Submitted by Automa??o e Estat?stica (sst@bczm.ufrn.br) on 2016-05-03T22:56:48Z No. of bitstreams: 1 HeltonMaiaPeixoto_TESE.pdf: 4738937 bytes, checksum: ea09c631840db43949ce6fbc2d32045c (MD5) / Approved for entry into archive by Arlan Eloi Leite Silva (eloihistoriador@yahoo.com.br) on 2016-05-05T20:05:06Z (GMT) No. of bitstreams: 1 HeltonMaiaPeixoto_TESE.pdf: 4738937 bytes, checksum: ea09c631840db43949ce6fbc2d32045c (MD5) / Made available in DSpace on 2016-05-05T20:05:06Z (GMT). No. of bitstreams: 1 HeltonMaiaPeixoto_TESE.pdf: 4738937 bytes, checksum: ea09c631840db43949ce6fbc2d32045c (MD5) Previous issue date: 2015-07-31 / As prote?nas fluorescentes constituem uma ferramenta fundamental em v?rios campos da biologia, pois permitem enxergar o desenvolvimento de estruturas e processos din?micos de c?lulas em tecido vivo, com o aux?lio da microscopia de fluoresc?ncia. A Optogen?tica ? outra t?cnica que atualmente ganha destaque na Neuroci?ncias e que, de forma geral, permite ativar/desativar neur?nios a partir da irradia??o luminosa de certos comprimentos de onda sobre as c?lulas que possuem canais i?nicos sens?veis ? luz, e ainda pode ser utilizada concomitantemente com as prote?nas fluorescentes. Esta tese possui dois objetivos principais. Inicialmente, s?o estudados os efeitos da intera??o da luz e o c?rebro de camundongos para aplica??es em experimentos de Optogen?tica. Nesta etapa, s?o modelados, a partir de caracter?sticas do c?rebro de camundongos e utilizando a teoria de Kulbelka-Munk, os efeitos de absor??o e espalhamento da luz, em comprimentos de onda espec?ficos, em fun??o da dist?ncia de penetra??o no tecido cerebral. Al?m disso, s?o modeladas as varia??es de temperatura utilizando o m?todo dos elementos finitos na resolu??o da equa??o de bioaquecimento de Pennes, com o aux?lio do COMSOL Multiphysics Modeling Software 4.4, onde s?o simulados protocolos de estimula??o luminosa, tipicamente utilizados em Optogen?tica. Posteriormente, s?o desenvolvidos algoritmos computacionais capazes de reduzir a exposi??o das c?lulas nervosas ? irradia??o luminosa necess?ria ? visualiza??o da fluoresc?ncia emitida por elas. Nesta etapa, s?o descritas as t?cnicas de processamento digital de imagens desenvolvidas para uso em microscopia de fluoresc?ncia, com o intuito de reduzir a exposi??o das amostras de c?rebro ? luz cont?nua, respons?vel pela excita??o dos fluorocromos. As t?cnicas de processamento de imagens desenvolvidas e utilizadas s?o capazes de rastrear, em tempo real, uma regi?o de interesse (ROI) e substituir a fluoresc?ncia emitida pelas c?lulas por uma m?scara virtual, como resultado da sobreposi??o da ROI que est? sendo rastreada e a informa??o de fluoresc?ncia previamente armazenada, mantendo a localiza??o das c?lulas independentemente do tempo de exposi??o ? luz fluorescente. Em resumo, esta tese pretende entender os efeitos da irradia??o luminosa no c?rebro, no contexto da Optogen?tica, al?m de fornecer uma ferramenta computacional que possa auxiliar certos experimentos em microscopia de fluoresc?ncia na redu??o do desvanecimento (bleaching) das amostras e dos danos (photodamage) causados ao tecido devido ? intensa exposi??o das estruturas fluorescentes ? luz. / The fluorescent proteins are an essential tool in many fields of biology, since they allow us to watch the development of structures and dynamic processes of cells in living tissue, with the aid of fluorescence microscopy. Optogenectics is another technique that is currently widely used in Neuroscience. In general, this technique allows to activate/deactivate neurons with the radiation of certain wavelengths on the cells that have ion channels sensitive to light, at the same time that can be used with fluorescent proteins. This dissertation has two main objectives. Initially, we study the interaction of light radiation and mice brain tissue to be applied in optogenetic experiments. In this step, we model absorption and scattering effects using mice brain tissue characteristics and Kubelka-Munk theory, for specific wavelengths, as a function of light penetration depth (distance) within the tissue. Furthermore, we model temperature variations using the finite element method to solve Pennes? bioheat equation, with the aid of COMSOL Multiphysics Modeling Software 4.4, where we simulate protocols of light stimulation tipically used in optogenetics. Subsequently, we develop some computational algorithms to reduce the exposure of neuron cells to the light radiation necessary for the visualization of their emitted fluorescence. At this stage, we describe the image processing techniques developed to be used in fluorescence microscopy to reduce the exposure of the brain samples to continuous light, which is responsible for fluorochrome excitation. The developed techniques are able to track, in real time, a region of interest (ROI) and replace the fluorescence emitted by the cells by a virtual mask, as a result of the overlay of the tracked ROI and the fluorescence information previously stored, preserving cell location, independently of the time exposure to fluorescent light. In summary, this dissertation intends to investigate and describe the effects of light radiation in brain tissue, within the context of Optogenetics, in addition to providing a computational tool to be used in fluorescence microscopy experiments to reduce image bleaching and photodamage due to the intense exposure of fluorescent cells to light radiation.

Made available in DSpace on 2014-12-17T14:56:03Z (GMT). No. of bitstreams: 1 ValnaideGB.pdf: 1369975 bytes, checksum: 404710d72240200cbd30a9116933d340 (MD5) Previous issue date: 2005-11-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Nowadays, classifying proteins in structural classes, which concerns the inference of patterns in their 3D conformation, is one of the most important open problems in Molecular Biology. The main reason for this is that the function of a protein is intrinsically related to its spatial conformation. However, such conformations are very difficult to be obtained experimentally in laboratory. Thus, this problem has drawn the attention of many researchers in Bioinformatics. Considering the great difference between the number of protein sequences already known and the number of three-dimensional structures determined experimentally, the demand of automated techniques for structural classification of proteins is very high. In this context, computational tools, especially Machine Learning (ML) techniques, have become essential to deal with this problem. In this work, ML techniques are used in the recognition of protein structural classes: Decision Trees, k-Nearest Neighbor, Naive Bayes, Support Vector Machine and Neural Networks. These methods have been chosen because they represent different paradigms of learning and have been widely used in the Bioinfornmatics literature. Aiming to obtain an improvment in the performance of these techniques (individual classifiers), homogeneous (Bagging and Boosting) and heterogeneous (Voting, Stacking and StackingC) multiclassification systems are used. Moreover, since the protein database used in this work presents the problem of imbalanced classes, artificial techniques for class balance (Undersampling Random, Tomek Links, CNN, NCL and OSS) are used to minimize such a problem. In order to evaluate the ML methods, a cross-validation procedure is applied, where the accuracy of the classifiers is measured using the mean of classification error rate, on independent test sets. These means are compared, two by two, by the hypothesis test aiming to evaluate if there is, statistically, a significant difference between them. With respect to the results obtained with the individual classifiers, Support Vector Machine presented the best accuracy. In terms of the multi-classification systems (homogeneous and heterogeneous), they showed, in general, a superior or similar performance when compared to the one achieved by the individual classifiers used - especially Boosting with Decision Tree and the StackingC with Linear Regression as meta classifier. The Voting method, despite of its simplicity, has shown to be adequate for solving the problem presented in this work. The techniques for class balance, on the other hand, have not produced a significant improvement in the global classification error. Nevertheless, the use of such techniques did improve the classification error for the minority class. In this context, the NCL technique has shown to be more appropriated / Atualmente, a classifica??o estrutural de prote?nas, que diz respeito ? infer?ncia de padr?es em sua conforma??o 3D, ? um dos principais problemas em aberto da Biologia Molecular. Esse problema vem recebendo a aten??o de muitos pesquisadores na ?rea de Bioinform?tica pelo fato de as fun??es das prote?nas estarem intrinsecamente relacionadas ?s suas diferentes conforma??es espaciais, que s?o de dif?cil obten??o experimental em laborat?rio. Considerando a grande diferen?a entre o n?mero de seq??ncias de prote?nas conhecidas e o n?mero de estruturas tridimensionais determinadas experimentalmente, ? alta a demanda por t?cnicas automatizadas de classifica??o estrutural de prote?nas. Nesse contexto, as ferramentas computacionais, principalmente as t?cnicas de Aprendizado de M?quina (AM), tornaram-se alternativas essenciais para tratar esse problema. Neste trabalho, t?cnicas de AM s?o empregadas no reconhecimento de classes estruturais de prote?nas: ?rvore de Decis?o, k-Vizinhos Mais Pr?ximos, Na?ve Bayes, M?quinas de Vetores Suporte e Redes Neurais Artificiais. Esses m?todos foram escolhidos por representarem diferentes paradigmas de aprendizado e serem bastante citados na literatura. Visando conseguir uma melhoria de desempenho na solu??o do problema abordado, sistemas de multiclassifica??o homog?nea (Bagging e Boosting) e heterog?nea (Voting, Stacking e StackingC) s?o aplicados nesta pesquisa, usando como base as t?cnicas de AM anteriormente mencionadas. Al?m disso, pelo fato de a base de dados de prote?nas considerada neste trabalho apresentar o problema de classes desbalanceadas, t?cnicas artificiais de balanceamento de classes (Under-sampling Aleat?rio, Tomek Links, CNN, NCL e OSS) s?o utilizadas a fim de minimizar esse problema e melhorar o desempenho dos classificadores. Para a avalia??o dos m?todos de AM, um procedimento de valida??o cruzada ? empregado, em que a acur?cia dos classificadores ? medida atrav?s das m?dias da taxa de classifica??o incorreta nos conjuntos de testes independentes. Essas m?dias s?o comparadas duas a duas pelo teste de hip?tese a fim de avaliar se h? diferen?a estatisticamente significativa entre elas. Com os resultados obtidos, pode-se observar, entre os classificadores base, o desempenho superior do m?todo M?quinas de Vetores Suporte. Os sistemas de multiclassifica??o (homog?nea e heterog?nea), por sua vez, apresentaram, em geral, uma acur?cia superior ou similar a dos classificadores usados como base, destacando-se o Boosting que usou ?rvore de Decis?o em sua forma??o e o StackingC tendo como meta classificador a Regress?o Linear. O m?todo Voting, apesar de sua simplicidade, tamb?m mostrou-se adequado para a solu??o do problema considerado nesta disserta??o. Em rela??o ?s t?cnicas de balanceamento de classes, n?o foram alcan?ados melhores resultados de classifica??o global com as bases de dados obtidas com a aplica??o de tais t?cnicas. No entanto, foi poss?vel uma melhor classifica??o espec?fica da classe minorit?ria, de dif?cil aprendizado. A t?cnica NCL foi a que se mostrou mais apropriada ao balanceamento de classes da base de dados de prote?nas

Made available in DSpace on 2014-12-17T15:01:21Z (GMT). No. of bitstreams: 1 RobertaVAC_DISSERT.pdf: 2111666 bytes, checksum: 5b317150d6db63bae04e852173ace9d2 (MD5) Previous issue date: 2009-12-14 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37?C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies / A leishmaniose visceral, causada pela esp?cie Leishmania chagasi, tamb?m conhecida como calazar, apresentou, no per?odo de 1990 a 2005, taxa de incid?ncia no Brasil variando entre 1 e 3 casos por 100 mil habitantes. A regi?o Nordeste, que at? o ano de 2000 contribuiu com quase 90% dos casos registrados, est? reduzindo sua participa??o na d?cada atual, chegando a 56% em 2005. O alto custo e toxicidade das drogas usadas no tratamento convencional dessa leishmaniose levaram a busca de tratamentos alternativos, com interesse especial na pesquisa e produ??o de ant?genos por microrganismos recombinantes para desenvolvimento de vacinas e kits de diagn?stico. A bact?ria Escherichia coli tem sido o microrganismo mais estudado e usado como hospedeiro para produ??o de prote?nas recombinantes. Nesse contexto, este trabalho tem como objetivo estudar a influ?ncia da indu??o no crescimento celular e a verifica??o do tipo de express?o (intra ou extracelular) de ant?genos de Leishmania chagasi atrav?s de cultivo de dois clones de E. coli recombinante (kmp11 e P36) em incubador rotativo (shaker) usando tr?s meios diferentes (2xTY, TB, FASS+EL). Para isso, foram realizados ensaios em condi??es estabelecidas na literatura para E. coli (37?C ? 200 rpm) e os meios suplementados com antibi?ticos para garantir somente o crescimento de c?lulas competentes. Na primeira etapa, foram realizados ensaios sem indu??o a fim de se verificar o comportamento cin?tico dos dois clones (crescimento e consumo de substrato) nos diferentes meios. Na segunda etapa, a indu??o foi realizada atrav?s da adi??o de IPTG (concentra??o final de 1mM), na primeira hora de cultivo. Foi observada que a express?o das prote?nas foi intracelular para todos os clones e meios testados, entretanto o maior n?vel foi verificado nitidamente pela densidade (intensidade) da banda nos g?is de eletroforese para o meio 2xTY e prote?na kmp11. Apesar de o meio 2xTY conter uma menor concentra??o de substratos, conseq?entemente, uma concentra??o celular reduzida quando comparada aos outros meios, pareceu que esta combina??o de meio e clone ? mais indicada para produ??o das prote?nas recombinantes testadas neste trabalho. O objetivo desse trabalho foi alcan?ado j? que a prote?na de interesse foi produzida. Conseq?entemente, este resultado motiva novos estudos para otimiza??o da produ??o usando diferentes estrat?gias de cultivo

Made available in DSpace on 2014-12-17T15:32:21Z (GMT). No. of bitstreams: 1 ClarissaFD_DISSERT.pdf: 1047575 bytes, checksum: 1162ac27915144b24bb631aefd3e421b (MD5) Previous issue date: 2012-02-15 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The Epidermoid Carcinoma (EC) is the most common lesions located in the region of the head and neck and, despite advances in treatment modalities, the prognosis is still poor. The malignant cells show an increase in glucose uptake, process mediated by glucose transporters (GLUTs). Increased expression of GLUT 1 and GLUT 3 is related to the aggressive behavior of this lesion. The aim of this study was to evaluate, through immunohistochemistry, the expression of GLUTs 1 and 3 in EC of the lower lip. The sample consisted of 40 cases of EC of the lower lip, of which 20 had regional lymph node metastasis and the remaining 20 with absence of metastasis. The percentages of immunostained cells in front of tumor invasion and in the center of tumor were evaluated. These results were related to the presence and absence of lymph node metastasis, TNM classification and histological grading. The percentage of cytoplasmic/membranous expression of GLUT 1 ranged from 77.35% to 100%, while for GLUT 3 this value ranged from 0.79% to 100%. As for nuclear staining for GLUT 1, this percentage ranged from 0 to 0.42%, however. GLUT 3 showed only one case with nuclear staining. Despite the significant expression of tumor cells related to the proteins studied, we observed no statistically significant relationship between the variables and the antibodies analyzed, regardless of the region evaluated. However, there was a moderate positive correlation between cytoplasmic/membranous immunoexpressions of GLUT 1 in invasion front and in the tumor center (r = 0.679, p <0.001). Similarly, moderate positive correlation was found between the nuclear immunoexpressions of GLUT 1 in the invasion front and in the tumor center (r = 0.547, p <0.001). For GLUT 3, was also observed a moderate statistically significant positive correlation between cytoplasmic/membranous expression in tumor invasion front and in tumor center (r = 0.589, p <0.001). We also observed that the immunoreactivity for GLUT 1 was higher than GLUT 3 expression in invasion front (p <0.001) and tumor center (p <0.001). From these results, this study suggests that tumor hypoxia is a remarkable characteristic of the EC of the lower lip and GLUT 1 may be primarily responsible for glucose uptake into the interior of the malignant cells / O Carcinoma epiderm?ide (CE) est? entre as les?es mais comuns localizadas na regi?o de cabe?a e pesco?o e apesar dos avan?os nas modalidades de tratamento, o progn?stico ainda ? pobre. As c?lulas malignas apresentam um aumento na absor??o de glicose, processo esse mediado pelos transportadores de glicose (GLUTs). O aumento da express?o de GLUT 1 e GLUT 3 encontra-se relacionado com o comportamento agressivo dessa les?o. O objetivo desse estudo consistiu em avaliar, atrav?s de estudo imuno-histoqu?mico, a express?o dos GLUTs 1 e 3 em CE de l?bio inferior. A amostra foi composta por 40 casos de CE de l?bio inferior, dos quais 20 apresentavam met?stase linfonodal regional e os 20 restantes com aus?ncia de met?stase. Foram avaliados os percentuais de c?lulas imunomarcadas no front de invas?o e no centro do tumor, esses resultados foram relacionados com a presen?a e aus?ncia de met?stase linfonodal, estadiamento cl?nico TNM e grada??o histol?gica. O percentual de marca??o citoplasm?tica/membranar para GLUT 1 variou de 77,35% a 100%, j? para GLUT 3 esse valor variou de 0,79% a 100%. Quanto ? marca??o nuclear para GLUT 1, este percentual variou de 0 a 0,42%, no entanto, GLUT 3 apresentou apenas um caso com marca??o nuclear. Apesar da expressiva marca??o das c?lulas tumorais frente aos marcadores estudados, n?o foi poss?vel observar rela??o estatisticamente significativa entre as vari?veis estudadas e os marcadores analisados, independente da regi?o avaliada. Contudo, foi observada moderada correla??o positiva estatisticamente significativa entre as imunoexpress?es citoplasm?tica/membranar de GLUT 1 no front de invas?o e no centro do tumor (r=0,679; p<0,001). De forma similar, foi constatada moderada correla??o positiva entre as imunoexpress?es nucleares de GLUT 1 no front de invas?o e no centro tumoral(r=0,547; p<0,001). Para GLUT 3, tamb?m foi observada moderada correla??o positiva entre as imunoexpress?es membranar/ citoplasm?tica dessa prote?na no front de invas?o e no centro tumoral (r=0,589; p<0,001). Foi observado, ainda, que a imunoexpress?o de GLUT 1 em rela??o a GLUT 3 foi maior tanto no front de invas?o (p < 0,001) como no centro do tumor (p<0,001). A partir desses resultados, conclui-se que a hip?xia tumoral ? uma caracter?stica marcante no CE de l?bio inferior e GLUT 1 pode ser o principal respons?vel pela capta??o de glicose para o interior das c?lulas malignas

Made available in DSpace on 2014-12-17T15:32:22Z (GMT). No. of bitstreams: 1 MarceloABN_DISSERT.pdf: 4711000 bytes, checksum: ea3602bfcefe0d8448e9d402030634a2 (MD5) Previous issue date: 2014-02-14 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / BMPs are components superfamily ligands transformation growth fator-&#946; (TGF-&#946;) secreted into the extracellular environment, with mechanisms of intercellular communication through specific ligands and receptors in various target cells, being recognized for its influence in osteogenic induction, also play an important role in tissue homeostasis, cell proliferation, differentiation control , in addition to being present in the development of various malignancies. The aim of this study was to compare the immunohistochemical expression of BMP-2, BMP-4 and its receptors BMPRIA and BMPRII in cases of ameloblastoma and adenomatoid odontogenic tumor. The sample consisted of 20 cases of solid ameloblastoma (SA), 10 cases of ameloblastoma unicystic (UA) and 16 cases of adenomatoid odontogenic tumor (AOT). The expression of BMPs and their receptors was evaluated in the parenchyma and stroma of lesions, establishing the percentage of immunopositive cells (0 - negative; 1-1 % to 10 % of cells positive; 2 - 11% to 25% of positive cells; 3 - 26% to 50% of cells positive; 4 - 51% to 75 % of positive cells; 5 - more than 75% positive cells). Analysis of the expression of BMP-2 revealed no statistically significant differences in parenchymal (p = 0.925) and stromal component (p = 0.345) between the groups, as well as BMP-4 (p = 0.873 / p = 0.131). In the epithelial component, SA and AOT had a higher frequency of score 5. In turn, all cases of UA were classified as score 5. The analysis of the stromal component showed no statistically significant difference between groups with respect to median scores BMPRIA positivity (p = 0.768) and BMPRII (p = 0.779). In the epithelial component of SA and UA, no statistically significant correlations between imunoexpression proteins analyzed were observed. In turn, the group of AOT, statistically significant positive correlations between the scores of expression of all studied proteins were found. In the stromal component, statistically significant positive correlations were found only in the SA group in BMP -4 and BMPRII (r = 0.476; p = .034), in the UA in BMP-4 and BMPRIA (r = 0.709; p = 0.022). The results of this study suggest that the BMPs and their receptors are involved in the development process odontogenic tumors. BMP-4, in turn, besides being present in odontogenic tumors have the capacity to form mineralized material. / As BMPs s?o componentes da superfam?lia de ligantes do fator transformador de crescimento-&#946; (TGF-&#946;), secretados no meio extracelular, com mecanismos de comunica??o intercelular por meio de ligantes e receptores espec?ficos em diversas c?lulas-alvo, sendo reconhecidas por sua influ?ncia na indu??o osteog?nica, tamb?m desempenhando importante papel na homeostase tecidual, prolifera??o celular, no controle de diferencia??o, al?m de estar presente no desenvolvimento de diversas neoplasias. O objetivo deste estudo foi comparar a express?o imuno-histoqu?mica da BMP-2, BMP-4 e seus receptores BMPRIA e BMPRII em casos de Ameloblastoma e Tumor odontog?nico adenomat?ide. A amostra foi constitu?da de 20 casos de Ameloblastoma s?lido (AS), 10 casos de Ameloblastoma unic?stico (AU) e 16 casos de Tumor odontog?nico adenomat?ide (TOA). A express?o das BMPs e seus receptores foi avaliada no par?nquima e estroma das les?es, estabelecendo-se o percentual de c?lulas imunopositivas (0 negativo; 1 - 1% a 10% das c?lulas positivas; 2 - 11% a 25% das c?lulas positivas; 3 - 26% a 50% das c?lulas positivas; 4 - 51% a 75% das c?lulas positivas; 5 - mais 75% de c?lulas positivas). A an?lise da express?o de BMP-2 n?o revelou diferen?as estatisticamente significativas no componente par?nquimatoso (p = 0,925) e estromal (p = 0,345) entre as les?es estudadas, assim como a BMP-4 (p = 0,873 / p = 0,131). No par?nquima, o AS e TOA apresentaram maior frequ?ncia do escore 5. Por sua vez, todos os casos de AU foram classificados como escore 5. A an?lise do componente estromal revelou n?o haver diferen?a estatisticamente significativa entre os grupos em rela??o ?s medianas dos escores de positividade para BMPRIA (p = 0,768) e BMPRII (p = 0,779). No par?nquima do AS e do AU, n?o foram observadas correla??es estatisticamente significativas entre as imunoexpress?es das prote?nas analisadas. Por sua vez, no grupo dos TOAs, foram constatadas correla??es positivas, estatisticamente significativas, entre os escores de express?o de todas as prote?nas avaliadas. No componente estromal, foram constatadas correla??es positivas, estatisticamente significativas, apenas no grupo do AS em BMP-4 e BMPRII (r = 0,476; p = 0,034) e do AU em BMP-4 e BMPRIA (r = 0,709; p = 0,022). Os resultados do presente estudo sugerem que as BMPs e seus receptores est?o envolvidos no processo de desenvolvimento de tumores odontog?nicos. A BMP-4, por sua vez, al?m de estar presente em tumores odontog?nicos possui a capacidade de forma??o de material mineralizado

Made available in DSpace on 2014-12-17T15:32:27Z (GMT). No. of bitstreams: 1 CristinaRFA.pdf: 1625942 bytes, checksum: 99d50db14437c5ea0e3022e4da576321 (MD5) Previous issue date: 2009-03-06 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Bone morphogenetic proteins (BMPs) are cytokines involved in proliferation and angiogenesis of many kind of human cancer. The present study analyzed the immunohistochemical expression of BMP-2, BMPR-II, BMPR-IA and endoglin (CD105) and their relationship with the biological behavior and local angiogenesis in tongue oral squamous cells carcinoma (SCC). The sample consisted of 25 cases of tongue SCC without metastasis, 25 tongue SCC with metastasis and 25 cases of Inflamatory Fibrous Hyperplasia (IFH).The histological grade of malignancy proposed by Bryne (1998), adapted by Miranda (2002) was used to classify all tongue SCC cases. Score 0 was attributed to absent-weak immunoexpression and score 1 for strong immunostaning and pattern of distribution was focal or diffuse. Microvessel counts (MVC) was established for CD105. Most of the patients with tongue SCC was male. The principal age in tongue SCC without metastasis was over 65 years and in tongue SCC with metastasis was between 45-65 years. There were predominance of stage II in TNM and in the specimens with high-grade, independent of studied group. For BMP-2, 56% of tongue SCC without metastasis and 72% tongue SCC with metastasis exhibited score 1 while the IFH showed secore 0 in 72% of the cases, with statistical association (p=0,007). Considering the BMPR-II, 52% of tongue SCC without metastasis exhibited score 0; 56% tongue SCC with metastasis and 60% IFH showed score 1. The majority cases of BMPR-IA demonstrated score 1 and 100% of CD105 exhibited strong immunoexpression in tongue SCC. Regarding the pattern distribution, it was noted a tendency to diffuse pattern for the proteins in all groups. The means of MVC were similar in tongue SCC without metastasis (32,91) and in tongue SCC with metastasis (32,05), however existed statistical difference with IFH (p<0,001). There was statistical association of BMP-2 expression with BMPR-II (p=0,008), BMPR-IA (p=0,006) and CD105 (p=0,046). An association between TNM and BMP-2 immunoexpression and their receptors was not detected, nevertheless this association was found with MVC (p=0,047) whose averages were higher for the stages II (35,97) e IV (35,69). No association between histological grading and these proteins was observed. This study suggests that the superexpression of BMP-2 signaling pathways acts on cell proliferation in tongue SCC and can be implicated with more invasive potential. Additionaly, the CD105 is a potent biological marker of neovascularization in this neoplasm and their association with BMP-2 and BMPR-IA receptor, showed that this type of cancer in BMP-2 is presented as pro-angiogenic in the metastatic process / As BMPs (prote?nas morfogen?ticas ?sseas) s?o citocinas relacionadas com a prolifera??o e angiog?nese em diversos tipos de c?ncer humano. Com este trabalho foi analisada a express?o imunoistoqu?mica das prote?nas BMP-2, BMPR-IA, BMPR-II e endoglina (CD105), correlacionando-a com o comportamento biol?gico e a angiog?nese local nos carcinomas epiderm?ides de l?ngua (CEL). A amostra foi composta de 25 casos de CEL sem met?stase (CELSM), 25 CEL com met?stase (CELCM) graduados segundo Bryne (1998) e adaptado por Miranda (2002), al?m de 25 casos de hiperplasia fibrosa inflamat?ria (HFI), utilizado como grupo controle. Foi utilizado escore 0 para marca??o ausente-fraca e 1 para forte; tipo de distribui??o focal ou difuso. Adicionalmente, para o CD105 foi realizada a contagem microvascular (MVC). A maior parte dos pacientes com CEL foi do sexo masculino, no grupo CELSM a faixa et?ria foi maior que 65 anos e o CELCM se encontrou entre 45-65 anos; houve predom?nio do est?gio II do TNM, assim como de esp?cimes de alto grau, independente do grupo estudado. Para BMP-2, 56% dos CELSM e 72% dos CELCM exibiram escore 1, enquanto a HFI exibiu 72% de escore 0, apresentando associa??o estat?stica (p=0,007). Para BMPR-II 52% dos CELSM exibiram escore 0; 56% CELCM e 60% da HFI escore 1 e no BMPR-IA ocorreu uma predomin?ncia de escore 1 e para o CD105 100% de marca??o forte nos CEL. Quanto ao tipo de distribui??o notou-se tend?ncia de distribui??o difusa de todas as prote?nas, em todos os grupos. Observaram-se, para MVC, m?dias muito semelhantes entre os CELSM (32,91) e os CELCM (32,05) exibindo, contudo, diferen?a estat?stica com as HFI (p<0,001).Observa-se uma associa??o estat?stica da BMP-2 com a BMPR-II (P=0,008), BMPR-IA (p=0,006) e o CD105 (0,046). N?o se observou associa??o entre o TNM e a imunoexpress?o da BMP-2 e seus receptores, por?m foi encontrada com a MVC (p=0,047), cujas maiores m?dias foram para os est?gios II (35,97) e IV (35,69), tal como n?o ocorreu associa??o entre a grada??o histol?gica e as prote?nas. Conclui-se que a superexpress?o da via de sinaliza??o da BMP-2 atua na prolifera??o celular, contribuindo para maior invasividade do CEL. O CD105 ? um potente marcador de neovasculariza??o deste neoplasma e sua associa??o com a BMP-2 e o receptor BMPR-IA, mostra que neste tipo de neoplasia a BMP-2 se apresenta como pr?-angiog?nico no processo metast?tico

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-04-27T15:00:50Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) mariana_aguiar_matos.pdf: 2901378 bytes, checksum: 40dbd704043d49a1eee587bb086c4eb4 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-05-16T19:24:00Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) mariana_aguiar_matos.pdf: 2901378 bytes, checksum: 40dbd704043d49a1eee587bb086c4eb4 (MD5) / Made available in DSpace on 2017-05-16T19:24:00Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) mariana_aguiar_matos.pdf: 2901378 bytes, checksum: 40dbd704043d49a1eee587bb086c4eb4 (MD5) Previous issue date: 2016 / 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) / O excesso de gordura corporal caracter?stico da obesidade est? relacionado a diversas altera??es metab?licas, que incluem a resist?ncia ? insulina. Dentre as medidas n?o farmacol?gicas empregadas para a melhora da sensibilidade ? insulina est? o treinamento f?sico aer?bio, como o treinamento intervalado de alta intensidade (HIIT, do ingl?s high intensity interval training). Sendo assim, esse estudo avaliou os efeitos do HIIT em componentes bioqu?micos, celulares e moleculares relacionados ? resist?ncia ? insulina em obesos. Indiv?duos obesos sens?veis (n=9) e resistentes ? insulina (n=8) foram submetidos a 8 semanas de HIIT, em cicloerg?metro, realizado 3 vezes por semana, com intensidade e volume progressivos (8 a 12 est?mulos; 80 a 110% da pot?ncia m?xima). Amostras de sangue venoso e do m?sculo vasto lateral foram obtidas antes e ap?s o programa de HIIT. Ap?s o programa de treinamento houve aumento da sensibilidade ? insulina nos obesos resistentes ? insulina, mas n?o houve redu??o da massa de gordura. A concentra??o de citocinas no soro, o estresse oxidativo sist?mico e frequ?ncia das c?lulas imunes n?o foram modificadas ap?s o treinamento. No m?sculo esquel?tico, o HIIT promoveu aumento da fosforila??o do substrato do receptor de insulina (IRS) (Tyr612), da Akt (Ser473) e da prote?na quinase dependente de c?lcio/calmodulina (CAMKII) (Thr286), e aumento do conte?do da ?-hidroxiacil-CoA desidrogenase (?-HAD) e citocromo C oxidase (COX-IV). Houve ainda, redu??o da fosforila??o da quinase regulada por sinal extracelular (ERK1/2) nos obesos resistentes ? insulina. Conclu?mos que 8 semanas de HIIT promoveram melhora da sensibilidade ? insulina, modificou componentes da via de sinaliza??o da insulina e do metabolismo oxidativo no m?sculo esquel?tico. Essas altera??es ocorreram independentes de mudan?as na gordura corporal total e de par?metros inflamat?rios sist?micos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Obesity is characterized by excess of body fat, and its development can lead to a variety of metabolic disorders, including insulin resistance. Exercise is recognized as a non-pharmacological approach to increasing skeletal muscle insulin sensitivity, although the mechanisms are not elucidated. Additionally, the understanding of high intensity interval training (HIIT, high intensity interval training) treat insulin resistance is less understood. Therefore, this study evaluated the effects of HIIT on biochemical, molecular, and cellular markers related to insulin resistance in sedentary obese individuals. Sensitive (n=9) and insulin resistant (n=8) obese individuals (body mass index ? 30 kg/m-2) were engaged in 8 weeks of HIIT using a cycle ergometer. The HIIT was performed 3 times a week, and its intensity and volume progressively increased throughout the training period (from 8 to 12 stimuli; from 80 to 110% of the maximum power). Venous blood and the vastus lateralis muscle samples were obtained before and after the HIIT. HIIT enhanced insulin sensitivity in insulin-resistant obese individuals without changing body fat mass. Cytokine concentration in serum, blood oxidative stress, and frequency of some immune cells were not altered by HIIT. In skeletal muscle, HIIT increased the phosphorylation of insulin receptor substrate (IRS) (Tyr612), Akt (Ser473), and protein kinase dependent calcium/calmodulin (CaMKII) (Thr286). HIIT also increased the expression of ?-hydroxyacyl-CoA dehydrogenase (?-HAD) and cytochrome C oxidase (COX-IV). A reduction of the kinase phosphorylation of extracellular signal-regulated (ERK1/2) was only seen in obese insulin resistant individuals. The results show that 8 weeks of HIIT enhanced insulin sensitivity, modified components of the insulin-signaling pathway, and improved skeletal muscle oxidative metabolism. These changes were independent of alterations in body fat and inflammatory parameters.

Made available in DSpace on 2014-12-17T14:13:22Z (GMT). No. of bitstreams: 1 SebastiaoDSF.pdf: 475002 bytes, checksum: d28adb2821adc67501d7e19f42e9b6a3 (MD5) Previous issue date: 2007-08-16 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Several clinic evaluations have been possible with radiobiocomplexes labeled with technetium-99m (99mTc). Some natural and synthetic drugs are capable of to interfere on the labeling of blood constituents with 99mTc, as well as on the biodistribution of radiobiocomplexes. Authors have also reported about the toxicity of several natural products. The aim of this study was to compare the effects of the Mentha crispa (hortel?) and of the Hypericum perforatum (hip?rico) in different experimental models. On the labeling of red blood cells (RBC) and plasma and cellular proteins with 99mTc, both extracts were capable of to decrease the radioactivity percentage on the cellular compartment and on the fixation on plasma and cellular proteins. On the morphometry of the RBC, only the hortel? was capable to alter the shape and the perimeter/area ratio of the RBC. On the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4), the hortel? increased the Na99mTcO4 distribution in the kidney, spleen, liver and thyroid, meanwhile the hip?rico decreased the Na99mTcO4 distribution in the bone, stomach, lungs and thyroid, and increased the Na99mTcO4 distribution in the pancreas. On the bacterial cultures survival, the hip?rico was capable of to protect the bacteria against the stannous chloride (SnCl2) effect. The hip?rico did not alter the topology of plasmidial DNA and did not protect the plasmidial DNA against the SnCl2 action. Probably, the effects presented by both extracts could be due to chemical compounds of the extracts that could alter the morphology of the RBC and the plasma membrane ions transport, and/or by phytocomplexes that could be formed with different effects dependent on the biological system considered / Avalia??es cl?nicas t?m sido poss?veis com radiobiocomplexos marcados com tecn?cio-99mTc (99mTc). Drogas naturais ou sint?ticas s?o capazes de interferir na marca??o de estruturas sangu?neas com 99mTc, assim como na biodistribui??o de radiobiocomplexos. Tamb?m tem sido descrita a toxicidade de v?rios produtos naturais. O objetivo deste estudo foi comparar o efeito dos extratos de Mentha crispa (hortel?) e de Hypericum perfloratum (hip?rico) em diferentes modelos experimentais. Na marca??o de estruturas sang??neas com 99mTc verificou-se que ambos os extratos foram capazes de diminuir a radioatividade no compartimento celular, nas prote?nas plasm?ticas e celulares. Na morfometria das hem?cias, apenas a hortel? foi capaz de alterar a forma e a rela??o per?metro/?reas das hem?cias. Na biodistribui??o do radiobiocomplexo pertecnetato de s?dio (Na99mTcO4) a hortel? aumentou a capta??o do Na99mTcO4 no rim, no ba?o, no f?gado e na tire?ide, enquanto que o hip?rico diminuiu a capta??o do Na99mTcO4 no osso, no est?mago, no pulm?o e na tire?ide, e aumentou no p?ncreas. Na sobreviv?ncia de culturas bacterianas o hip?rico foi capaz de proteger a bact?ria do efeito danoso do cloreto estanoso (SnCl2). O hip?rico n?o alterou a topologia nem protegeu o DNA plasmidial da a??o do SnCl2. Provavelmente os efeitos apresentados por ambos os extratos poderiam ser explicados por subst?ncias presentes nos extratos que poderiam alterar a morfologia das hem?cias, o transporte de ?ons pela membrana e/ou formar fitocomplexos. O estudo teve car?ter multidisciplinar com a participa??o das seguintes ?reas do conhecimento: Radiobiologia, Bot?nica, Endocrinologia, Fitoterapia e Hematologia

Made available in DSpace on 2014-12-17T14:13:23Z (GMT). No. of bitstreams: 1 TaniaSG.pdf: 391861 bytes, checksum: 1e1c65c6de1e587070af7bb8a918f265 (MD5) Previous issue date: 2007-08-17 / &#65279;Technetium-99m (99mTc) has been used to obtain several radiobiocomplexes utilized to aid in the diagnosis of diseases. Blood constituents, as red blood cells (RBC) and plasma proteins, have been labeled with 99mTc. Natural and synthetic drugs can alter the labeling of these constituents. The aim of this work was to investigate the possibility of a Buzhong YiQi Wan extract to alter (i) the labeling of blood constituents with 99mTc, (ii) the RBC morphology, and (iii) osmotic fragility of RBC withdrawn from Wistar rats. The data showed that the BYQW extract (i) could affect labeling of blood constituintes with 99mTc, (ii) could affect the membrane integrity decreasing the osmotic resistance and (iii) could not alter the shape of RBC. Probably, these findings would be associated &#65279;with properties of the substances present in the aqueous extract of BYQW. This study has multiple disciplinary aspects in knowledge areas: Radiobiology, Botanic, Phytotherapy and Haematology / &#65279;A pesquisa em Ci?ncias da Sa?de, assim como as avalia??es cl?nicas t?m sido favorecidas pelo uso de is?topos radioativos, sendo que o tecn?cio-99m (99mTc) tem sido o mais utilizado para obten??o de radiobiocomplexos com finalidade de diagn?stico. Diversas drogas naturais ou sint?ticas s?o capazes de interferir na marca??o de estruturas sangu?neas com 99mTc, assim como na biodistribui??o de outros radiobiocomplexos. Os procedimentos relacionados com a medicina tradicional chinesa tamb?m v?m ganhando destaque em todo o mundo. O objetivo deste estudo foi investigar o efeito da possibilidade de altera??es pelo extrato de Buzhong Yi Qi Wan (f?rmula magistral chinesa) nos constituintes do sangue marcados com 99mTc, (i) na marca??o de hem?cias e prote?nas plasm?ticas, (ii) na morfologia, e (iii) na fragilidade osm?tica de hem?cias de ratos Wistar. Foi observado, diminui??o significativa (p<0,05) na marca??o dos constituites sangu?neos com 99mTc, n?o altera??o da morfologia das hem?cias e modifica??o da curva de fragilidade das hem?cias (p<0,05). Esses efeitos poderiam estar associados com determinadas propriedades de compostos qu?micos presentes no extrato Buzhong Yi Qi Wan. O estudo tem car?ter multidisciplinar com a participa??o das seguintes ?reas do conhecimento: Radiobiologia, Bot?nica, Fitoterapia e Hematologia

Made available in DSpace on 2014-12-17T15:32:18Z (GMT). No. of bitstreams: 1 CyntiaCPC.pdf: 2470782 bytes, checksum: 4619c7ffcab85bffa54a732d30786d99 (MD5) Previous issue date: 2010-02-24 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Currently, bone morphogenetic proteins (BMPs) have effective participation in the growth of malignancies. Knowing that there are few studies involving BMPs and oral squamous cell carcinoma, this work constitutes an immunohistochemical study of BMP-2, BMPR IA and BMPR II in squamous cell carcinomas (SCC) of the lower lip relating to the clinical and pathological aspects of this lesion. The sample consisted of 40 cases of SCC of the lower lip, being 20 cases of SCC of the lower lip with regional metastasis and 20 cases without metastasis. We evaluated the intensity of expression (score 1 to mark absent / weak, score 2 for high ) and was found the percentage of labeled cells, where the score was 1 cases with 0 to 50% of positive cells, score 2 with 51 to 75% of positive cells, and score 3 more than 75% of positive cells. The sample comprised 72.5% of men with a mean age of 65.8 years, there was a predominance of stage II and 52.5% of the carcinomas were classified as low grade, being carcinoma with metastasis presenting most cases (70%) as carcinomas of high malignancy grade (p = 0.004). The largest number of cases of SCC of the lower lip that were in stages I / II (61, 9%) were classified as carcinomas of low grade malignancy and carcinomas in stages III / IV were classified as high-grade tumors (p = 0, 024). The BMP-2 showed strong intensity of immunostaining in 82.5%, BMPR-IA showed 55% of cases with an intensity of immunostaining absent / weak and BMPR-II showed 85% of cases with an intensity of immunostaining absent / weak. Only the protein BMPR-IA were significantly associated with all clinic-pathological parameters studied, metastasis (p <0.001), TNM (p <0.001) and histological grade of malignancy with (p = 0.028). The percentage of positive cells, all markers showed the highest number of cases with more than 75% of positive cells (score 3) and only BMPR-II showed statistical difference when related to the presence and absence of metastasis (p = 0.049 ). We conclude that there is disturbance in the BMP signaling pathway in EC-mediated lower lip and that high expression of BMP-2 associated with the expression of BMPR-IA and BMPR-II are associated with metastasis in carcinoma / Atualmente as prote?nas morfogen?ticas do osso (BMPs) t?m efetiva participa??o no crescimento de neoplasias malignas. Sabendo que s?o escassos os trabalhos envolvendo BMPs e o carcinoma epiderm?ide oral, este trabalho realizou um estudo imunoistoqu?mico da BMP-2, BMPR IA e BMPR II em carcinomas epiderm?ides (CE) de l?bio inferior relacionando com os aspectos clinico-patol?gicos desta les?o. A amostra constou de 40 casos de CE de l?bio inferior, sendo 20 casos de CE de l?bio inferior com met?stase linfonodal regional e 20 casos sem met?stase. A grada??o histol?gica de malignidade foi realizada no front invasivo da les?o. Foi avaliada a intensidade de express?o (escore 1 para marca??o ausente/ fraca e escore 2 para marca??o forte), bem como foi verificado a porcentagem de c?lulas positivas, onde o escore 1 era os casos com 0 a 50% das c?lulas positivas; escore 2 com 51 a 75% das c?lulas positivas; e escore 3 com mais de 75% das c?lulas positivas. A amostra foi composta por 72,5% de homens com a m?dia de idade de 65,8 anos, houve um predom?nio do est?gio II e 52,5% dos carcinomas foram classificados como de baixo grau, sendo os carcinomas com met?stase regional apresentando a maioria dos casos (70%) como carcinomas de alto grau de malignidade (p =0,004). O maior n?mero de casos de CE de l?bio inferior que estavam nos est?gios I/ II (61, 9%) foi classificado em carcinomas de baixo grau de malignidade e os carcinomas nos est?gios III/ IV foram classificados em alto grau de malignidade (p =0, 024). A BMP-2 apresentou intensidade da imunomarca??o forte em 82,5%, BMPR-IA observou-se 55% dos casos com intensidade de imunomarca??o ausente/ fraca e a BMPR-II revelou 85% dos casos com intensidade de imunomarca??o ausente/ fraca. Apenas a prote?na BMPR-IA apresentou associa??o estatisticamente significante com todos os par?metros clinico-patol?gicos estudados, met?stase (p<0,001), TNM (p<0,001) e grada??o histol?gica de malignidade com ( p=0,028). Quanto ? porcentagem de c?lulas positivas, todos os marcadores apresentaram o maior n?mero de casos com mais de 75% das c?lulas positivas (escore 3) e apenas a BMPR-II apresentou diferen?a estat?stica quando relacionada com a presen?a e aus?ncia de met?stase (p=0,049). Conclui-se que existe dist?rbio na via de sinaliza??o BMP-mediada no CE de l?bio inferior e que a alta express?o da BMP-2 associada com a express?o da BMPR-IA e BMPR-II est?o relacionadas com a met?stase neste carcinoma