Caracterização de dois cDNAs homológos e uma AP endonuclease em cana-de-açúcar

The genome of all organisms is subject to injuries that can be caused by endogenous and environmental factors. If these lesions are not corrected, it can be fixed generating a mutation which can be lethal to the organisms. In order to prevent this, there are different DNA repair mechanisms. These mechanisms are well known in bacteria, yeast, human, but not in plants. Two plant models Oriza sativa and Arabidopsis thaliana had the genome sequenced and due to this some DNA repair genes have been characterized. The aim of this work is to characterized two sugarcane cDNAs that had homology to AP endonuclease: scARP1 and scARP3. In silico has been done with these two sequences and other from plants. It has been observed domain conservation on these sequences, but the cystein at 65 position that is a characteristic from the redox domain in APE1 protein was not so conservated in plants. Phylogenetic relationship showed two branches, one branch with dicots and monocots sequence and the other branch with only monocots sequences. Another approach in order to characterized these two cDNAs was to construct overexpression cassettes (sense and antisense orientation) using the 35S promoter. After that, these cassettes were transferred to the binary vector pPZP211. Furthermore, previously in the laboratory was obtained a plant from nicotiana tabacum containing the overexpression cassette in anti-sense orientation. It has been observed that this plant had a slow development and problems in setting seeds. After some manual crossing, some seeds were obtained (T2) and it was analyzed the T2 segregation. The third approach used in this work was to clone the promoter region from these two cDNAs by PCR walking. The sequences obtained were analyzed using the program PLANTCARE. It was observed in these sequences some motives that may be related to oxidative stress response

Aplicação de sensoriamento remoto e sistema de informação geográfica na determinação da vulnerabilidade natural e ambeintal do Município de Guamaré(RN): simulação de risco às atividades da indústria petrolífera

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Terra e trabalho: permanência de uma política pública na Fazenda Mundo Novo, em Caicó-RN

This is study about the Project Vazantes, as a case of stay policy on access to land upstream of the weir humid New World, in Caicó-RN. It includes social and economic aspects of vazanteiros, that in their experiences with the land, recreate structures, forms and functions. Theoretically, the work is based on the ideas miltonianas (1986, 1988, 1996, 1997) on the production and reproduction of space, as seen this system of actions that is articulated to the public policies and the achievements solidarity, to the extent that production, in vazantes there is virtually no technology investment, technical assistance and rural credit. Regarding the search procedures, we conducted interviews and questionnaires with vazanteiros applied but did photographic records and research into documentary sources Agricultural Research Company's Rio Grande do Norte S / A (EMPARN), the Union of Rural Workers of Caicó (STRC ), In newspapers and magazines, as well as other sources of information related to the object of study.Such procedures, coupled with theoretical and methodological benchmarks, helped in the understanding of the relationship that takes place in the New World Farm, in order that space produced by actors (EMPARN and vazanteiros) comprises a structure of power that involves multiple interests in extent that land and labor are appropriate mechanisms as dialetizantes

Caracterização de uma nova exonuclease identificada em uma biblioteca metagenômica

A abordagem metagenômica tem permitido o acesso ao material genético de microrganismos não cultivados e tem sido usada para identificação de novos genes. Apesar da importância dos mecanismos de reparo de DNA para a manutenção da integridade genômica nosso conhecimento sobre mecanismos de reparo de DNA é baseado em organismos modelo como E. coli e pouco é conhecido sobre os organismos de vida livre e não cultivados. Neste trabalho, a abordagem metagenômica foi aplicada para descobrir novos genes envolvidos com a manutenção da integridade genômica. Um clone positivo foi identificado por replicar a biblioteca metagenômica em meio seletivo contendo H2O2. O clone metagenômico foi capaz de complementar parcialmente a deficiência em reparo de DNA de cepas simples e duplo-mutantes de E.coli (recA e xthA nfo, respectivamente) submetidas ao estresse gerado por H2O2 e MMS.A análise de sequência mostrou uma ORF codificando para uma proteína hipotética membro da superfamília Exo_Endo_Phos (PF03372) e, a filogenia indicou que a mesma não está inclusa em nenhuma das subfamílias EEP. Assim, uma nova nuclease foi identificada e experimentalmente caracterizada in vivo e in vitro. Ensaios específicos utilizando a nuclease purificada e oligonucleotideos fluorescentemente marcados revelaram sua atividade 3´-5´exonuclease, em substratos simples e dupla-fita, dependente de Magnésio e sensível a EDTA. Uma vez que este é o primeiro relato e caracterização de uma enzima obtida a partir de abordagem metagenômica mostrando uma atividade exonuclease, foi nomeada EXOMEG1

Alterações morfofisiológicas e moleculares em resposta a estresses abióticos em mamona (Ricinus communis L.)

The use of fossil fuels has been considered one of reason for the increase of pollution in the atmosphere and it may be related to the climate changes. Then, the research of the new sources of fuels will be important. Considering this, the use of biodiesel has been considered not as bad as petrol. The castor bean (Ricinus communis L.) is an important oilseed, which belongs to Euphorbiaceae family, and the oil found in the seed has important characteristics for biodiesel. This plant is considered as “rustic” as it does not need so much water for its development and oil production. Due to this, this plant has been considered to be ideal in semi-arid regions, such as the Northeast of Brazil. The aim of his study is to better understand the responses to abiotic stresses (drought and salinity) from castor bean plants using morphological, physiological and molecular tools. In order to do this, the castor bean plants were subjected to salt stress (50, 100, 150 and 200 mM NaCl) in a controlled environment and drought stress (5, 10, 15 days and 10 days cyclic). After these treatments, these plants were subjected to different analyzes: a) the expansion and retention of water from leaves; b) anatomy using leaves and roots. Based on these results, we found that castor suffered decrease in leaf area with increase drought stress, however restricted water loss, probably by accumulation of compatible solutes in the leaves. The anatomy data showed modifications in the vascular system. These modifications observed suggested that castor bean plant may be resistant to stress as it was verified in 5 days of drought as well as in 100 mM NaCl. In both conditions, these plants were fine. Probably these plants keep some solutes in the cell and then maintain the cell tugor. The data obtained in this study gave a better idea how castor bean plant responds to abiotic stress conditions - drought and salt stress

Metagenoma de comunidades microbianas expostas à radiação natural e a metais

Many species have specialized to live in the most varied existing environments showing the remarkable adaptability of the microbial world the most diverse physicochemical conditions. Environments exposed to natural radiation and metals are scarce around the world, presenting a microbiota still unknown. With a total number estimated between 4 and 6 x 1030 microrganisms on earth, they constitute an enormous biological and genetic pool to be explored. Metagenomic approach independent of cultivation, provides a new form to access to the potential genomic environmental samples becoming a powerful tool for the elucidation of ecological functions, metabolic profiles, as well as to identify new biomolecules. In this context, the genetic material of environmental soil and water samples from Açude Boqueirao Parelhas-RN, under the influence of natural radiation and the presence of metals, was extracted, pirosequencing and the generated sequences were analyzed by bioinformatics programs (MG-RAST and STAMP). Taxonomic comparative profiles of both samples showed high abundance of Domain Bacteria, followed by a small portion attributable to Eucaryota Domains, Archaea and Viruses. Proteobacteria, Actinobacteria and Bacterioidetes phyla showed the greater dominance in both samples. Important genera and species associated with resistance to various stressors found in region were observed. Sequences related to oxidative and heat stress, DNA replication and repair, and resistance to toxic compounds were observed, suggesting a significant relationship between the microbiota and their metabolic profile, influenced by regional environmental variables. The results of this study add valuable and unpublished data on the composition of microbial communities in these regions

Determinantes genéticos e ambientais das doenças hipertensivas da gravidez

The development of complex diseases such as preeclampsia are determined by both environmental and genetic factors, but there is also interaction among these factors. Preeclampsia is a pregnancy-specific disorder characterized by de-novo hypertension and proteinuria after 20th week of gestation. There is a broad spectrum of clinical presentations related to hypertensive disorders of pregnancy (HDP) that can range from mild preeclampsia to eclampsia (seizures) or HELLP syndrome (Hemolysis, Elevation of Liver enzymes, Low Platelets). Those clinical outcomes might be linked to different pathological mechanisms. Our work aims to identify factors (i.e. genes and environmental) associated with the HDP’s clinical spectrum. Using a case-control approach, we selected a total of 1498 pregnant women for epidemiological and genetic studies, encompassing 755 normotensive (control); 518 preeclampsia; 84 eclampsia; and 141 HELLP. Women were genotyped for 18 SNPs across 5 candidate genes (FLT1, ACVR2A, ERAP1, ERAP2 and LNPEP). For the environmental factors, we found maternal age, parity status and pre-gestational body mass index as important risk factors associated with disease. Genes were associated in a phenotype-specific manner: ACVR2A with early preeclampsia (rs1424954, p=0.002); FLT1 with HELLP syndrome (rs9513095, p=0.003); and ERAP1 with eclampsia (rs30187, p=0.03). Our results suggest that different genetic mechanisms along with specific environmental factors might determine the clinical spectrum of HDP. In addition, phenotype refinement seems to be an essential step in the search for complex disease genes

“Currículo encarnado”: cartografia simbólica e afinidades pós-coloniais

This thesis has as main objective to reflect about the defining conceptual elements of the embodied curriculum concept, ident ified curriculum conception from the analysis about the rationality employed in the doctoral work of teacher Antonio Fernando Gouvê a da Silva, entitled The construction of the curriculum in popular critical perspective: the significant words to the context ualized practices , written and defended in the Postgraduate Program in Education: Curriculum, in the Pontifícia Catholic University of São Paulo, in 2004. We looked, also, on the problematization of the affinities between postcolonial theories – analytical perspectives towards the discussion about coloniality and their effect on contemporary social weaving – and the embodied curriculum concept. We argue that the reflections present in the aforementioned work bring an articulated curriculum concept based on three conceptual symbiotic elements, namely: negativity, dialogue and praxis, which, by endorsing the possibility of a curricular ingrained practice in the life context of the individuals, have some post - colonial inclinations that lead to the problematizat ion about the neo - colonial manifestations in curricular sphere, outlining a critique of the modus operandi of the colonialism, particularly, in its cultural and epistemic dimension from which the education is inseparable. For that, we used as methodologica l procedure the symbolic cartography, knowledge building strategy systematized by Boaventura de Sousa Santos, which allowed us to construct interpretive maps that enabled the symbolization of the universe which we longed to understand, that is, the concept ual categories mentioned above, which, in our view, underlie the concept of embodied curriculum. In this direction, we anchored ourselves in a meaningful dialogue with the theoretical approach of Paulo Freire and some of his interpreters regarding the disc ussion on curriculum, especially the reflections dev eloped by Antonio Fernando Gouvê a da Silva, and authors whose theoretical developments resonate in prospects for humanization, social justice and empowerment, among which we highlight: Theodor Adorno, Hug o Zemelman, Wilfred Carr, Adolfo Sánchez Vázquez, among others. We seek, in the same manner, on the contributions of authors considered post - colonial, as Hugo Achugar, Gayatri Spivak, Boaventura de Sousa Santos and Enrique Dussel, to name a few, the reason s why we consider the embodied curriculum as a place of political - pedagogical enunciation, conducive to an educational praxis that engages in a curricular work of reality translation in order to see what overwhelms it to, then, elicit the construction of a school curriculum as an awareness project for releasing in relation to what is unjust and inhumane. We consider, finally, that the success of this curriculum embodied translation implies a larger number of speakers mobilized in the production of knowledge that yearns for social emancipation and contribute to the enrichment of human capabilities as the maintenance of the life and the dignity of people.

A Experiência como ordenação da realidade: uma estratégia orgânica para a educação científica

Knowledge is understanding. According to the philosopher Gaston Bachelard our immediate contact with the reality is only worth as confusing and provisional data. This phenomenological contact requires inventory and classification. For this reason our first reading on any phenomenon is limited to a basic levels of reality. Elements such as dynamics, functioning or detailed characteristics of what is observed can only be accessed at higher levels of reality, explains the physicist Werner Heisenberg. The ideas woven by these two great intellectuals oxygenates the notion that a well-made thinking does not require only observation and description of the nature, but assigns value and meaning to the knowledge. Based on these ideas and on the cognitive horizon brought by the complexity sciences, this research aims to nurture a reflection on our understanding of the world built from a rational perspective of experience, as an organic sequence of research. This arguments, over the study, describes how the experience is able to oxygenate a well-made thinking, as the concept created by Edgar Morin and expanded by Conceição Almeida. I argue that the experience as a path of investigative research allows one to ventures in the shadows of the unknown to access upper layers of reality. The experience is, therefore, an organic strategy for a well-made thinking - A nutritious mud that oxygenates, regulates, repairs and configures the quality of understanding. As a thread to discuss this ideas I've used my professional journey over a year and a half as a Natural Sciences' teacher on the Federal University of Rio Grande do Norte, where I could see how experiences helped on breaking a simplified understanding of the world. I chose to work with the research problems developed by 398 students over these three semesters. The problems were essential to the questioning of the phenomena that once seemed obvious or uninteresting, bringing out operational reasons and dynamics of the observed structures. Experience, in this sense, is the founder of dynamic thinking, as the need to deconstruct the phenomena's first impressions, assigning value and meaning to gestated knowledge.

Caracterização de um homólogo de HINT1 em cana-de-açúcar encontrado em bibliotecas subtrativas de CDNA para floração

The sugarcane is a monocot plant grown in tropical and subtropical regions, with Brazil being the largest producer. Despite its economic importance, little is known about the molecular flowering process in sugarcane. This physiological process can promote a loss up to 60% in sugar or bioethanol. Thus, this work had as objective characterize a HINT1 homologous gene previously identified in subtractive libraries of flowering. Genomic analysis of gene and promoter region structure allowed the observation that there are at least two distinct genes homologous to HINT on sugarcane. Bioinformatics analyses showed the conservation of the characteristic protein domain of HIT superfamily and indicate a phylogenetic relationship associated to cell location. Moreover, a possible relation with the SBTILISIN-like protein family through the information available in interatomas was observed. This suggests that the HINT gene of sugarcane can be related to plant development, there are several possibilities of interactions in the regulation of floral induction process, because the sequences present in regulatory regions indicate that differential expression of HINT was related to with climatic factors in the Northeast region of Brazil as well as to biotic stress and phytohormones. Furthermore, the sugarcane phenotypes indicate that the influence of HINT may happen due to product accumulation of its enzymatic activity. For these characteristics this gene can be used as a marker in the selection of new varieties.

Reservatório Tabatinga (Macaíba-RN): qualidade ambiental, conflitos e uso

The reservoirs are water sources built along the fluvial basins, between rivers and dams made by concrete or earth. In Brazil they are built for different purposes, standing out the generation of energy (hydroelectric power station), flowing regulation, water reserves and flooding control, therefore they have played and still play an important role in the modern society.In the Northeastern semiarid region, they are typically used to supply cities and as a source of food.In the state of Rio Grande do Norte, the large reservoirs are intended for the same purpose.The cities settled in the riverbanks, or which have river channels crossing them, face flooding related problems. In the city of Macaíba-RN, flooding occurred systematically during the rainy season, causing great inconvenience to the local population.As product of the collective claim Tabatinga Reservoir in Jundiaí river was built, upstream of the city. Facing this background, this thesis aimed to assess the sócio-environmental quality of this reservoir.To achieve this goal, methodologies pointed to assess water quality along with the aplication of a questionnaire were used aimed to verify the quality of water and to know the perception of the residents from urban and rural área settled near to the reservoir was performed. The results showed the existence of conflicts of residents of rural communities and the presence of the reservoir, while for the city's population, the reservoir is considered not only the right solution to solve flooding in urban areas, but also as economic source for the rural population. Considering the water source assessment, this study concluded that the Tabatinga Reservoir is unfit for human use, due to the presence of metals of toxicological significance with the potential to elicit damage to the genetic material of individuals that use water from this reservoir, leading to cause serious risks to health population.

Coordenação dos sistemas antioxidantes da catalase e da ascorbato peroxidase durante o envelhecimento acelerado de sementes de cártamo e girassol

World consumption of vegetable oils has increased in recent years because of its application in food, chemical, pharmaceutical and, more recently, energy industry. However, oilseeds, which these oils are extracted, have low viability, affecting the cultivation and productivity of these species. The aim of this study was to analyze the effect of aging on the coordination of catalase (CAT) and ascorbate peroxidase (APX) antioxidant systems in safflower and sunflower. . Therefore, seeds were subjected to accelerated aging for 3, 6 and 9 days and grown in moistened paper towel for 72 hours. Additionally, before accelerated aging, sunflower seeds were pretreated by osmopriming with 10 mM ascorbate (ASC) or 3 amino 1,2,4 triazol (3-AT), a specific inhibitor of CAT activitie. The method of artificial aging used was efficient in both species, because it caused a decrease in germination, seedling development and growth, especially in safflower. The aging caused inhibition of CAT activity for both species and to compensate for such inhibition , sunflower increased mRNA expression of this enzyme , while safflower mobilized over the activity of APX. Analysis of the expression of malate synthase and sugar content demonstrated that sunflower seeds consumes lipid reserves in quiescent state, while the safflower is more dependent on carbohydrate. Pretreatment with 3-AT inhibited CAT activity and stimulated the APX, though with ASC acted reverse on these systems. None of the treatments recovered the physiological decline aging. It is concluded that aging change the oilseeds antioxidant metabolism, despite interspecies variations in response to this process, the depletion of the CAT antioxidant system was common. Because of this we propose that the measurement of CAT activity can be used to identify aging seed lots.

Polpa de jambolão (Eugenia jambolana Lam.) fresca e desidratada: características físico-químicas, bioativas e funcionais, efeitos biológicos em Caenorhabditis elegans e uso para produção de frozen yogurt caprino probiótico

This work evaluated the fresh, spray dried (with 10 % of Arabic Gum) and freeze dried jambolan pulp (Eugenia jambolana Lam.) in regard to physicochemical (pH, moisture, water activity, average particle diameter, solubility and color), bioactive [total phenolic content (TPC), monomeric anthocyanin, pronathocyanidin (PA), total elagic acid (TEA), myricetin and cyanidin] and in vitro functionality (antioxidant, antienzymatic and antimicrobial activities]. In addition, the in vivo functionality of jambolan pulp was investigated using the Caenorhabditis elegans model for insulin signaling, longevity and induced neurodegeneration (Alzheimer’s disease and Parkinson’s disease related symptoms). The dried jambolan pulp presented TPC retention (50% to 75%), PA (90% to 98%), TEA (31% to 83%), myricetin (40% to 84%), cyanidin (72% to 84%) and antioxidant activity (15%). The fresh jambolan pulp, the freeze dried pulp and the spray dried jambolan pulp presented high enzymatic inhibitory activity against pancreatic lipase (4,4 to 5,8 mg/mL), alpha-glycosidase (10,3 to 13,8 mg/mL) and alpha-amylase (8,9 to 11,2 mg/mL). They also were active inhibitors against the pathogen S. aureus. The dried jambolan experimental samples were able to increase the expression of several genes linked to the insulin signaling pathways (SIR-2.1, PPTR-1, DAF-16, SOD-3, e CTL) and increased the lifespan in C. elegans (18,07 % - 24,34 %), besides decreasing the amyloid AB1-42 aggregation induced paralysis and MPP+ (1-methyl-4-phenylpyridinium) induced neurodegeneration. Based on that, the jambolan pulp and the spray dried jambolan pulp were further selected for the production of caprine frozen yogurt with the addition of Bifidobacterium animalis subsp. lactis BI-07. The final product were evaluated in regard to their physicochemical (pH, acidity, total solids, protein, total reducing sugars, fat, ashes, overrun, melting test), bioactive (TPC and monomeric anthocyanin, antioxidant activity, probiotic viability and sensory analysis (sensory acceptance). The results showed that samples with probiotic had lowest pH and higher acidity, TPC, anthocyanin and antioxidant activity. It was also observed low overrun (14.2% to 22.6%). vi Samples with probiotic had lower flavor scores. Overall, this research presents the jambolan as a highly functional bioactive-rich fruit with the potential to modulate important biological pathways, extend lifespan and retard the development of neurodegenerative diseases. Jambolan is an underexploited exotic fruit with a high colorant potential and this thesis shows for the first time in the literature important technological, biological and scientific data about this fruit that could be used towards the development of health-oriented food products.

Análise fitoquímica e avaliação das atividades anti-inflamatória, antipeçonhenta e citotóxica de extratos aquosos de Aspidosperma pyrifolium e Ipomoea asarifolia

Envenomation caused by venomous animals, mainly scorpions and snakes, are a serious matter of public health. Tityus serrulatus is considered the most venomous scorpion in South America because of the high level of toxicity of its venom. It is responsible for causing serious accidents, mainly with kids. The species Bothrops jararaca is a serpent that has in its venom a complex mixture of enzyme, peptides and other molecules. The toxins of the venom of B. jararaca induce local and systemic inflammatory responses. The treatment chosen to serious cases of envenomation is the intravenous administration of the specific antivenom. However, the treatment is not always accessible to those residents in rural areas, so that they use medicinal plant extracts as the treatment. In this context, aqueous extracts, fractions and isolated compounds of Aspidosperma pyrifolium (pereiro) and Ipomoea asarifolia (salsa, salsa-brava), used in popular medicine, were studied in this research to evaluate the anti-inflammatory activity in the peritonitis models induced by carrageenan and peritonitis induced by the venom of the T. serrulatus (VTs), and in the local oedema model and inflammatory infiltrate induced by the venom of the B. jararaca, administrated intravenously. The results of the assays of cytotoxicity, using the MTT, showed that the aqueous extracts from the plant species presented low toxicity to the cells that came from the fibroblast of the mouse embryo (3T3).The chemical analysis of the extracts by High Performance Liquid Chromatography revealed the presence of the rutin flavonoid, in A. pyrifoliu, and rutin, clorogenic acid and caffeic acid, in I. asarifolia. Concerning the pharmacological evaluation, the results showed that the pre-treatment using aqueous extracts and fractions reduced the total leukocyte migration to the abdominal cavity in the peritonitis model caused by the carrageenan and in the peritonitis model induced by the T. serulatus venom. Yet, these groups presented anti-oedematous activity, in the local oedema model caused by the venom of the B. jararaca, and reduced the inflammatory infiltrate to the muscle. The serum (anti-arachnid and anti-bothropic) specific to each venom acted inhibiting the inflammatory action of the venoms and were used as control. The compounds identified in the extracts were also tested and, similar to the plant extracts, showed meaningful anti-inflammatory effects, in the tested doses. Thus, these results are indicating the potential anti-inflammatory activity of the plants studied. This is the first research that evaluated the possible biological effects of the A. pyrifolium and I. asarifolia, showing the biological potential that these species have.

Mapeamento cromossômico de DNAs repetitivos com fins biotecnológicos em peixes marinhos e interesse comercial - Rachycentridae e Lutjanidae

The Rachycentron canadum species, commonly known as beijupirá or cobia is the only representative of Rachycentridae family which has been increasingly used in marine fish farming, in intensive cultivation. As advantageous features it has easy adaptation, prolific behavior, early growth in captivity and high commercial value. Additionally, specie of Lutjanidae family (Lutjanus synagris, Lutjanus jocu, Lutjanus analis, Lutjanus alexandrei and Ocyurus chrysurus) represents an important fisheries resource in all areas of its occurrence. In Brazil, the commercial exploitation of Lutjanidae which begun in the 60's and 80's, already has showed a decline in catch volumes. This fact suggests that the snappers must have a conservative management. Despite the economic potential, little is known about the genetic and cytogenetic characteristics of these species, especially with respect to repetitive DNA analysis, which represents the major part of the eukaryotes genome, playing important evolutionary roles in the fish genome. Cytogenetic data is increasingly being used in population studies and biotechnological purposes in fishes. The cytogenetical analyzes were performed using classical methods such as Giemsa staining, C-banding and Ag-NORs, fluorochromes base-specific staining (DAPI and MM) and physical mapping of repetitive sequences among which, telomeric sequences, transposons (Tol2), retrotransposons (Rex1 and Rex3), repetitive DNA (microsatellites and Cot-1) and transcriptionally active regions of the 18S and 5S ribosomal genes and histone (H3 and H2BA) by in situ hybridization with fluorescent probes (FISH). The chromosomal patterns obtained contributed to the organization of repetitive sequences in the genome of the species, as well as karyotypical differentiation. Unusual patterns of histone sequences expansion depict the first occurrence in marine fishes. The obtained data provided subsides to the genetic knowledge of the important fisheries resource represented by the species here analyzed, seeking the marine pisciculture improvement.