Made available in DSpace on 2014-12-17T14:13:36Z (GMT). No. of bitstreams: 1 AnnaCQM.pdf: 331321 bytes, checksum: d4329ae02e1718e26e150e9d4894a746 (MD5) Previous issue date: 2008-06-20 / Although many studies point to alterations in the organic concentrations of zinc in elderly patients, the mechanisms by which aging might cause changes in the metabolism of this nutrient remain unclear. Thus, we assessed the changes in plasma zinc, Zinc Binding Capacity to Plasma Protein (ZnBCPP) and Saturation Index (SI), comparing elderly individuals and young adults. The zinc analyses were performed by atomic absorption spectrophotometry. A statistically significant difference (p < 0.001) was found between the two groups, in relation to plasma zinc and SI, but the ZnBCPP did not differ between the younger and older subjects. In agreement with this result, it was shown in the young group that 76% (R2 = 0.760) of the ZnBCPP variations are explained by the variations in plasma zinc and SI. In the elderly group this measure decreased to 30.5% (R2 = 0.305). We conclude, therefore, that aging may be a factor associated to changes in control mechanisms and in zinc homeostasis, and could even alter ZnBCPP response patterns and other zinc-related indicators of nutritional status. / Embora muitos estudos apontem altera??es nas concentra??es org?nicas de zinco em pacientes idosos, os mecanismos pelos quais o envelhecimento poderia implicar em mudan?as no metabolismo deste nutriente ainda permanecem obscuros. Assim, procuramos avaliar as mudan?as relativas ao zinco plasm?tico, Capacidade de Liga??o do Zinco ? Prote?na Plasm?tica (CLPPZn) e ?ndice de Satura??o (IS). As an?lises de zinco foram realizadas por espectrofotometria de absor??o at?mica, comparando idosos e adultos jovens. Foi encontrada diferen?a estaticamente significante (p<0,001), entre os dois grupos, em rela??o ao zinco plasm?tico e IS, sendo que a CLPPZn n?o diferiu entre os idosos e adultos jovens. Corroborando este resultado, foi demonstrado que, no grupo jovem as varia??es na CLPPZn s?o explicadas em 76% (R2= 0,760) pelas varia??es no zinco plasm?tico e IS. J? no grupo de idosos esta medida diminui para 30,5% (R2= 0,305). Conclu?mos ent?o que o envelhecimento pode ser um fator associado ?s mudan?as nos mecanismos de controle e homeostase do zinco, alterando inclusive padr?es de resposta relativos a CLPPZn e demais indicadores do estado nutricional relativo ao zinco.

Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-04T16:04:23Z No. of bitstreams: 2 mariana_aguiar_matos.pdf: 3238462 bytes, checksum: 3f1d0300b61c8980e15bd5468c0ae135 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2016-01-04T16:05:17Z (GMT) No. of bitstreams: 2 mariana_aguiar_matos.pdf: 3238462 bytes, checksum: 3f1d0300b61c8980e15bd5468c0ae135 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-01-04T16:05:17Z (GMT). No. of bitstreams: 2 mariana_aguiar_matos.pdf: 3238462 bytes, checksum: 3f1d0300b61c8980e15bd5468c0ae135 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2012 / Funda??o de Amparo ? Pesquisa do estado de Minas Gerais (FAPEMIG) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / A maior quantidade de ?cidos graxos livres e de citocinas pr?-inflamat?rias plasm?tica presentes na obesidade, podem desencadear a resist?ncia a insulina, dentre outros fatores, pela fosforila??o inibit?ria do substrato do receptor de insulina 1 (IRS-1), via ativa??o de quinases relacionadas ao estresse, como a quinase C-jun N-terminal (JNK). Em obesos, a resist?ncia a insulina correlaciona-se com altera??es do sistema imune, e com a baixa express?o da prote?na de choque t?rmico de 72kDa (Hsp72) e aumento da ativa??o da JNK no m?sculo esquel?tico. Considerando que o exerc?cio f?sico aer?bico promove melhora da sensibilidade a insulina e tem um efeito anti-inflamat?rio. O objetivo desse estudo foi avaliar os efeitos de uma sess?o de exerc?cio f?sico aer?bico na express?o da HSP70, ativa??o da JNK e fosforila??o do IRS-1 no res?duo de serina 612 (IRS-1 ser612) no m?sculo esquel?tico de obesos. Al?m disso, foi avaliada a frequ?ncia dos linf?citos T auxiliares (CD4+) e citol?ticos (CD8+) e das subpopula??es de mon?citos cl?ssicos (CD14++CD16-), intermedi?rios (CD14++CD16+) e n?o cl?ssicos (CD14+CD16++). Os participantes do estudo (n=27) foram alocados em tr?s grupos experimentais (eutr?ficos sens?veis a insulina, obesos sens?veis a insulina, obesos resistentes a insulina) de acordo com a classifica??o do estado nutricional, de acordo com o ?ndice de massa corporal e presen?a ou n?o de resist?ncia a insulina, definida pelo modelo de avalia??o da homeostase (HOMA1-IR). Amostras de sangue venoso e do m?sculo vasto lateral foram obtidas antes e ap?s uma sess?o de exerc?cio f?sico aer?bico realizado a 60% do VO2pico,em cicloerg?metro, com dura??o de 60 minutos. Para avaliar a frequ?ncia das diferentes popula??es de mon?citos e linf?citos T circulantes utilizou-se a citometria de fluxo. As an?lises da express?o da HSP70, ativa??o da JNK e fosforila??o do IRS-1 ser612 no m?sculo esquel?tico foram feitas pelo procedimento de western blot. Nossos resultados demonstraram que obesos resistentes a insulina apresentam uma maior frequ?ncia de mon?citos intermedi?rios e maior fosforila??o do IRS-1 ser612 comparado aos eutr?ficos, maior ativa??o da JNK e menor express?o da HSP70 em rela??o aos demais grupos. Ap?s 1 hora do t?rmino da sess?o de exerc?cio aer?bico houve redu??o da frequ?ncia dos mon?citos intermedi?rios (CD14++CD16+) e dos linf?citos auxiliares (TCD4+) circulantes. Adicionalmente, a sess?o de exerc?cio induziu no m?sculo esquel?tico maior express?o da HSP70, redu??o da fosforila??o do IRS-1 ser612 nos grupos de indiv?duos obesos e menor atividade da JNK nos obesos resistentes a insulina. Conclui-se que uma sess?o de exerc?cio aer?bico promove altera??es que caracterizam redu??o da inflama??o e/ou estresse celular, que podem contribuir para a modula??o da sensibilidade a insulina promovida pelo exerc?cio f?sico. / Disserta??o (Mestrado) ? Programa Multic?ntrico de P?s-gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2012. / ABSTRACT The great amount of free fatty acids and proinflammatory cytokines present in obese individuals may trigger insulin resistance, through the inhibitory phosphorylation of insulin receptor substrate 1 (IRS-1), via activation of kinases related to stress, such as kinase C-Jun N-terminal (JNK). In obese individuals, insulin resistance correlates with alterations of the immune system and lower expression of the heat shock protein of 72kDa (Hsp72) and increased activation of JNK in skeletal muscle. Considering that aerobic exercise improve insulin sensitivity and has an anti-inflammatory effect, the purpose of this study was to evaluate the effects of a bout of aerobic exercise in the expression of HSP70, activation of JNK and phosphorylation of IRS-1 in serine residue 612 (IRS-1 ser612) in skeletal muscle of obese patients. Furthermore, we assessed the frequency of helper T lymphocytes (CD4 +) and cytolytic (CD8 +) and subpopulations of classical (CD14 + + CD16-), intermediate (CD14 + + CD16 +) and non-classical (CD14 + CD16 + +) monocytes. Study participants (n = 27) were divided into three experimental groups (eutrophic sensitive to insulin, insulin-sensitive obese, insulin-resistant obese) according to the classification of nutritional status, according to the body mass index, and the presence or absence of insulin resistance, defined by the homeostasis assessment model (HOMA1-IR). Venous blood and vastus lateralis samples were obtained before and after a bout of aerobic exercise performed at 60% of VO2peak on a cycle ergometer, lasting 60 minutes. To assess the frequency of the different populations of monocytes and T lymphocytes circulating we used flow cytometry. Analyses of of HSP70 expression, JNK activation and IRS-1 phosphorylation of ser612 in skeletal muscle were performed by western blot. Our results showed that obese insulin resistant subjects have an increased frequency of intermediate monocytes and higher phosphorylation of IRS-1 ser612 compared to normal weight individuals, and greater activation of JNK and lower expression of HSP70 than the other two groups. 1 hour after the exercise bout, we observed reductions on the frequency of intermediate circulating monocytes (CD14 + + CD16 +) and helper cells (CD4 +). Additionally, the exercise bout induced in the skeletal muscle higher expression of HSP70, decreased phosphorylation of IRS-1 ser612 in the obese groups and lower activity of JNK in the obese insulin resistant individuals. It is concluded that a bout of aerobic exercise promotes changes that characterize reduction of inflammation and / or cellular stress, which may contribute to the modulation of insulin sensitivity promoted by exercise.

Made available in DSpace on 2015-03-03T14:02:15Z (GMT). No. of bitstreams: 1 TerezaNSB_TESE.pdf: 1549121 bytes, checksum: b4b539cbd0ea200212814064938aaefb (MD5) Previous issue date: 2011-11-28 / Universidade Estadual do Rio Grande do Norte / O objetivo desse trabalho foi investigar o eosin?filo e a prote?na cati?nica eosinof?lica (ECP) na urina de pacientes com L?pus Eritematoso Sist?mico (LES), com e sem nefrite l?pica, como poss?veis marcadores de inflama??o renal. Foram estudados 74 pacientes com LES 20 com evid?ncia cl?nica e laboratorial de nefrite l?pica (LN grupo) e 54 sem envolvimento renal (n?o-LN grupo) quanto ? eosinofil?ria e ECP urin?ria (uECP). A eosinofil?ria foi observada atrav?s da colora??o de Hansel e as concentra??es de ECP urin?ria foram obtidas por fluoroenzimaimunoensaio e em seguida corrigidas pela creatinina urin?ria (uECP/uCr). As vari?veis do estudo foram comparadas com a hemat?ria glomerular, rela??o prote?na/creatinina urin?ria (uPr/uCr), creatinina s?rica, clearance de creatinina estimado, anti-dsDNA, n?veis s?ricos dos complementos C3 e C4, rela??o IL-5 urin?ria/creatinina e com o ?ndice de atividade da doen?a LES (Mex-SLEDAI). A avalia??o preditiva da eosinofil?ria e uECP foi observada atrav?s da curva ROC e o n?vel de signific?ncia do estudo foi p valor<0,05. Os resultados mostraram que a eosinofil?ria e as concentra??es da uECP e uECP/uCr foram mais elevadas nos pacientes do LN grupo em rela??o ao n?o-LN grupo (p<0,001 para todos). Essas vari?veis mostraram uma correla??o estatisticamente significativa com a hemat?ria, dismorfismo eritrocit?rio glomerular, cilindr?ria, rela??o uPr/uCr, creatinina s?rica, clearance de creatinina estimado, anti-dsDNA, rela??o IL-5 urin?ria/creatinina e com o Mex-SLEDAI (p<0,05). Os resultados da curva ROC mostraram uma melhor performance (?rea sob a curva-AUC) para a uECP/uCr, usando como vari?vel de classifica??o a uPr/uCr (AUC=0,94) e o clearance de creatinina estimado (AUC=0,84), p<0,0001. Conclui-se que dentre as vari?veis do estudo, a uECP/uCr pode servir como um novo marcador de inflama??o renal em pacientes com LES

Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2015-12-15T17:45:12Z No. of bitstreams: 1 SinaraCarlaDaSilvaAraujo_DISSERT.pdf: 1885448 bytes, checksum: b4a8226a4f1778e4f626da3394657f9a (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2015-12-29T20:51:37Z (GMT) No. of bitstreams: 1 SinaraCarlaDaSilvaAraujo_DISSERT.pdf: 1885448 bytes, checksum: b4a8226a4f1778e4f626da3394657f9a (MD5) / Made available in DSpace on 2015-12-29T20:51:37Z (GMT). No. of bitstreams: 1 SinaraCarlaDaSilvaAraujo_DISSERT.pdf: 1885448 bytes, checksum: b4a8226a4f1778e4f626da3394657f9a (MD5) Previous issue date: 2014-08-22 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / Os microrganismos apresentam uma imensa diversidade gen?tica e est?o presentes em toda biosfera, no entanto cerca de 1% das esp?cies pode ser cultivada por t?cnicas laboratoriais padr?o. A metagen?mica tornou poss?vel o acesso direto ao genoma microbiano derivado de amostras ambientais, utilizando t?cnicas independentes de cultivo. A metodologia permite obter informa??o funcional de prote?nas, assim como a identifica??o de potenciais produtos com interesse biotecnol?gico e de novos recursos biol?gicos industrialmente explor?veis, a exemplo novas solu??es para impactos ambientais. ?reas contaminadas com petr?leo s?o caracterizadas por um grande ac?mulo de hidrocarbonetos e surfactantes s?o utilizados para sua biorremedia??o. Sendo assim, a abordagem metagen?nima foi utilizada para selecionar genes envolvidos no processo de degrada??o e biossurfacta??o de hidrocarbonetos. Em um trabalho anterior, o DNA ambiental (eDNA) foi extra?do de amostras de solo coletadas em duas diferentes ?reas (Caatinga e rio salino) do Rio Grande do Norte (Brasil), as bibliotecas metagen?micas foram constru?das e analisadas funcionalmente. Os clones capazes de degradar o ?leo foram avaliados quanto ? capacidade de sintetizar biossurfactante. O clone foi sequenciado e an?lise da sequencia revelou uma ORF com 897 pb, 298 amino?cidos e prote?na de peso molecular pr?ximo a 34 kDa. A busca por homologia no GenBank revelou similaridade com a sequ?ncia que codifica uma prote?na hipot?tica de representantes da fam?lia Halobacteriaceae, que foram mostradas recentemente como produtoras de biossurfactantes. A presen?a da sequ?ncia codificante inserida e do fen?tipo adquirido foram confirmadas. Primers foram desenhados e suas ORFs amplificadas por PCR. Em seguida, foram subclonadas em vetor de express?o pETDuet-1, contendo uma cauda de histidina, para express?o e posterior purifica??o da prote?na de interesse. Os testes foram realizados para confirma??o da atividade de biossurfactante e degrada??o de hidrocarbonetos e apresentaram resultados positivos. O ensaio de imunodetec??o (western blot) com a utiliza??o do anticorpo monoclonal Anti-His? confirmou a presen?a da prote?na ambiental. Esse estudo foi o primeiro a relatar uma poss?vel prote?na com atividade biossurfactante obtida a partir de uma abordagem metagen?mica. / The microorganisms have a vast genetic diversity and they are present throughout the biosphere, however, only about 1% of the species can be cultivated by traditional cultivation techniques. Within this diversity there is a huge pool genetic and biological being explored. The metagenomics has enabled direct access to microbial genome derived from environmental samples using independent methods of cultivation. The methodology enables to obtain functional information about the proteins, as well as identify potential products with biotechnological interest and new industrially exploitable biological resources, such as new solutions to environmental impacts. Oil-contaminated areas are characterized by a large accumulation of hydrocarbons and surfactants may be used for bioremediation. Thus, the metagenomic approach was used in this study in order to select genes involved in the degradation and hydrocarbon emulsification. In a previous work, the environmental DNA (eDNA) was extracted from soil samples collected from two different areas (Caatinga and Saline River) of Rio Grande do Norte (Brazil), the metagenomic libraries were constructed and functionally analyzed. The clone able to degrade the oil was evaluated for the ability to synthesize biosurfactants. The sequence analysis revealed an ORF with 897 bp, 298 amino acids and a protein with around 34 kDa. The search for homology in GenBank revealed sequence similarity with a hypothetical protein of representatives Halobacteriaceae family, who were recently shown as strains producing biosurfactants. The presence of the inserted coding sequence and the acquired phenotype was confirmed. Primers were designed and the ORF amplified by PCR. The ORF was subcloned into pETDuet-1 expression vector for subsequent purification of the protein of interest containing a histidine tail. The tests performed to confirm the biosurfactant activity and the ability of hydrocarbon degradation showed positive results. The immunodetection test (western blot) using the monoclonal AntiHis? confirmed the presence of the environmental protein. This study was the first to report a possible protein with biosurfactant activity obtained from a metagenomic approach

Made available in DSpace on 2014-12-17T15:01:13Z (GMT). No. of bitstreams: 1 MichelleRFV.pdf: 525804 bytes, checksum: d94754591de23bce5b0b460d3e10d382 (MD5) Previous issue date: 2008-02-25 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / With advent of the technology of the recombinant DNA, the recombinant protein expression becomes an important tool in the studies of the structure, function and identification of new proteins, mainly with therapeutical purposes. The Escherichia coli has been procarioto predominant in the studies of genetic engineering due to wealth of information regarding its metabolism. Despite the expressivo advance of the studies of molecular biology and the immunology of the infections, it does not exist, currently, no prophylactic drug capable to prevent calazar. Of this form, it exists a great necessity of specific antigen identification for the vaccine development and kits for disgnostic against the visceral Leishmaniose. In this context, this work objectified to study the recombinant antigen expression of the Leishmania chagasi during the culture of Escherichia coli in shaker. A first set of assays was carried through with the objective of if knowing the kinetic behavior of the growth of two clones recombinant proteins (eIF, LACK) in two different compositions of culture medium (2xTY, TB) supplemented by antibiotics, without IPTG addition. In the second stage of the assays, the procedure of induction for IPTG was carried through, in order to verify the influence of the composition of the ways tested in the expression them recombinant proteins. On the basis of the gotten results, can be observed that the high complexity of culture medium favored the kinetic one of growth of clones recombinant (eIF, LACK), however, to if to deal with the assays submitted to the procedure of induction for IPTG, the raised complexity of culture medium did not favor the expression of recombinant proteins. On the other hand, they had been gotten resulted positive for all clones recombinant (eIF, LACK) tested, confirmed through the eletrofor?tico profile / Com advento da tecnologia do DNA recombinante, a express?o de prote?nas recombinantes torna-se uma ferramenta importante nos estudos da estrutura, fun??o e identifica??o de novas prote?nas, principalmente com finalidades terap?uticas. A Escherichia coli tem sido o procarioto predominante nos estudos da engenharia gen?tica devido ? riqueza de informa??es a respeito do seu metabolismo. Apesar do avan?o expressivo dos estudos da biologia molecular e da imunologia das infec??es, n?o existe, atualmente, nenhuma droga profil?tica capaz de prevenir o calazar. Desta forma, existe uma grande necessidade de identifica??o de ant?genos espec?ficos para o desenvolvimento de vacinas e kits para diagn?sticos contra a Leishmaniose visceral. Neste contexto, este trabalho objetivou estudar a express?o de ant?genos recombinantes da Leishmania chagasi durante o cultivo de Escherichia coli em incubador rotativo (shaker). Um primeiro conjunto de ensaios foi realizado com o objetivo de se conhecer o comportamento cin?tico do crescimento dois clones recombinantes (eIF, LACK) em duas diferentes composi??es de meios (2xTY, TB) suplementados por antibi?ticos, sem adi??o de IPTG. Na segunda etapa dos ensaios, foi realizado o procedimento de indu??o por IPTG, a fim de verificar a influ?ncia da composi??o dos meios testados na express?o das prote?nas recombinantes. Com base nos resultados obtidos, pode-se observar que a elevada complexidade do meio de cultivo favoreceu a cin?tica de crescimento dos clones recombinantes (eIF,LACK), no entanto, ao se tratar dos ensaios submetidos ao procedimento de indu??o por IPTG, a elevada complexidade do meio de cultivo n?o favoreceu a express?o das prote?nas recombinantes. Por outro lado, foram obtidos resultados positivos para todos os clones recombinante (eIF, LACK) testados, confirmada atrav?s do perfil eletrofor?tico

Made available in DSpace on 2014-12-17T15:31:02Z (GMT). No. of bitstreams: 1 CandiceFRB.pdf: 345545 bytes, checksum: a87f0c9c7b61e16a29dfad599e006ac3 (MD5) Previous issue date: 2007-10-29 / INTRODUCTION: The high sensitivity C-reactive protein (hsCRP) constitutes an inflammatory mediator used as predictor of cardiovascular risk that comes being researched as indicative relation factor between cardiovascular and periodontal diseases. PROPOSITION: To compare serumals levels of C-reactive protein between patients with and without generalized severe chronic periodontitis. METHODOLOGY: A seccional study was realized using a sample with 62 patients, being 31 participants carriers of periodontal diseases (Group I) and 31 without periodontal diseases (Group II), grouped to the pairs by age and sex. As inclusion criterio were selected patients with diagnosis of generalized severe chronic periodontitis, being preculeds, individuals which presented systemic disease, recent infection history, historical of CVA or stroke, smokers, pregnants and lactants. The research consisted of two stages, a clinc and other biochemist. The clinical stage is constituted of periodontal examination and the biochemist stage, of the peripheral blood collection for determination hsCRP levels and a hemogram to inquire any panel which could suggest infectious and/or inflammatory process. RESULTS: Periodontal disease group presented a average of 0,36mg/dL, while the group without disease presented 0,17 mg/dL, do not existing significant difference statistically between the averages (p = 0,061). The cardiovascular risk for the group I was classified high for 27,6% of participants and low for 72,4% of them. In the group II, 6,45% presented high risk e 93,5% low risk, being this significant relation statistically gotten for Fisher s Test (p = 0,042) presenting OR = 5,33; IC = 95% (1,02 27,4). The independets variables reseacred do not presented significant association statistically with the levels of hsCRP. CONCLUSION: The study indicated that despite of carriers patients of periodontal diseases do not present differents serumals levels of hsCRP from the other group, the periodontal disease was considered as risk factor for hsCRP plasmatic levels elevation / INTRODU??O: A prote?na C-reativa ultra-sens?vel (PCR-US) constitui um mediador inflamat?rio utilizada como preditor do risco cardiovascular que vem sendo pesquisado como fator indicativo da rela??o entre doen?as cardiovasculares e periodontais. PROPOSI??O: Comparar os n?veis s?ricos de prote?na C-reativa entre pacientes com e sem periodontite cr?nica severa generalizada. METODOLOGIA: Foi realizado um estudo seccional, utilizando uma amostra de 62 pacientes, sendo 31 participantes com doen?a periodontal (Grupo I) e 31 sem doen?a periodontal (Grupo II), selecionados segundo idade e sexo. Como crit?rios de inclus?o foram selecionados pacientes com diagn?stico de periodontite cr?nica severa generalizada, sendo exclu?dos, indiv?duos que apresentassem doen?a sist?mica, hist?ria de infec??o recente, hist?rico de AVC ou infarto, fumantes, gestantes e lactantes. A pesquisa constou de duas etapas, sendo uma cl?nica e outra bioqu?mica. A etapa cl?nica consistiu no exame periodontal e a bioqu?mica na coleta de sangue perif?rico para determina??o dos n?veis de PCR-US e um hemograma para averiguar algum quadro que pudesse sugerir processo infeccioso e/ou inflamat?rio. RESULTADOS: O grupo com doen?a periodontal apresentou uma m?dia de 0,36 mg/dL, enquanto o sem doen?a de 0,17 mg/dL, n?o existindo diferen?a estatisticamente significativa entre as m?dias (p=0,061). O risco cardiovascular para o grupo I foi classificado como alto para 27,6% dos participantes e baixo para 72,4%. No grupo II, 6,45% apresentavam alto risco e 93,5% baixo risco, sendo essa rela??o estatisticamente significativa obtida pelo teste de Fisher (p=0,042), apresentando um OR=5,33; IC95%(1,02-27,4). As vari?veis intervenientes pesquisadas n?o apresentaram associa??o estatisticamente significativa com os n?veis de PCR-US. CONCLUS?O: O estudo indicou que apesar dos pacientes portadores de periodontite severa cr?nica generalizada n?o apresentarem n?veis s?ricos de PCR-US diferentes dos sem doen?a, a doen?a periodontal atuou como fator de risco para eleva??o dos n?veis plasm?ticos de PCR-US

Submitted by PPG Biologia Celular e Molecular (bcm@pucrs.br) on 2017-10-24T11:23:13Z No. of bitstreams: 1 BRUNA_FERREIRA_LEAL_TES.pdf: 9428836 bytes, checksum: 9ef80f04ac10ad1e26b54ce86ddd80ce (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-10-26T15:45:53Z (GMT) No. of bitstreams: 1 BRUNA_FERREIRA_LEAL_TES.pdf: 9428836 bytes, checksum: 9ef80f04ac10ad1e26b54ce86ddd80ce (MD5) / Made available in DSpace on 2017-10-26T15:54:38Z (GMT). No. of bitstreams: 1 BRUNA_FERREIRA_LEAL_TES.pdf: 9428836 bytes, checksum: 9ef80f04ac10ad1e26b54ce86ddd80ce (MD5) Previous issue date: 2017-03-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Rhipicephalus microplus is a tick that infests preferably bovines and cause a great damage in the livestock. The main control method against ticks is the use of acaricides, which lead to the emergence of resistant populations, in addition to increasing costs and risk of contamination of meat, milk and environment. Thus, vaccine antigens have been identified and characterized as an alternative to acaricides. Salivary molecules of hematophagous parasites can inhibit responses associated to homeostasis and modulate the host immune system and, therefore, are options in the search for anti-tick vaccines. This work characterized a glycine rich-protein and the paramyosin of R. microplus. Glycine rich-proteins are secreted in tick saliva and are essential in the attachment and blood feeding. Paramyosin is able to evade the host immune system and proved to be an important allergen of mites and vaccine antigen against helminths. The cDNA and amino acid sequences of the R. microplus glycine rich-protein (RmGRP) were identified and analyzed, revealing two distinct portions of the protein (glycine residues presented dispersed in the N-terminal region and in repeated patterns along the C-terminal region). Cloning, expression and purification of RmGRP were performed and the recombinant protein was tested for recognition by sera from naturally and experimentally infested bovines, indicating their antigenicity, but also displaying a high heterogeneity in protein recognition between individuals and among infestations in the same individual. In addition, recombinant RmGRP was recognized by sera from rabbits immunized with saliva and salivary gland from partially and fully engorged females and eggs, but not from sera from rabbits immunized with gut. Expression of the gene encoding RmGRP (rmgrp) in different female tissues and tick development stages was evaluated by qRT-PCR. Gene transcription was found in eggs, larvae, adult males, salivary glands, fat bodies and ovaries of partially and fully engorged females, with the highest expression levels in 1-day-old larvae and salivary glands of fully engorged females. rmgrp was not expressed in 3- and 6-day-old eggs and in guts of partially and fully engorged females, corroborating to the aforementioned result. Effects of RNAm silencing corresponding to RmGRP and RmPRM were evaluated, which resulted in egg laying reduction in the group in which the RmPRM gene was silenced and in the reduction of larval hatching rate in the two groups. Therefore, it is suggested that both proteins are important during larval development, but only RmPRM is required in egg formation and/or laying. Cloning of the coding DNA regions and expression of N-terminal, internal and Cterminal fragments of R. microplus paramyosin (RmPRM) in Escherichia coli and of the complete RmPRM in Pichia pastoris were confirmed by SDS-PAGE and western-blot. From the data obtained, both RmGRP and RmPRM have characteristics that make them possible candidates to compose a cocktail vaccine against R. microplus. / O Rhipicephalus microplus ? um carrapato que infesta preferencialmente bovinos, causando um grande preju?zo ? pecu?ria. O principal m?todo de controle ? o uso de acaricidas, os quais selecionam popula??es resistentes, al?m de aumentar o custo e o risco de contamina??o da carne, do leite e do ambiente. Desta forma, ant?genos vacinais t?m sido identificados e caracterizados como alternativa aos acaricidas. Mol?culas salivares de parasitos hemat?fagos podem inibir respostas associadas ? homeostase e modular o sistema imune do hospedeiro e, portanto, s?o op??es na busca por vacinas anti-carrapato. Neste contexto, este trabalho caracterizou uma prote?na rica em glicinas e a paramiosina do R. microplus. Prote?nas ricas em glicinas s?o secretadas na saliva e s?o essenciais na fixa??o e na alimenta??o do parasito. J? a paramiosina ? capaz de evadir o sistema imune do hospedeiro e mostrou-se um importante alergeno em ?caros e ant?geno vacinal contra helmintos. As sequ?ncias de cDNA e amino?cidos da prote?na rica em glicinas de R. microplus (RmPRG) foram identificadas e analisadas, revelando duas por??es distintas da prote?na (com glicinas isoladas ao longo da regi?o N-terminal e padr?es repetidos contendo glicinas ao longo da regi?o C-terminal). A clonagem, express?o e purifica??o da RmGRP foram realizadas e a prote?na recombinante foi testada quanto ao reconhecimento por soros de bovinos naturalmente e experimentalmente infestados, indicando a sua antigenicidade, mas tamb?m uma alta heterogeneidade no reconhecimento da prote?na entre indiv?duos e entre infesta??es no mesmo indiv?duo. Al?m disso, a RmGRP recombinante foi reconhecida pelos soros de coelhos imunizados com saliva e gl?ndula salivar de f?meas parcialmente e totalmente ingurgitadas e ovos, mas n?o por soros de coelhos imunizados com intestino. A express?o do gene codificante para a RmGRP (rmgrp) em diferentes tecidos de f?meas e fases do desenvolvimento do carrapato foi avaliada por qRT-PCR. A transcri??o do gene foi encontrada em ovos, larvas, machos adultos, al?m de gl?ndulas salivares, corpos gordurosos e ov?rios de f?meas parcialmente e totalmente ingurgitadas, apresentando os maiores n?veis de express?o nas larvas de 1 dia e nas gl?ndulas salivares de f?meas totalmente ingurgitadas. rmgrp n?o foi expresso nos ovos de 3 e 6 dias e no intestino de f?meas parcialmente e totalmente ingurgitadas, corroborando o resultado anterior. Ainda foram avaliados os efeitos do silenciamento do RNAm correspondente ? RmGRP e ? RmPRM, o que resultou na redu??o da taxa de postura de ovos no grupo em que o gene da RmPRM foi silenciado e da eclos?o de larvas em f?meas tratadas nos dois grupos. Portanto, sugere-se que as duas prote?nas s?o importantes durante o desenvolvimento larval, mas s? a RmPRM seja necess?ria na forma??o e/ou postura dos ovos. Tamb?m foi realizada a 8 clonagem da regi?o codificante e express?o dos fragmentos N-terminal, interno e C-terminal da paramiosina de R. microplus (RmPRM) em E. coli e da RmPRM completa em P. pastoris, sendo confirmadas por SDS-PAGE e western-blot. A partir dos dados obtidos, ambas RmGRP e RmPRM apresentam caracter?sticas que fazem delas poss?veis candidatas a comporem uma vacina coquetel contra o R. microplus.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-10-18T11:49:54Z No. of bitstreams: 1 2014 - Ramon de Sousa Rego.pdf: 1491511 bytes, checksum: 746021e21f53b13713de3e8e94d03121 (MD5) / Made available in DSpace on 2016-10-18T11:49:54Z (GMT). No. of bitstreams: 1 2014 - Ramon de Sousa Rego.pdf: 1491511 bytes, checksum: 746021e21f53b13713de3e8e94d03121 (MD5) Previous issue date: 2014-08-26 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico - CNPq / The protein quality of milk of goats and digestibility of the lipid fraction are important factors that stand out when compared to milk from cows. The ?-lactoglobulin is the higher abundant protein in whey ruminants being produced in the mammary gland during lactation. It may represent up to 12% of the total protein. We seek to present work was to evaluate the gene ?-Lactoglobulin (BLG) with their genetic polymorphisms in the promoter regions 5 'and 3' UTR and associate them the characteristics of milk production in experimental goat herd. For this 150 goats (Capra hircus) of Saanen and Alpine, genotyped for polymorphisms of the BLG gene were used. For genotyping 10 mL of blood per animal were collected. The blood was used for DNA extraction and amplification of two fragments of the BLG gene by means of polymerase chain reaction (PCR). The amplified fragments were submitted to electrophoresis on polyacrylamide gel and evaluated by by restriction fragment length polymorphism (PCR-RFLP). The SmaI and SacII enzymes were used for the promoter (promoter + exon 1 region) region and the region of exon 7 (exon 7 + region 3 '), respectively. The differences in cutting patterns were visualized on a polyacrylamide gel. In the promoter region of the single nucleotide polymorphism (SNP) at position -60 (C/T) was identified and was associated with the percentage of protein in milk. This result suggests a relationship between genotypes of the promoter region of the BLG gene and the protein level of the milk. The presence of two polymorphisms +4641 (I2/I3) and +4601 (A/G) was observed in the region of exon 7. The I2-I3 variation is characterized by a repeat sequence of 10 bp, varying in two and three times, and it being possible to identify by electrophoresis. The I3 variation was often very low, and there was no association between this polymorphism and any production trait. Polymorphism +4601 (G/A) was identified by enzyme SacII digestion. Alleles were identified frequencies close to those reported by other authors and were associated with the percentage of fat in milk goats, and the animals with the S1S2 genotype had higher fat percentage than S1S1 animals and S2S2. The increased production of lipid content may be related to the characteristic of transporting fatty acids ?-lactoglob / A qualidade proteica do leite de cabras e a digestibilidade da fra??o lip?dica s?o importantes fatores que o destacam, quando comparado ao leite de vacas. A ?-Lactoglobulina ? a prote?na de maior abundancia no soro do leite em ruminantes sendo produzida na gl?ndula mam?ria durante o per?odo de lacta??o. Ela pode representar at? 12% do total proteico. Buscamos com o presente trabalho, avaliar o gene da ?-Lactoglobulina (BLG) com seus polimorfismos gen?ticos nas regi?es promotoras 5? e 3? UTR e associ?-los as caracter?sticas de produ??o de leite no rebanho caprino experimental. Para isso foram utilizadas 150 cabras (Capra hircus) das ra?as Saanen e Alpinas, genotipadas para os polimorfismos do gene BLG. Para a genotipagem foram coletados 10 mL de sangue por animal. O sangue foi utilizado para a extra??o de DNA e amplifica??o de 2 fragmentos do gene BLG por interm?dio da rea??o em cadeia da polimerase (PCR). Os fragmentos amplificados foram ent?o submetidos ? eletroforese em gel de poliacrilamida e avaliados pelo polimorfismo no tamanho do fragmento por restri??o (PCR-RFLP). Foram utilizadas as enzimas SmaI e SacII, para a regi?o promotora (regi?o promotora + ?xon 1) e para a regi?o do ?xon 7 (?xon 7 + regi?o 3?), respectivamente. As diferen?as nos padr?es de corte foram visualizadas em gel de poliacrilamida. Na regi?o promotora o polimorfismo de base ?nica (SNP) na posi??o -60 (C/T) foi identificado e apresentou associa??o com a percentagem de prote?nas no leite. Esse resultado sugere uma rela??o entre os gen?tipos da regi?o promotora do gene BLG e o n?vel proteico do leite. Foi observado na regi?o do ?xon 7 a presen?a de dois polimorfismos +4641 (I2/I3) e +4601 (A/G). A varia??o I2-I3 ? caracterizada pela repeti??o de uma sequ?ncia de 10 pb, variando em duas e tr?s vezes, e sendo poss?vel identifica-la por eletroforese. A varia??o I3 teve frequ?ncia muito baixa, e n?o houve associa??o entre esse polimorfismo e nenhuma caracter?stica de produ??o. O polimorfismo +4601 (A/G) foi identificado por meio da digest?o da enzima SacII. Os alelos identificados tiveram frequ?ncias pr?ximas ?s relatadas por outros autores e apresentaram associa??o com a percentagem de gordura no leite cabras, sendo que os animais com o gen?tipo S1S2 apresentaram maior percentagem de gordura que animais S1S1 e S2S2. A maior produ??o de conte?do lip?dico pode estar relacionada com a caracter?stica de transporte de ?cidos graxos da ?-Lactoglobulina

Submitted by Verena Bastos (verena@uefs.br) on 2015-09-21T13:18:11Z No. of bitstreams: 1 DISSERTACAO FINAL ALINE THOMAZ.pdf: 1680438 bytes, checksum: 85bf2d7886a394df745618b6fd50d435 (MD5) / Made available in DSpace on 2015-09-21T13:18:11Z (GMT). No. of bitstreams: 1 DISSERTACAO FINAL ALINE THOMAZ.pdf: 1680438 bytes, checksum: 85bf2d7886a394df745618b6fd50d435 (MD5) Previous issue date: 2015-09-27 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The vitamin D receptor (VDR) is a cytoplasmic transcription factor and when activated by its ligand, translocates to the nucleus and interacts with DNA in the promoter regions with which has affinity, acting as a modulator of gene transcription and thereby producing multiple biological effects. Its main activities are the regulation and maintenance of plasma levels of calcium and phosphorus, as well as presenting immunomodulatory activities, such as the suppression of T cell activation, formation of secretion patterns of cytokines, modulation of proliferation and interference in the apoptosis. So this receptor is an important target for drugs that can help in the discovery of new immunomodulators. The present work had the objective to produce the recombinant vitamin D receptor in its soluble form for conducting future assays of drug screening of potential immunomodulators and drug-receptor interaction studies. For this, we used initially Escherichia coli expression system transformed with the plasmid HS_VDR_EC1-PQE T7, but it was only possible to obtain the protein in the insoluble fraction, even varying the temperature, time of induction and IPTG concentration. In an attempt to obtain soluble VDR, we used a eukaryotic expression system in insect cells using the baculovirus as a vector. It was built a vector, pFASTBacHT_VDR, which had the sequence of this protein cloned from pCMX.VDR. From there, it was possible to obtain recombinant bacmids used in transfection of insect cells, generating recombinant baculovirus, to then proceed with the expression of VDR. / O receptor de vitamina D (VDR) ? um fator de transcri??o g?nica citoplasm?tico e, quando ativado pelo seu ligante, transloca-se para o n?cleo e interage com regi?es promotoras no DNA com a qual apresenta afinidade, atuando como fator modulador da transcri??o g?nica e assim produzindo m?ltiplos efeitos biol?gicos. Suas principais atividades s?o a regula??o e a manuten??o dos n?veis plasm?ticos de c?lcio e f?sforo, e apresenta atividades imunomoduladoras, como a supress?o da ativa??o de c?lulas T, forma??o de padr?es de secre??o de citocinas, a modula??o da prolifera??o e interfer?ncia na apoptose. Sendo assim, esse receptor representa um importante alvo de drogas que pode contribuir na descoberta de novos f?rmacos com a??o imunomoduladora. O presente trabalho teve, como objetivo, produzir o VDR recombinante na forma sol?vel para a realiza??o de futuros ensaios de triagem de potenciais drogas com a??o imunomoduladora e estudos de intera??o droga-receptor. Para isso, foi utilizado, inicialmente, o sistema de express?o em Escherichia coli, utilizando o plasm?deo HS_VDR_EC1-PQE T7, por?m s? foi poss?vel obter a prote?na na fra??o insol?vel, mesmo variando a temperatura, o tempo de indu??o e a concentra??o de IPTG. Na tentativa de obter o VDR sol?vel, foi utilizado um sistema de express?o eucari?tico em c?lulas de inseto, utilizando como vetor o baculov?rus. Foi constru?do um vetor pFASTBacHT_VDR, o qual teve a sequ?ncia desta prote?na clonada a partir do pCMX.VDR. A partir da?, foi poss?vel obter bacm?deos recombinantes, utilizados na transfec??o de c?lulas de inseto, gerando baculov?rus recombinantes, para, ent?o, seguir com a express?o do VDR.

Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-02-27T13:30:33Z No. of bitstreams: 5 30.pdf: 544436 bytes, checksum: 4dca3d1781581491c5f88c4947e642ce (MD5) license_url: 52 bytes, checksum: 3d480ae6c91e310daba2020f8787d6f9 (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) license.txt: 2109 bytes, checksum: aa477231e840f304454a16eb85a9235f (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2015-02-27T18:23:54Z (GMT) No. of bitstreams: 5 30.pdf: 544436 bytes, checksum: 4dca3d1781581491c5f88c4947e642ce (MD5) license_url: 52 bytes, checksum: 3d480ae6c91e310daba2020f8787d6f9 (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) license.txt: 2109 bytes, checksum: aa477231e840f304454a16eb85a9235f (MD5) / Made available in DSpace on 2015-02-27T18:23:54Z (GMT). No. of bitstreams: 5 30.pdf: 544436 bytes, checksum: 4dca3d1781581491c5f88c4947e642ce (MD5) license_url: 52 bytes, checksum: 3d480ae6c91e310daba2020f8787d6f9 (MD5) license_text: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) license_rdf: 23898 bytes, checksum: e363e809996cf46ada20da1accfcd9c7 (MD5) license.txt: 2109 bytes, checksum: aa477231e840f304454a16eb85a9235f (MD5) Previous issue date: 2011 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / Empresa de Pesquisa Agropecu?ria de Minas Gerais (EPAMIG) / Objetivou-se com o presente trabalho avaliar a produtividade de massa seca, a composi??o qu?mico-bromatol?gica e a extra??o de minerais do capim-braqui?ria em diferentes idades de rebrota??o ap?s a colheita do milho, arranjos, clones de eucalipto e locais de amostragem, na integra??o lavoura-pecu?ria-floresta. O experimento foi conduzido na Fazenda Experimental de Santa Rita/EPAMIG, Prudente de Morais, MG, em esquema de parcelas subsubdivididas, no delineamento em blocos casualizados, com tr?s repeti??es. Nas parcelas distribu?ram-se as idades de rebrota??o da forrageira (10, 17, 24, 31, 38, 45 e 52 dias ap?s a colheita do milho para silagem). As subparcelas corresponderam aos arranjos de eucalipto, em linhas duplas: (3x2) + 20 m e (2x2) + 9 m, e em linha simples: 9 x 2 m (experimento 1); e clones de eucalipto, GG 100, I 144 e VM 58 (experimento 2). As subsubparcelas consistiram nos locais de amostragem, no centro da entrelinha e sob a copa de eucalipto. No experimento 1 o arranjo (3x2) + 20 m proporciona maior produtividade de massa seca do capim-braqu?ria, no centro da entrelinha. Entretanto, as produtividades de massa seca s?o baixas, mesmo com elevadas altura de plantas. Os arranjos (2x2) + 9 m e 9 x 2 m proporcionam melhor composi??o bromatol?gica, no centro da entrelinha, com maior teor de PB e menor teor de FDN. Os teores de prote?na bruta s?o mais altos sob a copa de eucalipto, relativamente ao centro da entrelinha. O arranjo (3x2) + 20 m proporciona maior extra??o de N, K, Ca e Mg no centro da entrelinha do que sob a copa de eucalipto. Maior extra??o de P ocorreu no arranjo de (3x2) + 20 m, no centro da entrelinha. As extra??es de N, K, Ca, Mg e S aumentam com o avan?o da idade de rebrota??o. No experimento 2, Os clones de eucalipto n?o afetam a produtividade de massa seca do capim-braqui?ria que apresenta maior disponibilidade no centro da entrelinha do que sob a copa de eucalipto. O capim-braqui?ria cultivado junto com o clone I 144 apresenta melhor composi??o bromatol?gica, com mais elevados teores de PB, N, P, Ca, Mg e S e mais baixo teor de FDN. As extra??es de N, P, Ca, Mg e S aumentam com o avan?o da idade de rebrota??o do capim-braqui?ria, sendo menores sob a copa de eucalipto para P e Mg. O clone VM 58 proporciona maior extra??o de K pelo capim-braqui?ria e com mais alta efici?ncia de resposta, no centro da entrelinha de eucalipto. / Disserta??o (Mestrado) ? Programa de P?s-Gradua??o em Produ??o Vegetal, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2011. / ABSTRACT This study aimed to evaluate the productivity of dry mass, chemical bromatological composition and mineral extraction of signal grass at different sprout ages after maize harvest, eucalyptus arrangements, clones, and sampling sites, cultivated in crop-livestock-forest. A trial was carried out at Santa Rita Experimental Farm/EPAMIG, Prudente de Morais, MG, the experimental design was a randomized complete block (RBD) in a split split plot, with three replications. The ages of forage sprouts (10, 17, 24, 31, 38, 45 and 52 days after silage maize harvest) were distributed in the main plots. The eucalyptus arrangements, in double rows: (3 x 2) + 20 m and (2x2) + 9 m, and simple row: 9 x 2 m (experiment 1), and eucalyptus clones, GG 100, I 144 and VM 58 (experiment 2) in the subplots. The sub sub plots consisted of forage sampling sites, in the inter-row (center) and under the canopy of eucalyptus. In the experiment 1, the arrangement (3x2) + 20 m produced higher dry mass of signal grass, in the inter-row (center). However, yields of dry mass are low, even with high plant height. Arrangements (2x2) + 9m and 9 x 2 m provided better chemical composition, in the inter-row (center), with higher CP and lower NDF content. The crude protein levels are higher under the canopy of eucalyptus, for the inter-row (center). The arrangement (3x2) + 20 m provides better extraction of N, K, Ca and Mg in the inter-row (center) than under the canopy of eucalyptus. Greater extraction of P occurred in the arrangement (3x2) + 20 m in the inter-row (center). The extraction of N, K, Ca, Mg and S increase with advancing age sprouts. In the experiment 2, the eucalyptus clones do not affect the productivity of dry mass of signal grass and has greater availability in the inter-row (center) than under the canopy of eucalyptus. The signal grass grown with the clone I 144 has better chemical composition, with higher concentrations of CP, N, P, Ca, Mg e S and lower NDF content. The extractions of N, P, Ca, Mg and S increase with advancing maturity of signal grass, and it is smaller under the canopy of eucalyptus for P and Mg. The signal grass cultived with the VM 58 clone present higher extraction of K and highest efficiency of response, in the inter-row.