Neste trabalho realizamos um estudo do comportamento do momento magnético e do deslocamento isomérico de uma impureza intersticial de Fe em hospedeiros metálicos de valências 4 (Ti, Zr), 3 (Sc, Y). Investigamos também impurezas intersticiais e substitucionais de Mo e Fe em hospedeiros divalentes. Para realizar este estudo fizemos cálculos de estrutura eletrônica utilizando o RS-LMTO-ASA (\"Real Space - Linear Muffin-Tin Orbital - Atomic Spherical Approximation\"), um método de primeiros princípios dentro da aproximação do funcional densidade local, implementado no espaço real. Calculamos o momento magnético no sítio da impureza nos sistemas acima e constatamos que a impureza intersticial de Fe é não magnética nos hospedeiros de valências 4 e 3, e que tanto as impurezas intersticiais como as substitucionais podem apresentar momento magnético nos hospedeiros divalentes. Mostramos que para os sistemas divalentes o momento magnético depende fortemente da relaxação. Os nossos resultados são explicados através de um modelo simples, baseado no modelo de Wolff. Investigamos também o comportamento do deslocamento isomérico no sítio da impureza de Fe nesses vários sistemas. Constatamos que os nossos resultados concordam razoavelmente bem com os dados experimentais e explicam o comportamento das tendências observadas. / In the present work, we have studied the magnetic moments and the behavior of the isomer shift at the interstitial Fe impurity site in Ti, Sc, Zr and Y hosts. We have also investigated interstitial and substitutional Fe and Mo impurities in Ca, Sr and Yb hosts. To perform the calculations, we have used the RS-LMTO-ASA scheme, a first principles method, within the local spin density approximation, implemented in real space. We calculated the magnetic moments at the impurity site in the above systems and all the substitucional impurities are found to be magnetic. The results show that interstitial Fe is non-magnetic in the tri- and tetravalent hosts, but interstitial Fe and Mo impurities could develop local magnetic moment in divalent hosts. \'We show that the magnetic moment at the impurity site in these divalent hosts is strongly dependent on lattice relaxation. The results can be explained using simple arguments based on Wolff model. We have investigated in a systematic way the behavior of the isomer shift of Fe impurities in these systems. We observed that our results are in generally good agreement with experiment and lead to better understanding of the observed trends in terms of the volume occupied by the Fe in each host.

Este trabalho tem como objetivo o estudo da influência do boro na fase αNb5Si3 (Fase T2) a baixas temperaturas analisando suas propriedades elétricas e magnéticas. Para o estudo deste tema as amostras foram preparadas, seguindo a estequiometria Nb5Si3-xBx, via metalurgia do pó e também por fusão a arco, com x dentro do intervalo limitado por 0  X  1,0. Estas amostras foram analisadas através de difratometria de raios x, microscopia eletrônica de varredura, magnetização, transporte elétrico e medidas calorimétricas em baixa temperatura. As medidas das propriedades elétricas e magnéticas destas fases mostram que a substituição de boro por silício induz supercondutividade na fase T2. O caráter volumétrico da transição supercondutora é confirmado pela medida de capacidade calorífica. Assim, este trabalho é o primeiro a mostrar uma nova família de materiais supercondutores que cristalizam na estrutura protótipo Cr5B3. / This work to study the influence of boron during low temperature αNb5Si3 (T2 phase) analyzing their electrical and magnetic properties. For the study of this subject the samples were prepared, following the stoichiometry Nb5Si3-xBx via powder metallurgy and also by arc melting process, with x within the range limited by 0X1.0. These samples were analyzed by X-ray diffraction, scanning electron microscopy, magnetization, electrical transport and calorimetric measurements of low temperature. Measurements of electrical and magnetic properties of these phases show that the substitution of boron by silicon induces superconductivity at the T2 phase. The bulk nature of superconducting transition is confirmed by heat capacity measurement. This study is the first to show a new family of superconducting materials that crystallize in the Cr5B3 prototype structure.

Apesar de bem estabelecido em termos morfológicos, a repercussão funcional do envolvimento das pequenas vias aéreas nas pneumopatias intersticiais fibrosantes (PIF) permanece controversa. O presente estudo avaliou de maneira invasiva e não-invasiva (espirometria, volume de fechamento, variação da complacência dinâmica com a freqüência respiratória) a função das pequenas vias aéreas em portadores de PIF, comparando com grupo controle e correlacionando com índices morfométricos de biópsias. Os testes funcionais não diferenciaram portadores da doença dos controles quanto ao acomentimento das pequenas via aéreas e não se correlacionaram com os dados morfométricos / Although well known involvement of small airway in fibrosing interstitial lung disease based on morfologic issues, its functional consequences remains controversial. We present an invasive and non-invasive physiologic study for small airway function (pulmonary function tests, closing volume and frequency dependence of dynamic compliance) in patients with lung fibrosis. The physiological data were compared with normal controls and correlated with morfometric measurements from biopsies. Specific small airway function data could not show obstructive pattern when compared with normals, and did not correlate with morofmetric data

O excesso de adenina na dieta (ADE) promove precipitação intrabular de cristais, levando a uma nefrite intersticial progressiva com perda de função renal. Estudo recente demonstrou que esse processo requer ativação do sistema NF-kB. No presente estudo investigamos o possível envolvimento de outros componentes da imunidade inata, além do NF-kB. Verificamos também a hipótese de que a nefropatia associada aos cristais continua a progredir mesmo depois de cessada a sobrecarga de adenina. Foram estudados ratos Munich-Wistar machos e adultos sem tratamento (C) ou recebendo 0.5% de ADE na dieta. Após 1 semana, a ADE foi removida da dieta e os animais foram seguidos por 4 ou 24 semanas. A administração de ADE por 1 semana promoveu uma inflamação intersticial aguda, com perda de função renal, alteração da pressão caudal, sem alterações glomerulares. Os mediadores da imunidade inata, como TLR2, TLR4, inflamassoma NLRP3, IL1beta e IL-6, apresentaram-se ativados sem, no entanto, ativar o sistema NF-kB. Após cessada a sobrecarga de ADE, a inflamação persistiu, com infiltração por macrófagos, expressão elevada de AngII, deposição progressiva de colágeno e, na fase mais tardia, glomeruloesclerose, caracterizando um processo inflamatório crônico, autônomo, que não contou com a participação do eixo NLR/IL1beta. Em contraste, o sistema NF-kB foi ativado, sendo um dos possíveis estímulos a produção intra-renal de AngII. Dois mecanismos patogênicos podem ser identificados neste estudo: 1) agudo, associado à ativação do eixo NLR-IL1beta; 2) crônico, associado à produção de AngII renal e à ativação do sistema NF-kB / Excess adenine in the diet (ADE) promotes intratubular crystal precipitation, leading to progressive interstitial nephritis and loss of renal function. A recent study has shown that this process requires activation of the NF-kB system. In the present study we investigated the possible involvement of other components of innate immunity, in addition to NF-kB, as well as whether nephropathy associated with excess adenine continues to progress even after dietary cessation. Male Munich-Wistar rats without treatment (C) or receiving 0.5% of ADE in the diet were studied. After 1 week, ADE was removed from the diet and the animals were followed for 4 or 24 weeks. Administration of ADE for 1 week promote acute interstitial inflammation, with loss of renal function, alteration of caudal pressure, without glomerular changes. Mediators of innate immunity, such as TLR2, TLR4, NLRP3 inflamassome, IL1beta and IL-6 , were shown to be activated, with no apparent activation of the NF-kB system. In the late phases of the model, the inflammation persisted, with significant infiltration by macrophages, high expression of AngII, progressive collagen deposition and glomerulosclerosis, characterizing a chronic, autonomic inflammatory process that did not involve the participation of the NLR/IL1beta axis. By contrast, the NF-kB system was activated, with intra-renal AngII production as a possible stimulus. Two mechanisms operated this study: 1) an acute one, associated with activation of the NLR-IL1beta axis; 2) a chronic one, associated with intrarenal AngII production and NF-kB activation

INTRODUÇÃO: A inflamação desempenha um papel central no desenvolvimento da asma, que é considerada uma doença alérgica com um perfil inflamatório clássico Th2. No entanto, as citocinas de perfil IL-17 tem sido estudadas para melhor compreender sua participação na fisiopatologia desta doença. Pacientes asmáticos graves apresentam exacerbações freqüentes, sendo a causa infecciosa um dos principais desencadeantes e que podem perpetuar as respostas inflamatórias. A resposta Th17 pode estar claramente associada a esses eventos. OBJETIVO: Este estudo avaliou os efeitos da terapia com anticorpo monoclonal anti-IL-17 nas alterações presentes nos septos alveolares em um modelo experimental de inflamação pulmonar alérgica crônica exacerbado por LPS. MÉTODOS: Foram utilizados 60 camundongos macho da espécie BALB/c, sendo sensibilizados com ovoalbumina intraperitoneal e repetidamente expostos à inalação com ovoalbumina, seguidos por tratamento com ou sem anti-IL-17. Vinte e quatro horas antes do final do protocolo experimental de 29 dias, dois grupos receberam LPS intratraqueal (0,1 mg/ml, sendo os grupos OVA-LPS e OVA-LPS anti-IL-17). Posteriormente, avaliamos o fluido do lavado broncoalveolar (FLBA), por morfometria quantificamos o recrutamento das células apresentadoras de antígenos (FOXP3 e células dendríticas), de eosinófilos, a expressão celular de citocinas próinflamatórias (TNF-alfa, IL-2, IL-4, IL-5, IL-6, IL13 e IL-17), anti-inflamatórias (IL-10), quimiocina (TARC), além da quantificação de IL-6 por RT-PCR. Avaliamos também elementos da matriz extracelular (fibras colágenas tipo I e III, MMP-9, MMP-12, TIMP-1, TGF-beta, actina, decorina, lumicam, biglicano, fibronectina e integrina), edema dos septos alveolares, resposta das vias de estresse oxidativo através dos marcadores iNOS e 8-iso-PGF2alfa e as vias sinalizadoras NF-kB e Rho quinase, além da avaliação do diâmetro alveolar médio. RESULTADOS: Os animais do grupo OVA-LPS apresentaram uma potencialização de todas as respostas (p < 0,05) comparativamente ao grupo OVA, exceto para: expressão de IL-17, TNF-alfa, NF-kB, TIMP-1 e na fração de volume de fibras colágenas tipo I, decorina, lumicam e actina. No grupo OVA-anti-IL17 houve atenuação de todos os parâmetros quando comparado ao grupo OVA (p < 0,05). Nos animais do modelo de inflamação alérgica crônica exacerbado pelo LPS e tratado com anti-IL-17 (grupo OVA-LPS anti-IL-17) houve diminuição comparativamente ao grupo OVA-LPS dos seguintes parâmetros: número de células totais do lavado broncoalveolar, de células positivas para FOXP3 e células dendríticas, número de CD4+ e CD8+, de células positivas para IL-10 anti-inflamatória e citocinas pró-inflamatórias (TNF-alfa, IL-2, IL-4, IL-5, IL-6, IL-13 e IL-17); quimiocinas (TARC), expressão gênica de IL-6, edema nos septos alveolares; elementos da matriz extracelular (fração de volume de fibras colágenas tipo I e III, actina, decorina, biglicano, lumicam, fibronectina, integrina e expressão de células positivas para TGF-beta, MMP-9, MMP-12 e TIMP-1), da resposta das vias de estresse oxidativo: (células positivas para iNOS e fração de volume de isoprostano PGF-2alfa; da expressão celular de NF-kB, e das proteínas Rho quinase 1 e 2 (p < 0,05). Não houve diferenças no diâmetro alveolar médio entre todos os grupos experimentais. CONCLUSÃO: Esses dados sugerem que a inibição da IL-17 pode ser uma via terapêutica promissora para o tratamento da inflamação alérgica crônica, mesmo durante uma exacerbação e pode contribuir para o controle da inflamação Th1/ Th2/Th17, da expressão de quimiocinas, do remodelamento da matriz extracelular e do estresse oxidativo. As vias sinalizadoras de NF-kB e Rho-quinase estão envolvidas no controle dessas respostas neste modelo de inflamação alérgica crônica exacerbado pelo LPS / INTRODUCTION: Inflammation plays a central role in the development of asthma, which is considered an allergic disease with a classic Th2 inflammatory profile. However, IL-17 profile cytokines have been studied to better understand their involvement in the pathophysiology of this disease. Severe asthmatic patients have frequent exacerbations, the infectious cause being one of the main triggers and that can perpetuate the inflammatory responses. The Th17 response may be clearly associated with these events. OBJECTIVE: This study evaluates the effects of anti-IL-17 monoclonal antibody therapy on alveolar septa in an experimental model of chronic allergic lung inflammation of asthma exacerbated by LPS. METHODS: Sixty male BALB / c mice were used, being sensitized with intraperitoneal ovalbumin and repeatedly exposed to inhalation with ovalbumin, followed by treatment with or without anti-IL-17. Twenty-four hours before the end of the 29-day experimental protocol, two groups received intratracheal LPS (0.1 mg / ml, OVA-LPS and anti-IL-17 OVA-LPS groups). Afterwards, we evaluated bronchoalveolar lavage fluid (BALF), by morphometry we quantified the recruitment of antigen-presenting cells (FOXP3 and dendritic cells), eosinophils, the cellular expression of proinflammatory cytokines (TNF-alpha, IL-2, IL IL-5, IL-6, IL-13 and IL-17), anti-inflammatory cytokines (IL-10), chemokine (TARC), and quantification of IL6 by RT-PCR. We also evaluated elements of the extracellular matrix (collagen fibers type I and III, MMP-9, MMP-12, TIMP-1, TGF-beta, actin, decorin, lumicam, biglican and fibronectin), alveolar septum edema, oxidative stress through the marker iNOS and 8-iso-PGF2? and the signaling pathways NF-kB and Rho kinase. In addition, we evaluated of the mean alveolar diameter. RESULTS: The animals of the OVA-LPS group presented a potentiation of the all responses compared to OVA (p < 0.05), except for: expression of IL-17, TNFalpha, NF-kB, TIMP-1 and in the volume fraction of type I collagen fibers, decorin, lumicam and actin. Treatment with anti-IL-17 (OVA-anti-IL17 group) attenuated all parameters compared to OVA group (p < 0.05). The animals of the chronic allergic inflammation model exacerbated by LPS and treated with anti-IL-17 (OVA-LPS anti-IL-17 group) had a decreased of the following parameters compared to OVA-LPS: total bronchoalveolar lavage cells number, FOXP3 and dendritic positive cells, CD4 + and CD8 + numbers, of cells positive for IL-10 antiinflammatory, and pro-inflammatory cytokines (TNF-alfa, IL-2, IL-4, IL-5, IL-6, IL- 13 and IL-17); chemokine (TARC), genic expression IL6; edema in the alveolar septum; extracellular matrix elements (volume fraction of collagen fibers type I and III, actin, decorin, biglican, lumicam, fibronectin, and expression of TGF-beta, MMP-9, MMP-12 and TIMP-1 positive cells) of the oxidative stress pathways response (iNOS positive cells and isoprostane volume fraction 8-isoPGF-2alfa) NFkB and Rho kinase 1 and 2 positive cells (p < 0.05). There were no differences in mean alveolar diameter among all experimental groups. CONCLUSION: These data suggest that inhibition of IL-17 may be a promising therapeutic pathway for the treatment of chronic allergic inflammation, even during an exacerbation, and may contribute to the control of Th1 / Th2 / Th17 inflammation, chemokine expression, extracellular matrix remodeling and oxidative stress. Signaling pathways of NF-kB and Rho-kinase are involved in the control of these responses in this model of chronic allergic inflammation exacerbated by LPS

Foi estudado, através de ensaio de tração, o Nb policristalino com teor de hidrogênio de 0 a 50 partes por milhão em peso (ppm-p) nas temperaturas de 223, 273 e 293 K. Os ensaios de tração a velocidades constantes foram realizados com taxas iniciais de deformaçao de 4,2 X 10-5 e 42 X 10-5 s-1 , e os parâmetros de ductilidade e resistência mecânica foram analisados em função da concentração de hidrogênio. Foram também obtidos o coeficiente de sensitividade a taxa de deformação (m) e o volume de ativação (V) através de ensaios de tração, alternando-se abruptamente a taxa de deformação entre os valores acima mencionados, em sucessivos pontos da curva tensão-deformação. Para o cálculo destes dois últimos parâmetros foi proposta uma análise detalhada, considerando-se os efeitos elásticos e o encruamento durante a deformação plástica uniforme. Todo o estudo foi acompanhado por análise fratográfica Que permitiu verificar três comportamentos de ruptura da liga Nb-H, dependendo do teor de hidrogênio e da temperatura. A 223K foi observado que há uma forte redução de ductilidade do Nb para teores de hidrogênio até 10 ppm-p, sendo que para este teor o seu comportamento foi totalmente frágil / Tensile testing at 223, 273 and 293 K was carried out on polycrystalline Nb dopped from 0 to 50 parts per million in weight (ppm-wt) of Hydrogen. The tensile testing at constant velocity was done at 4,2 X 10 -5 and 42 X 10-5 s-1 of initial strain rate, and the ductility and strength parameters was analysed as a function of the hydrogen content. It was also obtained the strain rate sensitivity (m) and the activation volume (V), from tensile testing, cycling between the two above specified strain rates, at several points of the stress-strain curve. For the calculation of this two last parameters it was proposed a detailed analysis, considering the elastic effect and the work hardening during the uniform plastic deformation. All these studies was followed by fratographic analysis that alowed the identification of three rupture behavior for the Nb-H alloy, depending of the temperature and the hydrogen content. At 223 K, it was observed that there is a strong embrittlement of Nb for hydrogen content up to 10 ppm-wt, and for this value the behavior was completely brittle

Die Rationale für immuntherapeutische Ansätze zur Behandlung maligner Neoplasien geht davon aus, daß Tumore über spezifische Tumorantigene verfügen. Dendritische Zellen als die wichtigsten antigenpräsentierenden Zellen sind in der Lage, Tumorantigene naiven T-Zellen zu präsentieren und spezifische zytotoxische T-Zellen zu stimulieren. In der vorliegenden Arbeit wurden dendritische Zellen durch Stimulation mit Interleukin-4 (IL-4) und Granulozyten/ Makrophagen Koloniestimulierender Faktor (GM-CSF) aus peripheren mononukleären Blutzellen gesunder Spender und an Tumoren des gastroenteropankreatischen Systems erkrankter Patienten generiert. Mit den dendritischen Zellen cokultivierte immunologische Effektorzellen (Zytokin- induzierte Killerzellen, CIK-Zellen) wurden im Zytotoxizitätstest gegen kolorektale und pankreatische Karzinomzellen eingesetzt. CIK-Zellen sind zytototoxische Zellen, die durch Stimulation mit Zytokinen aus peripheren Blutlymphozyten erzeugt werden. Durch die Cokultivierung der Effektorzellen mit dendritischen Zellen konnte eine signifikante Steigerung der unspezifischen zytotoxischen Wirkung der CIK-Zellen bewirkt werden. Zur Steigerung der spezifischen Zytotoxizität wurden dendritische Zellen mit dem Gesamtprotein der tumor- assoziierten Antigene cancer associated antigen (CA 19-9) und carcinoembryonic antigen (CEA) gepulst. Effektorzellen zeigten nach der Cokultur mit gepulsten dendritischen Zellen zytotoxische Wirkung gegen Targetzellen, die das zum Pulsen verwendete Tumorantigen auf der Zelloberfläche exprimieren. Die Antigenspezifität der zytotoxischen Wirkung konnte durch eine signifikant verminderte Zellyse nach Blockade des Tumorantigens auf den Targetzellen belegt werden. Erstmals beschrieben ist hier das Pulsen dendritischer Zellen mit sowohl autologen als auch allogenen Seren von Patienten mit erhöhten Tumormarkerspiegeln. Eine Kultivierung dendritischer Zellen in tumormarkerhaltigem Serum bewirkte dosisabhängig eine verstärkte zytotoxische Wirkung cokultivierter Effektorzellen gegen Tumorzellen. Die verstärkte Zellyse zeigte sich unabhängig vom allogenem oder autologem Charakter des Serums. Der immunstimulierende Effekt des Patientenserums konnte durch eine vorhergehende Hitzeinaktivierung des Serums neutralisiert werden. Die höchsten Zellysen wurden durch eine Kultivierung dendritischer Zellen in tumormarkerhaltigem Serum und zusätzlichem Pulsen mit exogenem Tumorantigen erreicht. Untersuchungen an komplett autologen Systemen reproduzierten die an Zellkulturen erhobenen Befunde. Hierfür wurden erfolgreich Primärkulturen kolorektaler Tumore etabliert.Aus dem Blut von Tumorpatienten wurden dendritische Zellen generiert, die mit autologem Serum kultiviert wurden. Die cokultivierten autologen Effektorzellen erwiesen sich im Zytotoxizitätstest gegen autologe Tumorzellen als zytotoxisch. Die Cokultivierung der Effektorzellen mit den dendritischen Zellen bewirkte bei beiden Zellpopulationen Veränderungen. Dendritische Zellen zeigten nach der Cokultur eine verstärkte Expression antigenpräsentierender und costimulatorischer Moleküle. Bei den CIK-Zellen kam es zu einem Anstieg der Proliferationsrate. Bei Untersuchungen zur Antigenspezifität von T-Zellrezeptoren konnte vermehrt antigenspezifischer T-Zellrezeptor nachgewiesen werden. Des weiteren stieg das Verhältnis zwischen zytotoxischen T-Zellen und T-Helferzellen zugunsten der zytotoxischen T-Zellen. In ELISpot-Untersuchungen wurde eine Zunahme Interferon-gamma sezernierender CIK-Zellen nachgewiesen. Dendritische Zellen ließen sich erfolgreich mit inaktiviertem Adenovirus, an das kovalent Poly-L-Lysin gekoppelt ist, transfizieren. Die für den adenoviralen Gentransfer benötigten Oberflächenstrukturen konnten auf dendritischen Zellen nachgewiesen werden. Zur Verbesserung der Zytotoxizität wurden dendritische Zellen erfolgreich mit dem Gen für den Transaktivator CIITA transfiziert. CIITA- transfizierte dendritische Zellen exprimierten vermehrt MHC Klasse II-Moleküle. Die transduzierten dendritischen Zellen induzierten bei cokultivierten Effektorzellen eine erhöhte unspezifische Zytotoxizität. Mit Tumorantigen gepulste dendritische Zellen können bei der Entwicklung immuntherapeutischer Protokolle bei malignen Neoplasien von Bedeutung sein. / The immunotherapeutic approach against malignant neoplasias appreciates that tumours encode tumour rejection antigens, that enable them to induce protective immunity. Dendritic cells are major antigen-presenting cells and are able to present tumour antigens to naive T-cells and stimulate cytotoxic T-cells in a specific manner. In the present graduation-manuscript dendritic cells were generated in the presence of Interleukin-4 and granulocyte/macrophage colony-stimulating factor (GM-CSF) from peripheral mononuclear blood cells of healthy donors and tumour-patients. Immunological effector cells termed cytokine-induced killer cells (CIK cells) were co-cultured with dendritic cells and tested for their cytotoxic capacity against colorectal and pancreatic cancer cell-lines in a LDH-release assay. CIK cells are cytotoxic lymphocytes generated by incubation of peripheral blood lymphocytes with different cytokines. Co-culture of effector cells with dendritic cells led to a significant increase of the cytotoxic effect of CIK cells. For a further increase of specific cytotoxicity dendritic cells were pulsed with total protein of the tumour-associated antigens cancer associated antigen CA 19-9 and carcinoembryonic antigen (CEA). Co-cultured effector cells showed an increase in cytotoxicity against tumour-antigen expressing target cells, after co-culture with pulsed dendritic cells. The specificity of the cytotoxic effect could be shown by blocking the tumour-antigens with a monoclonal antibody. Autologous and allogenec untreated serums from patients with elevated tumour-marker levels were also used for pulsing of dendritic cells. Similar to the results when using total protein for pulsing, a cultivation in serum of patients with elevated tumour marker levels caused an intensified cytotoxic effect of effector cells against tumour cells in a dose-dependent manner. The intensified cytotoxicity was seen independent of the allogenec or autologous character of the serum. The immuno-stimulating effect of the patient serum could be neutralized by preceding heat inactivating. The highest cytotoxicity was achieved by a cultivation of dendritic cells in serum from patients with elevated tumour marker levels and additional pulsing with exogenous tumour antigen. Experiments with completely autologous systems reproduced the results made with cell-lines. Primary cultures of colorectal tumours were established. Dendritic cells were generated from the blood of tumour patients and were cultivated in autologous serum. Co-cultured autologous effector cells showed cytotoxicity when used against autologous tumour cells. Co-culturing of effector cells with dendritic cells caused modifications at both cell populations. Dendritic cells showed an increase expression of antigen-presenting and co-stimulatory molecules. CIK cells showed a higher proliferation-rate when co-cultured. They express more antigen-specific T-cell receptor, and the cytotoxic T-cells to T-helper cells ratio increased. ELISpot-assays showed an increase of interferon gamma producing cells. Dendritic cells were successfully transduced by using an inactivated adenovirus, which covalently binds poly-L-lysine. Dendritic cells express the molecules that enables adenoviral gene delivery on their surface. For the improvement of cytotoxicity dendritic cells were transduced with the gene encoding for the transactivator CIITA. CIITA transduced dendritic cells increases expression of MHC class II molecules. Cytotoxicity experiments with transduced dendritic cells resulted in an increased induction of non-specific cytolysis from co-cultured effector cells. DC pulsed with tumour-antigens may have a major impact on immunotherapeutic protocols for cancer patients.

La thérapie photodynamique (PDT) appliquée aux tumeurs cérébrales est évaluée comme une stratégie complémentaire par rapport aux thérapies conventionnelles. De nombreux travaux mettent en exergue le rôle prépondérant joué par l'effet vasculaire de la PDT dans l'éradication tumorale. Ainsi, une accumulation sélective du photosensibilisateur au niveau des néo-vaisseaux tumoraux favorise cet effet et donc, l'efficacité du traitement photodynamique. La stratégie vasculaire consistant à coupler un photosensibilisateur à un peptide ligand pour cibler le récepteur neuropiline-1 (NRP-1) surexprimé par les cellules endothéliales angiogéniques a été validée, démontrant également l'induction de l'expression du facteur tissulaire immédiatement après PDT. Grâce à l'utilisation de nanoparticules multifonctionnelles, des améliorations ont été apportées à la stratégie initiale pour une PDT interstitielle (iPDT) guidée par l'imagerie. Fonctionnalisées par le peptide ligand, vecteur du photosensibilisateur et d'un agent de contraste puis rendues furtives, les nanoparticules sélectionnées présentent les propriétés originales requises pour une action combinée en IRM et PDT ciblée. Les nano-objets sont affins pour NRP-1 et conservent leur caractéristique photo-activable. Les essais sur rats nude xénogreffés en orthotopique par un modèle de gliome malin humain, valident la faisabilité du concept de iPDT guidée par l'IRM en temps réel. Après injection des nanoparticules par voie intraveineuse, un rehaussement positif du signal IRM est observé au niveau de la zone tumorale pour optimiser l'implantation de la fibre optique. Les résultats obtenus par IRM de perfusion et, l'expression protéique de NRP-1 au niveau du tissu et des berges tumorales, valident la sélectivité des nanoparticules fonctionnalisées. La combinaison des techniques d'imagerie non-invasives (IRM, SRM, TEP/CT) a permis le suivi thérapeutique / Photodynamic therapy (PDT) for brain tumors appears to be complementary to conventional treatments. Number studies show the major role of the vascular effect in the tumor eradication by PDT. To promote this vascular effect, a selective targeting of neuropilin-1 (NRP-1), mainly over-expressed by tumor angiogenic vessels, was investigated using a photosensitizer coupled to a ligand peptide. We validated the interest of using this active-targeting strategy to promote this vascular effect by the induction of tissue factor expression immediately post-PDT. For interstitial PDT (iPDT) of brain tumors guided by real-time imaging, multifunctional nanoparticles consisting of a surface-localized tumor vasculature targeting NRP-1 and encapsulated PDT and imaging agents, have been developed. The selected nanoparticles are favourable to a photosensitizer targeting strategy for iPDT combined with MRI.Characterization studies of the nanoparticles reveal a photodynamic efficiency and demonstrate a molecular affinity of the functionalized nanoparticle to NRP-1 target. After intravenous injection of the multifunctional nanoparticles into rats with intracranial glioma, we demonstrate a positive contrast enhancement of the tumor tissue by MRI, allowing the optimization of the optical fiber implantation. Perfusion MRI data and NRP-1 protein expression of the tumor and brain adjacent to tumor tissues check selectivity of the functionalized nanoparticle. The combination of non-invasive techniques of imaging (MRI, MRS, PET/CT) validates this concept of iPDT guided by MRI

Les limitations rencontrées aujourd'hui dans le traitement du glioblastome (GBM) concernent notamment la qualité de l'exérèse dont dépend le pronostic et le manque de contrôle local de la croissance tumorale, sachant que les récidives apparaissent dans plus de 80% des cas dans le volume cible de radiothérapie. Dans ce contexte, la thérapie photodynamique interstitielle (iPDT) se présente comme un outil complémentaire prometteur qui permettrait d'améliorer le contrôle local de la tumeur. La première partie de ce travail de thèse a porté sur le suivi longitudinal par Imagerie par Résonance Magnétique (IRM) de la réponse tumorale post-iPDT sur un modèle de rat nude xenogreffé en orthotopique par un modèle de GBM humain. Le suivi par IRM et Spectroscopie par Résonance Magnétique (SRM) a fourni des indicateurs précoces de l'efficacité du traitement, permettant de discriminer dès un jour post-iPDT les animaux répondeurs des non-répondeurs. Cependant, une des limitations de la PDT, demeure la faible profondeur de pénétration de la lumière visible utilisée pour activer le photosensibilisateur et induire les réactions de photo-oxydations. La seconde partie de ce travail a porté sur l'évaluation d'un nouveau concept appelé "PDTX" permettant de coupler l'effet photodynamique à celui de la radiothérapie pour une radiothérapie photodynamique, en jouant notamment sur la complémentarité des espèces réactives de l'oxygène générées et des effets RX-induits. Pour cela, nous avons validé l'intérêt d'une nanoparticule hybride de type AGuIX® composée de terbium et de porphyrine, le terbium étant le scintillateur capable d'être excité par les rayons X et d'émettre des photons à une longueur d'onde appropriée pour activer le photosensibilisateur. Le transfert d'énergie par FRET (Förster Resonance Energy Transfer) entre le terbium et la porphyrine a été mis en exergue. Les résultats in vitro démontrent le potentiel thérapeutique de ce nouveau nano-objet à basse énergie / The limitations encountered today in the treatment of glioblastoma (GBM) involve the quality of the resection on which depends prognosis and the lack of local control of the tumor, knowing that relapses occur in 80% of cases in the radiotherapy target tumor volume. In this context, interstitial photodynamic therapy (iPDT) is a promising additional tool that would allow to improve local control of the tumor. The first part of this thesis focused on the longitudinal follow-up by Magnetic Resonance Imaging (MRI) of the post-iPDT tumor response in a nude rat model of orthotopic xenograft of human GBM cell line. MRI and Magnetic Resonance Spectroscopy (MRS) monitoring provided early indicators of the effectiveness of treatment, for discriminate from one day post-IPDT non-responders from responders animals. However, one of the limitations of PDT remains the low penetration of visible light used to activate the photosensitizer and induce reactions of photo-oxidation. This is why the second part of this research focused on the evaluation of a new concept called "PDTX" for coupling the photodynamic effect with radiotherapy effect for a photodynamic radiotherapy, playing especially on the complementarity of reactive species of oxygen generated and RX-induced effects. For this, we validated the interest of an AGuIX®-type hybrid nanoparticle composed of terbium and porphyrin, terbium being the scintillator capable of being excited by X-rays and emits photons at an appropriate wavelength in order to activate the photosensitizer. The energy transfer FRET (Förster Resonance Energy Transfer) between terbium and porphyrin was highlighted. In vitro results demonstrate the therapeutic potential of this new nano-object at low-energy

INTRODUÇÃO: A pneumonite de hipersensibilidade (PH) é uma doença intersticial causada pela inalação de antígenos orgânicos específicos ou substâncias de baixo peso molecular em indivíduos geneticamente suscetíveis. A PH crônica representa seu estágio final, na qual a exposição antigênica prolongada leva à fibrose. Na PH crônica, o envolvimento das pequenas vias aéreas (PVA) é proeminente; entretanto, uma avaliação detalhada através de métodos funcionais e de avaliação quantitativa e automatizada pela tomografia computadorizada (TC) não foi realizada previamente. MÉTODOS: estudo transversal de 28 pacientes com PH crônica, com avaliação através de provas de função pulmonar (PFPs); oscilometria forçada (FOT); análise automatizada do volume pulmonar através da TC, incluindo quantificação de aprisionamento aéreo; e teste cardiopulmonar de exercício (TCPE) incremental em cicloergômetro para avaliar performance ao exercício, incluindo medidas seriadas da capacidade inspiratória e hiperinsuflação dinâmica (HD). Foram incluídos pacientes entre 18 a 75 anos, com diagnóstico confirmado pela combinação de achados tomográficos, exposição antigênica e biópsia compatível e/ou LBA com linfocitose. Foram excluídos pacientes com CVF e/ou VEF1 < 30% predito, tabagismo > 20 anos-maço, uso de oxigênio suplementar; diagnóstico prévio de asma ou DPOC, diagnóstico de hipertensão arterial pulmonar ou impossibilidade de realizar TCPE. Os dados foram comparados com controles saudáveis. RESULTADOS: 28 pacientes (16 mulheres; idade média 56 +- 11 anos; CVF 57 +- 17% predito) foram avaliados, e todos apresentavam padrão ventilatório restritivo sem resposta broncodilatadora. Na FOT, 4 pacientes apresentaram resistência aumentada a 5 Hz (R5), enquanto todos apresentaram baixa reactância (X5), sendo que nenhum apresentou resposta broncodilatadora significativa. Pacientes com PH crônica tiveram menor capacidade de exercício com menor O2 de pico, diminuição da reserva ventilatória, hiperventilação, dessaturação de oxigênio e escores de dispneia (Borg) aumentados quando comparado aos controles. A prevalência de HD foi encontrada em apenas 18% da coorte. Ao comparar pacientes com PH crônica com O2 normal e baixo (< 84% predito, LIN), o último grupo apresentou maior hiperventilação (slope E/CO2), um menor volume corrente e menores escores de capacidade física na avaliação do questionário de qualidade de vida (SF-36). A análise da curva ROC mostrou que volumes pulmonares reduzidos (CVF%, CPT% e DLCO%) foram preditores de baixa capacidade ao exercício. Na TC, a PH crônica teve aumento de áreas com alta densidade em unidades Hounsfield, inferindo maior extensão de opacidades em vidro fosco e fibrose em relação aos controles saudáveis. A extensão das áreas de atenuação reduzida (AAR) e aprisionamento aéreo em relação ao volume pulmonar total é pequena, e não se correlaciona com índices funcionais obstrutivos; entretanto, pacientes com maior percentual dessas áreas apresentam menos fibrose e função pulmonar mais preservada. CONCLUSÃO: a PH crônica se caracterizou por um acometimento eminentemente restritivo, e não de obstrução de vias aéreas, nos diferentes métodos diagnósticos aplicados. A redução da capacidade de exercício foi prevalente devido à limitação ventilatória e de troca gasosa, a exemplo de outras doenças intersticiais pulmonares, e não pela HD. Redução dos volumes pulmonares foram bons preditores das respostas ventilatórias durante o exercício / INTRODUCTION: Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by the inhalation of specific organic antigens or low molecular weight substances in genetically susceptible individuals. Chronic HP represents its final stage, in which prolonged antigenic exposure causes fibrosis. In chronic HP, small airway involvement is prominent; however, a detailed characterization through functional evaluation and through automatic quantitative evaluation of computed tomography (CT) has not been previously assessed. METHODS: Cross-sectional study with 28 chronic HP patients, with evaluation by pulmonary function tests (PFTs), forced oscillometry (FOT), automated lung volume analysis through CT, including quantification of air trapping (AT); and incremental cardiopulmonary exercise testing (CPET) on a cycle ergometer to evaluate exercise performance, including serial measurements of inspiratory capacity to establish dynamic hyperinflation (DH). Inclusion criteria: patients aged 18 to 75 years, with a chronic HP diagnosis confirmed by the combination of CT findings, known antigenic exposure and compatible biopsy and / or BAL with lymphocytosis. Exclusion criteria: FVC and / or FEV1 < 30% predicted, smoking > 20 pack-years, supplemental oxygen use; previous diagnosis of asthma or COPD; pulmonary arterial hypertension, or medical conditions that could interfere with CPET. Data were compared with healthy controls. RESULTS: All patients (16 women; mean age 56 +- 11 years; FVC 57 +- 17% predicted) had restrictive ventilatory pattern without bronchodilator response. In FOT, 4 patients had increased resistance at 5 Hz (R5), all patients presented low reactance (X5) values, and none presented a significant bronchodilator response. Chronic HP patients had reduced exercise performance with lower peak V?O2, diminished breathing reserve, hyperventilation, oxygen desaturation and augmented Borg dyspnea scores when compared with controls. The prevalence of DH was only found in 18% of patients. When comparing chronic HP patients with normal and low peak VO2 (< 84%predicted, LLN), the later exhibited higher hyperventilation (VE/VCO2 slope), lower tidal volumes, and poorer physical functioning scores on Short-form-36 health survey. ROC curve analysis showed that reduced lung volumes (FVC%, TLC% and DLCO%) were high predictors of poor exercise capacity. On CT, chronic HP is characterized by increased pulmonary densities (Hounsfield Units) inferring the extension of ground glass opacities and fibrosis when compared with healthy subjects. The extension of low attenuation areas (LAA) and AT in relation to the hole lung volume is low and does not correlate with PFT indexes of obstruction; however, patients with greater extension of these areas had less fibrosis and more preserved PFTs. CONCLUSIONS: Chronic HP was characterized by an imminently restrictive lung disorder, and not by airway obstruction, according to the different diagnostic methods applied in this study. Reduction of exercise capacity was prevalent due to ventilatory and gas exchange limitation, similarly to other fibrotic interstitial lung diseases, rather than due to DH. Reduced lung volumes were good predictors of ventilatory responses during exercise