Made available in DSpace on 2014-12-17T15:18:10Z (GMT). No. of bitstreams: 1 CarolinaCSO.pdf: 673645 bytes, checksum: 65edb3dd5d260d66ee98b34da41f4185 (MD5) Previous issue date: 2009-02-27 / Gluconacetobacter diazotrophicus ? uma alfa-proteobact?ria Gram-negativa, tolerante a meios ?cidos, fixadora de nitrog?nio atmosf?rico e foi a primeira bact?ria diazotr?fica endof?tica isolada da cana-de-a??car. Por sua vez, Gluconobacter oxydans, tamb?m alfa-proteobact?ria Gram-negativa, possui a capacidade de oxidar incompletamente alco?is e carboidratos. Ambas de interesse biotecnol?gico e industrial, essas bact?rias tiveram seus genomas seq?enciados completamente em 2007. Desta forma, foi de interesse desse trabalho analisar e comparar os genes de reparo do DNA devido sua import?ncia na manuten??o da integridade gen?mica. Sendo assim, as vias de reparo presentes nos dois organismos foram identificadas, utilizando como base uma terceira alfa-proteobact?ria, a Caulobacter crescentus, cujos genes de reparo foram descritos por um trabalho anterior e tamb?m os genes bem estabelecidos para o reparo do DNA em Escherichia coli. Para esse estudo, um banco de dados contendo ort?logos para os genes de reparo de DNA encontrados nos organismos foi criado e an?lises comparativas por similaridade usando o pacote Blast e o software Clustal foram feitas. Este estudo demonstrou que as principais vias de reparo ao DNA reparos por excis?o, reparo direto, reparo recombinacional e reparo pelo sistema SOS est?o presentes nos organismos analisados, demonstrando, na maioria das vezes, boa similaridade com E. coli. Interessantemente, foram encontradas duplica??es g?nicas nos quais uma das c?pias estava presente no cromossomo e a outra, no plasm?deo, como no caso de UvrD, DnaE e Ssb, possivelmente caracterizando eventos de transfer?ncia lateral. Por fim, uma grande novidade foi a identifica??o de ort?logos para RecB em G. diazotrophicus e G. oxydans e de ort?logos duplicados de RecD em G. diazotrophicus. At? o momento, n?o havia sido relatada a presen?a de membros da via de inicia??o RecBCD do reparo recombinacional em alfaproteobact?rias

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-02-21T21:54:36Z No. of bitstreams: 1 GabrielleMacedoPereira_TESE.pdf: 7916343 bytes, checksum: 47f4f16d292c186e81c42187080b1343 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-02-23T21:04:51Z (GMT) No. of bitstreams: 1 GabrielleMacedoPereira_TESE.pdf: 7916343 bytes, checksum: 47f4f16d292c186e81c42187080b1343 (MD5) / Made available in DSpace on 2018-02-23T21:04:51Z (GMT). No. of bitstreams: 1 GabrielleMacedoPereira_TESE.pdf: 7916343 bytes, checksum: 47f4f16d292c186e81c42187080b1343 (MD5) Previous issue date: 2017-08-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O bioma Caatinga ? exclusivamente brasileiro, marcado pelo clima semi?rido. Este bioma se destaca pelo alto ?ndice de endemismo e desconhecimento cient?fico ao seu respeito. Para sobreviver neste ambiente, as plantas desenvolveram mecanismos intr?nsecos de percep??o dos sinais ambientais externos e suas respectivas respostas metab?licas. Dentre as esp?cies de ocorr?ncia no bioma Caatinga, como fonte de alcaloides bioativos, destaca-se Erythroxylum pungens, que apresenta potencial pouco explorado. Assim, o objetivo deste trabalho ? investigar o proteoma total e o perfil metab?lico de E. pungens, aliado ? bioprospec??o de alcaloides trop?nicos com potencial citot?xico. Para isso, no cap?tulo 1 identificou-se, por espectrometria de massas, sete alcaloides trop?nicos conhecidos; 3-(2-metilbutiriloxi)tropan-6, 7-diol como tamb?m 3-(2-metilbutiriloxi)nortropan-6, 7-diol foram isolados e caracterizado por RMN 1D e 2D pela primeira vez. N,N-Dimetil-1-H-indol-3-etanamina foi isolado e caracterizado a partir de ra?zes de E. pungens. Al?m disso, verificou-se que alcaloides isolados de E. pungens, frente a cinco linhagens celulares, apontaram potencial seletivo de 3-(2-metilbutiriloxi)tropan-6, 7-diol contra c?lulas de tumorais de pr?stata. Para melhor conhecer a esp?cie e, com vistas na produ??o de alcaloides trop?nicos, trabalhou-se o proteoma e perfil alcalo?dico no cap?tulo 2. Dessa forma foi poss?vel identificar 1746 prote?nas nas folhas, 1779 no caule e 1026 na raiz de E. pungens. Atrav?s da recupera??o dos termos no Gene Ontology, observou-se os processos nos quais estas prote?nas est?o envolvidas, verificando o mesmo perfil de processos para os tr?s ?rg?os de E. pungens, com destaque para respostas a estresses bi?ticos, abi?ticos, qu?micos, al?m das respostas ao estresse oxidativo. Foram observados n?veis significativos de prote?nas envolvidas na fotoss?ntese das folhas de E. pungens, fotorrespira??o, al?m prote?nas relacionadas ?s respostas ao estresse: prote?nas da fam?lia 14-3-3, peroxidases, catalases, super?xido-dismutase e prote?nas de choque t?rmico, que tamb?m podem atuar no turnover prote?co. Ainda foram identificadas cinco prote?nas hom?logas a tropinona redutase na folha, e produ??o de alcaloide em todos os ?rg?os. No cap?tulo 3, por meio do experimento com esp?cimes de E. pungens submetidas a estresse h?drico por suspens?o de rega em casa de vegeta??o, notou-se que esta esp?cie investe no aumento de prote?nas relacionadas ?s respostas ao estresse h?drico, aumenta significativamente o teor de prolina livre no citosol. Ademais, mat?m a produ??o de alcaloides que parecem estar mais relacionado a fase de desenvolvimento da planta. Com isso, os resultados deste trabalho busca contribuir para a fitoqu?mica e fisiologia de E. pungens, assim como promove o uso racional da esp?cie, e consequentemente a conserva??o do bioma Caatinga. / Caatinga biome is exclusive brazilian, marked by semi-arid climate. This biome stands out for the high endemism index and scientific unfamiliarity.To survive in this environment with peculiar edaphoclimatic conditions, plants show evolution of intrinsic mechanisms of external environmental signals perception and their respectives metabolic responses. Among the occurrence genera in Caatinga biome, as bioactive alkaloids source, the Erythroxylum pungens species, which has unexplored potential, is outstanding. Thus, the objective of this work is to investigate total proteome and metabolic fingerprint of E. pungens, together with tropane alkaloids bioprospection with cytotoxic potential. For this, in chapter 1 it was possible to identify, by mass spectrometry, seven known tropane alkaloids; 3-(2-methylbutyryloxy)tropan-6, 7-diol as well as 3-(2-methylbutyryloxy)tropan-6, 7-diol were isolated and characterized by 1D and 2D NMR for the first time. N,N-Dimethyl-1-H-indol-3-ethanamine was isolated and characterized from roots of E. punctures. In addition, it was found that alkaloids isolated from E. pungens, against five cell lines, indicated selective potential of 3-(2-methylbutyryloxy)tropan-6, 7-diol against prostate tumor cells. In chapter 2, it was possible to identify 1746 proteins in the leaves, 1779 in the stem and 1026 in the root of E. pungens. Through the recovery of the GO terms, it was possible to evaluate processes in which these proteins are involved, verifying the same process profile for three organs of E. pungens, with emphasis on biotic, abiotic, chemical stress responses, as well as responses to oxidative stress. Were observed significant proteins levels involved in photosynthesis of E. pungens leaves, photorespiration, as well as many proteins related to stress responses: proteins from 14-3-3 family, peroxidases, catalases, superoxide dismutase and shock proteins, which can also act on protein turnover. Were still identificated five proteins homologous to tropinone reductase, and alkaloid production in all organs suggesting that even under chronic stress conditions, E. pungens maintains the production of these metabolites. In chapter 3, through specimens submission experiment of E. pungens to water stress by irrigation suspension in greenhouse, it was observed that this species invest in the increase of proteins related to the responses to water stress, it significantly increases free proline content in cytosol which functions as an important osmoprotector. Furthermore, maintains alkaloids production that appear to be more related to development stage of the plant than stress condition in which it is inserted. The results of this work seek to contribute to knowledge of E. pungens phytochemistry and physiology, as well as to promote the species and consequently conservation of Caatinga biome.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-03-21T11:48:14Z No. of bitstreams: 1 JonathasDiegoLimaSantos_TESE.pdf: 8935214 bytes, checksum: 86433d89608ea11b99ac2a3911f14149 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-03-23T11:23:51Z (GMT) No. of bitstreams: 1 JonathasDiegoLimaSantos_TESE.pdf: 8935214 bytes, checksum: 86433d89608ea11b99ac2a3911f14149 (MD5) / Made available in DSpace on 2018-03-23T11:23:51Z (GMT). No. of bitstreams: 1 JonathasDiegoLimaSantos_TESE.pdf: 8935214 bytes, checksum: 86433d89608ea11b99ac2a3911f14149 (MD5) Previous issue date: 2017-12-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Chromobcterium violaceum (C. violaceum) ? uma bact?ria Gram-negativa, encontrada em regi?es tropicais e subtropicais, considerada organismo modelo de vida livre. Alguns estudos prote?micos realizados com esta bact?ria demonstraram sua capacidade de adapta??o a desafios ambientais, como alta concentra??o de ferro e exposi??o ao estresse oxidativo. No entanto, nenhum estudo foi feito com esta esp?cie submetida ? microgravidade simulada (MGS), ou seja, em condi??es em que a gravidade ? artificialmente reduzida para menos de 1xg. Estudos MGS podem ser importantes para entender as modula??es em n?vel molecular sofridas pelos organismos vivos. Portanto, o objetivo deste estudo foi caracterizar a resposta de C. violaceum, como organismo modelo de vida livre, cultivado em MGS, usando t?cnicas de prote?mica para entender como a bact?ria responde a esse estresse. A MGS foi conseguida por meio da rota??o do vessel ao redor do eixo horizontal perpendicular ao vetor gravitacional nos sistemas de cultura de c?lulas rotativas - Rotating Cell Culture Systems ? (RCCS4). MGS foi conduzido a uma velocidade de 40 rpm por um per?odo de 12 horas para obter a curva de crescimento a cada 2 horas. Prote?nas totais foram extra?das de amostras de cultura de c?lulas bacteriana coletada ?s 5 e 12 horas, correspondendo as fases exponenciais inicial (MG5) e tardia (MG12), respectivamente, tendo a curva de crescimento como refer?ncia. Ap?s a tripsiniza??o, as amostras foram analisadas em espectr?metro de massas Q-TOF. Como resultado um total de 212 prote?nas durante ?s 5h de crescimento e 192 ?s 12h foram detectadas, das quais 144 delas foram comuns em ambos os per?odos. No proteoma de C. violaceum obitido em 5h de crescimento 195 prote?nas foram identificadas em gravidade normal (GN5) e 155 prote?nas foram identificadas em MG5, sendo 18 upreguladas, 19 downreguladas e 17 expressas somente na condi??o de MG5. No proteoma obtido em 12h de crescimento, 165 prote?nas foram identificadas em gravidade normal (GN12) e 173 em MG12, das quais, 17 foram upreguladas, 22 downreguladas e 28 expressas somente na condi??o de MG12 Al?m disso, foi poss?vel identificar 25 prote?nas com fun??o desconhecida em MG5 e MG12. Utilizando ferramentas computacionais foi poss?vel construir redes de intera??es prote?na-prote?na (PPI) e as sub-redes contendo grupo de prote?nas com fun??es correlacionadas, analisar vias metab?licas, processos biol?gicos, contexto gen?mico e busca por dom?nios conservados e sequ?ncias hom?logas de prote?nas com fun??o desconhecida. Como resultado, identificamos sub-redes relacionadas com a bioss?ntese de prote?nas, regula??o da transcri??o e tradu??o, resposta ao estresse e metabolismo energ?tico, conclu?mos que as respostas celulares associadas ? express?o diferencial induzida pela MGS levaram ? diminui??o do crescimento de C. violaceum, acompanhado pela regula??o negativa de prote?nas relacionadas com processos transcricionais, traducionais e libera??o de energia por vias aer?bicas e pela regula??o positiva de prote?nas envolvidas no estresse oxidativo, na via anaer?bica e na sobreviv?ncia celular. / Chromobcterium violaceum (C.violaceum) is a Gram-negative bacteria, which has been found at tropical and subtropical regions, considered a free living model organism. Some proteomic studies performed with this bacterium have demonstrated its ability to adapt to environmental challenges such as high iron concentration and oxidative stress exposure. However, no study was made with this specie submitted to simulated microgravity (SMG), that is, under conditions in which the gravity is artificially reduced to less than 1xg. MGS studies may be important in understanding the molecular-level modulations undergone by living organisms. Therefore, the aim of this study was to characterize the response of C. violaceum, as free-living model organism, cultured at MGS, using proteomics techniques in order to understand how this bacterium response to this stress. The SMG was achieved by rotating the vessel around the horizontal axis perpendicular to the gravitational vector in Rotating Cell Culture Systems (RCCS4). SMG was conducted at a speed of 40 rpm for a period of 12 hours to obtain the growth curve every 2 hours. Total protein extraction was made in two times: 5 and 12 hours, corresponding to early (MG5) and late (MG12) exponential phase, respectively, taking the growth curve as a reference. After trypsinization, samples were analyzed with Q-TOF mass spectrometer. As a result a total of 212 proteins during 5h of growth and 192 at 12h were detected, of which 144 of them were common in both periods. We detected 155 proteins during MG5, from which 18 proteins were upregulated, 19 down-regulated and 17 proteins were exclusive when compared to GN5. In the proteome obtained in 12h of growth, 165 proteins were identified in normal gravity (GN12) and 173 in MG12, of which, 17 were upregulated, 22 were downregulated and 28 expressed only in MG12 condition. In addition, it was possible to identify 25 proteins with unknown function in MG5 and MG12. Using computational tools it was possible to construct networks of protein-protein interactions (PPI) and sub-networks containing group of proteins with correlated functions, to analyze metabolic pathways, biological processes, genomic context and search for conserved domains and homologous sequences of proteins with unknown function . As a result, we identified sub-networks related to protein biosynthesis, transcription regulation and translation, stress response and energetic metabolism, we conclude that the cellular responses associated with differential expression induced by MGS led to a decrease in the growth of C. violaceum, accompanied by the negative regulation of proteins related to transcriptional, translational and energy release by aerobic pathways and by the positive regulation of proteins involved in oxidative stress, anaerobic pathway and cell survival.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-04-02T15:02:41Z No. of bitstreams: 1 AdrianaMarinaESilvaParente_DISSERT.pdf: 1386035 bytes, checksum: 87910526a122893f94a9f3045c8c9c4f (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-04-05T13:08:04Z (GMT) No. of bitstreams: 1 AdrianaMarinaESilvaParente_DISSERT.pdf: 1386035 bytes, checksum: 87910526a122893f94a9f3045c8c9c4f (MD5) / Made available in DSpace on 2018-04-05T13:08:05Z (GMT). No. of bitstreams: 1 AdrianaMarinaESilvaParente_DISSERT.pdf: 1386035 bytes, checksum: 87910526a122893f94a9f3045c8c9c4f (MD5) Previous issue date: 2017-11-28 / Tityus stigmurus corresponde ? esp?cie de escorpi?o predominante na regi?o Nordeste do Brasil, sendo considerado um dos principais causadores de acidentes escorpi?nicos. Na pe?onha do T. stigmurus, que ? composta de uma mistura complexa de mol?culas de alta e baixa massa molecular, foi identificado e caracterizado um pept?deo antimicrobiano denominado Stigmurina (FFSLIPSLVGGLISAFK-NH2). Pept?deos antimicrobianos s?o pequenas mol?culas consideradas a primeira linha de defesa contra micro-organismos, apresentando amplo espectro de a??o antimicrobiana. A literatura reporta tamb?m a atividade desses pept?deos contra c?lulas cancer?genas. Sequ?ncias nativas de pept?deos bioativos como prot?tipo para a obten??o de novas mol?culas t?m sido utilizadas com o intuito de potencializar a sua atividade e reduzir a toxicidade. Nesse contexto, realizou-se a caracteriza??o estrutural in silico e por dicro?smo circular, bem como a avalia??o da atividade antimicrobiana, antiparasit?ria, antiproliferativa e hemol?tica de dois pept?deos an?logos obtidos ? partir da Stigmurina, denominados StigA3 e StigA4. A an?lise da conforma??o tridimensional in silico do StigA3 e StigA4 demonstrou uma estrutura helicoidal, sendo este resultado confirmado por dicro?smo circular. O aumento da carga superficial e do momento hidrof?bico de ambos os pept?deos an?logos quando comparado com a Stigmurina resultaram na potencializa??o da atividade antimicrobiana e antiparasit?ria. Os pept?deos StigA3 e StigA4 apresentaram a??o antiproliferativa semelhante ao pept?deo nativo, com exce??o para a c?lula normal, para qual os pept?deos an?logos se mostraram menos t?xicos. Portanto, estes resultados indicam uma potencial aplica??o terap?utica destes pept?deos an?logos, demonstrando a efici?ncia do desenho racional de f?rmacos para a obten??o de novos agentes antiproliferativos e anti-infecciosos. / Tityus stigmurus it?s the predominant scorpion specie in Northeast region from Brazil, it?s considered one of the main cause of scorpion accidents in this region. In T. stigmurus venom, which is a complex mixture of high and low molecular mass molecules, was identified an antimicrobial peptide denominated Stigmurin. Antimicrobial peptides are small molecules considered the first line of defense against microorganisms, they show broad spectrum action. Many authors have proved that these peptides can also be effective on cancerous cells. Mutation in these molecules sequence has been held aiming the increase in the activity maintaining its low toxicity. Therefore, it is proposed the characterization of the structure in silico and by circular dichroism, as well the antimicrobial, antiparasitic, anti-proliferative and hemolytic activities of two analog peptides from Stigmurin, denominated StigA3 and StigA4. We performed an in silico analysis for these peptides, where we were able to observe ?-helix structure, which was confirmed by circular dichroism. We were also able to find that the analog peptides net charge and hydrophobic moment were higher than Sitgmurin?s, which can explain the increase in antimicrobial and antiparasitic activity. The peptides StigA3 e StigA4 showed activity on cancerous cells similiar to the native peptide, except when tested on a normal cell we were able to find that they?re less toxic. Therefore, these results indicate a potential biotechnological application for the analogs peptides, even as prototype to new therapeutic agents.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-04-02T15:02:43Z No. of bitstreams: 1 GustavoHenriqueOliveiraCavalcante_DISSERT.pdf: 14332647 bytes, checksum: 07f54cbd966599c411e90b07dbb86ba8 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-04-05T13:57:34Z (GMT) No. of bitstreams: 1 GustavoHenriqueOliveiraCavalcante_DISSERT.pdf: 14332647 bytes, checksum: 07f54cbd966599c411e90b07dbb86ba8 (MD5) / Made available in DSpace on 2018-04-05T13:57:34Z (GMT). No. of bitstreams: 1 GustavoHenriqueOliveiraCavalcante_DISSERT.pdf: 14332647 bytes, checksum: 07f54cbd966599c411e90b07dbb86ba8 (MD5) Previous issue date: 2018-02-23 / O papilomav?rus humano (HPV) ? um pequeno v?rus de DNA de dupla fita circular, caracterizado como um dos mais comuns agentes sexualmente transmiss?veis no mundo, cuja detec??o e genotipagem acuradas s? s?o poss?veis por meio de t?cnicas de biologia molecular. Diferentes propriedades biol?gicas t?m sido reportadas dentre os mais de 170 tipos j? caracterizados, de maneira que um grupo particular de HPVs est? fortemente relacionado a infec??es persistentes, les?es intraepiteliais de diferentes graus e progress?o para c?nceres tais como cervical, anal, vulvar, vaginal, orofar?ngeo e de p?nis. No presente trabalho foram realizadas an?lises de variabilidade gen?tica e evolu??o molecular nos genomas dos principais HPVs de import?ncia cl?nica. Reconstru??es filogen?ticas e an?lises dos perfis de assinatura gen?tica nos genomas de cada gen?tipo sugeriram a presen?a de subgrupos de HPVs definidos por diferen?as nas sequ?ncias dos genes E1, E6, L1 e L2. Testes de evolu??o em n?vel de DNA revelaram uma atua??o mais forte da sele??o natural em c?dons espec?ficos, mais frequentemente nos genes E1, E2, L1 e L2. A partir dos dados obtidos nas an?lises de variabilidade, foi desenhado um novo conjunto de primers para a detec??o e genotipagem dos HPVs de import?ncia cl?nica por meio da t?cnica de rea??o em cadeia da polimerase (PCR). O gene E1 foi escolhido como alvo molecular devido a presen?a de uma regi?o conservada com tamanho vari?vel entre gen?tipos. O sistema proposto teve sua efici?ncia avaliada in vitro e foi comparado ao protocolo de PCR mais utilizado para detec??o do HPV em amostras cl?nicas. Utilizando a amplifica??o de ?cido nucleico de forma semianinhada (seminested), o sistema proposto foi capaz de detectar com boa sensibilidade alguns dos principais HPVs de alto risco oncog?nico e mostrou melhor especificidade em rela??o aos primers gen?ricos GP5+/6+, mesmo aplicando uma temperatura de anelamento consideravelmente maior. A an?lise do tamanho dos fragmentos amplificados usando a separa??o por eletroforese em gel de agarose pode favorecer a identifica??o do tipo de HPV presente nas amostras, permitindo a discrimina??o entre aqueles mais prevalentes na popula??o e a redu??o do tempo e do custo necess?rios para a identifica??o do agente. Opcionalmente, a separa??o dos produtos em matrizes de alta resolu??o e o sequenciamento direto podem ser usados para a tipagem, possibilitando a identifica??o de uma ampla variedade de gen?tipos de HPV descritos. / Human papillomavirus (HPV) is a small circular double-stranded DNA virus, characterized as one of the most common sexually transmitted agents in the world, whose accurate detection and genotyping is only possible through molecular biology techniques. Different biological properties have been reported among the more than 170 types already characterized, so that a particular group of HPVs is strongly related to persistent infections, intraepithelial lesions of different degrees and progression to cancers such as cervical, anal, vulvar, vaginal, oropharyngeal and penis. In the present work, analyzes of genetic variability and molecular evolution were performed in the genomes of the main clinically important HPVs. Phylogenetic reconstructions and analyzes of genetic signature profiles in the genomes of each genotype suggested the presence of subgroups of HPVs defined by differences in the E1, E6, L1 and L2 gene sequences. Evolution tests at DNA level have shown a stronger acting of natural selection at specific codons, more often in the E1, E2, L1 and L2 genes. From the data obtained in the analyzes of variability, a new set of primers was designed for the detection and genotyping of HPVs of clinical importance by polymerase chain reaction (PCR) technique. E1 gene was chosen as the molecular target due to the presence of a conserved region of variable size among genotypes. The proposed system had its efficiency evaluated in vitro and was compared to the most used PCR protocol for HPV detection in clinical samples. Using the seminested nucleic acid amplification, the proposed system was able to detect some of the major oncogenic HPVs with good sensitivity and showed a better specificity than the generic primers GP5+/6+, even applying a considerably higher annealing temperature. The analysis of the size of the amplified fragments using agarose gel electrophoresis may favor the identification of the HPV type present in the samples, allowing the discrimination between those more prevalent in the population and the reduction of the time and cost necessary for the identification of the agent. Optionally, the separation of products into high resolution matrices and direct sequencing can be used for typing, enabling the identification of a wide variety of HPV genotypes described.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2018-04-02T15:02:43Z No. of bitstreams: 1 LucianaFentanesMouraDeMelo_DISSERT.pdf: 1510669 bytes, checksum: f402172547ca541fafef3500e5ce47a8 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2018-04-05T14:12:02Z (GMT) No. of bitstreams: 1 LucianaFentanesMouraDeMelo_DISSERT.pdf: 1510669 bytes, checksum: f402172547ca541fafef3500e5ce47a8 (MD5) / Made available in DSpace on 2018-04-05T14:12:02Z (GMT). No. of bitstreams: 1 LucianaFentanesMouraDeMelo_DISSERT.pdf: 1510669 bytes, checksum: f402172547ca541fafef3500e5ce47a8 (MD5) Previous issue date: 2018-03-08 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O uso de plantas medicinais ? uma pr?tica comum da humanidade desde a antiguidade. Tal conhecimento baseado na experi?ncia tem sido repassado por muitas comunidades e grupos ?tnicos, e serve como ponto de partida para estudos bioqu?micos e farmacol?gicos dos princ?pios ativos presentes nas plantas. Para isto, neste trabalho foram avaliados o potencial antioxidante e tamb?m biol?gico, utilizando sistemas in vitro e in vivo dos diferentes extratos obtidos de folhas da planta Coccoloba alnifolia. Foram obtidos inicialmente seis extratos: hexano (EH), clorof?rmio (EC), etan?lico (EE), metan?lico (EM), ?gua final (EAF) e aquoso (EA) a partir das folhas frescas da planta. Posteriormente, foram avaliados o teor de a??car, prote?nas e compostos fen?licos. Os resultados obtidos na triagem fitoqu?mica sugerem a presen?a de fen?is, flavonoides, saponinas, n?cleo triterp?nicos e esteroidais e n?cleo esteroidal insaturado, j? com a metodologia de cromatografia em camada delgada (CCD), foi observado uma abund?ncia de flavonoides. Foram realizados quatro ensaios: Teste do Poder Redutor, Quela??o de ?ons Metais, Quela??o de Cobre e Atividade Sequestradora de Radical Hidroxila. Estes ensaios permitiram verificar que os extratos apresentaram atividade antioxidante nas concentra??es avaliadas de 100, 250 e 500 ?g/mL. Foi verificado principalmente para os extratos EE, EM e EA as atividades antioxidante nos ensaios poder redutor e sequestro do radical hidroxila. A correla??o de Pearson mostrou que estes dados est?o associados aos compostos fen?licos e a??cares. E com o modelo das linhagens celulares foi observado que a maioria dos extratos avaliados neste trabalho n?o mostrou-se citot?xico para as linhagens celulares. Os extratos tamb?m n?o inibiram o crescimento da levedura Candida parapsilosis. Os extratos obtidos da folha de C. alnifolia apresenta um potencial antioxidante interessante, e os resultados obtidos tanto nos modelos in vitro, como as linhagens celulares, refor?am que s?o necess?rios mais estudos para compreender o potencial destes extratos in vivo e identificar quais s?o as biomol?culas associadas com estas atividades e seus respectivos potenciais biotecnol?gicos. / The use of medicinal plants has been a common practice in folk medicine. This knowledge has been based on the experience that it has been passed though many generations. However, many times this knowledge do not have any scientific base. The molecules responsive for these activities are the secondary metabolic produce as a response to environmental conditions (biotic or abiotic stress). The Mata atl?ntica biome has a rich flora, which needs to be studied for its biological activities, and its potential to identify new biomolecules. In order to do this, six extracts (HE, CE, EE, ME, WEE e WE) were obtained from Coccoloba alnifolia leaves and then it was evaluated the antioxidant and the biological potential using different in vitro and in vivo systems. First, it was measured the sugar content, proteins and phenolic compounds. Based on these results, qualitative experiments were carried out to determine the secondary metabolites present in these extracts. It was detected the presence of phenols, flavonoids, saponins, triterpenic, and steroidal nuclei and unsaturated steroidal nuclei. After that, it was evaluated the antioxidant potential by four in vitro assay: Redution Power test, Metal Ion Chelation, Copper Chelation and Hydroxyl Radical Scavenging Activity. These assays allowed us verify that the extracts presented antioxidant activity at extract concentrations: 100, 250 and 500 ?g/mL. It was observed the ethanolic, methanolic and aqueous extracts had activity in reducing power and scavenging of the hydroxyl radical. The Pearson?s correlation showed that these results may be associated to the presence of phenolic compounds and sugars. The third in vitro model used in this work was cell culture. When it was used the 3T3 lineage it was observed that these extracts were not cytotoxic. However, it was not observed any antiproliferative activity in tumor cell lineages. In sumary, the data obtained here showed that using in vitro assays that these extracts in special ethanolic and aqueous had an interesting antioxidant potential and it will be important to go further in this research.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-01-10T15:32:21Z No. of bitstreams: 1 KarlaSamaraRochaSoares_TESE.pdf: 2883085 bytes, checksum: 02667bee72d9ca986a7f49f759803663 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-01-11T15:56:53Z (GMT) No. of bitstreams: 1 KarlaSamaraRochaSoares_TESE.pdf: 2883085 bytes, checksum: 02667bee72d9ca986a7f49f759803663 (MD5) / Made available in DSpace on 2017-01-11T15:56:53Z (GMT). No. of bitstreams: 1 KarlaSamaraRochaSoares_TESE.pdf: 2883085 bytes, checksum: 02667bee72d9ca986a7f49f759803663 (MD5) Previous issue date: 2016-05-31 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Acidentes com animais pe?onhentos representam um s?rio problema de sa?de p?blica em diversos pa?ses, destacando-se os acidentes of?dicos e escorpi?nicos. No Brasil, as serpentes do g?nero Bothrops e os escorpi?es pertencentes ao g?nero Tityus, s?o as de maior relev?ncia cl?nica. O tratamento para o envenenamento consiste em administra??o de soros antiof?dico e antiescorpi?nico. Vacinas que utilizam ant?genos puros e vias de administra??o alternativa requerem o uso de adjuvantes potentes e um sistema de entrega de ant?geno eficaz. Nanossistemas v?m sendo investigados como sistemas de libera??o para macromol?culas terap?uticas. A quitosana, devido as suas propriedades, tem sido extensivamente investigada na formula??o de nanocarreadores, particularmente de genes e prote?nas. Este estudo teve como objetivo a obten??o de nanopart?culas de quitosana (CNP) com base na gelifica??o i?nica para carrear prote?nas/pept?deos terap?uticos utilizados na imunoterapia e avalia??o do potencial imunoadjuvante dessas nanopart?culas na produ??o de soros antivenenos. CNP foram obtidas por gelifica??o i?nica, com tamanho m?dio de 160 nm, caracterizadas f?sico-quimicamente e o perfil de libera??o avaliado, demonstrando se tratar de um sistema de libera??o modificado. A estabilidade dos sistemas foi avaliada por 7 semanas, observando-se uma maior estabilidade dos sistemas associados aos venenos. Animais experimentais foram imunizados durante 6 semanas com 100 ?L de veneno das serpentes atrav?s inje??es subcut?neas, em diferentes concentra??es (5,0 ou 10,0%), encapsuladas em CNP ou associados ao hidr?xido de alum?nio (HA). Os resultados demonstram que os t?tulos de anticorpos obtidos para os animais vacinados com os nanossistemas foram equivalentes ou maiores aos obtidos para os animais vacinados com o HA, com a vantagem desses serem menos inflamat?rios que o HA, exigindo uma menor quantidade de ant?geno a ser administrada, por se tratar de um sistema de liberta??o modificada, revelando a obten??o de um novo sistema nanoparticulado com potencial aplica??o na soroterapia. / Accidents with venomous animals represent a serious public health problem in many countries of the world, with emphasis to snake and scorpion accidents. In Brazil, Bothrops snakes are the most clinically relevant. Regarding scorpions, the genus Tityus includes the species of higher medical importance. Treatment for envenomation consists mainly in the administration of antivenom sera. Vaccines using pure antigens in alternative administration routes require the use of potent adjuvants and an effective antigen delivery system. Nanosystems are being investigated as delivery systems for therapeutic macromolecules. Chitosan, due to its properties, has been extensively investigated in nanocarriers formulations, particularly for genes and proteins. This study aimed to obtain chitosan nanoparticles (CNP) based on ionic gelation for the delivery of therapeutic proteins/peptides used in immunotherapy, as well as to evaluate the immunoadjuvant potential of these nanoparticles in the production of antivenom serums. CNP were obtained by ionic gelation, with an average size of 160 nm. Physicochemical characterization and evaluation of the release profile demonstrated that it is a modified release system. The stability of different systems was evaluated for 7 weeks, observing a greater stability of the systems associated with venoms. Experimental animals were immunized subcutaneously for 6 weeks with 100 uL snake venoms at different concentrations (5.0 or 10.0%), either encapsulated in CNP or associated with aluminum hydroxide (AH). The results demonstrate that the antibody titers observed for animals vaccinated with the studied nanosystems were equivalent or higher than those observed for animals vaccinated with HA. Further advantages of the nanosystems were to be less inflammatory, and to require smaller amounts of antigen to be administered, because it is a modified-release system, with potential application in anti-venom serum therapy.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-01-27T13:32:41Z No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-01-31T13:32:42Z (GMT) No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) / Made available in DSpace on 2017-01-31T13:32:42Z (GMT). No. of bitstreams: 1 FabianaLimaBezerra_TESE.pdf: 4985146 bytes, checksum: ce1c1005e6775f052912c875b1d39910 (MD5) Previous issue date: 2016-06-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A dengue ? a mais importante arbovirose que afeta o homem e constitui-se em um grave problema de sa?de p?blica. A doen?a ? end?mica em regi?es tropicais e subtropicais, abrangendo mais de 100 pa?ses, correspondendo ? metade da popula??o mundial. As epidemias s?o recorrentes e estima-se que os v?rus da dengue causem 390 milh?es de infec??es a cada ano, sendo uma importante causa de morbidade. Apesar desse cen?rio, atualmente, ainda n?o se disp?e de um antiviral contra a dengue. Neste contexto, diversos estudos relatam a atividade antiviral dos polissacar?deos sulfatados obtidos das algas marinhas contra v?rus envelopados, cuja a??o est? associada ? interfer?ncia nas etapas iniciais do processo de infec??o viral. Neste estudo, avaliou-se a potencial atividade antiviral de extratos brutos ricos em polissacar?deos sulfatados obtidos das algas marrons Dictyota menstrualis (EBDMens) e Dictyota mertensii (EBDM) e das algas verdes Codium isthmocladum (EBCI) e Caulerpa sertularioides (EBCS) contra o v?rus dengue 2 (DENV-2), em linhagem de c?lulas Vero, usando diferentes estrat?gias metodol?gicas (tratamento simult?neo, tratamento p?s-infec??o, pr?-tratamento da c?lula, pr?tratamento do v?rus, adsor??o, p?s-adsor??o e penetra??o). A citotoxicidade dos extratos foi analisada pelo ensaio de redu??o do MTT. O estudo da atividade antiviral foi determinado pela quantifica??o do n?mero de c?pias do RNA viral por RT-qPCR ap?s 120 h de infec??o. Nenhum extrato exibiu toxicidade para as c?lulas Vero no ensaio de redu??o do MTT na concentra??o de 100 ?g/mL. Todos os extratos exibiram atividade antiviral quando adicionados durante os primeiros 90 minutos de infec??o, demonstrando redu??o significativa no n?mero de c?pias do RNA viral ap?s 120 horas. Os extratos EBDMens e EBDM foram mais eficazes nos ensaios de pr?-tratamento da c?lula e do v?rus, e o primeiro exerceu atividade antiviral superior ao EBDM durante a adsor??o viral. Os extratos EBCI e EBCS apresentaram atividade antiviral semelhante no ensaio de pr?-tratamento da c?lula. O EBCI foi mais eficaz na redu??o da adsor??o do DENV-2 em c?lulas Vero. Por?m, o EBCS se mostrou mais eficiente nos ensaios de pr?-tratamento do v?rus e penetra??o. Dentre os extratos avaliados, o EBCS parece ser o mais efetivo no combate ao DENV-2. Estes resultados revelam o potencial desses extratos ricos em polissacar?deos sulfatados como compostos com a??o antiviral, sugerindo que eles atuam nos est?gios iniciais da infec??o por DENV-2. / Dengue is considered the most important human arboviruses, and is a serious public health problem. This disease is endemic in tropical and subtropical regions, reaching more than 100 countries, which means the half of world population, and those epidemies have been appellant. It is estimated that Dengue virus could cause 390 millions of infections each year, and is an important cause of morbidity. Although this scenario, actually doesn?t exist a dengue antivirals still. Sorts of studies have showed antivirus activity of seaweed sulfated polysaccharides against enveloped viruses, which action seems to be associated to initials steps of infection process. In this study, was evaluated the potential antiviral activity of crude extracts rich in sulfated polysaccharides obtained from the brown algae Dyctiota menstrualis (EBDMens) and Dyctiota mertensii (EBDM), and green algae Codium isthmocladium (EBCl) and Caulerpa sertularioides (EBCS), tested against Dengue virus 2 (DENV-2) in Vero cell line, by using different methodological strategies (simultaneous treatment, post-infection treatment, cell pre-treatment, virus pre-treatment, adsorption, post-adsorption and penetration). The extracts cytotoxicity was evaluated by MTT reduction assay. The study of antiviral activity was determinated by quantifying viral RNA load using RT-qPCR, after 120 hours of infection. None extract has showed toxicity against Vero Cells in MTT reduction assay with 100 ?g/mL concentration. All extracts have shown antiviral activity when added during the first 90 minutes of infection, and a significant reduction of viral RNA number of copies after 120 hours. The EBDMens and EBDM extracts were more efficient to cell and virus pretreatment assays, and the first has showed higher antiviral activity than EBDM during viral adsorption. The EBCI and EBCS extracts have presented antiviral activity very similar in cell pre-treatment assay. EBCI was more efficient in the reduction of DENV-2 adsorption in Vero cell. However, EBCS has showed more efficient in cell pre-treatment and penetration assays. Between the extracts evaluated, EBCS seems to be more effective against DENV-2. These results have showed potential action of extracts rich in sulfated polysaccharides with antivirus activity, suggesting that they act in the initials steps of infection process of DENV-2.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-02-10T16:42:15Z No. of bitstreams: 1 CristianeSantosSanzioGurgel_TESE.pdf: 4190612 bytes, checksum: 773e7fec68bad460acc07eb6d8f76c81 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-02-10T21:19:07Z (GMT) No. of bitstreams: 1 CristianeSantosSanzioGurgel_TESE.pdf: 4190612 bytes, checksum: 773e7fec68bad460acc07eb6d8f76c81 (MD5) / Made available in DSpace on 2017-02-10T21:19:07Z (GMT). No. of bitstreams: 1 CristianeSantosSanzioGurgel_TESE.pdf: 4190612 bytes, checksum: 773e7fec68bad460acc07eb6d8f76c81 (MD5) Previous issue date: 2016-06-28 / Este estudo objetivou avaliar o estado de vitamina A de pu?rperas atendidas durante o parto na cidade de Natal/RN. Foram recrutadas no estudo 793 mulheres, 60.1% (n=485) da rede p?blica e 39.0% (n=310) a rede privada. Amostras de soro (n=619) e leite colostro (n=656) foram coletadas em ambiente hospitalar, ap?s jejum noturno. O leite maduro (n=154) foi coletado trinta dias ap?s o parto, em visita domiciliar. Os indicadores bioqu?micos (retinol no soro e leite materno) foram avaliados por local de moradia (capital vs interior) e por rede de atendimento em sa?de (p?blico vs privado). O consumo de vitamina A foi avaliado referente ao ?ltimo trimestre gestacional. Para avaliar as diferentes formas de suplementa??o materna com vitamina A e suas associa??es com os indicadores bioqu?micos (soro e leite materno) formaram-se subgrupos baseados nas suplementa??es que ocorreram durante a gesta??o: GC, F1, F2, F3 e no p?s-parto: GM. O retinol das amostras foi quantificado por cromatografia l?quida de alta efici?ncia (CLAE). Para o total de mulheres, a concentra??o m?dia de retinol s?rico foi de 41.8 ? 12.9?g/dL e a preval?ncia da DVA foi de 5.3% (n= 33) com retinol (<20 ?g/dL), com diferen?a significativa entre o retinol s?rico das mulheres provenientes da capital e do interior (p<0,01). Em Natal, as preval?ncias de defici?ncia encontradas nas regi?es norte, sul, leste e oeste foram respectivamente: 4.3% (n=6), 5.6% (n=7), 2.9% (n=3) e 11.9% (n=8). A m?dia de retinol no colostro no grupo total foi de 95,3+ 53.7?g/dL, entretanto 27.3% (n=179) apresentaram valores inadequados (<60 ?g/dL). Os valores m?dios estimados de retinol fornecido aos rec?m-nascidos atrav?s do colostro, n?o atingiram a recomenda??o m?nima de 400?g/RAE/dia da AI (Adequate Intake) para rec?m-nascidos, considerando a ingest?o de 396mL/dia. Houve diferen?a significativa entre o retinol no colostro das mulheres da capital e aquelas provenientes do interior (p<0.01). Ambos os grupos n?o forneceram a AI de vitamina A para o rec?m-nascido e tamb?m o mesmo foi observado com as lactantes das regi?es norte e oeste da cidade de Natal. No leite maduro, nenhum dos grupos de mulheres das diferentes regi?es atingiu a recomenda??o, considerando a ingest?o de 780mL/dia pelos rec?m-nascidos. Ao avaliar as pu?rperas separadamente por rede de atendimento em sa?de (p?blico vs privado) foi encontrada diferen?a significativa entre o retinol s?rico e retinol no colostro (p<0.0001), mas n?o houve diferen?a para o leite maduro (p>0.05). Na estimativa do fornecimento de retinol atrav?s do colostro e leite maduro, as mulheres da rede p?blica n?o forneceram vitamina A dentro da recomenda??o m?nima para o rec?m-nascido (AI=400?g/RAE/dia), ao contr?rio das mulheres da rede privada, que forneceram. O consumo diet?tico m?dio total de vitamina A das parturientes foi de 987.1 + 674.4 ?gRAE/dia, sendo 872.2 + 639.2 ?gRAE/dia na da rede p?blica e 1169.2 + 695.2 ?gRAE/dia na rede privada, com diferen?a altamente significativa (p<0,00001). Na avalia??o individual, 38.4% (n=100) e 17.3% (n=28) das mulheres das redes p?blica e privada tinham ingest?o abaixo da ideal. Ao se estudar as diferentes formas de suplementa??o com vitamina A, n?o foram encontrados casos de DVA nos grupos suplementados com F1, F2 e F3. Ao se analisar o efeito da suplementa??o sobre o retinol do colostro, o grupo F2 (betacaroteno) apresentou mais casos de inadequa??o (40%). Os grupos F2 e GM n?o forneceram a quantidade de retinol m?nima recomendada pela AI aos rec?m-nascidos. No retinol do leite maduro n?o houve diferen?a entre os grupos GC, F1, F2, F3 e GM e com percentuais de inadequa??o mais baixos no GM (14.3%) e os grupos GC e F2 n?o forneceram a quantidade de retinol m?nima recomendada pela AI para os rec?m-nascidos. Concluiu-se que a preval?ncia de DVA entre as pu?rperas atendidas em Natal foi considerada um problema "leve" de sa?de p?blica na popula??o em geral. Os grupos de alto risco neste estudo viviam em cidades do interior, eram atendidos na rede p?blica de sa?de e n?o tomavam vitamina A, como o suplemento regular durante a gesta??o. / This study aimed to evaluate the vitamin A status of women who delivered in Natal/RN. A total of 795 women were enrolled in the study. Serum (n=619) and colostrum (n=656) samples were collected in the hospital after an overnight fast. Mature milk samples (n=15) were collected at the women?s house thirty days after delivery. Biochemical indicators were evaluated according to home location (capital city vs. country towns) and type of health care system (public vs. private). Vitamin A intake was assessed using a food-frequency questionnaire (FFQ) corresponding to the last trimester of pregnancy. In order to evaluate the different forms of maternal supplementation with vitamin A and their associations with biochemical markers (maternal serum and breast milk), subgroups were formed based on the type of supplementation that occurred during pregnancy: GC, F1, F2, F3 and postpartum: GM. Retinol concentrations in the biological samples were quantified by high performance liquid chromatography (HPLC). For the total sample, the mean serum retinol concentration was 41.8 ?12.9?g/dL and the prevalence of VAD was 5.3% (n=33) of women presenting retinol concentrations (<20 ?g/dL), evidencing a significant difference in serum retinol concentrations between women from the capital city and from the countryside (p <0.01). In Natal, the prevalence of disability found in the north, south, east and west were, respectively, 4.3% (n= 6), 5.6% (n= 7), 2.9% (n= 3) and 11.9% (n= 8). The overall mean retinol concentration in colostrum was 95.3 ? 53.7?g/dL; however, 27.3% (n=179) of women presented inadequate values (<60 ?g/dL). The average estimated amounts of retinol provided to newborns through colostrum did not meet the minimum recommendation of 400 ?g/RAE/day of AI (Adequate Intake) for newborns in both groups, considering the intake of 396mL/day. It was found a significant difference in colostrum retinol concentrations between women from the capital city and from the countryside (p<0.01). In Natal, colostrum milk of women from the northern and western regions did not meet the AI. For mature milk, none of the groups from the different regions met the recommendation, considering the intake of 780mL/day. Evaluating the sample separately by childbirth care system (public vs. private), it was found a significant difference in serum and colostrum retinol concentrations between the groups (p <0.0001); there was no difference for the mature milk (p>0.05). Estimating the retinol supply through colostrum and mature milk, women attending the public health system did not provide the minimum vitamin A amount recommended for newborns (AI= 400?g/RAE/day), unlike women's private network, which provided. The average total dietary vitamin A intake of pregnant women was 987.1 ? 674.4 ?gRAE/day, was 872.2 + 639.2 ?gRAE/day for women attending the public health system and 1169.2 + 695.2 ?gRAE/day for those attending the private system, evidencing a highly significant difference (p<0.001). Individually assessing the participants, 38.4% (n=100) e 17.3% (n=28) of women in the public and private systems had vitamin A intakes below the ideal. There was no difference in serum retinol concentrations between the CG, F1, F2, F3 and MG groups (p<0.05), although only the supplemented groups F1, F2 and F3 had no cases of VAD. Regarding colostrum retinol levels, the F2 group (beta-carotene) presented the highest number of inadequate cases (40%). The F2 and MG groups did not provide the minimum amount of retinol recommended by AI for newborns. Regarding retinol concentrations in mature milk, there was no difference between the CG, F1, F2, F3 and MG groups, and the MG group presented the lowest percentage of inadequacy (14.3%), while the CG and F2 groups did not provide the minimum amount of retinol recommended by the AI for infants. It was concluded that the prevalence of VAD among mothers who delivered in Natal was considered a ?mild? public health problem in the overall population. High-risk groups in this study lived in towns, were attended in the public health system and did not take vitamin A as regular supplement during pregnancy.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2017-02-17T19:43:31Z No. of bitstreams: 1 DanielChavesDeLima_TESE.pdf: 22514409 bytes, checksum: 31f3c8faf388b52b979837aa634b9a04 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2017-02-20T18:50:56Z (GMT) No. of bitstreams: 1 DanielChavesDeLima_TESE.pdf: 22514409 bytes, checksum: 31f3c8faf388b52b979837aa634b9a04 (MD5) / Made available in DSpace on 2017-02-20T18:50:56Z (GMT). No. of bitstreams: 1 DanielChavesDeLima_TESE.pdf: 22514409 bytes, checksum: 31f3c8faf388b52b979837aa634b9a04 (MD5) Previous issue date: 2016-09-23 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Chromobacterium violaceum is a ?-proteobacteria commonly found around tropical and subtropical regions throughout the globe. It produces many metabolites with biotechnological properties such as antitumoral peptides, antibiotics and polymers that have potential to replace the oil-based ones. Although it has been extensively studied over the past 40 years, there are many aspects of C. violaceum that remains unclear until today. We have conducted a biochemical study on the homologous recombination (HR) machinery of C. violaceum, mainly in RecA and its paralog, RadA/Sms. We performed in vitro assays from initial and late steps of HR such as D-loop formation and branch migration, respectively, with their corresponding molecular actors and how RadA/Sms influenced each one. We observed cvRadA/Sms influences negatively D-loop formation promoted by cvRecA and through pull-down assay we have observed an interaction between these two proteins. We also observed the DNA-binding preference of cvRadA/Sms and cvRecA and observed that this protein binds preferentially to dsDNA instead ssDNA, unlike cvRecA. No involvement of cvRadA/Sms on branch migration reactions was detected. In this work, we also described, for the first time, the isolation, sequencing and annotation of a new plasmid from C. violaceum, which we named ChVi1 and has 44,236 base pairs, 39 predicted open reading frames (ORFs) and, possibly, two origins of replication. Most of the ORFs codes for hypothetical and structural bacteriophage proteins. By using restriction digestion and Next-generation sequencing (NGS) we also looked for the presence of a similar plasmid in other seven C. violaceum strains isolated from amazon region. Our analysis suggest the presence of a plasmid similar to ChVi1 in two of these strains. The present work describes for the first time a biochemical characterization of RadA/Sms and RecA from C. violaceum which have different roles in HR. Moreover, the discovery of ChVi1 opens a path to further explore C. violaceum?s biology.