As espécies do gênero Viguiera Kunth, recentemente transferidas para Aldama La Llave (Asteraceae, Heliantheae), ainda possuem problemas de delimitação taxonômica. Tal gênero possui 35 espécies distribuídas por todo o território brasileiro, principalmente no cerrado. Análises químicas têm demonstrado seu potencial em auxiliar na resolução de problemas em vários níveis taxonômicos com base em grupos químicos de metabólitos secundários. Além disso, análises fitoquímicas tem revelado que determinadas substâncias de Aldama possuem potencial para serem estudadas tendo vista a investigação de mecanismos moleculares anti-inflamatórios. Desta forma, foi proposto neste trabalho realizar a abordagem da metabolômica não-direcionada auxiliada por quimiometria, visando-se fornecer dados químicos tanto para contribuições quimiotaxonômicas quanto para encontrar substâncias bioativas. Por meio de tal abordagem, análises multivariadas não supervisionadas (PCA e HCA), bem como supervisionadas (OPLS-DA), com dados provenientes de UHPLC-DAD-Orbitrap, demonstraram que o gênero Aldama é quimioconsistente e, deste modo, determinadas substâncias químicas discriminantes foram sugeridas. Além disso, espécies que apresentam problemas taxonômicos tiveram os seus posicionamentos infragenéricos explicado do ponto de vista quimiotaxonômico. Quanto às análises para encontrar substâncias bioativas (fitoquímica direcionada), os alvos inflamatórios pesquisados foram as enzimas pró-inflamatórias COX-1 e 5-LOX, com as quais se realizou ensaios in vitro verificando-se a atividade positiva de extratos de espécies de Aldama. A. trichophylla foi a espécie selecionada para realizar a fitoquímica direcionada, uma vez que apresentou inibição dupla contra as enzimas e por se tratar de uma espécie muito pouco estudada do ponto de vista fitoquímico. Os melhores modelos de inibição de COX-1 e 5-LOX criados com dados químicos de Aldama foram selecionados a partir de diversas combinações de algoritmos, softwares de processamento (MZmine, MetAlign e MSClust) e técnicas-hifenadas, tais como UHPLC-Orbitrap e HPLC-TOF. Deste modo, determinados picos de íons foram apontados pela quimiometria como sendo discriminantes para a atividade de inibição dupla. Tais picos foram desreplicados com base em seus perfis de UV e padrões de fragmentação via HCD-Orbitrap e Ion-Trap. Deste modo, foi possível desreplicar três prováveis substâncias químicas inibidoras de COX-1 e 5-LOX: kaempferol-3-O-glucuronideo, quercetina-3-O-metil-7-glucuronideo e kaempferol-3-O-(6\"-malonil-glucosideo). Por fim, foi realizada a fitoquímica direcionada. O fracionamento cromatográfico permitiu isolar tais substâncias e uma análise preliminar de RMN 1H foi realizada com o intuito de realizar a identificação estrutural. Tais substâncias terão a suas atividades confirmadas na inibição das duas enzimas em um futuro próximo. / The species of the genus Viguiera Kunth, recently transferred to the genus Aldama La Llave (Asteraceae, Heliantheae), still show problems of taxonomic boundaries. This genus has 35 species distributed throughout the Brazilian territory, mostly in the cerrado. Chemical analyses has demonstrated its potential to help in the solution of problems at various taxonomic levels based on chemical groups of secondary metabolites. Furthermore, phytochemical analyses have shown that certain compounds from Aldama have the potential to be studied aiming to investigate anti-inflammatory molecular mechanisms. Thus, in this work it was proposed to carry out the untargeted metabolomic approach aided by chemometrics, aiming to provide chemical data either for chemotaxonomic contributions and to find bioactive compounds. Through this approach, both unsupervised (PCA and HCA) and supervised multivariate analysis (OPLS-DA) combined with data from UHPLC-DAD-Orbitrap analyses showed that the genus Aldama is \"chemoconsistent\" and thus certain discriminant compounds were suggested. Additionally, some species with taxonomic problems had their infrageneric positioning explained from the chemotaxonomic point of view of. With regards to the analyses carried out to find bioactive compounds (targeted phytochemistry), the inflammatory targets investigated in this study were the COX-1 and LOX-5 pro-inflammatory enzymes with which in vitro assays were made and positive activity of extracts from species of Aldama was observed. A. trichophylla was the selected species for the targeted phytochemistry because it showed dual inhibition activity against both enzymes and it is still little investigated from the chemical point of view. The best models for inhibition of COX-1 and 5-LOX obtained with chemical data of Aldama were selected by means of various combinations of algorithms, processing software (MZmine, MetAlign and MSClust) and hyphenated techniques, such as HPLC-TOF and UHPLC-Orbitrap. Thus, certain peak ions were appointed by chemometrics as discriminant for the dual inhibition activity. These peaks were dereplicated based on their UV profiles and fragmentation patterns via HCD-Orbitrap and Ion-Trap. Thus, it was possible to dereplicate three probable chemical inhibitors of COX-1 and 5-LOX: kaempferol-3-O-glucuronide, quercetin-3-O-methyl-7-glucuronide and kaempferol-3-O-(6\"-malonyl-glucoside). Finally, the targeted phytochemistry was carried out. The chromatographic fractionation allowed us to isolate these three compounds and a preliminary 1H NMR analysis was performed in order to conclude their structural identification. In the near future, these substances will have their activity evaluated by the inhibition of the two enzymes.

Made available in DSpace on 2016-04-26T20:48:50Z (GMT). No. of bitstreams: 1 Andre Gal Mountian.pdf: 404598 bytes, checksum: 6a3572319e1f94ff0a3766a923762bee (MD5) Previous issue date: 2008-05-07 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aim of this work is to evaluate the theoretical foundations of the World Bank activities in relation to poverty, especially in the 90s decade. It aims to examine the extent in which the concept of poverty of the Bank and its strategies of poverty reduction were based in the liberal theoretical matrix, pointing out the limitations of these strategies. The research is inscribed in the field of Political Economy, examining the institution and its contradictions, in order to understand the theoretical and historical backgrounds of its conceptions. The methodology employed is the crtitical analysis of the studies of the Bank, especially of the World Development Report. The first conclusion of this study is that there is a strong identification between the general conceptions of the World Bank and Liberal Economy. From the preponderance of the market as the best distributor of the resources in the economy and the trust that this mechanism can generate benefits to the whole society, including the lower classes, the concept of the poor is constructed as that individual that does not achieve basic resources of survival via market. For this group of individuals, the solution would be then the development of social policies focused on their needs, while at the same time that the orientation of the economy to the market is intensified. The fist limitation that was verified of these strategies of reducing poverty of the World Bark is that the kernel of its concern is in relation to extreme poverty, a specific category of poor, that one that does not have the basic resources for survival. There are not, within the Bank proposal, an emphasis to diminish social inequality, unless when affecting the growth and efficiency of economy. Another dimension of these limitations refers to the subsidiary role of reducing poverty in relation to the central objective of the Bank: economic growth. Poverty reduction has to be compatible with a model of development of the Bank, contributing to its intensification. Furthermore, the social policies focused on poverty, together with the priority of arguments of economic efficiency around notions of universal social rights, come to empty and weaken the concept of citizenship / O objetivo deste trabalho é realizar uma avaliação das bases teóricas da atuação do Banco Mundial com relação à pobreza, especialmente a partir da década de 1990. Pretende-se verificar até que ponto o conceito de pobreza do Banco e suas estratégias de redução se basearam na matriz teórica liberal, e identificar as limitações dessas estratégias. A pesquisa se insere no campo da Economia Política, tratando a instituição sob o ponto de vista de suas contradições, buscando compreender as raízes teóricas e históricas de suas concepções. A metodologia utilizada é a análise crítica de estudos do Banco, especialmente o Relatório sobre o Desenvolvimento Mundial, assim como em bibliografia secundária. A primeira conclusão deste trabalho é que existe uma forte identificação entre as concepções gerais do Banco Mundial e o Liberalismo Econômico. A partir da aceitação do mercado como melhor alocador de recursos na economia e da confiança que esse mecanismo pode gerar benefícios a toda sociedade, inclusive às camadas mais baixas, o conceito de pobre do Banco Mundial é construído como aquele indivíduo que não consegue condições mínimas de sobrevivência, via mercado. Para esse grupo de indivíduos, o melhor remédio seria o desenvolvimento de políticas sociais focalizadas em suas necessidades, ao mesmo tempo em que a orientação da economia para o mercado é intensificada. A primeira limitação verificada das estratégias de combate à pobreza do Banco Mundial é que a essência de sua preocupação é com a pobreza extrema, uma categoria específica de pobre, aquela que não tem condições mínimas de sobrevivência. Não há, nas propostas do Banco, uma ênfase na diminuição das desigualdades sociais, a menos quando prejudicam o crescimento e a eficiência da economia. Outra dimensão de limitações diz respeito ao papel subsidiário do combate à pobreza com relação ao objetivo central do Banco: o crescimento econômico. O combate à pobreza deve ser compatível com o modelo de crescimento do Banco, contribuindo ainda para sua intensificação. Ainda, as políticas sociais focalizadas na pobreza, juntamente com a prioridade de argumentos de eficiência econômica sob noções de direitos sociais universais, acabam por esvaziar e enfraquecer o conceito de cidadania

Submitted by JOSIANE SANTOS DE OLIVEIRA (josianeso) on 2017-06-08T12:06:15Z No. of bitstreams: 1 Emanuelle Nava Smaniotto_.pdf: 445452 bytes, checksum: 5741b34057dbeb9c0f08c71040f4979b (MD5) / Made available in DSpace on 2017-06-08T12:06:15Z (GMT). No. of bitstreams: 1 Emanuelle Nava Smaniotto_.pdf: 445452 bytes, checksum: 5741b34057dbeb9c0f08c71040f4979b (MD5) Previous issue date: 2017-02-24 / Nenhuma / Este trabalho apresenta uma análise sobre o impacto da oferta de crédito no investimento privado no Brasil, entre 2001 e 2016. Para isso, é introduzida a metodologia de Mudança de Regimes Markovianos (MR-DR), de modo a capturar os diferentes ciclos existentes entre Crédito Livre, Crédito Direcionado e Investimento Privado. Ademais, é aplicado o modelo de Vetores Autorregressivos (VAR/VEC), com o intuito de analisar as causalidades e funções impulso-resposta. É possível comprovar o impacto positivo das séries de crédito sobre o investimento privado. Demonstra-se, também, a importância da concessão de crédito direcionado para a sustentação dos níveis de investimento privado nos períodos de instabilidade econômica. Desta forma, consolida-se junto à literatura mais um indício da importância de políticas macroeconômicas em períodos de instabilidade, com exemplo ao período pós crise de 2009, mediante intensificação de Recursos Direcionados, de modo a manter os níveis de Formação Bruta de Máquinas e Equipamentos no país. / This paper shows an analysis about the credit supply impact on private investment in Brazil, between 2001 and 2016. For this purpose, the Markovian Regime Change (MR-DR) methodology is introduce in order to capture the different cycles between Free Credit, Directed Credit and Private Investment. In addition, the Autoregressive Vectors (VAR / VEC) model is apply to analyze the causalities and impulse-response functions. It was possible to prove the positive impact of the credit series on private investment. Thus, is demonstrate the importance of directed credit concession for sustaining private investment levels in periods of economic instability. On another hand, an indication of the importance of macroeconomic policies in periods of instability, with an example to the post-crisis period of 2009, has been consolidated along with the literature, through intensification of Directed Resources, in order to sustain the levels aboutPrivate Investment (FBME) in country.

Les mastocytes (MCs) sont issus des cellules hématopoïétiques multipotentes non engagées, CD34+, et jouent un rôle important dans l’initiation de la réponse immunitaire innée et adaptative, ainsi que dans les réactions allergiques IgE-dépendantes. Les mastocytoses sont des néoplasies myéloïdes caractérisées par une accumulation anormale et l'activation fréquente de mastocytes dans divers organes. Les organes généralement atteints sont la moelle osseuse, la peau, le foie et le tractus gastro-intestinal. Elles sont caractérisées dans l’immense majorité des cas par la présence de mutations acquises de la structure du récepteur KIT (plus particulièrement KIT D816V) qui induisent l’activation constitutive de ce récepteur à activité tyrosine kinase intrinsèque. Le traitement actuel de ces pathologies est décevant car la mutation KIT D816V résiste à la plupart des inhibiteurs de tyrosine kinases (ITKs).Dès le début de ma thèse, nous avons pu établir, à partir de cellules souches hématopoïétiques de sang de cordon humain normal cultivées à long terme en présence de stem cell factor (SCF), une nouvelle lignée mastocytaire humaine stable, dénommée ROSA KIT WT, restant strictement dépendante pour sa survie et sa prolifération du SCF, exprimant le récepteur de haute affinité aux IgE (FcεRI), et présentant un récepteur KIT de structure normale. Cette lignée, facile à cultiver en grandes quantités, permet d’envisager l’étude approfondie des évènements intracellulaires menant à l’activation mastocytaire IgE-dépendante et la mise au point de tests de criblage à haut débit dans le domaine de la thérapeutique anti-allergique et/ou anti-inflammatoire.Par ailleurs, afin de pouvoir étudier le rôle transformant des mutants de KIT retrouvés au cours des mastocytoses, nous avons transfecté les cellules ROSA KIT WT par des vecteurs lentiviraux apportant une construction codant pour le KIT muté en D816V ou le KIT muté Delta 417-419 insY. Nous avons ainsi obtenu deux nouvelles lignées mastocytaires humaines SCF-indépendantes, ROSA KIT D816V et ROSA KIT Delta 417-419 insY, pour lesquelles nous avons montré qu’il existe une activation constitutive de KIT, mais aussi de STAT5 et d’AKT. Ces deux lignées de pourront être utilisées soit pour étudier l’impact des mutations de KIT sur la signalisation intracellulaire, soit pour le criblage molécules à activité antiproliférative potentielle dirigées soit contre KIT muté, soit contre l'une ou l’autre des molécules intracellulaires impliquées dans la transduction du signal KIT muté / Mast cells are cells with ubiquitous tissue distribution, derived from CD34 + multipotent hematopoietic cells. These cells play a fundamental role in the initiation of innate and adaptive immune response and in IgE-dependent allergic reactions or in various inflammatory reactions.Mastocytosis is defined as a myeloproliferative neoplastic disorder, caused by an abnormal accumulation of mast cells in one or more organ systems. Mastocytosis presents in cutaneous and systemic forms. In patients with systemic mastocytosis, the most frequent point mutation is KIT D816V, whereas in pediatric patients, where the disease is usually restricted to the skin, different KIT defects have been detected, mostly in the extracellular portion of KIT.In a primary culture of mast cells made from precursors of one cord blood, we have successfully isolated a new human mast cell line called ROSA KIT WT with a phenotype and reactivity comparable to those of normal mast cells. This cell line is dependent on SCF for growth and expresses the KIT receptor wild. It is easy to grow in large quantities, to freeze and to activate by IgE-anti IgE couple or a couple of allergen and corresponding specific IgE. This cell line can be used to study pathophysiologic mechanisms of allergy and to develop and use high-throughput screening tests of molecules in search of anti-allergic properties.In addition, we transfected these cells by lentiviral vectors providing constructs encoding the mutated KIT D816V or the mutated KIT Delta 417-419 insY, two KIT abnormalities encountered in the course of mastocytosis. This allowed us to establish two new cell lines independent of SCF for proliferation, ROSA KIT D816V and ROSA KIT Delta417-419 insY, which are particularly easy to grow in large quantities, and whose phenotype is similar to that of abnormal mast cells during mastocytosis. These two cell lines can be used for pathophysiologic studies on mastocytosis and for high throughput screening of molecules in search of antiproliferative effects specifically directed against the mutated KIT or against one or other of the intracellular molecules involved in signal transduction induced by mutant KIT.

Os tumores de musculatura liso do útero se desenvolvem a partir do miométrio e podem apresentar carcterísticas clínicas malignas e benignas. Dentre eles, o leiomiossarcoma (LMS) é o tumor maligno mais comum, com altas taxas de metástase e recidiva, mesmo sendo diagnosticado em estágios iniciais. Já os leiomiomas (LM) são os tumores benignos mais frequentes em mulheres em idade reprodutiva. Ambos possuem mesma diferenciação celular, porém com comportamentos clínico e biológico bastante distintos, e até o momento não se dispõe de tratamento específico ou curativo. Nesse contexto, a busca por novos alvos moleculares pode contribuir não só para um melhor entendimento dessas neoplasias, como também para a descoberta de novas terapias. Em estudo prévio foi observada a expressão aumentada de FOXO3a nos sarcomas uterinos, em comparação aos LMs e ao miométrio adjacente (MM). Além disso, sua expressão foi crescente de acordo com o potencial de malignidade do tumor. Assim, o objetivo do presente estudo foi avaliar in vitro o efeito de terapia alvo específica para FOXO3a em células de LM e LMS. Para isto, linhagens celulares de MM (ATCC PCS-460-011), LM (THESCs - CRL-4003) e LMS (SK-UT-1 - HTB-114) foram caracterizadas quanto à expressão basal de FOXO3a (gene e proteína) e submetidas a tratamento com Genisteína e Metformina ou inativação do gene por siRNA. Os efeitos dos tratamentos foram avaliados por PCR em tempo real, Western Blot, imunocitoquímica, ensaios de proliferação, migração e apoptose. Nossos resultados mostraram que todos os tratamentos realizados interferiram na capacidade de proliferação e migração das células, com maior inibição após as 48 horas nos LMS e 72h nos LM. O efeito obtido na transfecção com siRNA apresentou maior eficiência após 48 h da transfecção nos LMS e 72h nos LM. Os efeitos da inibição de FOXO3a foram maiores na proliferação e migração dos LM, porém os resultados não foram estatisticamente significativos. Dentre as substâncias testadas, a Metformina apresentou maior efeito sobre a proliferação, migração e viabilidade das linhagens celulares. A Genisteína também apresentou efeito inibitório nas células, porém o controle com veículo também apresentou o mesmo efeito citotóxico. De modo geral, os efeitos obtidos com os fármacos, foram tempo e concentração dependentes. Em conjunto, nossos resultados sugerem um relevante do FOXO3a nos tumores de musculatura lisa uterinos, além de apresentá-lo como potencial alvo para terapia específica / Smooth muscle tumors of the uterus develop from the myometrium and may present benign and malignant clinical features. Among them, leiomyosarcoma (LMS) is the most frequent malignant tumor, with high rates of metastasis and relapse, even when diagnosed in early stages. On the other hand, leiomyomas (LM) are the most frequent benign tumors in women of reproductive age. Both have the same cellular differentiation, but with very different clinical and biological behaviors, and so far no specific or curative treatment is available. In this context, the search for new molecular targets can contribute not only for a better understanding of these neoplasms, but also for the discovery of new therapies. In a previous study, increased expression of FOXO3a in uterine sarcomas was observed, compared to LMs and adjacent myometrium (MM). In addition, its expression was increasing according to the malignancy potential of the tumor. Thus, the aim of the present study was to evaluate in vitro, the effect of specific targeted therapy for FOXO3a on LM and LMS cells. For this, MM (ATCC PCS-460-011), LM (THESCs-CRL-4003) and LMS (SK-UT-1-HTB-114) cell lines were characterized for basal expression of FOXO3a (gene and protein) and subsequently submitted to treatment with metformin and genistein, or silencing of FOXO3a by siRNA. The effects of the treatments were evaluated by real-time PCR, Western Blot, immunocytochemistry, proliferation, migration and apoptosis assays. Our results showed that all treatments interfered in the proliferation and migration capacity of the cells, with greater inhibition after 48 hours for LMS and 72 hours LM. The effect obtained in the transfection with siRNA showed higher efficiency after 48 hours of transfection in LMS and 72 hours in LM. The effects of inhibition of FOXO3a were greater in the proliferation and migration of the LM, but the results were not statistically significant. Among the substances tested, Metformin had a greater effect on proliferation, migration and viability of the cell lines. Genistein also had an inhibitory effect on the cells, but the control with the vehicle also presented the same cytotoxic effect. In general, the effects obtained with the drugs were time and concentration dependent. Together, our results suggest a relevant role of FOXO3a in uterine smooth muscle tumors, in addition to presenting it as a potential target for specific therapy

O sarcoma de Ewing (SE) é um dos mais agressivos tipos de câncer pediátrico. Apesar dos significativos avanços no tratamento dessa doença, ainda há uma grande necessidade no aumento das taxas de cura, redução da toxicidade quimioterápica e redução da resistência ao tratamento. Tem sido proposto que SE provém de precursores neuronais, podendo ter sua fisiologia afetada, pois, por neurotrofinas (NTs). Examinamos a influência de receptores de NTs (Trks) em SE. Foram avaliadas a expressão proteica de NTs (NGF e BDNF) e seus receptores (TrkA e TrkB, respectivamente) em amostras de tumores de pacientes com SE, e a expressão de mRNA nas linhagens celulares RD-ES e SK-ES-1. O tratamento das linhagens com o pan-inibidor de Trks (K252a) modificou a morfologia celular e diminuiu a expressão de mRNA de NGF, TrkA, BDNF e TrkB. Ainda, a inibição de Trks diminuiu drasticamente a proliferação e capacidade clonogênica celular. Efeitos sinérgicos foram observados quando as células foram tratadas em conjunto com baixas doses de quimioterápicos, tanto em células selvagens de SE, quanto nas quais induzimos quimiorresistência. Esse estudo sugere, pela primeira vez, que a inibição de Trks reduz a proliferação e sobrevivência celular em SE, além de aumentar a sensibilidade ao tratamento quimioterápico. / Ewing's sarcoma (ES) is one of the most aggressive types of pediatric cancer. Despite significant advances in the treatment of this disease, there is still a great need in increasing cure rates, reducing chemotherapy toxicity and treatment resistance. It has been proposed that ES might derive from neuronal precursors and may be influenced, therefore, by neurotrophins (NTs). We have examined the influence of Trk neurotrophin receptors in ES. Protein expression of NTs (NGF and BDNF) and their receptors (TrkA, and TrkB, respectively) was detected in tumor samples from patients with ES, and mRNA expression was analyzed in the RD-ES, SK-ES-1 cell lines. Treating cells with a Trk Pan-inhibitor (K252a) altered cell morphology and decreased the mRNA expression of NGF, TrkA, BDNF, and TrkB. In addition, Trk inhibition dramatically decreased cell proliferation and clonogenic capacity. Synergistic effects were observed when cells were treated in combination with low doses of cytotoxic chemotherapeutics, both in normal ES cells and cells in which chemoresistance was induced. The results suggest for the first time that Trk inhibition can reduce the proliferation and survival of ES cells and sensitize them to cytotoxic chemotherapy.

Les lymphomes anaplasiques à grandes cellules (LAGC) sont des lymphomes non-hodgkiniens dits de type T ou nul, représentant la majorité des lymphomes T pédiatriques (20 à 30%). Dans plus de 80% des cas, une translocation chromosomique réciproque aboutissant à l'expression anormale de protéines chimères de type X-ALK qui arborent constitutivement et de manière anormale l'activité tyrosine-kinase ALK (Anaplastic Lymphoma Kinase) est le moteur de la tumorigenèse (LAGC dits "ALK+ "). L'une des particularités de ces lymphomes, mise en évidence par mon équipe, est le fait que B-Cell Lymphoma-2 (BCL-2) demeure indétectable dans les cas ALK+ contrairement aux cas ALK-. Ce point est d'autant plus surprenant que BCL-2, oncogène largement établi comme prototype de protéines anti-apoptotiques ainsi que régulateur clé de l'autophagie, est fortement surexprimé dans la majorité des lymphomes. A l'inverse, Human Antigen R (HuR) est surexprimée dans les LAGC (comme dans la plupart des cancers). Il a été démontré que cette protéine de liaison aux ARN participait au maintien du phénotype tumoral, et que sa localisation subcellulaire et ses fonctions dépendaient étroitement de son statut de phosphorylation, lequel est régulé par ALK dans les LAGC ALK+. Au niveau du cytoplasme, HuR permet de stabiliser et d'augmenter la traduction d'ARNm possédant, dans leur région 3'-UTR, des séquences riches en adénine et uridine (AU-rich elements, "ARE"). De manière plus générale, HuR a la capacité de dialoguer avec les microARNs (miARN), soit en empêchant leur action par compétition, soit à l'inverse en coopérant avec ces derniers et en promouvant ainsi leur fonction de régulateur négatif sur certains transcrits cibles communs. Le transcrit Bcl-2, dont l'expression est réprimée dans les LAGC ALK+, fait partie des cibles potentielles de HuR. Au cours de ma thèse, j'ai ainsi cherché à comprendre les mécanismes moléculaires mis en jeu dans la répression de l'expression de Bcl-2, en me focalisant sur le rôle de HuR et de miARN "partenaires" dans ce processus. Mes données semblent indiquer que ce mécanisme implique le recrutement par HuR du miR-34a sur l'ARNm Bcl-2, conduisant à la mise en silence de ce dernier. A l'inverse, quand l'activité tyrosine- kinase de ALK est inhibée, l'interaction entre HuR et le transcrit Bcl-2 diminue, ce qui limite le recrutement de miR-34a et conduit à une restauration de l'expression de cette oncogène majeur dans les cellules lymphomateuses. Dans le contexte des essais cliniques d'inhibiteurs ciblant l'activité tyrosine kinase de ALK tels que le Crizotinib, la question de cette ré-expression de BCL-2 éclaire d'une lumière nouvelle certains échecs thérapeutiques subis par cette molécule pourtant prometteuse. Je me suis donc également consacré, pendant ma thèse, à l'étude des conséquences de cette ré-expression de BCL-2 sur les LAGC ALK+ traités par le Crizotinib. Les résultats que j'ai obtenus in vitro et in vivo montrent que contrecarrer, par interférence à l'ARN, l'élévation du taux de BCL-2 consécutive au traitement par le Crizotinib, permet de potentialiser les effets de la drogue : cela se traduit en particulier par une potentialisation de la mort par apoptose induite par le traitement mais aussi, de manière fascinante, par une conversion de la réponse autophagique initialement cytoprotectrice et pro-tumorale en une autophagie incontrôlée et délétère, qui participe alors à l'effet thérapeutique accru de la drogue. De manière globale, ce travail permet d'envisager de nouvelles combinaisons et alternatives thérapeutiques pour les patients souffrants de LAGC ALK+, et illustre la complexité des régulations croisées entre processus apoptotiques et autophagiques. / Anaplastic large cell lymphoma (ALCL) are T/-null non-hodgkin lymphoma representing most of childhood T-cell lymphoma (up to 30%). More than 80% of cases bear reciprocal chromosomic translocation responsible for abnormal expression and constitutive activation of X-ALK type (Anaplastic Lymphoma Kinase) chimeric proteins (ALK+ ALCL). A striking characteristic of this lymphoma is that B-Cell Lymphoma-2 (BCL-2) remains undetectable in ALK+ cases compared to ALK- cases. This is all the more surprising as the BCL-2 oncogene, which is firmly established as a prototypic anti-apoptotic factor as well as a key autophagy regulator, has been shown to be overexpressed in a majority of lymphomas. On the other hand, the RNA-binding protein HuR (Human Antigen R) is overexpressed in ALCL (as in most cancers). It has been demonstrated that this protein was involved in the sustainability of the tumoral phenotype, and that its subcellular localization and functions were closely related to its phosphorylation status, which in turn heavily depends on ALK activity in ALK+ ALCL. In the cytoplasm, HuR has the ability to bind adenine and uridine-rich elements (ARE) located on the 3'-UTR of target mRNAs, and both protect them from degradation and increase their translation. From a general point of view, HuR is able to establish an interplay with microRNAs (miRNAs), either blocking them through competition, or actually cooperating with them and thus promote their function of negative regulators of gene expression on common target transcripts. The BCL-2 transcript, which expression seems to be silenced in ALK-expressing ALCL, has been described as a potential target of HuR. During my PhD work, I dedicated myself to understand the molecular mechanism at work in the silencing of BCL-2 expression with a focus on HuR and collaborating miRNA. The data I obtained point at a cooperation between HuR and miR-34a leading to the silencing of the BCL-2 transcript. However, when the ALK tyrosine kinase activity is inhibited, it appears the interaction between the BCL-2 mRNA diminishes, which limitates the miR-34a 's access to this transcript and ultimately results in a re-expression of the BCL-2 oncogene in these lymphoma cells. In the current context of clinical trials for ALK-targeting inhibitors, such as the Crizotinib, this BCL-2 re-expression observed upon ALK inhibition shed light on potential reasons behind some therapeutic failures that have recently been reported. Indeed, during my PhD work, I also studied the consequences of the BCL-2 re-expression observed in Crizotinib-treated cells. The data I obtained in vitro and in vivo show that, by blocking this re-expression using RNA interference, the Crizotinib anti-tumoral efficiency can be greatly potentiated. This potentiation took the form of an increase of apoptotic cell death induction and, interestingly, also affected the autophagic response triggered by the drug, making it switch from a cytoprotective- type, protumoral autophagic flux to an enhanced, deletary-type and tumor suppressive flux, adding to the therapeutic effect of the drug. This work in general provides insights for new therapeutic combinations that could potentially benefit to ALK+ ALCL patients, and illustrates the complex cross-regulations between apoptotic and autophagic pathway.

Les méthodes de contrôle de vibrations passives basées sur des absorbeurslinéaires ont été largement étudiées et ils ont aujourd'hui une vaste gammed'applications. Cependant, les absorbeurs linéaires n’étant efficaces quelorsqu'ils sont accordés à la fréquence que l'on veut contrôler, ils présententdes limites considérables quand ils sont appliqués à des systèmes possédantdes incertitudes sur les paramètres modaux ou ayant une fréquence propredépendante de la force extérieure. Dans cette thèse la réduction des vibrations dans les systèmes mécaniques à l'aide d'un absorbeur Nonlinear Energy Sink est étudiée. Le phénomène qui gouverne la physique de ce dispositif est appelé pompage énergétique (Targeted Energy Transfer) et il consiste en un transfert irréversible d'énergie du système principal vers le NES, où l’énergie est dissipée. Ce transfert d'énergie peut se produire pour une large gamme de fréquences et sans besoin que le NES ne soit accordé _a une fréquence spécifique.La dynamique d'un premier type de NES appelé Vibro-Impact NonlinearEnergy Sink (VI-NES) est investiguée expérimentalement grâce à unoscillateur linéaire (OL) à un degré de liberté forcé harmoniquement auquelle VI-NES est attaché. Le pompage énergétique du OL vers le VI-NESest observé expérimentalement, ce qui a permis d'obtenir une importanteréduction du pic de résonance du système principal. Le système est étudiéanalytiquement à l'aide de la méthode Multi-Echelles et le comportementnon-linéaire observé est expliqué théoriquement. Le deuxième type de NES présenté est le Magnetic-Strung NES avec récupération d'énergie. Cette étude ajoute l'aspect lié à la récupération d'énergie au domaine de recherche des absorbeurs non-linéaires. Le système consiste en un oscillateur linéaire (OL) à un degré de liberté forcé harmoniquement auquel le MS-NES est appliqué. La force non-linéaire de rappel peut être modulée grâce à une force magnétique introduite judicieusement, ce qui permet au NES d'avoir plusieurs configurations possibles. Lesystème résultant est un système électromécanique où l'énergie vibratoire dusystème principal est absorbée par le NES et est ensuite dissipée en partiepar l'amortissement visqueux et convertie en partie en puissance électrique.Les études numérique et expérimentale analysent les performances du MSNESen tant qu'absorbeur d'énergie et en tant que récupérateur d'énergie.Finalement, les idées et les perspectives issues de cette étude sont traitéeset les directions pour les travaux futurs sont fournies. / Passive vibration control methods using linear dampers have been largelystudied and investigated, and they have nowadays a broad range of applications.However, linear dampers are efficient when tuned to the specificfrequency to control but present substantial limitations when applied to primarysystems with uncertainties on the modal parameters or to systemshaving a natural frequency that may vary with external forcing.In this thesis the vibration mitigation in mechanical systems by meansof a Nonlinear Energy Sink absorber is studied. The phenomenon governingthe physics of this kind of device is referred to as Targeted Energy Transferand it consists in an irreversible energy transfer from the primary systemto the NES where the energy is then dissipated. This energy transfer mayoccur over a broad range of frequencies with no need for the NES to betuned to a specific one.The dynamics of a first type of NES called Vibro-Impact Nonlinear EnergySink (VI-NES) is experimentally investigated via a harmonically forcedsingle-degree-of-freedom linear oscillator to which a VI-NES is attached. ATargeted Energy Transfer from the LO towards the VI-NES is experimentallyobserved and a significant reduction of the primary system's resonancepeak is obtained. The system is analytically studied by means of the MultipleScales method and the nonlinear behavior experimentally observed istheoretically explained. The second type of NES presented is the Magnetic-Strung NES withenergy harvesting. This study adds the energy harvesting aspect to the researchon nonlinear vibration absorbers. The system consists in a harmonicallyforced single-degree-of-freedom linear oscillator to which the MS-NESis applied. The type of nonlinearity used can be shaped thanks to a magneticforce aptly introduced, allowing the NES to have several possible configurations.The resulting system is an electro-mechanical system in which thevibration energy of the primary system is absorbed by the NES and subsequentlypartially dissipated by the viscous damping and partially convertedinto electrical power. The numerical and experimental studies analyze theperformances of the MS-NES both as an energy absorber and as an energyharvester.

Le travail présenté dans cette thèse est dédié à l’étude d’un absorbeur bistable continu basé sur le principe du "Nonlinear Energy Sink" (NES) et son utilisation pour l’atténuation des vibrations d’un système mécanique à plusieurs degrés de liberté sous excitation acoustique. Le modèle analytique du comportement linéaire de l’absorbeur ainsi que le modèle numérique complet ont été présentés, analysés et validés par des séries d’expériences. Le complexité du transfert énergétique ciblé ("Targeted Energy Transfer" ou TET) entre l'absorbeur et le système primaire à contrôler n’a pas permis une description analytique simple. Nous avons donc choisi de concentrer cette étude sur l’exploration expérimentale et numérique de l’absorbeur couplé à des systèmes mécaniques sous excitations harmonique et aléatoire ainsi que sur l’identification des mécanismes de transfert d’énergie. Le système couplé a montré une dynamique très riche du fait de différents régimes de TET qui ont été décrits dans la littérature pour d’autres types de NES. Ce projet a été financé par Saint-Gobain. L’absorbeur a été adapté pour l’application prévue par la direction industrielle de la thèse: contrôle des vibrations de la double paroi sous excitation acoustique afin d’améliorer l’isolation acoustique fournie par le système.Les connaissances qualitatives sur la dynamique de l’absorbeur obtenues à partir des résultats expérimentaux et numériques, ainsi que l’analogie avec les autres types de NES, ont permis la création d’un absorbeur qui répond à la problématique posée. Les moyens pour l’optimisation et le développement de l’absorbeur ont été identifiés et les simulations préliminaires ont été fournies. / The work presented in this thesis is dedicated to the study of a continuous bistable absorber based on the principle of Nonlinear Energy Sink (NES) and its use for the vibration mitigation of a many-degree-offreedom mechanical systems under acoustic excitation. The analytical model of the linear behavior of the absorber and its complete numerical model were presented, analyzed and validated by series of experiments. The complexity of the Targeted Energy Transfer (TET) between the absorber and the primary system did not allow a simple analytical description. We have chosen to concentrate this study on the experimental and numerical exploration of the absorber coupled to mechanical systems under harmonic and random excitations, as well as on the identification of the mechanisms of energy transfer. The coupled system have shown very rich dynamics as it possessed different regimes of TET, which were earlier described in literature for other types of NES. This project was funded by Saint-Gobain. The absorber was adapted for the application foreseen by the industrial supervisors of the PhD: the vibration control of partitioning double walls under acoustic excitation so that to improve the acoustic isolation provided by the system. The qualitative knowledge on the absorber dynamics obtained from the experimental and numerical results, as well as the analogy with the other types of NES, permitted the creation of an absorber which corresponds to the problematic. The ways for the further optimization and development of the absorber were identified and preliminary simulations were provided.

Lanthanide phosphate (LnPO4) and lanthanide vanadate (LnVO4) nanoparticles (NPs) are promising platforms for theranostic applications because of their chemical stability, low solubility, low toxicity, and unique luminescence and magnetic properties. Motivated by the high radiation resistance and ability to host actinides of naturally occurring lanthanide-based compounds, LnPO4 and LnVO4 NPs were studied as radionuclide carriers for targeted radionuclide therapy using in vivoα-generators, 223Ra, 225Ac, and 227Th. The implementation of these radionuclides has shown potential for the treatment of micrometastases and solid tumors as well as challenges in the retention of decay daughters at the target site to minimize unwanted radiotoxicity. LnPO4 and LnVO4 core-shell NPs doped with either 156Eu, a “cocktail” of 85, 89Sr and 156Eu, or in vivo α-generators 223Ra, 225Ac, and 227Th were synthesized in aqueous media. In vitro retention of radionuclides was investigated by dialyzing the radionuclide-doped NPs suspensions against deionized water and quantifying the activity in dialysate aliquots over time. The crystal structure, morphology, physical stability, luminescence and magnetic properties were evaluated. Partial retention of 156Eu (~70–95%) and 85, 89Sr (>80%) was evidenced in LnPO4 core NPs, while 227Th and decay daughters were quantitatively retained in LaPO4 core + 2 shells NPs (>99%). Gd0.8Eu0.2VO4 and GdVO4 core-shell NPs showed partial retention of 223Ra (~75%), 225Ac (75–95%), 227Th (>96%), and decay daughters. Radionuclide retention was influenced by the lanthanide concentration, crystal structure, and number of shells. The partial retention of radionuclides in both LnPO4 and LnVO4 core-shell NPs may enhance the treatment efficacy while minimizing unwanted toxicity. LnVO4 core and core-shell NPs have potential as carriers of short-lived radionuclides for both diagnostic and therapeutic applications. Emission intensities were higher for LnVO4 with respect to LnPO4 NPs, whereas no significant difference was observed in the magnetic susceptibilities. GdVO4 core NPs displayed enhancement of the signal intensity in T1-weighted images. This work evidences the promising application of both LnPO4 and LnVO4 NPs as platforms for targeted radionuclide therapy and multimodal molecular imaging.