Idealizou-se o presente ensaio com o objetivo de padronizar o perfil hematológico, bioquímico, eletroforético (proteinograma sérico) e quantificação das células linfocitárias CD4, CD5, CD8 por citometria de fluxo de cães da raça Golden Retrivier normais (grupos GR) e de cães distróficos (grupos GRMD). Para tanto, considerou-se a divisão dos grupos de acordo com a idade dos animais, desde o nascimento até a idade adulta, compondo assim seis grupos, sendo eles GR I, II, III e GRMD I, II, III. No presente projeto realizamos os estudos eletroforéticos de cães pertencentes a todos os grupos, estudos hematológicos e bioquímicos dos cães pertencentes aos grupos GR II, III, GRMD II e III, imunofenotipagem linfocitária dos grupos GR III e GRMD III. Os resultados eritroleucométricos e trombométricos obtidos para os cães pertencentes aos grupos GR II e GR III apresentaram valores médios dentro normalidade. Com relação aos cães pertencentes aos grupos GRMD III, observamos que o eritrograma se encontra dentro dos valores de referência. Contudo, considerando o leucograma, os valores médios (3,79/ mm3) referentes à mensuração de basófilos apresentaram-se acima dos valores de normalidade, variando de 0 a 159/mm3. Ademais, considerando os valores máximos, alguns animais pertencentes ao grupo em questão, apresentaram leucocitose com neutrofilia sem desvio à esquerda, além de trombocitose, monocitose e linfopenia, no momento das coletas. As dosagens bioquímicas séricas de todos os cães afetados apresentaram valores médios acima dos valores de normalidade para a dosagem de AST, ALT e CK. Para o estudo das proteínas séricas, a técnica SDS-PAGE permitiu o fracionamento de vinte e três proteínas, cujos pesos moleculares variaram de 16.000 a 260.000 daltons em todos os grupos estudados. Destas, foi possível identificar nominalmente doze frações protéicas: IgA (PM 142.000 Da), proteína C-reativa (PM122.000 Da), ceruloplasmina (PM 110.000 Da), fosforilase (PM 95.000 Da), transferrina (PM 82.000 Da), hemopexina (PM 78.000 Da), albumina (PM 66.000 Da), alfa1 antitripsina (PM 62.000 Da), IgG de cadeia pesada (PM 55.000 Da), haptoglobina (PM 45.000 Da), glicoproteína ácida (PM 40.000 Da) e IgG de cadeia leve (PM 23.000 Da). As demais proteínas foram identificadas pelos respectivos pesos moleculares. Não foi possível identificar nominalmente as frações protéicas de pesos moleculares 260.000 Da, 230.000 Da, 180.000 Da, 165.000 Da, 158.000 Da, 91.000 Da, 35.000 Da, 30.000 Da, 28.000 Da e 16.000 Da. Dentre essas proteínas foi possível observar que a proteína de peso molecular 91.000 Da foi encontrada apenas nos grupos GR I e GRMD I não sendo identificada nos outros grupos estudados. Considerando as alterações encontradas com relação às proteínas de fase aguda, evidenciamos as respostas de fase aguda frente às lesões musculares que ocorrem progressivamente em cães distróficos, principalmente em animais distróficos jovens, podendo inferir que os mesmos apresentaram alterações protéicas perante a injúria tecidual como é relatado em vários processos inflamatórios relacionados com outras patologias. Da mesma forma, os animais afetados pela distrofia, possuem alterações imunoglobulínicas quando comparados a animais normais. Com relação a imunofenotipagem linfocitária foi confeccionado um histograma das populações de linfócitos CD4+, CD5+ e CD8+ a partir da região do gate de linfócitos. Todos os animais afetados pela distrofia apresentaram porcentagem significativamente maior com relação à população linfocitária CD4+ e CD5+ quando comparados aos cães normais. Desta forma, podemos inferir que o processo progressivo da distrofia, esta relacionado com alterações nas populações celulares que compõe o sistema imune dos pacientes, pois devido a ausência de distrofina o músculo fica mais susceptível á lesões, sendo que na sua ocorrência, há liberação de citocinas que estimulam as células hepáticas a secretarem as proteínas de fase aguda e a promoverem uma co-estimulação de linfócitos T. Assim, podemos sugerir que os resultados encontrados em nosso trabalho são conseqüências da resposta inflamatória gerada pela lesão muscular. Esperamos com esse estudo, fornecer mais informações sobre a fisiopatogenia da doença, bem como promover um maior entendimento sobre a avaliação imunológica e resposta inflamatória neste modelo de estudo. Ademais, que os valores de base encontrados possam auxiliar na avaliação de possíveis reações nos testes pré-clínicos, facilitando assim, o entendimento das reações benéficas ou adversas para validar e explicar o mecanismo de ação de uma terapia celular, gênica ou mesmo medicamentosa / Devised to test this in order to standardize the hematological, biochemical, electrophoretic (Serum protein) and quantification of CD4 lymphocyte cells, CD5, CD8 by flow cytometry of Golden Retriever dogs normal (group GR) and dogs dystrophic (GRMD groups). To this end, we considered the division of groups according to age of animals, from birth to adulthood, thus composing six groups, as Gr I, II, III and GRMD I, II, III. In this project we performed electrophoretic studies of dogs belonging to all groups, haematological and biochemical studies of dogs belonging to the groups GR II, III, II and III GRMD, lymphocyte immunophenotyping in groups III and GR GRMD III. The results obtained for trombometric, erythometric and dogs belonging to the groups and GR II GR III showed mean values within normal limits. With respect to dogs belonging to groups III GRMD, we observed that the erythrocyte is within the reference values. However, considering the WBC, the mean (3.79 / mm3) relating to the measurement of basophils were above normal values, ranging from 0 to 159/mm3. Moreover, considering the maximum, some animals belonging to the group in question had leukocytosis with neutrophilia without left shift, and thrombocytosis, monocytosis and lymphopenia at the time of collection. The biochemical serum of all affected dogs had mean values above the normal range for the determination of AST, ALT and CK. For the study of proteins, SDS-PAGE technique allowed the fractionation of twenty-three proteins whose molecular weights ranged from 16,000 to 260,000 daltons in all groups. These could be identified by name twelve fractions: IgA (PM 142,000 Da), C-reactive protein (PM122.000 Da), ceruloplasmin (PM 110,000 Da), phosphorylase (95,000 Da PM), transferrin (82,000 Da PM), hemopexin (PM 78 000 Da), albumin (66,000 Da PM), alpha1 antitrypsin (PM 62,000 Da), IgG heavy chain (55,000 Da PM), haptoglobin (PM 45,000 Da), glycoprotein (PM 40,000 Da) and IgG light chain (PM 23 000 Da). The other proteins were identified by their molecular weights. We could not identify by name the protein fractions of molecular weight 260,000 Da, 230,000 Da, 180,000 Da, 165,000 Da, 158,000 Da, 91,000 Da, 35,000 Da, 30,000 Da, 28,000 Da and 16,000 Da. Among these proteins was observed that the protein molecular weight of 91,000 Da was found only in groups GR I and GRMD I was not identified in other groups. Considering the changes found with relation to acute phase proteins, we noted the acute phase response in the face of muscle injuries that occur gradually in dystrophic dogs, especially in young dystrophic animals, which may infer that they had abnormal protein before tissue injury as reported in many inflammatory-related pathologies. Likewise, animals affected by muscular dystrophy, alterations immunoglobulin when compared to normal animals. With respect to lymphocyte immunophenotyping was made a histogram of the populations of CD4, CD5 and CD8 from the region of the gate of lymphocytes. All animals affected by muscular dystrophy showed significantly higher percentage with respect to CD4 and CD5 lymphocyte population compared to normal dogs. Thus, we can infer that the process of progressive muscular dystrophy, is associated with changes in cell populations that make up the immune system of patients, because due to the absence of dystrophin, the muscle is more susceptible to damage, and its occurrence , a release of cytokines that stimulate liver cells to secrete acute phase proteins and promote a co-stimulation of T lymphocytes Thus, we suggest that the findings in our study are consequences of the inflammatory response generated by muscle injury.We hope that this study, provide more information about the pathophysiolgy of the disease, and promote a greater understanding of the immune and inflammatory responde assessment in this study model. Moreover, the base values found could help in assessing possible responses in preclinical testing, aiding the understanding of the beneficial or adverse reactions to validate and explain the mechanism of action of a cell therapy, gene or drug treatments

L’interleukine-4, cytokine clef du système immunitaire, est l’une des composantesprincipales de la réponse humorale. Un variant d’épissage de l’IL-4 humaine, nommé IL-4δ2est caractérisé par une délétion de l’exon 2. L’expression d’isoformes de cytokines peut êtrespécifique d’un tissu, d’un stimulus ou d’un état pathologique. Un intérêt particulier leur estaccordé car leur présence ou leur niveau d’expression peut en faire des biomarqueurspotentiels de certains stades pathologiques. Ainsi, ce projet a été consacré à l’étude del’expression et de la fonctionnalité de l’IL-4δ2 dans le but de mieux caractériser son rôle ausein de la réponse immunitaire.Une étude cinétique des niveaux d’expression de l’interleukine-4 et de son variant,réalisée chez des donneurs sains, montre une expression de ces deux variants dépendante dudonneur. L’étude a été poursuivie pour déterminer le type cellulaire capable de produire l’IL-4δ2. Ainsi, l’IL-4δ2 ne semble pas être exprimée par les cellules CD4+ et CD8+ à l’inversedes granulocytes.La fonction agoniste ou antagoniste à l’IL-4, de l’isoforme δ2, sujette à controverse, ajustifié une exploration de sa fonctionnalité. Nous avons ainsi évalué la capacité de l’IL-4δ2 àactiver les voies de signalisation de l’IL-4. Une absence d’activation de mécanismescellulaires similaires à l’IL-4 nous suggère un potentiel rôle inhibiteur de ce variant.Au cours du travail sur l’IL-4δ2, un nouveau variant d’épissage de l’IL-4 a étédécouvert chez l’homme. Par épissage alternatif, un nouvel exon est retenu dans l’ARNm dece variant. L’étude de celui-ci nous a permis de proposer, pour cet ARNm, une dégradationpar le mécanisme de Nonsens-Mediated Decay (NMD). Cette découverte apporte un niveausupplémentaire dans la compréhension du système de régulation de l’IL-4.Ce sujet d’étude apporte de nouveaux éléments quant à l’expression et la fonction del’IL-4δ2. De plus, l’identification d’un nouveau variant d’épissage enrichit la connaissancesur la régulation de l’expression du gène de l’Il4. D’une façon générale, la prise en comptedes variants d’épissage des cytokines devrait permettre de mieux caractériser la réponseimmunitaire, essentielle dans un contexte de vaccinologie. / Interleukin-4 (IL-4) is a key cytokine driving the humoral component of the immunesystem. An alternative splice variant of human IL-4, deleted of the second exon and so calledIL-4δ2 has been described. The expression of alternative splice variants is known to be tissuespecific,dependent of a particular stimulus or a pathological state. Their potential asbiomarkers is of increasing interest. Thus, this project was dedicated to the functionality ofIL-4δ2, improving characterization of the immune response.A kinetic study on the expression levels of IL-4 and its spliced variant, conducted onhealthy donors has shown to be donor-specific. The determination of the cell type able toproduce IL-4δ2 indicated that, CD4+ and CD8+ cells were not expressing the isoform, incontrast to granulocytes.The controversial agonist or antagonist function of IL-4δ2 was discussed throughfunctionality. The ability of IL-4δ2 to induce the signaling pathways of IL-4 was evaluated.An absence of similar profile of activation to IL-4 suggests a potential inhibitory role of IL-4δ2.During the study on IL-4δ2, a new alternatively spliced variant of IL-4 was discoveredin humans. Upon splicing, a new exon is retained in this variant. Its functional outcome as asubstrate for Nonsens-Mediated Decay (NMD) allowed bringing a new insight in thecomprehension of IL-4 regulatory system.Our work brought novel elements in the expression and functionality of IL-4δ2.Moreover, the discovery of a new alternatively spliced variant enriched the knowledge on theregulation pathways of Il4 gene expression. A focus on alternatively spliced variants ofcytokines is likely to clarify the complex regulation of the immune system.

Abstract Peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) and AMP-activated protein kinase (AMPK) are major factors regulating energy homeostasis. In this study, we aimed to investigate how energy flux affects several hepatic functions mediated by these factors. We define a novel role of PGC-1α and AMPK as modulators of the immune system in the liver. We show that PGC-1α is involved in the regulation of a cluster of genes related to the immune system, most importantly Interleukin 1 receptor antagonist (IL1Rn). Since PGC-1α is responsive to energetic stress associated with fasting or physical exercise, the same stimuli promote IL1Rn in hepatocytes. We identify AMPK as an independent inducer of IL1Rn and hypothesise that it could account for the anti-inflammatory effect of the antidiabetic drug metformin. We also demonstrate that metformin reduces expression of Sirtuin 3 (SIRT3) in hepatocytes and promotes acetylation of mitochondrial protein. We suggest that this mechanism, in spite of increased mitochondrial biogenesis, contributes to reduced ATP synthesis in metformin-treated samples. In addition, we demonstrate that Pregnane X receptor (PXR) is induced in the liver during fasting and by PGC-1α in hepatocytes. Furthermore, we describe a negative regulatory mechanism involving SIRT1, activated by pyruvate and interfering with PXR signaling. We show that SIRT1 attenuates PGC-1α-mediated co-activation of PXR and its target genes, i.e. Cyp3a11, with possible implications for drug and xenobiotic metabolism. In conclusion, we demonstrate how energetic stress affects various hepatic functions mediated by PGC-1α and AMPK. Moreover, we describe SIRT1 and metformin as factors capable of modulating this response. / Tiivistelmä Peroksisomiproliferaattori-aktivoituvan reseptori gamman koaktivaattori 1α (PGC-1α) ja AMP:n aktivoima proteiinikinaasi (AMPK) ovat keskeisiä energiametabolian säätelijöitä. Tässä tutkimuksessa oli tavoitteena selvittää kuinka energiataso vaikuttaa useisiin, näiden tekijöiden säätelemiin maksan toimintoihin. Osoitamme että PGC-1α ja AMPK tekijöillä on ennestään tuntematon merkitys immuunijärjestelmän säätelyssä maksassa. Näytämme myös, että PGC-1α säätelee joukkoa geenejä, joiden tehtävä liittyy immuunijärjestelmään, tärkeimpänä Interleukiini 1 reseptori antagonistia (IL1Rn). Paastoon ja fyysiseen aktiivisuuteen liittyvä energiastressi aktivoi PGC-1α:aa ja näiden samojen stimuluksien havaittiin lisäävän myös IL1Rn tasoa hepatosyyteissä. Havaitsimme AMPK:n olevan itsenäinen IL1Rn indusori ja hypoteesimme mukaan tämä voi välittää diabeteslääkkeenä käytettävän metformiinin anti-inflammatorisia vaikutuksia. Osoitamme myös, että metformiini alentaa Sirtuiini (SIRT) 3:n ekspressiota maksasoluissa ja lisää mitokondriaalisten proteiinien asetylaatiota. Uskomme tämän mekanismin, huolimatta lisääntyneestä mitokondrioiden biogeneesistä, myötävaikuttavan vähentyneeseen ATP synteesiin metformiinikäsitellyissä näytteissä. Lisäksi osoitamme, että paasto ja PGC-1α indusoivat Pregnaani X reseptorin (PXR) ilmentymistä maksasoluissa. Kuvaamme myös PXR signalointiin vaikuttavan ja pyruvaatin aktivoiman, SIRT1:n välitteisen, negatiivisen säätelymekanismin. SIRT1 estää PGC-1α välitteistä PXR koaktivaatiota ja kohdegeenien, kuten Cyp3a11, aktivaatiota, millä voidaan olettaa olevan merkitystä lääkeaineiden ja vierasaineiden metaboliaan. Yhteenvetona osoitamme, että energiastressi PGC-1α:n ja AMPK:n välittämänä vaikuttaa useisiin maksan toimintoihin. Lisäksi näytämme, että SIRT1 ja metformiini voivat moduloida näitä vaikutuksia.

L’extraction de l’uranium a eu pour conséquence une remobilisation de cet actinide au niveau des écosystèmes avoisinants les mines. L’utilisation de sels métalliques lors de la réhabilitation des sites miniers et la présence naturelle de métaux ont accentué les niveaux de la contamination métallique dans les hydrosystèmes soumis aux rejets miniers uranifères.Des expériences in situ ont été conduites au niveau de deux anciens sites miniers uranifères français. L’encagement de l’épinoche à trois épines a été employé pour connaître les effets sublétaux de ce mélange métallique, ainsi que sur la susceptibilité de ce poisson envers un stress biologique.Cette pollution, caractérisée par de plus importantes concentrations métalliques (notamment en uranium), a entraîné un stress oxydant chez l’épinoche visible sur plusieurs biomarqueurs, ainsi que d’autres effets dépendants du site d’exposition.La contamination polymétallique a occasionné une augmentation de la susceptibilité des épinoches au stress biologique, en empêchant leurs réponses phagocytaire antioxydante à ce stress. Ces travaux ont permis de renforcer l’intérêt de la technique d’encagement lors d’une étude environnementale, et celui des immunomarqueurs au sein d’une approche multi-biomarqueurs. / Uranium extraction has resulted in a remobilisation of this actinide into mine surrounding ecosystems. Uses of metal salts during mining site rehabilitation, and the natural presence of metals have increased the metal contamination in hydrosystems submitted to mine tailings.In situ experiments were conducted in two former French uranium mining sites. Three-spined stickleback caging was used to determine the sublethal effects of this metal mixture on this freshwater fish, as well as its effects on fish susceptibility to a sudden biological stress.This pollution, characterised by higher metal concentrations (especially for uranium), has led to an oxidative stress in sticklebacks visible through several biomarkers, and other effects dependent on the study site. The polymetallic contamination has modified the stickleback responses to the biological stress, by preventing their phagocytic and antioxidant responses. This work has reinforced the interest of the caging technique during environmental studies and that of immunomarkers in a multi-biomarker approach.

The main purpose of the natural immune system is to protect the body against any unwanted foreign cells that could infect the body and lead to devastating results. The nature immune system has different lymphocytes to detect and destroy these unwanted foreign patterns. The natural immune system can be modeled into an artificial immune system that can be used to detect any unwanted patterns in a non-biological environment. One of the main tasks of an immune system is to learn the structure of these unwanted patterns for a faster response to future foreign patterns with the same or similar structure. The artificial immune system (AIS) can therefore be seen as a pattern recognition system. The AIS contains artificial lymphocytes (ALC) that classify any pattern either as part of a predetermined set of patterns or not. In the immune system, lymphocytes have different states: Immature, Mature, Memory or Annihilated. Lymphocytes in the annihilated state needs to be removed from the active set of ALCs. The process of moving from one state to the next needs to be controlled in an efficient manner. This dissertation presents an AIS for detection of unwanted patterns with a dynamical active set of ALCs and proposes a threshold function to determine the state of an ALC. The AIS in the dissertation uses evolutionary computation techniques to evolve an optimal set of lymphocytes for better detection of unwanted patterns and removes ALCs in the annihilated state from the active set of ALCs. / Dissertation (MSc (Computer Science))--University of Pretoria, 2007. / Computer Science / unrestricted

Dans un contexte d'usage raisonné des antibiotiques lié au développement des résistances bactériennes, il est pertinent de chercher à optimiser les profils d'exposition plasmatique qui conduiraient à la meilleure efficacité de l'antibiotique sur les bactéries pathogènes. La charge bactérienne n'est pas stationnaire tout au long du développement d'une infection, mais elle augmente spontanément ou diminue avec un traitement antibactérien efficace. L'objectif de cette thèse était d'évaluer l'influence de la variation de la charge bactérienne sur l'efficacité des antibiotiques et du système immunitaire.Au cours d'un premier travail, nous avons montré que dans les conditions de réalisation de tests de sensibilité in vitro (détermination de CMI), une dégradation de certains antibiotiques se produisait, d'amplitude variable selon les molécules testées. Cette dégradation peut être responsable d'une augmentation des valeurs de CMI et de CMB. Les variations observées étaient cependant inférieures à une dilution au demi dans une gamme de concentrations. Cette dégradation ne devrait pas avoir d'impact significatif sur les résultats des tests de sensibilité aux antibiotiques concernés, excepté dans des cas particuliers comme des pathogènes à croissance très lente. Dans un deuxième travail, nous avons étudié in vitro chez E. coli et S. aureus l'effet de la taille de l'inoculum bactérien sur l'activité bactéricide de 2 céphalosporines, la céphalexine et le cefprozil. Nous avons observé une diminution de l'activité bactéricide des 2 céphalosporines avec l'augmentation de la taille de l'inoculum chez E. coli et S. aureus. Nous avons également montré une efficacité et une puissance moins importante des 2 céphalosporines sur S. aureus par rapport à E coli.Dans un troisième temps, nous avons développé un système incluant les trois composantes suivantes : une bactérie - S. aureus -, des cellules du système immunitaire - des macrophages murins issus de la moelle osseuse - et un antibiotique -la céphalexine-. Dans ce système, nous avons fait varier la taille de l'inoculum bactérien de départ ainsi les concentrations en antibiotique. Une augmentation de la phagocytose bactérienne et de la mortalité des macrophages ont été observées avec l'augmentation de la charge bactérienne. L'activité bactéricide des macrophages était saturable et en présence d'une charge bactérienne trop importante, une partie des macrophages sont devenus un réservoir de S. aureus phagocytés. En présence de la céphalexine, qui a une distribution exclusivement extracellulaire, les quantités de bactéries extracellulaires, " candidates " à la phagocytose, ont diminué. Ainsi, en présence de céphalexine et pour les charges bactériennes initiales les plus faibles, les capacités de survie et de bactéricidie des macrophages ont été préservées. Cette action n'a cependant plus été visible en présence de gros inocula bactériens pour lesquels l'action limitée de la céphalexine n'a pas permis de prévenir la saturation des macrophages et ses conséquences. Le modèle à trois composantes que nous avons développé constitue une première étape dans le développement de modèles in vitro qui associent des éléments de l'immunité innée aux modèles pharmacologiques classiques bactéries/antibiotiques, avec l'objectif d'optimiser l'évaluation préclinique de molécules antibactériennes. / As one of the current pre-eminent public health concerns is to reasonably use antibiotics in order to limit antibacterial resistance development, it appears relevant to determine the plasmatic exposition profile that would lead to the best efficiency of the antibiotic on pathogenic bacteria. The bacterial load is not stationary during an infection but it increases or decreases with an effective antibiotic treatment. The aim of this thesis was to evaluate the influence of variation of the bacterial load on antibiotic and immune system activity.First, we showed that during antibacterial sensibility tests, such as standard MIC determination, some antibiotics underwent abiotic degradation during incubation, with a magnitude depending on the drug tested. This degradation can increase MIC and MBC values. However, the observed discrepancy (less than one twofold dilution) suggests that this would only be clinically significant in special cases such as slow-growing bacteria.Then, we studied, with E. coli and S. aureus, the in vitro effect of the bacterial inoculum size on bactericidal activity of 2 cephalosporins, cephalexin and cefprozil. We observed a decrease of bactericidal activity of both cephalosporins with an increase of the initial inocula of E. coli and S. aureus. A decreased efficacy and potency of the 2 cephalosporins against S. aureus compared to E. coli was also found. Finally, we developed an in vitro 3-components model including a bacterium -S. aureus-, cells of the immune system -murine bone-marrow-derived macrophages- and an antibiotic -cephalexin-. Within this system, we tested several initial bacterial inoculum sizes and different antibiotic concentrations. Increased bacterial phagocytosis and macrophage mortality were observed with increasing bacterial inocula. Bactericidal activity of macrophages was saturable and faced to a large bacterial inoculum, some macrophages became a reservoir for living S. aureus. With cephalexin, which is an extracellular antibiotic, extracellular bacteria diminished over time implying a diminution of the bacteria to be phagocytosed by macrophages. Thus, macrophages bactericidal and survival abilities were preserved with cephalexin and small bacterial inocula. This effect of the antibiotic was no longer visible with highest bacterial inocula for which limited action of cephalexin did not allow to prevent macrophages bursting. The tripartite model we developed is a first step toward innovative in vitro models combining elements of innate immunity with classical bacteria/antibiotics pharmacological models, with the objective of optimising preclinical evaluation of antibacterial drugs.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-01-19T10:47:34Z No. of bitstreams: 1 felipedasilvaseta.pdf: 1053075 bytes, checksum: 8a24a576cad55e9b46efe4bde9405104 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-01-25T17:44:56Z (GMT) No. of bitstreams: 1 felipedasilvaseta.pdf: 1053075 bytes, checksum: 8a24a576cad55e9b46efe4bde9405104 (MD5) / Made available in DSpace on 2016-01-25T17:44:56Z (GMT). No. of bitstreams: 1 felipedasilvaseta.pdf: 1053075 bytes, checksum: 8a24a576cad55e9b46efe4bde9405104 (MD5) Previous issue date: 2015-08-10 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O presente trabalho propõe uma metodologia para a resolução do problema de reconfiguração ótima de sistemas de distribuição de energia elétrica utilizando uma representação mais realista de parâmetros com incertezas. O objetivo é avaliar o impacto de se representar incertezas dos sistemas no problema de reconfiguração em relação a modelos tradicionais determinísticos. O modelo de reconfiguração probabilística proposto visa minimizar as perdas totais de energia considerando incertezas sobre a demanda e sobre a geração distribuída a partir da energia eólica, além de diferentes níveis de carregamento dos sistemas. A metodologia proposta é baseada na técnica meta-heurística Sistema Imunológico Artificial. Os fundamentos da matemática intervalar são incorporados em um fluxo de potência intervalar que modela as incertezas da demanda provenientes principalmente de erros de previsão e medição, bem como incertezas na geração por fontes eólicas devido a intermitências nos regimes de ventos. Desta forma, as variáveis de entrada intervalares são as demandas ativas e reativas das barras do sistema e os valores de velocidade de vento nas regiões das usinas eólicas. As incertezas da entrada são propagadas para as variáveis de saída do fluxo de potência, como as tensões nodais. Como resultado, as perdas totais de energia a serem minimizadas também são determinadas na forma intervalar. Uma metodologia para comparação de intervalos baseada na média e no raio dos intervalos é utilizada para determinar a topologia ótima. Restrições de tensão, radialidade e conectividade da rede são consideradas. O algoritmo proposto é testado em sistemas conhecidos da literatura. / The present work proposes a methodology to solve the problem of optimal reconfiguration of power distribution systems by using a more realistic representation of uncertain parameters. The objective is to evaluate the impact of representing uncertainties in the reconfiguration problem in relation to traditional deterministic models. The proposed probabilistic reconfiguration model aims at minimizing the total energy loss considering uncertainties on the load demand and the distributed generation from wind energy, as well as different load levels. The proposed methodology is based on the meta-heuristic technique Artificial Immune System. The interval mathematics fundamentals are embedded in an interval power flow that models the uncertainties of load forecast and measurements, as well as uncertainties due to the intermittences of the wind. Therefore, the input interval variables are the active and reactive loads at the network nodes and the wind speed in the regions where the wind farms are installed. The input uncertainties are thus propagated to the output power flow variables as the nodal voltages. As a result, the total energy losses to be minimized are also given in interval form. A methodology for comparing intervals that is based on the interval average and size is used to determine the best topology. Voltage constraints, radial configuration and network connectivity are considered. The proposed algorithm is tested in systems known in the literature.

A esclerose múltipla (EM) é uma doença autoimune desmielinizante progressiva imunomediada por linfócitos T auto-reativos, que provocam uma cascata imunológica, amplificando a inflamação local. No Diabetes mellitus tipo 1 (DM1), existem linfócitos T auto reativos destroem as células beta do pâncreas, causando deficiência na produção de insulina. O desenvolvimento de terapêuticas específicas fica limitado pela etiologia indefinida destas doenças, apesar de avanços na terapêutica antiinflamatória e imunossupressora. Uma alternativa de tratamento atual para tais doenças é o transplante autólogo de células tronco hematopoéticas (TACTH). O presente estudo, observacional do tipo transversal, com a coleta de dados de caráter retrospectivo, tem como objetivo identificar as reações adversas manifestadas pelos pacientes diabéticos ou de esclerose múltipla, submetidos ao TACTH no período de 2004 a dezembro de 2010. Para a coleta de dados elaborou-se dois instrumentos que foram submetidos à validação aparente e de conteúdo por três juízes. A amostra final do estudo foi constituída pela obtenção dos dados de 72 prontuários, sendo 23 de pacientes diabéticos e 49 de pacientes com EM. Em relação aos pacientes diabéticos 16 pertenciam ao sexo masculino e a idade média foi 18,26 anos. Todos possuíam positividade para o anticorpo anti-carboxilase do ácido glutâmico (antiGAD65). Quanto aos pacientes com EM, trinta e três pertenciam ao sexo feminino e idade média foi de 37,2 anos. O subtipo da doença mais frequente foi o surto-remissivo em 21 (42,9%) pacientes. A escala expandida do estado de incapacidade (EDSS) variou entre 3,0 e 6,5. Em relação às reações adversas manifestadas pelos pacientes diabéticos foram mais frequentes os calafrios, febre, cefaléia, náusea e vômito e nos pacientes com esclerose múltipla foram retenção hídrica e cefaléia. As principais intervenções de enfermagem identificadas para os pacientes diabéticos e com EM foram monitorização dos sinais vitais, coleta de hemocultura, otimização da administração de medicamentos antieméticos, controle da infusão da globulina antitimocitária, orientações sobre alimentação e para reduzir o risco de queda. Os pacientes com DM1 apresentam reações mais agudas e necessitam de monitorização contínua. Já os pacientes com EM são mais dependentes dos cuidados de enfermagem, exigindo maior tempo de cuidados prestados pelo profissional. Embora o DM1 e a EM sejam doenças distintas, percebe-se que na prática clínica, exigem do enfermeiro uma excelência no cuidado, quer pelas particularidades do tratamento realizado ou pelas singularidades de cada uma delas. / Multiple sclerosis (MS) is a progressive demyelinating autoimmune disease, immune- mediated by auto-reactive T lymphocytes, which provoke an immunological cascade, enhancing the local inflammation. In type 1 diabetes mellitus (DM1), self-reactive T lymphocytes exist that destroy ? cells in the pancreas, causing insulin production deficiency. The development of specific therapeutics is limited by these diseases\' undefined etiology, despite advances in anti-inflammatory and immunosuppressive therapy. A current treatment alternative for these diseases is autologous hematopoietic stem cell transplantations (AHSCT). The aim of this observational and cross-sectional study with retrospective data collection is to identify the adverse reactions manifested by diabetic or MS patients who were submitted to AHSCT between 2004 and December 2010. For data collection, two instruments were elaborated, submitted to face and content validation with the help of three experts. The final study sample comprised data from 72 patient files, 23 from diabetic and 49 from MS patients. As for the diabetic patients, 16 were male and the mean age was 18.26 years. All were positive for the anti-glutamic acid decarboxylase (antiGAD65) antibody. Concerning MS patients, 33 were female and the mean age was 37.2 years. The most frequent disease subtype was relapsing-remitting in 21 (42.9%) patients. The expanded disability status scale (EDSS) score ranged between 3.0 and 6.5. As for the adverse reactions the diabetic patients manifested, shivers, fever, migraine, nausea and vomiting were the most frequent, while fluid retention and migraine were the most frequent among multiple sclerosis patients. The main nursing interventions identified for the diabetic and MS patients were vital sign monitoring, blood culture collection, optimization of anti-emetic drug administration, control of anti- thymocyte globulin infusion, dietary orientations and advice to reduce the risk of falls. DM1 patients present more acute reactions and need continuous monitoring. MS patients are more dependent on nursing care, demanding lower professional care time. Although DM1 and MS are distinct conditions, in clinical practice, they demand excellent care from nurses, whether due to the particularities of the treatment or the singularities of each disease.

Em anos recentes, muitos algoritmos bio-inspirados têm surgido para resolver problemas de classificação. Em confirmação a isso, a revista Nature, em 2002, publicou um artigo que já apontava para o ano de 2003 o uso comercial de Sistemas Imunológicos Artificiais para detecção de fraude em instituições financeiras por uma empresa britânica. Apesar disso, não observamos, a luz de nosso conhecimento, nenhuma publicação científica com resultados promissores desde então. Nosso trabalho tratou de aplicar Sistemas Imunológicos Artificiais (AIS) para detecção de fraude em cartões de crédito. Comparamos AIS com os métodos de Árvore de Decisão (DT), Redes Neurais (NN), Redes Bayesianas (BN) e Naive Bayes (NB). Para uma comparação mais justa entre os métodos, busca exaustiva e algoritmo genético (GA) foram utilizados para selecionar um conjunto paramétrico otimizado, no sentido de minimizar o custo de fraude na base de dados de cartões de crédito cedida por um emissor de cartões de crédito brasileiro. Em adição à essa otimização, fizemos também uma análise e busca por parâmetros mais robustos via multi-resolução, estes parâmetros são apresentados neste trabalho. Especificidades de bases de fraude como desbalanceamento de dados e o diferente custo entre falso positivo e negativo foram levadas em conta. Todas as execuções foram realizadas no Weka, um software público e Open Source, e sempre foram utilizadas bases de teste para validação dos classificadores. Os resultados obtidos são consistentes com Maes et al. que mostra que BN são melhores que NN e, embora NN seja um dos métodos mais utilizados hoje, para nossa base de dados e nossas implementações, encontra-se entre os piores métodos. Apesar do resultado pobre usando parâmetros default, AIS obteve o melhor resultado com os parâmetros otimizados pelo GA, o que levou DT e AIS a apresentarem os melhores e mais robustos resultados entre todos os métodos testados. / In 2002, January the 31st, the famous journal Nature, with a strong impact in the scientific environment, published some news about immune based systems. Among the different considered applications, we can find detection of fraudulent financial transactions. One can find there the possibility of a commercial use of such system as close as 2003, in a British company. In spite of that, we do not know of any scientific publication that uses Artificial Immune Systems in financial fraud detection. This work reports results very satisfactory on the application of Artificial Immune Systems (AIS) to credit card fraud detection. In fact, scientific financial fraud detection publications are quite rare, as point out Phua et al. [PLSG05], in particular for credit card transactions. Phua et al. points out the fact that no public database of financial fraud transactions is available for public tests as the main cause of such a small number of publications. Two of the most important publications in this subject that report results about their implementations are the prized Maes (2000), that compares Neural Networks and Bayesian Networks in credit card fraud detection, with a favored result for Bayesian Networks and Stolfo et al. (1997), that proposed the method AdaCost. This thesis joins both these works and publishes results in credit card fraud detection. Moreover, in spite the non availability of Maes data and implementations, we reproduce the results of their and amplify the set of comparisons in such a way to compare the methods Neural Networks, Bayesian Networks, and also Artificial Immune Systems, Decision Trees, and even the simple Naïve Bayes. We reproduce in certain way the results of Stolfo et al. (1997) when we verify that the usage of a cost sensitive meta-heuristics, in fact generalized from the generalization done from the AdaBoost to the AdaCost, applied to several tested methods substantially improves it performance for all methods, but Naive Bayes. Our analysis took into account the skewed nature of the dataset, as well as the need of a parametric adjustment, sometimes through the usage of genetic algorithms, in order to obtain the best results from each compared method.

Toll like receptors (TLRs) are found on B cells, macrophages, monocytes, and dendritic cells, and these cells belong to the innate immune system that recognizes antigens and induces multiple cell responses through the release of cytokines. TLR1, TLR2 and TLR6 function as heterodimers, either as TLR1/TLR2 or TLR2/TLR6 to recognize lipopeptides. TLR1/2 dimer activation releases inflammatory cytokines, while TLR2/TLR6 dimer activation releases immunomodulatory cytokines. Based on the size of the binding pocket between TLR2 and TLR6, it was hypothesized that lipopeptides, such as FSL1, could be simplified while keeping overall activity. FSL1 is a lipopeptide first isolated from Mycoplasma salivarum that activates macrophages at picomolar concentrations. It is expected that synthetic lipopeptides mimicking immunostimulatory molecules such as FSL1 will allow development of better ways to stimulate or modulate the immune system. Therefore, novel synthetic TLR2/6 ligands were synthesized replacing the polylysine chain with a polyamine chain showing activation of the immune cells in a manner like FSL1. Natural killer T-cell (NKT) antigens, such as α-galactosylceramide (α-GalCer), are carried through the body by lipid transfer proteins before they interact with the NKT cells. Not all the proteins involved in glycolipid transportation have been characterized. The synthesis of an α-GalCer analogue, termed CD1d-Triceps was designed to help find additional proteins involved in glycolipid trafficking. CD1d-Triceps has three functionalities: the first is the α-GalCer structure, and the other two are on C6 of the sugar: biotin, which helps tag the molecule for its purification, and a photoactive tag that, upon UV light activation, will cross-link with neighboring proteins. Antibiotic-resistant strains of Staphylococcus aureus (SA) are a growing health problem worldwide. Serotype 5 and 8 are the most common SA pathogens. Loading the serotype 5 or 8 disaccharides onto Qβ-particles that are linked to an NKT cell activator yield a vaccine that is expected to trigger adaptive immunity to the disaccharide. Previous similar studies showed production of antibodies with high affinity against Streptococcus pyogenes oligosaccharides in a similar vaccine.