Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-03-10T20:24:54Z No. of bitstreams: 1 MayaraSantaRosaLima_DISSERT.pdf: 1762065 bytes, checksum: 81b2ba7e141b5e4144aae1950f65a16e (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-03-17T00:16:56Z (GMT) No. of bitstreams: 1 MayaraSantaRosaLima_DISSERT.pdf: 1762065 bytes, checksum: 81b2ba7e141b5e4144aae1950f65a16e (MD5) / Made available in DSpace on 2016-03-17T00:16:56Z (GMT). No. of bitstreams: 1 MayaraSantaRosaLima_DISSERT.pdf: 1762065 bytes, checksum: 81b2ba7e141b5e4144aae1950f65a16e (MD5) Previous issue date: 2015-05-26 / A vitamina A ? um nutriente essencial em diversos processos fisiol?gicos, como crescimento e desenvolvimento, de modo que um adequado estado nutricional nesse nutriente ? fundamental na gesta??o e lacta??o. Mulheres lactantes e crian?as em aleitamento materno s?o consideradas grupos de risco para a defici?ncia de vitamina A e alguns fatores podem aumentar o risco de hipovitaminose, como a prematuridade. O objetivo deste trabalho foi avaliar a concentra??o de vitamina A em lactantes e rec?m-nascidos pr?-termo e a termo, por meio da determina??o do retinol no soro materno, no soro do cord?o umbilical e no leite materno coletado at? 72 horas p?s-parto. Foram recrutadas 182 parturientes, divididas em grupo pr?-termo (GPT; n=118) e grupo termo (GT; n=64). No grupo pr?- termo tamb?m foram analisadas amostras de leite de transi??o (7?-15? dia; n=68) e leite maduro (30?-55? dia; n=46). O retinol foi analisado por cromatografia l?quida de alta efici?ncia (CLAE). A concentra??o materna de retinol s?rico foi 48,6 ? 12,3 ?g/dL no GPT e 42,8 ? 16,3 ?g/dL no GT (p<0,01). O retinol no soro do cord?o umbilical foi 20,4 ? 7,4 ?g/dL no GPT e 23,2 ? 7,6 ?g/dL no GT (p>0,05). Entre os rec?m-nascidos, 43% dos prematuros e 36% dos a termo apresentaram baixos n?veis de retinol s?rico no cord?o umbilical (<20 ?g/dL). No colostro, lactantes pr?- termo e termo apresentaram m?dia de retinol de 100,8 ? 49,0 ?g/dL e 127,5 ? 65,1 ?g/dL, respectivamente (p<0,05). A m?dia de retinol no leite pr?-termo aumentou para 112,5 ? 49,7 ?g/dL na fase de transi??o e reduziu para 57,2 ? 23,4 ?g/dL no leite maduro, diferindo significativamente entre todas as fases (p<0,05). Ao comparar com a recomenda??o de ingest?o de vitamina A (400 ?g/dia) o leite colostro do GT atingiu a recomenda??o para lactentes, por?m no GPT a recomenda??o n?o foi atingida em nenhuma das fases. As m?es de rec?m-nascidos prematuros possu?am concentra??o s?rica de retinol superior ? de m?es a termo, entretanto, isso n?o foi refletido no retinol do soro do cord?o umbilical, uma vez que os prematuros apresentaram menor concentra??o da vitamina. Tal condi??o pode ser explicada devido ? menor hemodilui??o fisiol?gica materna e transfer?ncia placent?ria de retinol para o feto durante a gesta??o pr?-termo. A compara??o do retinol no colostro evidenciou menor concentra??o no GPT, no entanto na fase de transi??o houve um aumento importante do conte?do de retinol liberado pela gl?ndula mam?ria de lactantes pr?-termo. Essa situa??o evidencia uma adapta??o fisiol?gica pr?pria da prematuridade, provavelmente no sentido de contribuir mais para a forma??o das reservas hep?ticas de retinol dos lactentes prematuros. / Vitamin A is an essential nutrient for many physiological processes such as growth and development, so that their adequate nutritional state is essential during pregnancy and lactation. Lactating women and children in breastfeeding are considered risk groups for vitamin A deficiency and some factors may increase the risk of vitamin A deficiency, such as prematurity. The aim of this work was to evaluate the vitamin A concentration in preterm and term lactating women and newborns by determination of retinol in maternal serum, umbilical cord serum and breast milk collected until 72 hours postpartum. 182 mothers were recruited and divided into preterm group (GPT; n = 118) and term group (GT, n = 64). In preterm group were also analyzed transition milk (7th-15th day; n = 68) and mature milk (30th-55th day; n = 46) samples. Retinol was analyzed by high-performance liquid chromatography (HPLC). Maternal retinol concentration in serum was 48.6 ? 12.3 ?g/dL in GPT and 42.8 ? 16.3 ?g/dL in the GT (p <0.01). Cord serum retinol was 20.4 ? 7.4 ?g/dL in GPT and 23.2 ? 7.6 ?g/dL in GT (p> 0.05). Among newborns, 43% of premature and 36% of term had low levels of serum retinol in umbilical cord (<20 ?g/dL). In colostrum, the retinol in preterm and term groups had an average of 100.8 ? 49.0 ?g/dL and 127.5 ? 65.1 ?g/dL, respectively (p <0.05). The retinol average in preterm milk increased to 112.5 ? 49.7 ?g/dL in transition phase and decreased to 57.2 ? 23.4 ?g/dL in mature milk, differing significantly in all stages (p <0.05). When comparing with the recommendation of vitamin A intake (400 ?g/day) GT colostrum reached the recommendation for infants, but in GPT the recommendation was not achieved at any stage. Mothers of premature infants had higher serum retinol than mothers at term; however, this was not reflected in serum retinol of umbilical cord, since premature had lower concentration of retinol. Such condition can be explained due to lower maternal physiological hemodilution and placental transfer of retinol to the fetus during preterm gestation. Comparison of retinol in colostrum showed lower concentrations in GPT; however the transition phase there was a significant increase of retinol content released by the mammary gland of preterm mothers. This situation highlights a specific physiological adaptation of prematurity, likely to more contribute to formation of hepatic reserves of retinol in premature infants.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-03-22T19:30:53Z No. of bitstreams: 1 MairaConceicaoJeronimoDeSouzaLima_TESE.pdf: 6599799 bytes, checksum: 12ec2e74167618ab1f1b22a3284a60a5 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-03-23T23:43:41Z (GMT) No. of bitstreams: 1 MairaConceicaoJeronimoDeSouzaLima_TESE.pdf: 6599799 bytes, checksum: 12ec2e74167618ab1f1b22a3284a60a5 (MD5) / Made available in DSpace on 2016-03-23T23:43:41Z (GMT). No. of bitstreams: 1 MairaConceicaoJeronimoDeSouzaLima_TESE.pdf: 6599799 bytes, checksum: 12ec2e74167618ab1f1b22a3284a60a5 (MD5) Previous issue date: 2015-04-17 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Envenenamentos causados por animais pe?onhentos s?o um grave problema de sa?de p?blica, enquadrando-se nesse cen?rio, principalmente, os escorpi?es e as serpentes. Tityus serrulatus ? considerado o escorpi?o mais venenoso da America do Sul, devido ? alta toxicidade do seu veneno, respons?vel por causar acidentes graves, principalmente em crian?as. A esp?cie Bothrops jararaca ? uma serpente que possui no seu veneno uma mistura complexa de enzimas, pept?deos e outras mol?culas. As toxinas do veneno de B. jararaca induzem respostas inflamat?rias locais e sist?micas. O tratamento de escolha para os casos graves de envenenamento ? a administra??o intravenosa do antiveneno especifico. Por?m, nem sempre esse tratamento est? acess?vel para os moradores de zonas rurais, que fazem uso de extratos de plantas medicinais. Nesse contexto, extratos aquosos, fra??es e compostos isolados de Aspidosperma pyrifolium (pereiro) e Ipomoea asarifolia (salsa), usadas na medicina popular, foram objeto de estudo deste trabalho para avaliar a atividade anti-inflamat?ria nos modelos de peritonite induzida por carragenina e peritonite induzida pelo veneno de T. serrulatus (VTs), e nos no modelo de edema local e infiltrado inflamat?rio induzido pelo veneno de B. jararaca, administrados pela via intravenosa. Os resultados dos ensaios de citotoxidade utilizando o ensaio MTT demonstraram que os extratos aquosos das esp?cies vegetais apresentaram baixa toxicidade para c?lulas provenientes de fibroblasto de embri?o de camundongo (3T3). A an?lise qu?mica dos extratos por Cromatografia L?quida de Alta Efici?ncia revelou a presen?a do flavon?ide rutina, na A. pyrifolium, e rutina, ?cido clorog?nico e ?cido caf?ico, na I. asarifolia. Quanto ? avalia??o farmacol?gica, os resultados demonstraram que o pr?-tratamento com extratos aquosos e fra??es reduziram a migra??o de leuc?citos totais para a cavidade abdominal no modelo de peritonite causada por carragenina e no modelo de peritonite induzida por veneno de T. serrulatus. Ainda, esses grupos apresentaram atividade antiedematog?nica, no modelo de edema local causado pelo veneno de B. jararaca, e reduziram o infiltrado inflamat?rio para o m?sculo. Os soros (antiaracn?dico e anti-botr?pico) espec?ficos para cada veneno atuaram inibindo a a??o inflamat?ria dos venenos e foram utilizados como controles. Os compostos identificados nos extratos, rutina, ?cido clorog?nico e ?cido caf?ico, tamb?m foram testados e, assim como os extratos das plantas, exibiram efeitos anti-inflamat?rios significativos, nas doses testadas. Dessa forma, esses resultados d?o evid?ncias, ?s plantas estudadas, de potencial atividade anti-inflamat?ria. Esse ? o primeiro estudo que avaliou os poss?veis efeitos terap?uticos de A. pyrifolium e I. asarifolia, mostrando o potencial terap?utico que essas esp?cies possuem. / 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.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:12Z No. of bitstreams: 1 DiegoGomesTeixeira_DISSERT.pdf: 3901094 bytes, checksum: 8c230a0e1503d41ddbfe3f8758b2dbce (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T21:48:14Z (GMT) No. of bitstreams: 1 DiegoGomesTeixeira_DISSERT.pdf: 3901094 bytes, checksum: 8c230a0e1503d41ddbfe3f8758b2dbce (MD5) / Made available in DSpace on 2016-04-28T21:48:14Z (GMT). No. of bitstreams: 1 DiegoGomesTeixeira_DISSERT.pdf: 3901094 bytes, checksum: 8c230a0e1503d41ddbfe3f8758b2dbce (MD5) Previous issue date: 2015-07-02 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A Leishmaniose ? uma doen?a infecciosa que at? o momento n?o possui uma vacina capaz de eliminar o parasita do hospedeiro, sendo tratada apenas por meio de drogas pouco eficientes e de alto custo. Essa doen?a normalmente se apresenta formando ulcera??es na pele, mucosas e v?sceras. Os pa?ses mais atingidos por esse parasita encontram-se nas regi?es tropicais do planeta, e afeta aproximadamente dois milh?es de pessoas a cada ano. Acredita-se que no continente americano a esp?cie Leishmania infantum tenha sido introduzida pelos imigrantes durante o processo de coloniza??o dos pa?ses da Am?rica Central e do Sul. Nas ?ltimas d?cadas, estudos v?m mostrando que a expans?o urbana associada com o fluxo de pessoas para regi?es muito povoadas est?o contribuindo para uma expans?o no n?mero de reservat?rios para o parasita, influenciando uma maior varia??o de suas caracter?sticas g?nicas. Tais varia??es gen?ticas v?m sendo associadas ao perfil sintomatol?gico dos parasitas em seus hospedeiros, principalmente humanos. No estado do Rio Grande do Norte, Brasil, foram isolados e sequenciados o genoma de 20 cepas de L. infantum as quais apresentaram diferentes padr?es sintomatol?gicos (sintom?tico e assintom?tico) e com diferentes per?odos de isolamento, 5 na d?cada de 1990 e 15 nos anos recentes. Apesar dos diferentes padr?es cl?nicos o genoma dos isolados apresentaram alto grau de identidade entre si, at? mesmo os isolados da d?cada de 90 quando comparados com os recentes. Entretanto, as poucas varia??es foram suficientes para a identifica??o de padr?es de agrupamentos dos isolados por meio de an?lises de componentes principais, mostrando que os isolados dos anos recentes se encontram mais homog?neos na popula??o. An?lises de reconstru??o filogen?tica utilizando m?todos Bayesianos foram realizadas e observou-se a manuten??o nos padr?es de agrupamento j? vistos nos resultados anteriores, al?m disso foi gerado um expanded bayesian skyline plot por onde foi poss?vel constatar o crescimento da popula??o gen?mica de L. infantum quando comparado com a d?cada de 1990. Foram observadas altera??es no n?mero de c?pias cromoss?micas em todos os isolados, entretanto apenas o cromossomo 31 se apresentou como exclusivamente triss?mico. O presente trabalho apresenta ind?cios de padr?es nos genomas de isolados de L. infantum relacionando-os ?s caracter?sticas cl?nicas dos hospedeiros. / Leishmaniasis is a complex of diseases with no preventable or therapeutic vaccine available. There is only a small set of drugs for treatment of these diseases, which evolve with distinct clinical outcomes from skin ulcers, to mucosal involvement or visceralization. The type of clinical presentation usually is associated with the infecting species of Leishmania. Leishmania infantum is the visceralizing species, causing visceral leishmaniasis, in Latin America, Europe and northern Africa and was introduced in the Americas by Europeans during colonization. Visceral leishmaniasis used to be a sporadic disease found in rural areas Northeast of Brazil, however with urbanization, outbreaks have been reported in periurban areas of major cities. L. infantum infection can evolve with control of the parasite without apparent clinical symptoms, with only a minority developing symptomatic disease. The reasons why one evolve with selfresolution and others develop full blown disease is not entirely understood, but seems to include host genetic susceptibility, comorbidity, sand fly exposure and potentially genetic variation in the infecting Leishmania species. Our ongoing study, have shown structural genome variations in several clinical Leishmania isolates obtained from people infected in the State of Rio Grande do Norte. Despite the different clinical patterns, the isolates? genomes showed a high identity degree to each other, even the ones isolated from the 90 compared with the latest. However, the few found changes were sufficient for the isolation of the distinct groups by a principal component analysis, showing that the isolates from more recent years are more homogeneous among the population. Phylogenetic analyzes using Bayesian reconstruction methods have been made and supported the grouping patterns already seen in the foregoing results. Moreover by an Expanded Bayesian Skyline Plot it was possible to observe the growth of the genomic population of L. infantum compared with the 1990s. Changes were observed in the number of chromosome copies in all strains, but only chromosome 31 is presented as exclusively trisomic. This work presents evidence on patterns of L. infantum isolated genomes relating them to clinical characteristics of their hosts.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:13Z No. of bitstreams: 1 JonalsonNogueiraAraujo_DISSERT.pdf: 2283136 bytes, checksum: 83cd9344cc38a30060e338ebb45988cd (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T22:04:54Z (GMT) No. of bitstreams: 1 JonalsonNogueiraAraujo_DISSERT.pdf: 2283136 bytes, checksum: 83cd9344cc38a30060e338ebb45988cd (MD5) / Made available in DSpace on 2016-04-28T22:04:54Z (GMT). No. of bitstreams: 1 JonalsonNogueiraAraujo_DISSERT.pdf: 2283136 bytes, checksum: 83cd9344cc38a30060e338ebb45988cd (MD5) Previous issue date: 2015-06-26 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Sementes de plantas s?o reservat?rios de mol?culas com grande potencial para elabora??o de bioprodutos e por este motivo uma especial aten??o tem sido direcionada na busca de prote?nas bioativas com a??o antimetab?lica e propriedades farmacol?gicas. As lectinas de plantas s?o prote?nas com atividades biol?gicas relacionadas a defesa, mas que podem ser utilizadas, heterologamente, como interferentes no metabolismo de outros organismos. Uma lectina das sementes de Canavalia maritima (CML) foi purificada em dois passos, pelo fracionamento com sulfato de am?nio seguido pela cromatografia de afinidade em coluna Sephadex G- 50. CML foi analisada por SDS-PAGE e parte de sua sequ?ncia de amino?cidos determinada por espectrometria de massas. Os 20 res?duos identificados apresentaram 100% de identidade com ConM. ConM foi testada contra eritr?citos do sangue perif?rico humano e foi constatado a aus?ncia de toxicidade quando incubados com at? 1000 ?g/mL da prote?na. J? contra c?lulas mononucleares, a lectina n?o foi significativamente t?xica nas concentra??es de 1, 2,5 e 5 ?g/mL, mas o foi a partir da concentra??o 7 ?g/mL e contra a linhagem RAEC a prote?na n?o apresentou toxicidade significante com 25 ?g/ml da prote?na. Outros ensaios de citotoxicidade revelaram que a ConM ? t?xica para as linhagens tumorais A549, 786- 0, HT-29 e HeLa no intervalo entre 24 e 72 horas, principalmente nas concentra??es de 5, 7 e 10 ?g/ml. ConM foi capaz de aglutinar as formas promastigotas de Leishmania spp na concentra??o de 6,25 ?g/ml. Quanto a a??o contra Candida spp, nenhuma das concentra??es testadas (1 a 500 ?g/ml) foi capaz de interferir no crescimento. Quando avaliada a atividade bacteriost?tica, a ConM nas concentra??es variando de 0,39 a 400 ?g/ml n?o inibiu o crescimento de Escherichia coli, j? para Staphylococcus aureus a concentra??o de 25 ?g/ml se mostrou ativa, reduzindo o crescimento em 75 %. Com base nos dados obtidos, a lectina isolada corresponde a uma isolectina do tipo ConA, e suas propriedades anti-tumoral, bacteriost?tico e aglutinante para Leishmania precisam ser melhor investigados para que seu potencial biotecnol?gico seja explorado. / Plant seeds are reservoirs of molecules with great potential for development of bioproducts and for this reason special attention has been directed in the search of bioactive proteins with antimetabolic and pharmacological properties action. Plant lectins are proteins with biological activities related to defense, but that can be used, heterologously as interfering with the metabolism of other organisms. A Canavalia mar?tima seeds lectin (CML) was purified in two steps by ammonium sulfate fractionation followed by affinity chromatography on Sephadex G-50 column. CML was analyzed by SDS-PAGE and part of its amino acid sequence determined by mass spectrometry. The 20 resulting residues identified showed 100% identity with conm. ConM was tested against erythrocytes of human peripheral blood and it was observed absence of toxicity when incubated with up to 1000 ?g/mL of protein. Since against mononuclear cells, the lectin was not significantly toxic at concentrations of 1, 2.5 and 5 ?g/mL, but it was from concentration 7 ?g/mL. Against RAEC lineage protein showed no significant toxicity with 25 ?g/mL of protein. Other cytotoxicity assays revealed that CONM is toxic for the tumor cell lines A549, 786-0, HT-29 and HeLa in the range between 24 and 72 hours, particularly at concentrations of 5, 7 and 10 ?g/mL. ConM was still able to agglutinate promastigotes of Leishmania spp at concentrations of 6.25 ?g/mL. About action against Candida spp any of the tested concentrations (1 to 500 ?g/ml) was able to interfere with growth. When evaluated bacteriostatic activity, ConM in concentrations ranging from 0.39 to 400 ?g/mL did not inhibit the growth of Escherichia coli, but the concentration of 25 ?g/mL proved active aganist Staphylococcus aureus, reducing its growth by 75%. Based on these data, the lectin corresponds to a isolectin ConA-like, and its anti-tumor properties, bacteriostatic and binder for Leishmania need to be further investigated for its biotechnological potential exploited.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-04-26T19:16:13Z No. of bitstreams: 1 LuizaAraujoDaCostaXavier_DISSERT.pdf: 2033184 bytes, checksum: 2e016a1123ca6a8907a0176e6866a4a7 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-04-28T22:16:51Z (GMT) No. of bitstreams: 1 LuizaAraujoDaCostaXavier_DISSERT.pdf: 2033184 bytes, checksum: 2e016a1123ca6a8907a0176e6866a4a7 (MD5) / Made available in DSpace on 2016-04-28T22:16:51Z (GMT). No. of bitstreams: 1 LuizaAraujoDaCostaXavier_DISSERT.pdf: 2033184 bytes, checksum: 2e016a1123ca6a8907a0176e6866a4a7 (MD5) Previous issue date: 2015-07-31 / As fendas labiopalatinas n?o-sindr?micas (FL/PNS) s?o defeitos cong?nitos comuns em humanos, caracterizadas pelo desenvolvimento incompleto de estruturas que separam a cavidade nasal da cavidade oral, e s?o causadas devido a uma intera??o entre fatores gen?ticos e ambientais. Dados obtidos de estudos de caso-controle e interven??o diet?tica indicam que a suplementa??o materna com multivitaminas contendo ?cido f?lico previne o surgimento das fissuras orais. ? conhecido que o ?cido f?lico participa de fun??es celulares essenciais, como a s?ntese de nucleot?deos para o reparo do DNA, as quais contribuem para a prote??o da integridade do genoma contra eventos de danos gerados por fatores end?genos e/ou ex?genos. Inclusive, estudos revelam que a defici?ncia desta vitamina aumenta a forma??o de micron?cleos, estruturas citogen?ticas indicadoras de danos no DNA. Al?m disso, esta defici?ncia ? modulada pelos polimorfismos gen?ticos associados ao metabolismo do ?cido f?lico, comprometendo o desempenho das fun??es de estabilidade do genoma e, portanto, est?o sendo associados com o desenvolvimento de v?rios dist?rbios, dentre eles as fissuras orais. A partir deste contexto, foi conduzido um estudo transversal do tipo caso-controle n?o pareado com o objetivo de avaliar a frequ?ncia de biomarcadores de instabilidade gen?mica, sua rela??o com polimorfismos gen?ticos do metabolismo do folato, e se essas vari?veis est?o associadas com o desenvolvimento das FL/PNS em crian?as do Rio Grande do Norte, Brasil. Foram recrutados 48 pacientes fissurados e 18 crian?as controles, respectivamente, no Hospital de Pediatria Professor Heriberto Ferreira Bezerra (HOSPED)/UFRN e em escolas estaduais da cidade de Natal, RN. Com o devido consentimento dos participantes, realizou-se uma entrevista com os respons?veis para obten??o de dados epidemiol?gicos, como tamb?m procedeu-se a coleta sangu?nea das crian?as para a realiza??o dos testes. Foi executado o ensaio do micron?cleo com bloqueio da citocinese para calcular a frequ?ncia de micron?cleos (MN), pontes nucleoplasm?ticas (PN) e brotos nucleares (BN). A partir da extra??o do DNA gen?mico, avaliou-se os polimorfismos da metilenotetrahidrofolato redutase C677T e A1298C, metionina sintase A2756G, metionina sintase redutase A66G e do receptor de folato reduzido A80G pela t?cnica de rea??o em cadeia da polimerase associada ao polimorfismo de tamanho do fragmento de restri??o (PCR-RFLP). As crian?as com FL/PNS apresentaram maior frequ?ncia basal de MN, PN e BN que o grupo de crian?as controle (p < 0,001), e nenhum dos polimorfismos avaliados modificaram significativamente a frequ?ncia destes biomarcadores. Al?m disso, crian?as com FL/PNS apresentaram 2,3 vezes mais risco de exibir altas frequ?ncias de MN (p = 0,043) segundo modelo de regress?o log?stica bin?ria. A alta instabilidade do genoma nas crian?as com fissuras orais sugere que os eventos genot?xicos, em particular os que promovem quebras na dupla fita do DNA e n?o s?o devidamente reparados formando micron?cleos, representam fatores relevantes no desenvolvimento das fendas labiopalatinas n?o-sindr?micas. / The non-syndromic clefts of the lip and/or palate (NSCL/P) are common birth defects in humans characterized by an incomplete development of cellular structures that separate the nasal cavity from the oral cavity, and they are caused due to an interaction between genetic and environmental factors. Data from case-control and dietary intervention studies indicates that maternal supplementation with multivitamins containing folic acid prevents the formation of oral clefts. It is known that folic acid plays a role in essential cellular functions, such as nucleotides synthesis for DNA repair, which contribute to the protection of genome integrity from damage events generated by endogenous and/or exogenous factors. In fact, studies show that a deficiency in this vitamin increases micronuclei formation, a cytogenetic structure that indicates misrepair of damaged DNA. Furthermore, this deficiency is modulated by genetic polymorphisms associated with folic acid metabolism, affecting the performance of genome stability functions and therefore, it has been associated with the development of various disorders, including oral clefts. From this context, it was planned a unpaired case-control cross-sectional study in order to assess the frequency of genome instability biomarkers, their relationship with genetic polymorphisms in folate metabolism, and if these variables are associated with the development of NSCL/P in children from a Northeast region at Brazil. For this research, it was recruited 48 NSCL/P patients and 18 control children, respectively, at the Pediatric Hospital Professor Heriberto Ferreira Bezerra (HOSPED)/ UFRN and at schools in Natal city. With the participants consent, they were interviewed with a standard questionnaire to obtain epidemiological data, and the children underwent a blood sampling for the tests. It was performed the cytokinesis-block micronucleus assay to estimate the frequency of micronuclei (MN), nucleoplasmic bridges (NPB) and nuclear buds (NB). Also, from the genomic DNA extraction, it was evaluated by PCR-RFLP the polymorphisms of methylenetetrahydrofolate reductase C677T and A1298C, methionine synthase A2756G, methionine synthase reductase A66G and reduced folate carrier A80G. Children with NSCL/P had higher baseline frequency of MN, NPB and NB than the control group (p < 0.001), and none of the evaluated polymorphisms significantly modified the frequency of these biomarkers. In addition, children with clefts had 2.3 times more risk of displaying high frequency of MN (p = 0.043) according to the binary logistic regression model. The high genomic instability in children with oral clefts suggests that genotoxic events that promote double strand breaks on DNA and are not properly repaired, thus originating micronuclei, represent significant factors in the development of non-syndromic cleft lip/ palate.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-05-31T00:26:34Z No. of bitstreams: 1 RafaelBarrosGomesDaCamara_TESE.pdf: 5185586 bytes, checksum: ac9f52b2d0105b2f8e729ae0d148ebaa (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-06-02T23:03:58Z (GMT) No. of bitstreams: 1 RafaelBarrosGomesDaCamara_TESE.pdf: 5185586 bytes, checksum: ac9f52b2d0105b2f8e729ae0d148ebaa (MD5) / Made available in DSpace on 2016-06-02T23:03:58Z (GMT). No. of bitstreams: 1 RafaelBarrosGomesDaCamara_TESE.pdf: 5185586 bytes, checksum: ac9f52b2d0105b2f8e729ae0d148ebaa (MD5) Previous issue date: 2015-08-20 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / O presente estudo teve como objetivo extrair, caracterizar e realizar uma an?lise prospectiva de atividades farmacol?gicas de polissacar?deos sulfatados da macroalga verde Caulerpa prolifera. Sete fra??es (CP-0.3/CP-0.5/CP-0.7/CP-0.9/CP-1.1/CP-1.5/CP-2.0) foram obtidas de C. prolifera por prote?lise alcalina seguida de precipita??o sequencial em acetona. As an?lises f?sico-qu?micas indicaram que C. prolifera sintetiza diferentes popula??es de heteropolissacar?deos sulfatados. Na an?lise da atividade anticoagulante, todas as fra??es, exceto CP-0.3, apresentaram influ?ncia sobre a via intr?nseca da coagula??o. Todas as fra??es apresentaram atividade antioxidante em 6 ensaios diferentes, sendo mais pronunciadas no teste de sequestro de per?xido de hidrog?nio, com destaque para CP-0.3, CP-0.7 e CP-0.9 (quando se obteve at? 61% de sequestro), no teste de quela??o f?rrica (com destaque para CP-0.9, com 56% de quela??o) e no teste de quela??o c?prica (com destaque para CP-2.0, com quela??o de 78%). Com rela??o a a??o imunomoduladora, a presen?a de CP-0.3, CP-0.7 e CP-0.9 promoveu aumento da produ??o de ?xido n?trico (ON) em macr?fagos em at? 48, 142 e 163 vezes, respectivamente. De maneira oposta, a s?ntese de ON caiu em 73% quando macr?fagos ativados por LPS foram incubados concomitantemente com CP-2. A atividade anti-adipog?nica das fra??es tamb?m foi avaliada e o destaque ficou com CP-1.5, na presen?a desta fra??o (0,2 mg/mL) a diferencia??o de pr?-adip?citos (3T3-L1) em adip?citos foi reduzida em 60%. Vale salientar, que a viabilidade das c?lulas 3T3-L1 n?o foi afetada pela presen?a de CP-1.5. O que n?o ocorreu com c?lulas de c?ncer cervical humano (HeLa) e de adenocarcinoma renal humano (786-0), j? que a viabilidade dessas c?lulas caiu quando elas foram expostas as fra??es, o destaque foi para a redu??o da viabilidade em torno de 55% quando as c?lulas HeLa foram incubadas com CP-0.3, CP-0.5 e CP-0.9. J? com 786-0, o melhor resultado encontrado foi com o uso da fra??o CP-1.5, quando se obteve uma redu??o de ~75% da viabilidade celular. N?o se identificou atividade leishmanicida ou microbicida contra a Klebsiella pneumoniae Carbapenemase (KPC) com ao se testar as fra??es contra estes organismos. No entanto, a prolifera??o de Staphylococcus epidermidis foi diminu?da em 30% na presen?a de CP-1.5. Outro resultado relevante, foi a observa??o de que a forma??o de cristais de oxalato c?lcio na presen?a das fra??es era alterada. Destaca-se CP-0.3, CP-0.5 e CP-1.1, j? que na presen?a dessas fra??es identificou-se apenas a forma??o de cristais dihidratados e de tamanho m?dio 80% menor do que aquele verificado no grupo controle. Dados oriundos da avalia??o do potencial zeta dos cristais formados na presen?a das fra??es e da avalia??o de imagens de microscopia confocal de cristais formados na presen?a das fra??es marcadas fluorescentemente levaram a hip?tese de que os polissacar?deos presentes nas fra??es interagem com o c?lcio da rede cristalina e isso afeta o crescimento e a morfologia dos cristais formados. Os resultados aqui descritos levam a sugest?o de os principais agentes respons?veis pelas atividades observadas com as fra??es seriam os polissacar?deos sulfatados, portanto passos posteriores de purifica??o desses pol?meros, bem como a investiga??o de seus mecanismos de a??o devem ser investigados. Por fim, os dados aqui obtidos levam a observa??o de que as fra??es ricas em polissacar?deos obtidas da alga C. prolifera apresentam um potencial multiterap?utico por serem anticoagulantes, antioxidantes, imunomoduladores, citot?xicas contra c?lulas tumorais, anti-adipogenicos, antibacterianas e alterarem a forma??o de cristais de oxalato de c?lcio. / This study aimed to extract, characterize and conduct a prospective analysis of pharmacological activities of sulfated polysaccharides from green seaweed Caulerpa prolifera. Seven fractions (CP-0.3/CP-0.5/CP-0.7/CP-0.9/CP-1.1/CP-1.5/CP-2.0) were obtained from C. prolifera by alkaline proteolysis followed by sequential precipitation in acetone. The physicochemical analyzes indicated that C. prolifera synthesizes a homogalactan (CP-0.9) and different populations of sulfated heteropolysaccharides. In the analysis of anticoagulant activity, all fractions except CP-0.3, influenced the intrinsic coagulation pathway. All fractions showed antioxidant activity in six different assays being more pronounced in hydrogen peroxide scavenging assay, especially CP-0.3, CP-0.7 and CP-0.9 (which obtained 61% of hydrogen peroxide scavenging), in ferric chelation assay (especially CP-0.9 with 56% chelation) and cupric chelation assay (especially CP-2.0 with 78% chelation). With respect to immunomodulatory activity, the presence of CP-0.3, CP-0.7 and CP-0.9 showed an immunogenic potential, increasing the production of nitric oxide (NO) by 48, 142 and 163 times, respectively. Conversely, the NO synthesis fell 73% after the activation of macrophages by LPS, incubated concurrently with CP-2.0. The anti-adipogenic activity of the fractions was also evaluated and CP-1.5 was able to reduce the differentiation of pre-adipocytes (3T3-L1) into adipocytes by 60%, without affecting the cell viability. The fractions CP-0.3, CP-0.5 and CP-0.9 reduced the viability of the HeLa cells (human cervical adenocarcinoma) by 55% and CP-1.5 reduced the viability of the 786-0 cells (human renal adenocarcinoma) by 75%. Leishmanicidal activity and microbicide effect against Carbapenem-resistant Klebsiella pneumoniae (KPC) have not been identified. However, the viability of Staphylococcus epidermidis was reduced by 23.8% in the presence of CP -1.5. All fractions were able to change the formation of calcium oxalate crystals. CP-0.3, CP-0.5 and CP-1.1 only promoted the formation of COD type crystals with a very small size (1 ?m). Confocal microscopy and zeta potential data of crystals formed in the presence of the samples showed that the polysaccharides present in the fractions must interact with calcium ions present throughout the crystal lattice, affecting the growth and morphology of crystals The results described herein indicate that the fractions rich in polysaccharides obtained from the green seaweed C. prolifera present a multi therapeutic potential, and subsequent purification steps, as well as research on the mechanisms of action by which these polymers act should be investigated.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-06-10T19:08:01Z No. of bitstreams: 1 ArianeFerreiraLacerda_TESE.pdf: 2669795 bytes, checksum: 86dd6a13d7f026cc0544d3cb0e76e5f1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-06-13T20:26:33Z (GMT) No. of bitstreams: 1 ArianeFerreiraLacerda_TESE.pdf: 2669795 bytes, checksum: 86dd6a13d7f026cc0544d3cb0e76e5f1 (MD5) / Made available in DSpace on 2016-06-13T20:26:34Z (GMT). No. of bitstreams: 1 ArianeFerreiraLacerda_TESE.pdf: 2669795 bytes, checksum: 86dd6a13d7f026cc0544d3cb0e76e5f1 (MD5) Previous issue date: 2015-06-30 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A inefic?cia de pesticidas qu?micos no controle de fungos fitopatog?nicos na agricultura e a frequente incid?ncia de doen?as humanas causadas por bact?rias resistentes a antibi?ticos levam ? busca por compostos antimicrobianos alternativos. Neste contexto, defensinas vegetais constituem uma promissora ferramenta no controle de agentes patog?nicos tanto de plantas quanto de humanos. Defensinas de plantas s?o pept?deos cati?nicos com aproximadamente 50 res?duos de amino?cidos, ricas em ciste?nas e cuja estrutura tridimensional ? bastante conservada entre as diferentes esp?cies vegetais. Estas mol?culas de a??o antimicrobiana representam um importante componente inato da resposta de defesa vegetal contra pat?genos e s?o expressas em diversos tecidos da planta, como folhas, tub?rculos, flores, vagens e sementes. O presente trabalho teve por finalidade a avalia??o da atividade antimicrobiana de duas defensinas vegetais contra diferentes esp?cies de fungos fitopatog?nicos e bact?rias patog?nicas ao homem. A defensina Drr230a, cujo gene foi isolado de ervilha (Pisum sativum) e a defensina CD1, cujo gene foi identificado no transcriptoma de caf? (Coffea arabica) foram subclonadas em vetor de express?o de levedura e expressas em Pichia pastoris. O gene cd1 foi subclonado em duas formas recombinantes: CD1tC, contendo uma sequ?ncia codificante para seis histidinas (6xHis) na regi?o C-terminal do pept?deo e CD1tN, contendo sequ?ncia codificante para 6xHis na regi?o N-terminal. No caso da defensina Drr230a, a sequ?ncia codificante para 6xHis foi inserida apenas na regi?o N-terminal da prote?na. Ensaios de atividade antimicrobiana das prote?nas recombinantes purificadas rDrr230a e rCD1 contra Phakopsora pachyrhizi, agente causal da ferrugem asi?tica da soja, foram realizados para analisar a inibi??o da germina??o de esporos in vitro e a severidade da doen?a causada pelo fungo in planta. As duas defensinas recombinantes testadas foram capazes de inibir a germina??o de uredosporos de P. pachyrhizi, n?o havendo diferen?a entre a a??o antimicrobiana de CD1tC e CD1tN. Ademais, rDrr230a e rCD1 reduziram drasticamente a severidade da ferrugem asi?tica da soja, conforme demonstrado em ensaios in planta. Apesar de rCD1 n?o ter sido capaz de inibir a prolifera??o das bact?rias patog?nicas humanas Staplylococcus aureus e Klebsiella pneumoniae, rCD1 mostrou-se capaz de inibir o crescimento do fungo fitopatog?nico Fusarium tucumaniae, causador da s?ndrome da morte s?bita da soja. Os resultados obtidos mostram que tais defensinas vegetais s?o candidatas ?teis para serem utilizadas em programas de engenharia gen?tica de plantas para controlar doen?as f?ngicas impactantes na agricultura. / The inefficiency of chemical pesticides to control phytopathogenic fungi in agriculture and the frequent incidence of human diseases caused by bacteria which are resistant to antibiotics lead to the search for alternative antimicrobial compounds. In this context, plant defensins are a promising tool for the control of both plant and human pathogenic agents. Plant defensins are cationic peptides of about 50 amino acid residues, rich in cysteine and whose tridimensional structure is considerably conserved among different plant species. These antimicrobial molecules represent an important innate component from plant defense response against pathogens and are expressed in various plant tissues, such as leaves, tubers, flowers, pods and seeds. The present work aimed at the evaluation of the antimicrobial activity of two plant defensins against different phytopathogenic fungi and pathogenic bacteria to humans. The defensin Drr230a, whose gene was isolated from pea (Pisum sativum), and the defensin CD1,whose gene was identified within coffee (Coffea arabica) transcriptome, were subcloned in yeast expression vector and expressed in Pichia pastoris. The gene cd1 was subcloned as two different recombinant forms: CD1tC, containing a six-histidine sequence (6xHis) at the peptide C-terminal region and CD1tN, containing 6xHis coding sequence at the N-terminal region. In the case of the defensin Drr230a, the 6xHis coding sequence was inserted only at the N-terminal region. Assays of the antimicrobial activity of the purified recombinant proteins rDrr230a and rCD1 against Phakopsora pachyrhizi, causal agent of soybean Asian rust, were performed to analyze the in vitro spore germination inhibition and disease severity caused by the fungus in planta. Both recombinant defensins were able to inhibit P. pachyrhizi uredospore germination, with no difference between the antimicrobial action of either CD1tC or CD1tN. Moreover, rDrr230a and rCD1 drastically reduced severity of soybean Asian rust, as demonstrated by in planta assays. In spite of the fact that rCD1 was not able to inhibit proliferation of the human pathogenic bacteria Staplylococcus aureus and Klebsiella pneumoniae, rCD1 was able to inhibit growth of the phytopathogenic fungus Fusarium tucumaniae, that causes soybean sudden death syndrome. The obtained results show that these plant defensins are useful candidates to be used in plant genetic engineering programs to control agriculture impacting fungal diseases.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-06-15T00:21:54Z No. of bitstreams: 1 RomuloRodrigoDeSouzaAlmeida_DISSERT.pdf: 1782048 bytes, checksum: b2c3f99e10e637474fe9a749a13cc2be (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-06-17T23:35:49Z (GMT) No. of bitstreams: 1 RomuloRodrigoDeSouzaAlmeida_DISSERT.pdf: 1782048 bytes, checksum: b2c3f99e10e637474fe9a749a13cc2be (MD5) / Made available in DSpace on 2016-06-17T23:35:49Z (GMT). No. of bitstreams: 1 RomuloRodrigoDeSouzaAlmeida_DISSERT.pdf: 1782048 bytes, checksum: b2c3f99e10e637474fe9a749a13cc2be (MD5) Previous issue date: 2015-07-29 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / V?rios estudos tem estabelecido uma associa??o entre Diabetes Mellitus tipo 1 altera??es hep?ticas estresse oxidativo. No entanto os conhecimentos de como essas altera??es ocorrem no sistema antioxidante ainda n?o est? elucidado. Este estudo avaliou os efeitos de quatro semanas de diabetes tipo 1 no tecido hep?tico. Ratos machos wistar foram divididos em 2 grupos: Controle (C) e Diab?tico (D). A an?lise da atividade de enzimas antioxidantes (CAT e SOD); a express?o do mRNA de CAT, SOD1, SOD2, GPX1 e PRX4 foram avaliadas; marcadores de estresse oxidativo tamb?m foram avaliados (Peroxida??o de lip?deos, carbonila??o de prote?nas e conte?do tiol) e conte?do de H2O2 hep?tico. Como resultado a diabetes aumentou a atividade da SOD e a express?o g?nica da SOD2, enquanto diminuiu a express?o da SOD1. Assim como houve diminui??o da atividade da CAT e na express?o g?nica da CAT, GPX1 e PRX4. Houve tamb?m mudan?as em biomarcadores de estresse oxidativo. Nossos resultados sugerem que provavelmente as quatro semanas da diabetes, induzem altera??es precoces no sistema antioxidante no f?gado de ratos induzidos por STZ, agravando altera??es decorrentes do estresse oxidativo levando a forma??o de esp?cies reativas de oxig?nio, podendo contribuir para um preju?zo da fun??o hep?ticas. / The correlation between the type 1 diabetes mellitus and oxidative stress have been described in several studies, however its underlying mechanisms are not fully elucidated. The present work aimed to evaluate the effects of four weeks of streptozootocin-induced (STZ) diabetes in the redox homeostasis of rat hepatocytes. Thus, the liver of male Wistar rats from control and diabetic groups were collected and the activity and expression of antioxidant enzymes, as well the main markers of oxidative stress and content of H2O2 in these tissues were measured. The diabetes induced the activity of superoxide dismutase (SOD) and the gene expression of its mitochondrial isoform, SOD2. However, the expression of SOD1, the cytoplasmic isoform, was reduced by this disease. The activity and expression of catalase (CAT), as well the expression of glutathione peroxidase 1 (GPX1) and peroxiredoxin 4 (PRX4) were drastically reduced in the hepatocytes of diabetics rats. Even with this debility in the peroxidases mRNA expression, the content of H2O2 was reduced in the liver of diabetics rats when compared to the control group. The diabetes caused an increase of lipid peroxidation and a decrease of protein thiol content, showing that this disease causes distinct oxidative effects in different cell biomolecules. Our results indicate that four week of diabetes induced by STZ is already enough to compromise the enzymatic antioxidant systems of the hepatocytes.

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-07-21T17:14:40Z No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-07-22T11:33:09Z (GMT) No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) / Made available in DSpace on 2016-07-22T11:33:09Z (GMT). No. of bitstreams: 1 AmandaLarissaMarquesDeMedeiros_TESE.pdf: 2401701 bytes, checksum: 6c9f1744f688fa158feed1ba0fad56b1 (MD5) Previous issue date: 2015-10-16 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) / A cana-de-a??car ? uma importante cultura para nosso pa?s, que contribui com quase metade de toda a produtividade mundial. Muitos desafios s?o enfrentados pelas plantas, que muitas vezes n?o atingem todo o seu potencial gen?tico devido a presen?a de estresses bi?ticos e abi?ticos. Como consequ?ncia, esses estresses podem gerar esp?cies reativas de oxig?nio, que podem danificar o material gen?tico. Outra consequ?ncia do estresse ? a flora??o precoce, que diminui tamb?m a produtividade da cana-de-a??car. Neste contexto, este trabalho apresenta como objetivos caracterizar os genes scMUTM1 e scMUTM2, duas DNA glicosilases pertencentes ? via de reparo por excis?o de base (BER), al?m de identificar transcritos potencialmente relacionados com estresse e reparo de DNA, em duas variedades com fen?tipo contrastante para flora??o. A caracteriza??o das glicosilases incluiu a constru??o de cassetes de express?o de prote?nas, para posterior purifica??o e uso em ensaios enzim?ticos; constru??o de ?rvore filogen?tica e an?lise de promotor deste gene. Com a reconstru??o filogen?tica de MUTM foi poss?vel observar que h? o agrupamento desta sequ?ncia em um ramo com monocotiled?neas e outro com dicotiled?neas. Isto sugere que a duplica??o desta sequ?ncia pode ter ocorrido depois da diverg?ncia desses dois grupos. Com a an?lise do promotor deste gene, ? sugerido que, provavelmente, o gene ScMUTM1 pode estar sofrendo subfuncionaliza??o, uma vez que a regi?o promotora do gene ScMUTM2 apresenta uma maior quantidade de sequ?ncias regulat?rias relacionadas ao estresse. Por meio das an?lises de express?o g?nica utilizando microarranjos, se observou a superexpress?o de genes relacionados ao estresse de forma marcante em uma das condi??es de an?lise relacionadas com flora??o precoce. / Sugarcane is an important culture for Brazil that holds almost half of all worldwide productivity. Plants face many challenges, because of biotic and abiotic stresses presents in the production field, which could prevent plants from reaching their genetic potential. As consequence, those stresses can generate Reactive Oxygen Species ? ROS ? that can cause damages on DNA. Another consequence of stress is the early-flowering process, which contributes for a reduction on yield. In this context, the aim of this work is to characterize ScMUTM1 and ScMUTM2, two DNA glycosylases belonging to base excision repair pathway; and identify genes potentially related to stress and DNA repair in two sugarcane cultivars with contrasting flowering phenotypes. The characterization of the DNA glycosylases included the construction of vector to over express the recombinant proteins ScMUTM1 and ScMUTM2; they will be used in a near future to purification of these proteins and use in enzymatic assays. It was also made a phylogenetic reconstruction of this gene in plants and analysis of its promoter. With the phylogenetic analysis, it is possible to observe the presence of these genes grouped inside a branch with monocots and another one with dicots. This suggests that the duplication of this gene probably occurred after the separation of these two groups. The analysis of the promotor of MUTM shows of the presence of stress-related regulatory motifs at ScMUTM2 promoter, when compared with ScMUTM1. This may suggests that ScMUTM1 might be suffering sub functionalization process. After the analysis of microarrays data, it is observed an up-regulation from some stress-related genes in one of the conditions analyzed, related to early flowering process.

Made available in DSpace on 2014-12-17T15:18:09Z (GMT). No. of bitstreams: 1 JairMS.pdf: 604378 bytes, checksum: 8f3220e89d5c178954aadb5186b98f52 (MD5) Previous issue date: 2009-02-27 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Hybrids among transgenic plants and related species are expected to occur if they are sympatric and when there are not crossing barriers; as is the case, in Brazil, of cry1Ac transgenic cotton and Gossypium barbadense. This species has been maintained as dooryard plants, and should be preserved as a genetic resource. Hybrids were evaluated about traits related to fitness, leading to infer about its chances of survivor and selection. A barbadense genotype collected at the state of Mato Grosso was outcrossed to the variety DP 404, containing the gene cry1Ac, and to the isoline DP 404. All the F1 individuals and 122 among 170 F2 individuals expressed the toxin, and presented levels of resistance to pink bollworm (Pectinophora gossypiella) and cotton leafworm (Alabama argillacea) equivalent to the transgenic parent and superior to the isoline, barbadense or non transgenic hybrids. The percentage of germination and number of days to germinate did not differ among genotypes. Anthesis of the first flower and opening of the first cotton boll occurred earlier for herbaceous cotton and F1 hybrids than F2 population in average; all the populations presented a number of days to flower and opening of the first boll smaller then barbadense. The highest plants were barbadenses, and herbaceus the smallest, with F1 and F2 populations presenting intermediary heights. The number of seeds per plants were superior for F1 hybrids an herbaceous cotton, F2 populations were in average intermediary; the barbadense genotype produced the smallest number of seeds per plant. Pink bollworm, mainly, and also cotton leafworm, are important barbadense pests, so the transgene positive effect could favor the selection of hybrids, and hence G. hirsutum genome, against the maintenance of pure G. barbadense genome. The selection may be influenced by the plant uses: the smaller size of hybrids when compared to the barbadense may lead them to be differentiated from these parents to which medicinal properties are attributed; on the other hand, the greater boll production may favor hybrids maintenance with the purpose of producing lamp wicks, or use as an ornamental or swab / Espera-se que hibrida??es entre plantas transg?nicas e esp?cies pr?ximas ocorram sempre que haja simpatria e n?o existam barreiras de cruzamento, como ? o caso, no Brasil, do algodoeiro contendo o gene cry1Ac e a esp?cie Gossypium barbadense. Os barbadenses v?m sendo mantidos em quintais, e ? importante sua preserva??o como recurso gen?tico. H?bridos foram avaliados para caracteres relacionados com a adaptabilidade, inferindo sobre suas chances de sobreviv?ncia e sele??o. Para isto, obtiveram-se plantas F1 e F2 do cruzamento entre um algodoeiro barbadense coletado no estado de Mato Grosso com as cultivares DP404, contendo cry1Ac, e de sua isolinha DP4049. Todos os 37 indiv?duos F1 e 122 dentre 170 indiv?duos F2 avaliados expressaram cry1Ac, e apresentaram n?veis de resist?ncia ?s lagartas rosada (Pectinophora gossypiella) e curuquer?-doalgodoeiro (Alabama argillacea) equivalentes ao parental transg?nicos e superiores a isolinha, barbadense e h?bridos que n?o continham o transgene. N?o foram observadas diferen?as significativas para a porcentagem de germina??o ou quantidade de dias para germina??o. O florescimento e abertura do primeiro capulho ocorreram mais cedo nos algodoeiros herb?ceos e nos h?bridos que nas popula??es F2 em m?dia; todas as popula??es apresentaram um n?mero de dias para o florescimento e abertura do primeiro capulho significativamente mais baixo que os barbadenses. As plantas mais altas na matura??o eram os barbadenses, e os herb?ceos as mais baixas, sendo as popula??es F1 e F2 intermedi?rias. O n?mero de sementes por planta foi superior nos h?bridos F1 e algodoeiros herb?ceos, sendo as popula??es F2 intermedi?rias em m?dia, e os barbadenses aqueles que produziram menor n?mero de sementes. A lagarta rosada, principalmente, e tamb?m a curuquer?-do-algodoeiro s?o importantes pragas de G. barbadense, portanto o efeito positivo do transgene pode favorecer a sele??o de h?bridos contendo o transgene, e deste modo favorecer plantas que cont?m genoma de G. hirsutum, em detrimento da pureza do genoma de barbadense. A sele??o deve ser influenciada pelos usos da planta: o fato de os h?bridos serem mais baixos pode diferenci?-los dos pais barbadenses aos quais a propriedades medicinais s?o atribuidas; por outro lado, a produ??o bem maior de capulhos pode favorecer a manuten??o dos h?bridos quando se visa confec??o de pavios ou ornamenta??o de quintais e jardins