resection (IS: AUC = 54%, Sens = 60%, Spec = 71%; vs HFO: AUC = 84%, Sens = 100%, Spec = 80%). In this case, the algorithm identifies all seizure-free patients (N = 7) with two false positives. To conclude, we observed that the IS and HFO can be found in intra-operative ECoG record, despite the anesthesia and the short time of record. The possibility to classify the patients before any cortical resection suggest that ECoG can be important to decide the use of adjuvant pharmacological treatment or to change for tailored resection procedure. The mechanism responsible for this effect is still unknown, thus more studies are necessary to clarify the processes related to it
Resumo:
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Resumo:
Desde os descobrimentos pioneiros de Hubel e Wiesel acumulou-se uma vasta literatura descrevendo as respostas neuronais do córtex visual primário (V1) a diferentes estímulos visuais. Estes estímulos consistem principalmente em barras em movimento, pontos ou grades, que são úteis para explorar as respostas dentro do campo receptivo clássico (CRF do inglês classical receptive field) a características básicas dos estímulos visuais como a orientação, direção de movimento, contraste, entre outras. Entretanto, nas últimas duas décadas, tornou-se cada vez mais evidente que a atividade de neurônios em V1 pode ser modulada por estímulos fora do CRF. Desta forma, áreas visuais primárias poderiam estar envolvidas em funções visuais mais complexas como, por exemplo, a separação de um objeto ou figura do seu fundo (segregação figura-fundo) e assume-se que as conexões intrínsecas de longo alcance em V1, assim como as conexões de áreas visuais superiores, estão ativamente envolvidas neste processo. Sua possível função foi inferida a partir da análise das variações das respostas induzidas por um estímulo localizado fora do CRF de neurônios individuais. Mesmo sendo muito provável que estas conexões tenham também um impacto tanto na atividade conjunta de neurônios envolvidos no processamento da figura quanto no potencial de campo, estas questões permanecem pouco estudadas. Visando examinar a modulação do contexto visual nessas atividades, coletamos potenciais de ação e potenciais de campo em paralelo de até 48 eletrodos implantados na área visual primária de gatos anestesiados. Estimulamos com grades compostas e cenas naturais, focando-nos na atividade de neurônios cujo CRF estava situado na figura. Da mesma forma, visando examinar a influência das conexões laterais, o sinal proveniente da área visual isotópica e contralateral foi removido através da desativação reversível por resfriamento. Fizemos isso devido a: i) as conexões laterais intrínsecas não podem ser facilmente manipuladas sem afetar diretamente os sinais que estão sendo medidos, ii) as conexões inter-hemisféricas compartilham as principais características anatômicas com a rede lateral intrínseca e podem ser vistas como uma continuação funcional das mesmas entre os dois hemisférios e iii) o resfriamento desativa as conexões de forma causal e reversível, silenciando temporariamente seu sinal, permitindo conclusões diretas a respeito da sua contribuição. Nossos resultados demonstram que o mecanismo de segmentação figurafundo se reflete nas taxas de disparo de neurônios individuais, assim como na potência do potencial de campo e na relação entre sua fase e os padrões de disparo produzidos pela população. Além disso, as conexões laterais inter-hemisféricas modulam estas variáveis dependendo da estimulação feita fora do CRF. Observamos também uma influência deste circuito lateral na coerência entre potenciais de campo entre eletrodos distantes. Em conclusão, nossos resultados dão suporte à ideia de um mecanismo complexo de segmentação figura-fundo atuando desde as áreas visuais primárias em diferentes escalas de frequência. Esse mecanismo parece envolver grupos de neurônios ativos sincronicamente e dependentes da fase do potencial de campo. Nossos resultados também são compatíveis com a hipótese que conexões laterais de longo alcance também fazem parte deste mecanismo
Resumo:
Hebb proposed that synapses between neurons that fire synchronously are strengthened, forming cell assemblies and phase sequences. The former, on a shorter scale, are ensembles of synchronized cells that function transiently as a closed processing system; the latter, on a larger scale, correspond to the sequential activation of cell assemblies able to represent percepts and behaviors. Nowadays, the recording of large neuronal populations allows for the detection of multiple cell assemblies. Within Hebb’s theory, the next logical step is the analysis of phase sequences. Here we detected phase sequences as consecutive assembly activation patterns, and then analyzed their graph attributes in relation to behavior. We investigated action potentials recorded from the adult rat hippocampus and neocortex before, during and after novel object exploration (experimental periods). Within assembly graphs, each assembly corresponded to a node, and each edge corresponded to the temporal sequence of consecutive node activations. The sum of all assembly activations was proportional to firing rates, but the activity of individual assemblies was not. Assembly repertoire was stable across experimental periods, suggesting that novel experience does not create new assemblies in the adult rat. Assembly graph attributes, on the other hand, varied significantly across behavioral states and experimental periods, and were separable enough to correctly classify experimental periods (Naïve Bayes classifier; maximum AUROCs ranging from 0.55 to 0.99) and behavioral states (waking, slow wave sleep, and rapid eye movement sleep; maximum AUROCs ranging from 0.64 to 0.98). Our findings agree with Hebb’s view that neuronal assemblies correspond to primitive building blocks of representation, nearly unchanged in 10 the adult, while phase sequences are labile across behavioral states and change after novel experience. The results are compatible with a role for phase sequences in behavior and cognition
Resumo:
Os recentes avanços técnicos das duas últimas décadas para o registro de sinais neuroeletrofisiológicos foram essenciais para que se testassem hipóteses há muito propostas acerca de como células nervosas processam e armazenam informação. No entanto, ao permitir maior detalhamento dos dados coletados, as novas tecnologias levam inevitavelmente ao aumento de sua complexidade estatística e, consequentemente, à necessidade de novas ferramentas matemático-computacionais para sua análise. Nesta tese, apresentamos novos métodos para a análise de dois componentes fundamentais nas atuais teorias da codificação neural: (1) assembleias celulares, definidas pela co-ativação de subgrupos neuronais; e (2) o padrão temporal de atividade de neurônios individuais. Em relação a (1), desenvolvemos um método baseado em análise de componentes independentes para identificar e rastrear padrões de co-ativação significativos com alta resolução temporal. Superamos limitações de métodos anteriores, ao efetivamente isolar assembleias e abrir a possibilidade de analisar simultaneamente grandes populações neuronais. Em relação a (2), apresentamos uma nova técnica para a extração de padrões de atividade em trens de disparo baseada na decomposição wavelet. Demonstramos, por meio de simulações e de aplicação a dados reais, que nossa ferramenta supera as mais utilizadas atualmente para decodificar respostas de neurônios e estimar a informação de Shannon entre trens de disparos e estímulos externos.
Resumo:
The aim of this study is to understand the perception of medical students at the Federal University of Rio Grande do Norte (UFRN) about humanization in the context of their medical formation, using a qualitative approach. The focus group and participant observation techniques were used, involving a multidisciplinary team composed of professionals from the areas of anthropology and psychology, as well as professors from the medical course, who studied two groups of nine students in their final year. The data were analyzed using the categorical thematic content analysis technique, from which emerged three categories: student/patient relationship, teaching/learning and student/professor relationship. The first allows us to identify that student-patient contact is an essential experience for adopting a more humanized view of the disease process. The second category shows that unqualified professors in the pedagogic practices inherent to the teaching profession and the theory the practical dichotomy hinder the autonomous and holistic formation of knowledge. Similarly, the lack of practices outside the academic environment and the absence of multiprofessional stimulation interfere in the construction of an integral view of the individual. From the third category, the student/professor relationship, emerge two opposing subcategories (professor model and assymetric relationships), which reflect the importance of the professor`s ethical humanist position, as opposed to an authoritarian attitude, to form the professional attitude of the student. The results point important aspects of the medical formation that may open a discussion about humanization, in the context of new national curricular guidelines
Resumo:
No Brasil, a despeito das conquistas obtidas a partir da implantação do Sistema Único de Saúde e dos avanços legais e institucionais na atenção a diversos problemas relacionados ao gênero, a incorporação de temas relacionados aos direitos humanos, sexuais e reprodutivos no contexto da formação profissional em saúde permanece deficiente. Este trabalho teve como objetivos avaliar a inserção da temática saúde sexual e reprodutiva no currículo do curso de graduação em Medicina, por meio do emprego de avaliações de conteúdo cognitivo, procedimental e atitudinal. Trata-se de estudo de intervenção educacional envolvendo alunos do internato do curso de Medicina da UFRN. Foram utilizados os seguintes métodos avaliativos: prova escrita, exame clínico objetivo estruturado (OSCE) e Mini-CEX. Como variáveis explicativas foram consideradas o sexo, idade e participação prévia no componente curricular optativo Saúde Reprodutiva . A avaliação do processo constou da aplicação de questionários de satisfação e entrevistas acerca dos métodos avaliativos utilizados. Considerando os três métodos avaliativos empregados, 183 estudantes participaram do estudo, com média de idade de 24,5 ± 2,2 anos, sendo 52,5% do sexo masculino e 47,5% do sexo feminino. No contexto geral, observamos concordância entre os desempenhos dos estudantes nas avaliações de conteúdo cognitivo, procedimental e atitudinal. A participação dos estudantes no componente curricular eletivo Saúde Reprodutiva mostrou-se associada com melhor desempenho em algumas dimensões da avaliação cognitiva e na avaliação com o Mini-CEX, em relação às competências de anamnese, profissionalismo e qualidades humanísticas, relação médico paciente e desempenho global. A análise da fidedignidade entre os avaliadores na avaliação com o método OSCE mostrou-se adequada (alfa de Cronbach superior a 70%) em relação ao desempenho global e aos aspectos técnicos das competências avaliadas, observando-se baixa confiabilidade na avaliação da comunicação médico-paciente. O presente trabalho constitui-se numa experiência educacional inovadora e pioneira no âmbito da educação médica brasileira no que tange à inserção da temática de saúde sexual e reprodutiva na graduação, sugerindo-se um impacto positivo da iniciativa na formação do médico generalista na UFRN. A avaliação de conhecimentos, habilidades e atitudes em saúde sexual e reprodutiva na graduação de Medicina mostrou-se factível, com alta concordância entre os diferentes métodos empregados. Os métodos OSCE e Mini-CEX podem ser aplicados ao contexto da saúde sexual e reprodutiva, possibilitando a avaliação de competências clínicas relevantes para a formação do médico generalista e que habitualmente não são contempladas nas avaliações rotineiramente realizadas na graduação
Resumo:
Epilepsy is a chronic disorder of the central nervous system, most frequently characterized by abnormal electrical impulses in the brain. It is the oldest recorded neurological disease and has been surrounded by myths, mistaken beliefs and preconceptions. Three professionals work with epileptic patients on a daily basis: teachers, physical educators and physicians . This study aimed to analyze and compare the level of knowledge, preconceptions and attitudes of undergraduate students in the courses of Pedagogy, Physical Education and Medicine, future professionals who will deal with epileptic patients. This descriptive observational study was conducted with 286 university undergraduates of both sexes (women 99/34.6% and men 187/65.4%) from the first to fourth year of Pedagogy, Physical Education and Medicine courses at the Universidade Estadual Rio Grande do Norte (UERN), with mean age of 23 years. A validated and adapted 17-question questionnaire, divided into nine knowledge-based questions, five related to preconceptions and three on attitudes, was applied. Results allowed elaboration of a manuscript entitled Comparative Analysis of Knowledge, Attitude and Preconceptions in University Undergraduates of Pedagogy, Physical Education and Medicine Courses in Relation to Epilepsy. The findings of this article show that knowledge of epilepsy in the three groups is very satisfactory. The low level of preconception indicates that educational or clarifying information is being transmitted, albeit by television, a layman s information vehicle, which has undoubtedly contributed to reducing the stigma of epilepsy through educational information. Results also suggest a lack of knowledge on how to act during a seizure, especially in education professionals. Although there are limitations in our sample, the relevance and contribution of this study is to call attention to the importance and need for these future professionals to be informed and learn correct attitudes with respect to epilepsy during their academic formation. This is essential at a time when the disease is being brought out of the shadows. This research was supported by the Office of the Dean of Teaching, and Deans of Pedagogy, Physical Education and Medicine at UERN. The research was made possible by the multidisciplinary interaction among a physical educator, child neurologist and statistician, all contributing to achieving the aims set out here
Resumo:
Before the scenario full of criticism about a medical model that gives privilege to the diseases and not to the diseased, there are many arguments that defend the need of redeem the humanized relationship between doctor and patient. It became indispensable to mold during the medical graduation a professional capable of perform a special care, less instrumental and more humanized; however, even though the advances of the pedagogical program of the medical graduation, we still face numerous challenges in the process of molding. This study has as general goal to understand if the students medicine experience with the Integrative Community Therapy (TCI) at the Primary Attention – APS/Family Healthy Strategy-ESF, presents potential to configure itself while strategy of teaching-learning to the integral and humanized care. It was held a qualitative research with the students of the medical graduation from the tenth to the twelfth semester that had experience with the TCI, as part of the Boarding of Family and Community Medicine – MFC. We used interviews with script and we resorted to analyze the narratives to Gadamerian Hermeneutics. It was possible to find that before join the boarding of MFC, the students were unaware the TCI and their preconceptions lined up with depreciated character. The experience with the TCI enabled the reframing of the prejudices and the build of new concepts. Internship in ESF and participate of TCI revealed potential to learning of the humanized care by the practical exercise with experiences that privilege the built of ties; the autonomy of the patient; the fulfillment of the longitudinality at the care of the patient; the acknowledgment of the power of resilience of the patients, at the strength of the collective, at the pain sharing, at the strength of a good communication, at the gains of qualified listening exercise. The absence of models of what to do was replaced by experiences of pains and joys at the learning of becoming a doctor. The pains spoke of the structural difficulties (inputs), at the get along with the socials vulnerabilities of the users and the difficult of perform a good communication with the patients. The joys were experienced at the finding of the humanized care exercise. Questions as structural difficulties, low number of people with TCI degree, a shortly experience of with TCI, show up as limitations to its utilization as pedagogical tool. In turn, the reflective potential is capable of cause resignifications about the know-how before the pain of the other being very much present at the narratives, signaling the potential of the learning of TCI. Therefore, this study advocate that the participation of the students at the TCI, beyond the power of offering the students a teaching-learning strategy to the humanized care, represents the possibility of enlarge the horizons of those future doctors at a glance much more conscious of the difficulties and potential of a professional at the ESF, contributing to the graduation of more sensitized professionals and prepared to perform an integral and humanized approach of the person and his/her community, contributing to an APS/ESF more resolute and rewarding to everyone.
Resumo:
Currently, the diagnostic ultrasound is inserted in various areas of medical action and carried out by many medical professionals, from which militate directly in the image area, such as radiologists and sonographers, but also by gynecologists, pediatricians, neurologists, general practitioners, endocrinologists, angiologists, orthopedists, rheumatologists, urologists, general and vascular surgeons. It is well known that the medical professional, for the exercise of its mission, requires a broad set of skills, competencies and attitudes developed and exercised during their training period. Living with medical students over nearly 20 years in hospital environment, I noticed gaps in the learning process by the students about what is diagnostic ultrasound and its applications, demonstrating failures as understanding the basic acoustic ultrasound, difficulties in identifying of anatomical structures in ultrasound images and inability in requests examinations and interpretations of images and reports. Based on these findings, it was developed in this Professional Masters a multimedia digital book that exposes what the ultrasound as a diagnostic modality imaging, dealing with its historiography and its physical/acoustic concepts, relating the process of formation of the ultrasound image, discussing about the features of sonographic equipments and their embedded technologies and highlighting its diagnostic applications , the latter presented through videos which will be described aspects of captured ultrasound images. This book will be available for access in digital format, serving as a teaching tool in medical education since the beginning of the course, so that can be used in conjunction with the discipline of Gross Anatomy, offered in the basic cycle of the Medicine Undergraduate Course of the Federal University of Rio Grande do Norte (UFRN).
Resumo:
BACKGROUND: Among the wide range of skills displayed by a medical doctor is undoubtedly the need to use cohesive and well grounded clinical reasoning in order for medical care to be indeed effective. It is in this respect that conceptual maps emerge; these are a methodological innovation that allows a comprehensive, panoramic and associative outlook of theoretical content, making it more practical and applicable to the reality of clinical observation. Promoting learning, learning resources and a feedback system between professor and students, as well as assessing and monitoring the performance of students during their academic training, are the main features of this tool. OBJETIVE: Assess the use of conceptual maps as a teaching-learning tool in the training of undergraduate medical students at Universidade Federal do Rio Grande do Norte (UFRN). METHODOLOGY: Interventional, randomized, cross-sectional study conducted with students from the 3rd and 5th periods of the medical course at UFRN, during the second semester of 2014, totaling 86 participants, divided into two groups: GI (intervention – clinical case resolution with a conceptual map) and GII (control – clinical case resolution without a conceptual map) in each period. RESULTS: The use of conceptual maps to teach liver failure syndrome resulted in a statistically significant cognitive gain for G1 students from the 5th period (GI: 6.8±1.6 and 8.0±1.5, p = 0.024; GII: 7.2±2.1 and 8.0±1.7, p = 0.125, pre and post-intermediate means, respectively), a result not observed in the period 3rd (GI: 7.7±1.3 and 8.0±1.4, p = 0.501; GII: 6.7±1.8 and 7.8±1.8; p=0.068, pre and post-intermediate means, respectively). Students in the 3 rd period gave better responses to the first clinical case, with a larger number of suitable concepts and crosslinks, when they used conceptual maps (GI: 91.3±13.15 and GII: 64.84±22.84, p=0,002). Students in the 5th period exhibited better clinical reasoning and more complete responses using the tool (p=0,01). Most of the students were not aware of the tool (53.8% from the 3rd period and 65.3% from the 5th period). Among those who knew about conceptual maps, most (59.3%) had only used them during high school, 14.8% had never used them and only seven students (25.9%) used them during the medical course. Analysis of open responses, obtained in process assessment showed clear satisfaction and enthusiasm with learning about the new tool, and frequent suggestions to use it at other moments in the course. Assessment of learning profile, using the VARK questionnaire, showed that most students from both periods exhibited a multimodal style. CONCLUSION: Despite their scant knowledge regarding the tool, good acceptability and understanding was observed in the study participants. The conceptual maps allowed cognitive gains, better responses and clinical reasoning in teaching liver failure syndrome to 5th period students.
Resumo:
OBJETIVE: To evaluate the perception of Medical and Nursing students of Medicine and Nursing graduation courses at the Federal University of Rio Grande do Norte (UFRN), on obstetrics teaching and labor assistance in the context of the maternal care provided by the Maternity Hospital Januário Cicco (MEJC) and contribute to curricular updating planning of teaching obstetrics, in accordance with the principles of humanization. METHODS: It was conducted a study of cross-cutting approach and qualitative and quantitative descriptive method, with students in medicine and nursing of UFRN, who were attending or had been attended the midwifery disciplines medicine; and women's health in nursing. The data were collected through a questionnaire with objective and subjective questions, and stored in a database, spreadsheet software Excel / Office 2010 with all the variables. RESULTS: We interviewed 169 students, 118 of medical school and 51 from the nursing program, of which 46.75% were male and 53.25% female. The largest number of medical respondents is the 11th period (40.67%), and nursing, on 10, (43.15%). These students witnessed 1,073 births, and (61.8%) vaginal deliveries. The obstetricians were the most performed vaginal deliveries (40%). Nurses performed only (8.13%). The assistance provided to women during the process of labor and delivery, was configured as suitable for 87.58% of respondents and these glimpsed humane conduct. Students who performed deliveries, 76.27% were medical and 11.76% were nursing. All had guidance while doing them. A total of 19.50% medical students reported that there is a prejudice against vaginal delivery, particularly for the humanized birth, unlike all nursing students (100%) who reported that there is this prejudice. Most students (73%) showed preference for vaginal delivery, especially nursing students. On the knowledge of myths in relation to vaginal delivery, 60.35% answered that they know some. CONCLUSION: This study presents itself as a highly relevant, since the results may contribute to curricular changing and updating related to obstetrics teaching, but also serve as a resource for analysis of humanization practices that should be developed in educational institutions and which are recommended by the Ministry of Health.
Resumo:
The teaching of the lumbar puncture (LP) technique with simulator is not well systematized in the curricula of medical schools. Studies show that training in the simulator provides learning technical skills, acquisition and retention of knowledge, improve self-confidence of the learner and enables the transfer to clinical practice. We intend this study to introduce simulated training in LP in medical course at the Universidade Federal do Rio Grande do Norte evaluating the experience taking into account quantitative aspects (performance on standardized tests) and qualitative (perception of the students regarding the method and the teaching process learning). The study was conducted in two phases. In the first phase practical training in PL was introduced in the 3rd year of medical school. Seventy-seven students were trained in small groups, guided by a checklist developed in the model Objective Structured Assessment of Technical Skill (OSATS), at this moment they knew they were not under performance evaluation. They were also asked whether they had prior chances to make an LP in patients. At the end of the first phase the students evaluated training in the following areas: teaching technique, simulator realism, time available per group, number of participants per group and relevance to medical practice. In the second phase, two years later, 18 students trained in first stage performed a new LP on the mannequin simulator, and its performance was evaluated through the same checklist of training in order to verify the technical retention. In addition, they answered a multiple choice test about practical aspects of the LP technique. Each participant received individual feedback on their performance at the end of their participation in the study. In the first phase of the study we found that only 4% of students had performed a lumbar puncture in patients until the 3rd year. The training of LP technique with simulator mannequin was considered relevant and the teaching methods was thoroughly evaluated. In the second phase, all participants were successful in implementing the lumbar puncture on the mannequin simulator, compliance with the most steps in a reasonable time, suggesting that would be able to perform the procedure in a patient.
Resumo:
The uses of radiobiocomplexes labeled with technetium-99m contributed to health science advances. Stannous chloride (SnCl2) has been used as a reducing agent for the labeling process. Cytotoxic and genotoxic effect of the SnCl2 have been described in several studies and with this experimental models alterations in molecular and cellular level can be evaluated. In the last years the physicals therapists acquired new devices which emits electromagnetic radiation such us Extremely Low Frequency Pulsated Electromagnetic Fields (E.L.F. P.E.M.F.), radiofrequency, Intense Pulsed Light (I.P.L.) and others which emits sonic waves such us Biorresonance. Scientific evidence of the effects and dosage is important to protect public health and to reach exposition levels that result in significant biological effects. The aim of this project is to verify the effects of these physical agents in plasmid DNA and E. coli AB1157 cultures in presence or absence of SnCl2 and the effects in blood constituents labeled with technetium-99m. Wistar rats blood was exposed to the cited sources and the labelling of blood constituents with 99mTc was carried through. Cultures of E. coli AB1157 and plasmidial samples DNA had been also exposed the physical agents. The results suggest that these agents are capable of altering neither the survival of E. coli cells or plasmid DNA electrophoresis mobility. The multidiscipline character was clearly in this study due the interaction between Nuclear Medicine department of the UERJ and the Laboratory of Physical Agents of the Maimonides University in Argentina until the union between the teacher (biomedical and physiotherapist) and student (physiotherapist), besides collaborators of the area of Physics and Biology, promoting new ideas and perspectives and also adding the knowledge of different areas and origins
Resumo:
The decoction of Brazilian pepper tree barks (Schinus terebinthifolius, Raddi), is used in medicine as wound healing and antiinflamatory. Once extracts from this plant are used for acceleration of scar s process, it is important to study their mutagenic and genotoxic potential. In previous works in our laboratory, it was observed mutagenicity caused by the decoction when in high concentrations. Among the chemical compounds of this plant that could be able to induce mutation, the flavonoids were the only group that was referred to have either an oxidant or antioxidant potential. The flavonoids were isolated, purified and quantified by adsorptive column chromatography under silica gel, bacterial and in vitro genotoxic tests were realized to determine if the flavonoids were the responsible agents for this mutagenicity found. The tests realized with plasmidial DNA were indicative that the flavonoids are probably genotoxic, due to the presence of correlation between increase of the flavonoid concentration and in plasmidial DNA double strand breakage visualized in agarose gel, as well as they were capable to generated abasic sites shown by the in vitro treatment with exonuclease III. The same tests with plasmidial DNA in the presence of copper [10 µM] and of a Tris-HCl pH 7.5 [10 µM] buffer were realized with the isolated flavonoids to determine if there would be or not participation of reactive oxygen species (ROS). The transformation of plasmidial DNA in different bacterial strains proficient and deficient in DNA repair enzymes in the presence or not of a Tris-HCl buffer, suggests that the enzymes that repair oxidative lesions are necessary to repair the lesions generated by the flavonoids and that ROS are generated and are necessary to promote the lesions. Bacterial tests with Escherichia coli strains of the CC collection (deficient or not for DNA repair enzymes), showed that the flavonoids are able to increase the frequency of mutations, mainly in strains mutated in repair enzymes (MutM, MutY-glicosylases and double mutant), suggesting that these agents are responsible for the enhancement in the mutation rate. In order to determine the mutation spectrum caused by the flavonoids of the Brazilian pepper tree stem bark, plasmidial DNA previously treated with the flavonoids were transformed in bacterial strains deficient and proficient in the DNA repair enzymes, followed by a blue-white selection with X-gal, DNA amplification by PCR and sequencing the positive mutant clones. Analysis of the mutants obtained from strains CC104, CC104mutM, CC104mutY, CC104mutMmutY, BW9101, BW9109 indicated a predominance of some mutations like G:C to C:G that can be correlated with the origin of 8-oxoG, due to oxidative lesions caused by the flavonoids. So it can concluded that the flavonoid isolated or in fractions enriched on them are genotoxic and mutagenic, and their mutations are predominantly oxidative, mediated by ROS, and the lesions are recognized by the BER system. In this way it is proposed that the flavonoids can act in two different ways to generate the DNA lesion: 1. in a Fenton-like reaction, when the flavonoid are in the presence of metal ions and that together with the water generate ROS that promotes the DNA lesions; 2. in another way the lesions can be generated by the formation of ROS due to the internal chemical structure of the flavonoid molecule due to the quantity and location of hydroxyl groups, and so producing the DNA lesions, those lesions can be directly (suggested by the in vitro experiments) or indirectly done (supported by the experiments using the CC bacterial strains)