Construção de bibliotecas de cDNAs subtraídos a partir das sementes de Mamona submetidas à estresse hídrico e análise in silico de transcritos potencialmente expressos sob condições de estresse: Metalotioneina, proteína reprimida por auxina de 12.5 kDa e Glutelina tipo-A3

Estresses ambientais abióticos são fatores que causam respostas ao nível molecular, fisiológico e morfológico em plantas, dependendo também de sua intensidade e duração. É visto que algumas espécies apresentam tolerância a condições estressantes e ao mesmo tempo são fontes naturais de matéria prima para indústria. Nesse contexto encontra-se a mamona (Ricinus comunnis L.), principal fonte de óleo de rícino valorizado por suas aplicações farmacêuticas e principalmente industriais, vem sendo usada como cultura em regiões onde a disponibilidade de água é reduzida, usada como fonte de renda para agricultura da região nordeste brasileira. Visto que pouco se sabe sobre as respostas moleculares que levam essa planta a tolerar regiões secas e como as sementes, principais foco de interesse, respondem a essa escassez, nesse trabalho foram construídas duas bibliotecas de cDNAs, onde a partir de uma abordagem subtrativa, continham RNAs diferencialmente expressos em sementes de plantas mamona submetidas ao estresse hídrico durante 5 dias (biblioteca L7), e a outra RNAs diferencialmente expressos em sementes controle (biblioteca L5). A biblioteca L7 apresentou a maior variedade de transcritos com um total de 182. A maior parte das funções estabelecidas pelo sistema Gene Ontology - GO, foram direcionadas aos “Processos Metabólicos” (526), em segundo “Respostas a estímulos” (57), o terceiro termo mais abundante foram referentes a “Desenvolvimento”(26). Já na biblioteca L5, foram encontrados 91 transcritos, com maior parte de suas funções referentes a “Processos Metabólicos”(413), em segundo “Respostas a estímulos” (8) e em terceiro Regulação (6). Alguns dos transcritos da biblioteca L7 foram escolhidos para análise por repetirem-se mais de 3x e não aparecerem na biblioteca L5, o que indica uma possível regulação positiva sobre estresse. As análises sobre Metalotioneína (4x), mostraram que a sequência de proteica apresentava os domínios conservados que a caracterizava como tipo II, onde são encontrados dois domínios funcionais ricos em cisteína com posições altamente conservadas, desempenhando a função de ligar-se a metais pesados, correlacionadas assim como a atividade de eliminação EROs e defesa contra o estresse oxidativo, além de apresentar homologia com a sequência de Bruguiera gymnorhiza, uma planta de mangue adaptada a ambientes salinos. Analisamos também os transcritos da referente a proteína AUXIN-REPRESSED 12.5 KDA (3x), apontada como sendo reprimida pelo hormônio auxina e associada ao processo de dormência da semente, é descrito em uma família gênica onde vários membros pertencem as vias de resposta ao estresse. Por último, analisamos a proteína GLUTELIN TYPE-A 3 (5x), uma importante proteína de armazenamento com caráter hidrofílico, possivelmente direcionada para o vacúolo. Em nosso trabalho foi possível observar um aumento de transcritos em relação a subtração controle, possivelmente reflexo do aumento do metabolismo da semente, tanto para resposta defensiva ao estresse hídrico quanto para o amadurecimento rápido da semente onde foram observados transcritos referentes a resposta oxidativa, controle hormonal, proteínas de reserva e produção de óleo.

Avaliação da atividade antimicrobiana das defensinas recombinantes de ervilha (drr230a) e café (cd1) produzidas em Pichia pastoris

The inefficiency of chemical pesticides to control phytopathogenic fungi in agriculture and the frequent incidence of human diseases caused by bacteria which are resistant to antibiotics lead to the search for alternative antimicrobial compounds. In this context, plant defensins are a promising tool for the control of both plant and human pathogenic agents. Plant defensins are cationic peptides of about 50 amino acid residues, rich in cysteine and whose tridimensional structure is considerably conserved among different plant species. These antimicrobial molecules represent an important innate component from plant defense response against pathogens and are expressed in various plant tissues, such as leaves, tubers, flowers, pods and seeds. The present work aimed at the evaluation of the antimicrobial activity of two plant defensins against different phytopathogenic fungi and pathogenic bacteria to humans. The defensin Drr230a, whose gene was isolated from pea (Pisum sativum), and the defensin CD1,whose gene was identified within coffee (Coffea arabica) transcriptome, were subcloned in yeast expression vector and expressed in Pichia pastoris. The gene cd1 was subcloned as two different recombinant forms: CD1tC, containing a six-histidine sequence (6xHis) at the peptide C-terminal region and CD1tN, containing 6xHis coding sequence at the N-terminal region. In the case of the defensin Drr230a, the 6xHis coding sequence was inserted only at the N-terminal region. Assays of the antimicrobial activity of the purified recombinant proteins rDrr230a and rCD1 against Phakopsora pachyrhizi, causal agent of soybean Asian rust, were performed to analyze the in vitro spore germination inhibition and disease severity caused by the fungus in planta. Both recombinant defensins were able to inhibit P. pachyrhizi uredospore germination, with no difference between the antimicrobial action of either CD1tC or CD1tN. Moreover, rDrr230a and rCD1 drastically reduced severity of soybean Asian rust, as demonstrated by in planta assays. In spite of the fact that rCD1 was not able to inhibit proliferation of the human pathogenic bacteria Staplylococcus aureus and Klebsiella pneumoniae, rCD1 was able to inhibit growth of the phytopathogenic fungus Fusarium tucumaniae, that causes soybean sudden death syndrome. The obtained results show that these plant defensins are useful candidates to be used in plant genetic engineering programs to control agriculture impacting fungal diseases.

Caracterização de genes de reparo de DNA em cana-de-açúcar

Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Caracterização de genes de reparo de DNA em cana-de-açúcar

Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species – ROS – that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Utilização de sismos regionais para a determinação de um modelo 1D de velocidades da onda P na Província Borborema - NE do Brasil

In this work it was performed a study to obtain parameters for an 1D regional velocity model for the Borborema Province, NE Brazil. It was used earthquakes occurred between 2001 and 2013 with magnitude greater than 2.9 mb either from epicentres determined from local seismic networks or by back azimuth determination, when possible. We chose seven events which occurred in the main seismic areas in the Borborema Province. The selected events were recorded in up to 74 seismic stations from the following networks: RSISNE, INCT-ET, João Câmara – RN, São Rafael – RN, Caruaru - PE, São Caetano - PE, Castanhão - CE, Santana do Acarau - CE, Taipu – RN e Sobral – CE, and the RCBR (IRIS/USGS—GSN). For the determination of the model parameters were inverted via a travel-time table and its fit. These model parameters were compared with other known model (global and regional) and have improved the epicentral determination. This final set of parameters model, we called MBB is laterally homogeneous with an upper crust at 11,45 km depth and total crustal thickness of 33,9 km. The P-wave velocity in the upper crust was estimated at 6.0 km/s and 6.64 km/s for it lower part. The P-wave velocity in the upper mantle we estimated at 8.21 km/s with an VP/VS ratio of approximately 1.74.

Utilização de sismos regionais para a determinação de um modelo 1D de velocidades da onda P na Província Borborema - NE do Brasil

In this work it was performed a study to obtain parameters for an 1D regional velocity model for the Borborema Province, NE Brazil. It was used earthquakes occurred between 2001 and 2013 with magnitude greater than 2.9 mb either from epicentres determined from local seismic networks or by back azimuth determination, when possible. We chose seven events which occurred in the main seismic areas in the Borborema Province. The selected events were recorded in up to 74 seismic stations from the following networks: RSISNE, INCT-ET, João Câmara – RN, São Rafael – RN, Caruaru - PE, São Caetano - PE, Castanhão - CE, Santana do Acarau - CE, Taipu – RN e Sobral – CE, and the RCBR (IRIS/USGS—GSN). For the determination of the model parameters were inverted via a travel-time table and its fit. These model parameters were compared with other known model (global and regional) and have improved the epicentral determination. This final set of parameters model, we called MBB is laterally homogeneous with an upper crust at 11,45 km depth and total crustal thickness of 33,9 km. The P-wave velocity in the upper crust was estimated at 6.0 km/s and 6.64 km/s for it lower part. The P-wave velocity in the upper mantle we estimated at 8.21 km/s with an VP/VS ratio of approximately 1.74.

Processo identitário profissional: as experiências formativas de licenciandos do curso de Física - UFPI

In this research, we have as study object the identification process with the profession of students of the Undergraduate course for preparing professors for teaching Physics of the Federal University of Piauí. We start from the following question: - What relationships can be established among the senses given by the students to the group of the experiences lived in their initial formation and the identity construction as teacher? The presupposition is that the process of identification with the profession can contain formative experiences that are impossible to be reduced to the situations of disciplines learning. We considered that, in the development of the process of professional identification, the articulations and the connections that the students established with themselves, with the others - their formative teachers, managers - and with the world - the University, the public school - they should be understood in the interdependence relationships, consubstantiated in the experiences lived during the initial stage of their formation. Therefore, our objective is to understand the process of identification of the students with the teaching profession, based in the senses given to their formative experiences. In order to get this aim, we used the methodology of the Comprehensive Interview, and we chose as methodological and theoretical principles: the multi-referential theoretical framework, 'configuration and interdependence', the relationship among 'identity and alterity', the sensitive hearing and the 'intellectual craft'. The study is organized in two parts: the first, contains two chapters, that have introductory character. The second part is unfolded in five chapters, and it constitutes the development of the thesis. In the "paths", we revealed, starting from the actions and intentions that mobilized the students, processes involved in the choice of the course, and, this way, the initial identifications with the being an university student. We looked for to identify possible dissonances among the curriculum 'instituted and instituting', as well as the implications of its management of the formative experiences of future teachers. In short, we tried to capture, through the speeches of a group of students, the signs, the images and the senses of their formative experiences, which indicate as a vision of how their teaching profession is built. With this study, we sought to contribute for the researches on the field of degrees, taking as axis the formative processes that configure those courses. Finally, that the proposition of professor formation politics should be fundamentally based in internal demands, lived in the formative processes.

PROMOÇÃO DA CIDADANIA PELAS RÁDIOS COMUNITÁRIAS DO ABCD PAULISTA, SOB DESAFIOS E ENFRENTAMENTOS POLÍTICOS.

Esta pesquisa, intitulada “Promoção da cidadania pelas rádios comunitárias do ABCD Paulista, sob desafios e enfrentamentos políticos”, estuda 11 rádios comunitárias autorizadas pelo Ministério das Comunicações para funcionamento no Grande ABCD Paulista. Na região, cinco cidades das sete ali existentes abrigam rádios comunitárias, como Diadema (rádios “Navegantes” e “Nova Diadema”); Mauá (rádios “Mauá” e “Z”); Ribeirão Pires (rádio “Pérola da Serra”); Rio Grande da Serra (rádio “Esplanada”) e São Bernardo do Campo (rádios “Lírio dos Vales”, “Nova Riacho”, “Paraty”, “Princesa” e “Represa”). As outras duas cidades daquele território, Santo André e São Caetano do Sul, não registram emissoras comunitárias autorizadas para funcionamento. O objetivo deste estudo é o de revelar o perfil das mencionadas emissoras; a contribuição que oferecem aos processos da promoção de cidadania e inclusão social; seus problemas operacionais estruturais para sobrevivência e reações para superação. A metodologia utilizada consiste em pesquisa bibliográfica, pesquisa documental, entrevistas, visitas às rádios e estudo de programação. Estudou-se o histórico da região; os conceitos de cidadania; participação; radiodifusão comunitária e a própria trajetória das emissoras. Na sequência, houve a consulta em instituições oficiais para o conhecimento das rádios comunitárias autorizadas para funcionamento no ABCD. Posteriormente, seguiu-se a pesquisa com várias visitas de observação. As entrevistas tiveram características semiestruturadas com os radialistas e demais depoentes para este trabalho, especialistas na presente temática. Concluiu-se que existem inúmeras dificuldades que as 11 emissoras comunitárias do ABCD Paulista enfrentam para conseguir manter as rádios funcionamento. A manutenção das dificuldades se dá principalmente pela força da legislação responsável por tal segmento radiofônico comunitário, que o impede de obter apoio comercial e patrocínios.

Incidencia de la política pública para la superación de la pobreza extrema en el marco del desarrollo económico local en garzón Huila. Período 1994-2014.

El auge del estudio de políticas públicas tiene su apogeo en Estados Unidos en 1960 pero recobra importancia a nivel latinoamericano en 1980 y para el caso colombiano en los años de 1990 como lo afirma Salazar Vargas, (1995). Partiendo de este referente, la investigación, Incidencia de la política pública para la superación de la pobreza extrema en el marco del desarrollo económico local en el municipio de Garzón - Huila durante el período 1994-2014, propende por describir la política pública nacional para la superación de la pobreza extrema para el periodo referido y comprender como ha sido implementada a través de la arquitectura institucional en el marco de la dinámica económica local. El ejercicio investigativo se realiza a partir de un enfoque descriptivo e interpretativo que combino técnicas cualitativas y cuantitativas y permitió mediante procesos empíricos y críticos, dar respuesta a la pregunta de investigación ¿Cómo interactúa la política pública para superar la pobreza extrema, formulada en los últimos 20 años en Colombia, con los elementos determinantes del desarrollo económico local en el Municipio de Garzón - Huila? Así entonces, desde el enfoque cualitativo, fue posible contrastar información proveniente de fuentes primarias y secundarias; entre la información secundaria de tipo cualitativo están los planes de desarrollo nacional, departamental y municipal sistematizados y analizados a través de registros analíticos especializados -RAE´s-, y la información de tipo cuantitativa estuvo representada en las estadísticas e informes oficiales de las entidades del orden nacional, departamental y municipal. La información primaria de tipo cualitativo correspondió a la información generada a partir de una entrevista semi estructural implementada con un grupo de actores considerados relevantes en el territorio. Como resultado de la investigación se identifica un municipio con un bajo desempeño en la dinámica de sus indicadores en materia de pobreza y pobreza extrema, situación crítica considerando que fue el segundo municipio de importancia del departamento, y que actualmente ocupa el cuarto o quinto lugar con un escenario poco prometedor dada la pérdida de empleos, resultado de la puesta en operación de la represa el Quimbo, ameritando en términos de política pública para el desarrollo local, el diseño de estrategias en el corto y mediano plazo que generen empleos permanentes que absorban un porcentaje importante de mano de obra no calificada, la cual fácilmente podría ingresar en las estadísticas de pobres y pobres extremos.

Diseño e implementación de un sistema de mantenimiento de las obras civiles de la central hidroeléctrica de Saucay de propiedad de Elecaustro

En el presente trabajo se podrá encontrar un marco referencial en el que se indican los problemas que se presentan en el hormigón y el método de utilizado para realizar pruebas no destructivas que determinan su estado actual; características físicas, químicas y bacteriológicas del agua, proceso de desinfección del agua de consumo humano mediante el empleo de hipoclorito de calcio, así como también la clasificación vial establecida en el Ecuador, señalización vertical, estabilidad de taludes, diseño de muros de gaviones. Se indica también la ubicación, descripción, estado actual, trabajos de mantenimiento preventivo a ejecutarse y el listado de tareas correctivas de las obras civiles de la Central Hidroeléctrica de Saucay de propiedad de ELECAUSTRO S.A. Cuenta además con el proceso utilizado para implementar el sistema de mantenimiento de las obras civiles de la Central Hidroeléctrica de Saucay, mediante la utilización del programa SisMAC (Sistema de Mantenimiento asistido por computador). Finalmente se presenta un cronograma para la ejecución de las actividades de mantenimiento preventivo y el respectivo presupuesto referencial de estas actividades

Criação de um banco de dados para o centro de atendimento terapêutico e social - CATES

O presente trabalho apresenta o relatório das atividades desenvolvidas durante a disciplina de Estágio Supervisionado do curso de Arquivologia da UFSM, elaborado como requisito parcial para a obtenção do grau de bacharel em Arquivologia. As atividades foram desenvolvidas no Centro de Atendimento Terapêutico e Social (CATES). O relatório expõe a realidade arquivística encontrada na organização através do diagnóstico, onde ficou clara a necessidade de implementar um banco de dados para o gerenciamento da informação dos pacientes e atendimentos realizados no órgão. Posteriormente ao diagnóstico foi elaborada uma proposta de trabalho que proporcionou a modelagem de um banco de dados que atendesse as necessidades do CATES.

Os hábitos de leitura dos estudantes do ensino básico da Região Autónoma dos Açores

Tese de Doutoramento, Educação (Metodologia do Ensino da Língua Portuguesa), 15 de Julho 2013, Universidade dos Açores.

As questões de género nos materiais pedagógico-didáticos ao serviço do ensino de História e Geografia

Mestrado, Ensino de História e Geografia no 3.º Ciclo do Ensino Básico e Secundário, 7 de Julho de 2016, Universidade dos Açores (Relatório de Estágio).

Avaliação das propriedades farmacológicas de polissacarídeos do fungo Scleroderma nitidum

Several pharmacological properties have been attributed to isolated compounds from mushroom. Recently, have these compounds, especially the polysaccharides derived from mushrooms, modulate the immune system, and its antitumor, antiviral, antibiotic and antiinflammatory activities. This study assesses the possible pharmacological properties of the polysaccharides from Scleroderma nitidum mushroom. The centesimal composition of the tissue showed that this fungus is composed mainly of fibers (35.61%), ash (33.69%) and carbohydrates (25.31%). The chemical analysis of the polysaccharide fraction showed high levels of carbohydrates (94.71%) and low content of protein (5.29%). These polysaccharides are composed of glucose, galactose, mannose and fucose in the following molar ratios 0.156, 0.044, 0.025, 0.066 and the infrared analysis showed a possible polysaccharide-protein complex. The polysaccharides from Scleroderma nitidum showed antioxidant potential with concentration-dependent antioxidant activity compared to ascorbic acid. The analysis scavenging of superoxide radical and inhibition of lipid peroxidation showed that the polysaccharides from S. nitidum have an IC50 of 12.70 mg/ml and EC50 10.4 μg/ml, respectively. The antioxidant activity was confirmed by the presence of reducing potential of these polysaccharides. The effect of these polymers on the inflammatory process was tested using the carrageenan or histamine-induced paw edema model and the sodium thioglycolate or zymosan-induced model. The polysaccharides were effective in reducing edema (73% at 50 mg/kg) and cell infiltrate (37% at 10 mg/kg) in both inflammation models tested. Nitric oxide, a mediator in the inflammatory process, showed a reduction of around 26% at 10 mg/kg of body weight. Analysis of pro- and anti-inflammatory cytokines showed that in the groups treated with polysaccharides from S. nitidum there was an increase in cytokines such as IL-1ra, IL-10, and MIP-1β concomitant with the decrease in INF-γ (75%) and IL-2 (22%). We observed the influence of polysaccharides on the modulation of the expression of nuclear factor κB. Thus, polysaccharides from S. nitidum reduced the expression of NF-κB by up to 64%. The results obtained suggest that NF-κB modulation is one of the possible mechanisms that explain the anti-inflammatory effect of polysaccharides from the fungus S. nitidum.

Caracterização e comparação de genes expressos em ápices meristemáticos de cana-de-açúcar cultivados em SP e RN

In this work, we used sugarcane as a model due to its importance for sugar and ethanol production. Unlike the current plant models, sugarcane presents a complex genetics and an enormous allelic variation. Here, we report the analysis of SAGE libraries produced using the shoot apical meristem from contrasted genotypes by flowering induction (non-flowering vs. early-flowering varieties) grown under São Paulo state conditions. The expression pattern was analyzed using samples from São Paulo (SP) and Rio Grande do Norte (RN) states. These results showed that cDNAs identified by SAGE libraries had differential expression only in São Paulo state samples. Furthermore, the cDNA identified CYP (Citocrome P450) was chosen for in silico and genome characterization because it was found in SAGE libraries and subtractive libraries from samples from RN. Phylogenetic trees showed the relationship for these sequences. Furthermore, the qRT-PCR for CYP showed a potential role as flowering indutor for RN samples considering different isophorms. Considering the results present here, it can be consider that CYP gene may be used as molecular marker