Análise da Expressão de APE1 após Estresse Oxidativo Induzido em Células Proficientes e Deficientes na Via de Reparo por Excisão de Nucleotídeos

Riboflavin is a vitamin very important in aerobic organisms, as a precursor of many coenzymes involved in the electron transporter chain. However, after photosensitization of riboflavin with UV or visible light, it generates reactive oxygen species (ROS), which can oxidize the DNA. The repair of oxidative lesions on DNA occurs through the base excision repair pathway (BER), where APE1 endonuclease plays a central role. On the other hand, the nucleotide excision repair pathway (NER) repairs helix-distorting lesions. Recently, it was described the participation of NERproteins in the repair of oxidative damage and in stimulation of repair function fromAPE1. The aim of this research was to evaluate the cytotoxic effects of photosensitized riboflavin (RF*) in cells proficient and deficient in NER, correlating with APE1 expression. For this propose, the cells were treated with RF* and it was performed the cell viability assay, extraction of whole proteins, cells fractionation, immunoblotting, indirect immunofluorescence and analysis of polymorphisms of BER gens. The results evidenced that cells deficient in XPA and CSB proteins were more sensitive to RF*. However, XPC-deficient cells presented similar resistance to MRC5- SV cells, which is proficient in NER. These results indicate that XPA and CSB proteins have an important role on repair of oxidative lesions induced by RF*. Additionally, it was evidenced that single nucleotide polymorphisms (SNPs) in BER enzymes may influence in sensitivity of NER-deficient cell lines. Concerning the APE1 expression, the results showed that expression of this protein after treatment with RF* only changed in XPC-deficient cells. Though, it was observed that APE1 is recruited and is bound to chromatin in MRC5-SV and XPA cells after treatment with RF*. The results also showed the induction of DNA damage after treatment with RF*, through the analysis of-H2AX, since the treatment promoted an increase of endogenous levels of this phosphorylated protein, which acts signaling double strand-break on DNA. On the other hand, in XPC-deficient cells, regardless of resistance of RF*, the endogenous levels of APE1 are extremely reduced when compared with other cell lines and APE1 is not bound to chromatin after treatment with RF*. These results conclude that RF* was able to induce cell death in NERdeficient cells, where XPA and CSB cells were more sensitive when compared with MRC5-SV and XPC-deficient cells. This last result is potentially very interesting, since XPC-deficient cell line presents low levels of APE1. Additionally, the results evidenced that APE1 protein can be involved in the repair of oxidative damage induced by RF*, because APE1 is recruited and bound strongly to chromatin after treatment.

Ecologia do fitoplâncton impactado por cultivo de tilápia do Nilo, oreochromis niloticus, em tanques-rede no açude Ministro João Alves, Parelhas, Rio Grande do Norte

The dams are limnic ecosystems of great importance for its multiple uses, among them, water supply for the public and to culture of artisanal fish are most relevant. The aim of the present study is to evaluate the physical-chemical characteristics and the phytoplankton community in two chosen sites (Point 1 littoral zone of point source; Point 2 pelagic zone of non-point source) of the Minister João Alves dam, which is also known as Boqueirão de Parelhas/RN. This represents the spatial distribution of the phytoplankton species in order to understand any possible alterations of the water quality and the phytoplankton composition in relation to the water quality originating from the impact of the tilapia, Oreochromis niloticus, culture. The study period also encompasses temporal variations exhibited in two seasons of an annual cycle, one during the dry season (Oct, Nov and Dec of 2008 and Jan of 2009), and the other rainy season (Mar, Apr, May and June of 2008) to extend the observation. The physicalchemical parameters, such as pH, temperature, electrical conductivity, concentration of dissolved oxygen were measured in situ and the values of the inorganic nutrients (nitrate, ammonium and orto-phosfato) and chlorophyll in the laboratory. The quali-quantitative analyses of the phytoplankton had been carried through sedimentation technique and the enumeration of the random of 400 cells, colonies and filaments counted using Sedgwick-Rafter counting chamber. The results of pH varied widely from the acidic to alkaline range with the minimum of 5.8 (± 0.8) and the maximum of 9.2 (± 0.7-0.8), at point 1 and 2. The dissolved oxygen content was higher in the rainy period than that in the dry period. The maximum electrical conductivity was of 1409 μScm-1 in point 1 and 431 minim of μScm-1, in point 2. There was a considerable alteration in the levels of inorganic nutrients such as nitrate-nitrogen, ammoniacal nitrogen and orthophosphate during the two cycles of study period. Phytoplankton assemblages presented a picture of alternate dominance among species Cyanobacteria, Bacillariophyceae and Chlorophyceae. The trophic state index diagnosed to the category of mesotrophic, which is based on the values of chlorophyll, total phosphorus and Secchi-disc measurements. The wind driven turbulence of the water column and the fresh inflow of water (flushing and dilution) during rainy season acted as constraint and did-not allow an exaggerated growth of the species of cyanobacteria. On the basis of the present we conclude that the culture of tilapias in cage-culture fails to produce pollution load that could compromise the quality of the water of the dam, probably be due to small dimension of the culture in relation to the size, volume of the water and the reservoir capacity support its own environment

Caracterização funcional de dois cDNAs de cana-de- açúcar : PKCI e SHAGGY

Flowering is controlled by several environmental and endogenous factors, usually associated with a complex network of metabolic mechanisms. The gene characterization in Arabidopsis model has provided much information about the genetic and molecular mechanisms that control flowering process. Some of these genes had been found in rice and maize. However, in sugarcane this processe is not well known. It is known that early flowering may reduce its production up to 60% at northeast conditions. Considering the impact of early flowering in sugarcane production, the aim of this work was to make the gene characterization of two cDNAs previously identified in subtractive cDNA libraries: scPKCI and scSHAGGY. The in silico analysis showed that these two cDNAs presented both their sequence and functional catalytic domains conserved. The results of transgenic plants containing the overexpression of the gene cassette scPKCI in sense orientation showed that this construction had a negative influence on the plant development as it was observed a decrease in plant height and leaf size. For the scPKCI overexpression in antisense orientation it was observed change in the number of branches from T1 transgenic plants, whereas transgenic T2 plants showed slow development during germination and initial stages of development. The other cDNA analyzed had homology to SHAGGY protein. The overexpression construct in sense orientation did not shown any effect on development. The only difference observed it was an increase in stigma structure. These results allowed us to propose a model how these two genes may be interact and affect floweringdevelopment.

Análise de cDNAs identificados em bibliotecas substrativas de cDNA para floração de variedades de cana-de-açúcar cultivadas no Rio Grande do Norte(RN): fator de transição NAC, Calmodulina e Fosfatidiltransferase

The flowering is a physiological process that it is vital for plants. This physiological process has been well studied in the plant model Arabidopsis, but in sugarcane this process is not well known. The transition of the shoot apical meristem from vegetative to flowering is a critical factor for plant development. At Brazil northeastern region, the transition to flowering in sugarcane has an important effect as it may reduce up to 60% its production. This is a consequence of the sugar translocation from stalks to the shoot apical meristem which is necessary during the flowering process. Therefore, the aim of this work was to explore and analyze cDNAs previously identified using subtractive cDNA libraries. The results showed that these cDNAs showed differential expression profile in varieties of sugarcane (early x late flowering). The in silico analysis suggested that these cDNAs had homology to calmodulin, NAC transcription factor and phosphatidylinositol, a SEC14, which were described in the literature as having a role in the process of floral development. To better understand the role of the cDNA homologous to calmodulin, tobacco plants were transformed with overexpression cassettes in sense and antissense orientation. Plants overexpressing the cassette in sense orientation did not flowered, while plants overexpressing the cassette in the antissense orientation produced flowers. The data obtained in this study suggested the possible role from CAM sequence, SEC14 and NAC in the induction/floral development pathway in sugarcane, this is the first study in order to analyze these genes in the sugarcane flowering process.

Avaliação de parâmetros de estresse oxidativo em plantas de cana-de-açúcar tratadas com peróxico de hidrogênio

The genus Saccharum belongs to Poaceae family. Sugarcane has become important monocultures in Brazil due to their products: ethanol and sugar. The production may change between different regions from Brazil. This difference is related to soil, climatic conditions and temperature that promotes oxidative stress that may induce an early flowering. The aim of this work was to identify the effects of oxidative stress. In order to analyse this, sugarcane plants were submitted to oxidative stress using hydrogen peroxide. After this treatment, the oxidative stress were analyzed Then, the plant responses were analyzed under different approaches, using morphophysiological, biochemical and molecular tools. Thus, sugarcane plants were grown under controlled conditions and until two months they were subjected first to a hydroponics condition for 24 hours in order to acclimation. After this period, these plants were submitted to oxidative stresse using 0 mM, 10 mM, 20 mM and 30 mM hydrogen peroxide during 8 hours. The histomorphometric analysis allowed us to verify that both root and leaf tissues had a structural changes as it was observed by the increased in cell volume, lignin accumulation in cell walls. Besides, this observation suggested that there was a change in redox balance. Also, it was analyzed the activity of the SOD, CAT and APX enzymes. It was observed an increase in the SOD activity in roots and it was also observed a lipid peroxidation in leaves and roots. Then, in order to identify proteins that were differently expressed in this conditions it was used the proteomic tool either by bidimensional gel or by direct sequencing using the Q-TOF EZI. The results obtained with this approach identified more than 3.000 proteins with the score ranging from 100-5000 ions. Some of the proteins identified were: light Harvesting; oxygenevolving; Thioredoxin; Ftsh-like protein Pftf precusor; Luminal-binding protein; 2 cys peroxiredoxin e Lipoxygenase. All these proteins are involved in oxidative stress response, photsynthetic pathways, and some were classified hypothetical proteins and/or unknown (30% of total). Thus, our data allows us to propose that this treatment induced an oxidative stress and the plant in response changed its physiological process, it made changes in tissue, changed the redox response in order to survival to this new condition

Avaliação da genotoxicidade e da qualidade das águas do açude de Lucrécia/RN

The occurrence of human cancer in the municipality of Lucrécia (RN, Brazil) has shown high frequencies (INCA, 2007). Tests of micronucleus (MN) and nuclear abnormalities (NA) were performed to assess the genotoxic potential of water from the Lucrécia dam, which is located in a semi-arid region under the influence of crop irrigation and irregular rainfall. Water samples were collected in this source for analyzing the concentration of cyanobacteria, metals and radioactivity. Erythrocytes of Nile tilapia (Oreochromis niloticus) were collected in dam and cells of human oral mucosa in the urban area of this municipality for the bioassays of MN and NA. In fish were also analyzed concentrations of metals in samples of liver and gills. The genotoxicity tests with biological samples of fish and humans have shown significant increases in the frequencies of MN and NA (p ≤ 0.05) and are indicative of increased DNA damage in relation to the control groups. In conclusion, the results obtained from water samples and biological municipality of Lucrécia indicates that the presence of chemical and microbiological pollutants, and increase of genotoxic in human of this municipality. We suggest the implementation of advanced water treatment, to prevent further degradation of the aquatic environment and decrease in the life quality. This research of environmental quality assessment was performed in order to contemplate a multi and interdisciplinary character of this water resource and that can induce genotoxic damage in the organisms in this study region

O papel de interações positivas entre plantas na regeneração de áreas degradadas na caatinga

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Diálogos e reflexão

The present work highlights the importance of Dialogue in dealing with situations of crisis, and human conflicts. It has a starting point in some ideas of David Bohm, and in so doing, it presents itself with the challenge of testing the premise that dialogical strategies, of thought and action, can make the human choices inherent to those situations, more coherent and creative. They also allow us to elucidate and grasp them, towards broader comprehension and resolution, while keeping the framework of consistent assessment. However, we believe that said strategies are not, necessarily, problem solvers. With that in mind, we expanded the notion of Dialogue , systematizing dialogical visions of the world, derived from the ideas of David Bohm, Paulo Freire and Mikhail Bakhtin, in which they give priority to the Dialogue component, in all human relations within crucial contexts. From the elaboration of that systemic and dialogical interface, in which Dialogue appears as common theme, and pervasive tool, we detected the emerging of ancillary themes such as Liberty , Consciousness , Creativity , Ethics and Responsibility . We see these themes as ways of comprehending Life, as each one embodies interests, needs and shared human motivations. We have articulated them as a network, and added to that the need of Reflection in conjunction with Dialogue as a watchful call for noticing the incoherence

Universidade e educação ambiental

The society consists of the inter-relations between diverse symbolic fields of performance, where some agents if identify for values, habits and goals. The present thesis analyzed, taking for base the Theory of the Social Field of Pierre Bourdieu, the ambient field and the university, leaving of the principle that the university is one of the propitious places for the quarrel, study, knowledge elaboration, and that the ambient education is one of the ways of dissemination of knowledge and action in favor of the ambient question. It was looked to contextualize and to identify the main factors that configure the university while a social field, and to evidence the conflict and connector links with the ambientalism. It is perceived university as space of tensions, whose produced capital stock in the academic scope must be enough convincing in the direction of the social transformation, of the expansion of conscience, the critical matureness, and thus to appear as agent of social transformation. The joint between the diverse social fields, promoted for the university, will be able to contribute for knowledge formularization that can consist base for elaboration of politics and programs to answer to the problems of the society contemporary. It has the inter-relationship between the two fields, however, this relation still is fragile, needing that a continuity practical social that they make possible that the ambient question is part of the culture of the university. This will be able to give with the intensification of the actions and the inclusion of the ambient questions in the communication mechanisms and administrative management. To surpass discontinuities caused for changes of groups leading and for interests politician conjunctural are necessary that they create habitus, what passes for the institutionalization of practical and the constitution of a culture that the agents allow if to reorganize defending the interests of social transformation front to the economic interests, as much in the university how much in the ambient field

Educação e sustentabilidae :Assentamento Maria da Paz - João Câmara/RN

This work is a research and action in the field of enviromental education, oriented for the construction of the land s distribuction of the Maria da Paz Settling, in João Câmara/RN, as process of social learning, collecive production of new knowledge, values and attitudes related to the environment. It was consolidated through a partnership beteween the UFRN (GERAH/DARQ and GEPEM/DEPED), MST and INCRA/RN. The drawing that represents the way the space organization of the settling was made constructed through effort of many people, in a process of dicussion with the community had as technical support the environment inventory (soil, vegetation, water resources, and others) allowing the agro-ecology zoning of the settlers participation conditions, their contradiction and conflicts, the challenges that appear in the search for consensus and the factor that creat chages

Formação moral das crianças: construção de regras fundamentais aos valores do Movimento dos Trabalhadores Rurais Sem Terra (MST)

The acquiring process of morals on a person is one of the most important aspects of his Social Identity. The basis for his ethics and moral choices are built when he interacts with the world. A child that interacts with participants of Movimento Sem Terra (MST) movement that fights for the Land Reform and the transformation of the society may have the opportunities to acquire the culture, morals and ethics of this movement. Based on this understanding, this work intends to comprehend how children think and incorporate the rules that are the base of the values and principles of MST, considering the diversity of the situations, the limits and the possibilities to experience these values in their everyday life in the Movement. To understand how the process of cognitive construction of the rules takes place in a child, it is important to consider the theories of Jean Piaget. According to him, morals development follows a sequence: the anomie (0 to 2 years old), marked by the absence of rules; the heteronomy (2 to 6/7 years old), where takes place the adoption of rules due to exterior obedience, such as a relative, an institution or a movement; and the autonomy (from 6/7 years old on), in which rules are considered legitimate. All the children in this research have relatives working at MST. The research has two parts. We have first observed the behavior of three groups of children (beyond six years old) while they were involved on their normal activities (kindergarten) activities. On the second moment, we have interviewed 20 children (between 3 and 10 years old). We used flashcards containing scenes; we also told stories and asked moral questions involving the character s behavior. We have noticed the unilateral respect and extern coercion are between the definers of the moral decisions of a child. The empathy and the reduction of the egocentrism help seeing the situation of the point of view of other, although it doesn t mean that one is going to accept others point of view. In the taking decision of the child other factors are also considered such as the space of socialization (family, school). Though the children don t work or take part at MST activities, they have already opinions about involved people behaviors. The interaction with relatives and teachers is one of the most important aspects to encourage them elaborate moral understandings according to the ethics of this movement

Corporeidade, dialogicidade e virtualidade: desvelando a aprendizagem

Cette étude essaie d offrir des contributions pour la construction de connaissance concernant à la corporeité et dans l espace virtuel. À travers des processus interactifs elle propose la construction la production de nouvelles possibilités/stratégies pour la production de connaissance, en adoptant une pratique éducative spécifique: apprendre dans l éducation à distance (EAD). L apprentissage est le centre de ce travail, les acteurs les élèves sont sujets de l action et de la transformation selon la penseé de Paulo Freire et Corpo-vivido dans sa totalité, selon Marleau Ponty. Ce dialogue vif et révecu fait un parcours méthodologique à partir des interlocutions de Pierre-Lévy (1993,1996). On a réalisé une trajectoire pour se représenter, renvoyer, dialoguer et mettre en perspective, en ayant comme point de départ les attributs communicatifs et imagés des matériels d étude utilisés par l EAD, les changements socioculturels provoqués par des possibles dépassements paradigmatiques manifestés dans la société informations et de la communication, et les présuppositions théoriques qui sont ancréesdans les formulations des stratégies et/ou de propositions méthodologiques et comme élément intervenant dans le processus de construction de la connaissance à distance: le sujet de la corporeité. L espace empirique de la recherche est donné à l intérieur du cours TV na Escola e os desafios de hoje , et presenté dans la période de 2000 à 2004 dans l État du Pará. Le texte a été divisé à quatre chapitres. Le premier moment du texte parle de la métaphore de la Fleur et de l organisation du travail. Le deuxième moment systématise ce qui a été projeté pour donner une vision du travail dans l ensemble. Le troisième moment apporte des mots et des concrétisations qui deviennent réelles dans le Desvelando o corpo e o diálogo: a Flor da aprendizagem percebida , en reflétant sur la corporéification du mot s en allant à la rencontre et à la participation d'un apprentissage de dialogue. Et finalement le quatrième moment où nous parcourons la culture de la potentialité de la modalité de l'enseignement à distance, faisant le portrait en arrière plan de l'étude tout en mettant en évidence l'étude du phénomène dans le cours "TV na escola e os desafios de hoje" les paroles des sujets et leur interlocutions. A Flor numa totalidade englobe dans une totalité les moments qui le précedent et invite tous les autres moments à d autres révélations

A formação do professor em diferentes espaços socializadores: um olhar sobre os alunos do curso Pedagogia da Terra da UFRN

This work is an analysis about the teacher's formation starting from certain aspects of its culture. It is supposed that the teacher is constituted in an individual that acts starting from singular aspects, in an individual point of view, but at the same time it interacts with other individuals in an environment strongly marked by the culture. Those two dimensions are representatives of the socio-cultural characteristics and they can be seen in the perspective of the individual and collective identity. Based on those presuppositions we chose, as reference, four environment of formation: the family, the school, the work and the Movement of the Rural Workers Without-Soil/MST (MST: The without-soil-ones in Portuguese) observing as the teachers refers to those environment highlighting, mostly, the formative aspects stood out. The individuals researched are teachers involved in the education of settled communities, who are students of the Earth Pedagogy course of the Universidade Federal do Rio Grande do Norte/UFFRN. For our analyses, we used narrative texts, written by the teachers, as conclusion of the discipline History of the Brazilian Education in which they tell their life histories. The speeches show as the environments of reference influence in their world conceptions, attitudes and values that not just mix in an individual dimension, but also collective. The research make us reflect about the possibility to think on the educational action starting from that understanding of wide formation considering the present elements in the courses of teacher's life

Saberes e práticas em experiência de construção da sustentabilidade no meio rural nordestino

The purpose of this work is to analyze the knowledge relationships that articulate in projects of maintainable rural development construction for the paraiban semi-arid, analyzing the farmers daily practices and their relationship with the technological practices spread by ONGs. We took as empirical object the rural community of Lajedo de Timbaúba, municipal district of Soledade-PB, located in the very small region of paraiban Curimataú. It is a community where registers one of the first cases of maintainable rural development planning in the state of Paraíba. The analysis was centered on the farmers experiences of life in sustainability experience, trying to understand how they acquire new knowledge and how they interact with them. In methodological terms, it was considered feasible to place the knowledge interaction between the farmers and technicians from ONGs by placing the analysis according to Paulo Freire s questioning (2006): extension or communication? To understand the farmers daily practices, it was resorted to the theoretical contribution by Michel de Certeau (2008) in order to discern a microresistance movement of inversion/rejection/changing by the farmers in relation to their external knowledge. Just from the theoretical point of view and resorting to the imaginary social by Cornelius Castoriadis (1982), it was considered the way of living of the farmers researched, having as reference the experience in the material and symbolic production of their lives. It became indispensable, therefore, not to dissociate the knowledge relationships between farmers and technicians from ONGs from the sustainability concepts, maintainable rural development, and rural extension. The results of the study revealed that the farmers from Lajedo de Timbaúba while dealing with the technological practices proposed by ONGs that work in the community, express those practices from their daily logic, and they constitute them in survival strategies that are inserted in their own idiosyncrasy. It was verified, therefore, that the external knowledge presented by the proposals of sociability alternatives with the droughts in the perspective of maintainable development while placed in the farmers daily relationship, they are judged as advantageous or disadvantageous when they are confronted with their peculiar way of doing their daily work. The technological practices are incorporated, denied, or recreated starting from evaluative criteria related to the preservation of the soil and to the economical and social reproduction of the unit of production of family agriculture

Ensino de Entropia: um enfoque histórico e epistemológico.

The rational construction necessary to systematize scientific knowledge in physics, introduces difficulties of understanding in some of its concepts. One of these concepts which exemplify properly this difficulty in learning or teaching is entropy. This thesis propose the construction of a didactic route which constitute itself a historical and epistemological course to entropy, intending to contribute for teaching this concept as well as other physics concepts. The basic assumption to build this route is that through the historical review of the development of this concept in the way suggested by Bachelard s (1884-1962) epistemology it is possible to make subjects, to be taught and learned, more meaningful. Initially I composed a brief biographical note to give the reader an idea about the issues, interests and reflections, related to science, and how I dealt with them in my private and professional life, as well as the role they played to lead me to write this thesis. The strategy to construct the route to entropy was to split the usual contents of basic thermodynamics in three moments in a way they can constitute epistemological units , which can be identified by the way of thinking in the corresponding moments of scientific knowledge production: a technical and empiricist moment, a rationalist and positivist moment and a post-positivist rationalist one. The transition between each moment is characterized by a rupture with the former way of thinking; however the progress in the construction of knowledge in the area is evident. As the final part of this work I present an analysis based on elements of Bachelard s epistemology that are present in each moment. This analysis is the basic component of the didactic route that I propose myself to build. The way I made this route guide to entropy could contribute to the construction of other didactic routes in physics and other sciences, in a way to unveil hidden meanings and as a tool to humanize scientific knowledge.