Cancer is a worldwide health problem and despite advances in traditional treatments i.e. surgery, chemotherapy and radiotherapy, the cure rate remains disappointing for some cancers. Different novel therapeutic strategies are being developed. In this thesis two nontraditional cancer therapy approaches are studied: gene therapy using viral vectors and antitumour vaccination with dendritic cell - tumour cell (DC/TC) hybrids. We have developed a novel ELISPOT titration method for viral vectors that is based on the actual expression of the transgene in target cells. This method was developed with recombinant parvovirus MVM-IL2, but it should be adaptable for other vectors carrying expression cassettes for secreted transgene products for which antibodies are available. The ELISPOT titration method allows for faster and better quantification of transducing units present in vector stocks as opposed to titration by in situ hybridisation (annexe I). The MVMIL2 vector has shown an anti-tumour effect against melanoma in an immunocompetent mouse model (annexe IV). Previous work concerns photodynamic inactivation of adenoviral vectors for biosafety and an in vivo study in which a synergistic effect of antiangiogenesis gene therapy combined with radiotherapy could be shown (annexes V and VI). DC/TC hybrids have been proposed as cancer vaccines for their simultaneous expression of antigen presentation machinery and tumour associated antigens. Hybrids are classically generated by polyethylene glycol (PEG) or electrofusion. These methods however require special skills and equipment and cause rather high cell lethality. Fusion via the expression of viral fusogenic membrane glycoproteins (FMG), such as the vesicular stomatitis virus-G (VSV-G) (annexe III) or the Gibbon ape Leukemia Virus (GaLV) FMG, have recently been described. We have mainly focussed on the latter. Transduction of cells with GaLV-FMG proved to be a limiting step for an efficient generation of hybrids. On the other hand, constitutive expression of GaLV-FMG leads to lethal syncytia formation in human cells. Therefore we developed a novel fusion strategy for the generation of DC/TC cell hybrids that involves the use of a non-human fusogenic cell line that constitutively expresses the GaLV-FMG. With this method we were able to generate reproducible yields of DC/TC triparental hybrids. The formation of tri-parental hybrids via the fusogenic cell line is an interesting alternative to existing DC/TC fusion methods because of its simplicity and its flexibility in the choice of fusion partners, i.e. autologous or allogeneic DCs and tumour cells. Moreover, the tri-parent hybrid system offers the possibility to further enhance the immune response by the addition of transgenes that code for immuno-modulating factors to the fusogenic cell line (annexe II).

L’immunothérapie anticancéreuse est basée sur la capacité du système immunitaire à reconnaître les cellules tumorales comme étrangères et à les éliminer. Les stratégies immunothérapeutiques abordées dans ce travail, incluent l’activation du système immunitaire par l’expression de facteurs immunomodulateurs (l’interleukine-2) via l’utilisation d’un vecteur dérivé du parvovirus MVM, ou par présentation des antigènes tumoraux par la machinerie des cellules dendritiques (DC), via la génération d’hybrides entre DC et cellules tumorales (TC). L’intérêt majeur du parvovirus autonome MVM en tant que vecteur pour la thérapie génique du cancer vient de son expression préférentielle dans les cellules transformées (oncotropisme) et de son aptitude à lyser celles-ci (oncolyse). Les vecteurs générés au laboratoire conservent l’unité de transcription NS et expriment l’IL2 humaine sous contrôle du promoteur P38, à la place des protéines de capside. Malgré les améliorations apportées à la production de vecteurs recombinants, la faible concentration des stocks reste un problème. Il a été montré que, de nombreux virus sont mieux produits en conditions de faible tension en oxygène (hypoxie). Nous avons tenté d’améliorer les titres des vecteurs en les produisant sous faible tension d’oxygène mais sans y parvenir (annexe 1). Dans un modèle in vivo utilisant la lignée de mélanome K-1735 dans des souris immunocompétentes, des cellules tumorales infectées in vitro avant leur implantation en sous-cutané ont montré un effet anti-tumoral du vecteur MVM/IL2 (annexe 2). Afin de mettre en évidence l’apport de l’oncolyse parvovirale dans l’activité anti-tumorale, nous avons mis au point des expériences, dans le même modèle de tumeur, visant à comparer l’efficacité du vecteur MVM/IL2 à celle d’autres vecteurs, Ad/IL2 et Rétrovirus/IL2, ne possédant pas d’activité oncolytique. Dans le but de mettre en évidence une éventuelle réponse immune in vivo, nous avons utilisé le modèle de tumeur TC-1 mais ce modèle s’est montré moins sensible à l’effet du vecteur MVM/IL2 et nous n’avons pas pu démontrer d’activation de cellules cytotoxiques spécifiques de la tumeur. Il a été proposé d’utiliser des hybrides entre DC/TC pour la vaccination anti-tumorale pour optimaliser la présentation des antigènes tumoraux. Une lignée cellulaire exprimant la protéine fusogène du virus de la leucémie du Gibbon (GaLV-FMG, Gibbon ape leukemia virus) a été dérivée de la lignée cellulaire CHO (cellules ovariennes de hamster chinois) au laboratoire. Cette lignée CHO-FMG, utilisée comme partenaire intermédiaire, a permis la fusion entre cellules tumorales et dendritiques (annexe 3). Nous avons montré que l’expression transitoire après infection par un vecteur AAV-FMG ou après transfection transitoire ne génère pas un pourcentage significatif d’hybrides. En effet, le niveau d’expression ainsi que le pourcentage de cellules transduites exprimant FMG s’est révélé trop faible. Ceci a mis en valeur l’efficacité de la lignée stable CHO-FMG comme intermédiaire de la fusion. De plus, nous avons intégré dans la lignée fusogène, le gène de l’interleukine-2, qui devrait permettre d’augmenter l’efficacité de l’induction de la réponse immune. L’activation des cellules effectrices de l’immunité anti-tumorale via la présentation antigénique et/ou par des cytokines est au centre de ce travail. Une meilleure compréhension des mécanismes sous-jacents devrait permettre d’améliorer ces systèmes de vaccination non-conventionnels.

This thesis addresses the exponential growth of experimental data and the resulting computational complexity seen in two major scientific applications, which account for significant cycles consumed on today’s supercomputers. The first application concerns computation of the adaptive optics system in next-generation ground-based telescopes, which will expand our knowledge of the universe but confronts the astronomy community with daunting real-time computation requirements. The second application deals with emerging frequency-domain redatuming methods, e.g., Marchenko redatuming, which are game-changers in exploration geophysics. They are valuable to oil and gas applications and will soon be to geothermal exploration and carbon capture storage. However, they are impractical at industrial scale due to prohibitive computational complexity and memory footprint. We tackle the aforementioned challenges by designing high-performance algebraic and stochastic algorithms, which exploit the data sparsity structure of the matrix operator. We show that popular randomized algorithms from machine learning can also solve large covariance matrix problems that capture the correlations of wavefront sensors detecting the atmospheric turbulence for ground-based telescopes. Algebraic compression based on low-rank approximations that retains the most significant portion of the spectrum of the operator provides numerical solutions at the accuracy level required by the application. In addition, selective use of lower precisions can further reduce the data volume by trading off application accuracy for memory footprint. Reducing memory footprint has ancillary implications for reduced energy expenditure and reduced execution time because moving a word is more expensive than computing with it on today’s architectures. We exploit the data sparsity of matrices representative of these two scientific applications and propose four algorithms to accelerate the corresponding computational workload. In soft real-time control of an adaptive optics system, we design a stochastic Levenberg-Marquardt method and high-performance solver for Discrete-time Algebraic Riccati Equations. We create a tile low-rank matrix-vector multiplication algorithm used in both hard real-time control of ground-based telescopes and seismic redatuming. Finally, we leverage multiple precisions to further improve the performance of seismic redatuming applications We implement our algorithms on essentially all families of currently relevant HPC architectures and customized AI accelerators and demonstrate significant performance improvement and validated numerical solutions.

O objetivo deste trabalho é discutir a aplicação do Fluxo Unitário da Produção (FUP) no processo de fabricação do vestuário de moda, com ênfase em produtos de moda feminina. Para tanto, foi realizada uma pesquisa de campo, por meio de estudo de caso, em empresa do setor. O resultado da pesquisa mostra que, apesar das particularidades do setor, como o baixo volume de produção, curto ciclo de vida do produto e grande diversidade de modelos, a aplicação do Fluxo Unitário da Produção na manufatura do vestuário de moda indica uma possível otimização do processo produtivo, permitindo o balanceamento da linha de montagem e o dimensionamento da capacidade produtiva, fatores que minimizam os riscos de gargalos produtivos e auxiliam na elaboração do planejamento, programação e controle da produção. O presente estudo tem como objetivos específicos: i) pesquisar os processos de manufatura utilizada, ii ) focalizar as técnicas e processos produtivos aplicados nos produtos manufaturados; iii) caracterizar fisicamente a estrutura fabril e descrever os seus processos produtivos durante a manufatura dos produtos com a aplicação do fluxo unitário na Manufatura do Vestuário da Moda (MVM); iv) apontar sugestões de uso de novas ferramentas como a melhoria contínua do processo produtivo. As recomendações elaboradas são baseadas em levantamento bibliográfico e em estudo de caso na empresa fabricante de produtos por encomenda. Essas serão legitimadas por informações de especialistas no assunto e por funcionários da empresa estudada. Foi levantada até o momento presente a relação dos fornecedores externos e as suas capacidades instaladas que manufaturam os produtos vendidos, pois a Empresa A apontou a presença de gargalo produtivo por não conseguir entregar o volume produtivo solicitado. Embora o estudo esteja ocorrendo dentro do cronograma de execução, muitos dos conceitos e técnicas da produção enxuta são aplicáveis 7 nos ambientes produtivos de alta variedade da mesma família de produto, com baixo volume e pode contribuir de forma significativa para a melhoria do desempenho da empresa, para a melhoria contínua do processo, para a melhoria da competitividade e eliminação das perdas produtivas com indicadores de desempenhos nos setores envolvidos. Após a pesquisa com a aplicação do fluxo unitário da Manufatura do Vestuário da Moda na Empresa A, será verificada se a hipótese da aplicação do Fluxo Unitário da Produção, da indústria automobilística, é possível para a indústria do vestuário. Coletados os resultados dessa aplicação, após a análise dos dados e avaliação verificará se há aprovação da Manufatura do Vestuário de Moda (MVM) / The objective of this paper is to discuss the implementation of Flow Unit of Production in the manufacturing of fashion apparel process, with emphasis on women\'s fashion products. For this purpose, a field research was conducted through case study, in company of the sector. The survey results show that despite the particularities of the industry, such as production volume, short cycle product life and wide range of models, the application of Flow Unit of Production in the fashion garment manufacturing indicates a possible process optimization productive, allowing the assembly line balancing and capacity of the productive capacity, factors that minimize the risk of production bottlenecks and assist in the preparation of planning, scheduling and production control. This study has the following objectives: i) search the manufacturing processes used, ii) focus on the techniques and processes applied in manufactured products; iii) physically characterize the industrial structure and describe their production processes for the manufacture of products with the application of flow unit in Fashion Garment Manufacturing (MVM); iv) point out suggestions for the use of new tools such as the continuous improvement of the production process. The recommendations made are based on literature review and case study on the manufacturer of custom products. These will be legitimized by experts of information on the subject and by employees of the studied company. It has been raised to date a list of external vendors and their installed capacities to manufacture products sold, as the company \"A\" pointed out the presence of \"bottleneck\" productive for failing to deliver the production volumes requested. Although the study is taking place within the execution schedule, many of the concepts and techniques of lean manufacturing are applicable in production environments high variety of the same product family, with low volume and can contribute significantly to improving the company\'s performance, for continuous process improvement, to improve competitiveness and elimination of production 9 losses with performance indicators in the sectors involved. After research with the application of the unitary flow of Manufacture of fashionable clothes in the company in \"A\", will be checked the hypothesis of application of Flow Unit of Production, the automotive industry, it is possible for the clothing industry. Collected the results of that application, after the data analysis and evaluation check for approval of Manufacture of Fashionable Clothes

O objetivo deste trabalho é discutir a aplicação do Fluxo Unitário da Produção (FUP) no processo de fabricação do vestuário de moda, com ênfase em produtos de moda feminina. Para tanto, foi realizada uma pesquisa de campo, por meio de estudo de caso, em empresa do setor. O resultado da pesquisa mostra que, apesar das particularidades do setor, como o baixo volume de produção, curto ciclo de vida do produto e grande diversidade de modelos, a aplicação do Fluxo Unitário da Produção na manufatura do vestuário de moda indica uma possível otimização do processo produtivo, permitindo o balanceamento da linha de montagem e o dimensionamento da capacidade produtiva, fatores que minimizam os riscos de gargalos produtivos e auxiliam na elaboração do planejamento, programação e controle da produção. O presente estudo tem como objetivos específicos: i) pesquisar os processos de manufatura utilizada, ii ) focalizar as técnicas e processos produtivos aplicados nos produtos manufaturados; iii) caracterizar fisicamente a estrutura fabril e descrever os seus processos produtivos durante a manufatura dos produtos com a aplicação do fluxo unitário na Manufatura do Vestuário da Moda (MVM); iv) apontar sugestões de uso de novas ferramentas como a melhoria contínua do processo produtivo. As recomendações elaboradas são baseadas em levantamento bibliográfico e em estudo de caso na empresa fabricante de produtos por encomenda. Essas serão legitimadas por informações de especialistas no assunto e por funcionários da empresa estudada. Foi levantada até o momento presente a relação dos fornecedores externos e as suas capacidades instaladas que manufaturam os produtos vendidos, pois a Empresa A apontou a presença de gargalo produtivo por não conseguir entregar o volume produtivo solicitado. Embora o estudo esteja ocorrendo dentro do cronograma de execução, muitos dos conceitos e técnicas da produção enxuta são aplicáveis 7 nos ambientes produtivos de alta variedade da mesma família de produto, com baixo volume e pode contribuir de forma significativa para a melhoria do desempenho da empresa, para a melhoria contínua do processo, para a melhoria da competitividade e eliminação das perdas produtivas com indicadores de desempenhos nos setores envolvidos. Após a pesquisa com a aplicação do fluxo unitário da Manufatura do Vestuário da Moda na Empresa A, será verificada se a hipótese da aplicação do Fluxo Unitário da Produção, da indústria automobilística, é possível para a indústria do vestuário. Coletados os resultados dessa aplicação, após a análise dos dados e avaliação verificará se há aprovação da Manufatura do Vestuário de Moda (MVM) / The objective of this paper is to discuss the implementation of Flow Unit of Production in the manufacturing of fashion apparel process, with emphasis on women\'s fashion products. For this purpose, a field research was conducted through case study, in company of the sector. The survey results show that despite the particularities of the industry, such as production volume, short cycle product life and wide range of models, the application of Flow Unit of Production in the fashion garment manufacturing indicates a possible process optimization productive, allowing the assembly line balancing and capacity of the productive capacity, factors that minimize the risk of production bottlenecks and assist in the preparation of planning, scheduling and production control. This study has the following objectives: i) search the manufacturing processes used, ii) focus on the techniques and processes applied in manufactured products; iii) physically characterize the industrial structure and describe their production processes for the manufacture of products with the application of flow unit in Fashion Garment Manufacturing (MVM); iv) point out suggestions for the use of new tools such as the continuous improvement of the production process. The recommendations made are based on literature review and case study on the manufacturer of custom products. These will be legitimized by experts of information on the subject and by employees of the studied company. It has been raised to date a list of external vendors and their installed capacities to manufacture products sold, as the company \"A\" pointed out the presence of \"bottleneck\" productive for failing to deliver the production volumes requested. Although the study is taking place within the execution schedule, many of the concepts and techniques of lean manufacturing are applicable in production environments high variety of the same product family, with low volume and can contribute significantly to improving the company\'s performance, for continuous process improvement, to improve competitiveness and elimination of production 9 losses with performance indicators in the sectors involved. After research with the application of the unitary flow of Manufacture of fashionable clothes in the company in \"A\", will be checked the hypothesis of application of Flow Unit of Production, the automotive industry, it is possible for the clothing industry. Collected the results of that application, after the data analysis and evaluation check for approval of Manufacture of Fashionable Clothes

Pour mieux comprendre les structures de la couronne solaire de température élevée et leurs évolutions, des méthodes de vision à 3 dimensions à partir des images EUV de SOHO/EIT ont été développées. Des anaglyphes ont servi à mieux comprendre le rôle du champ magnétique et des intéractions entre champs<br />fermés et ouverts. Il est apparu que la reconnection entre des trous coronaux (CH) et des régions actives (AR) peut expliquer certaines disparitions de boucles coronales fermées. De plus, des analyses multi-instruments ont montré que la structuration des<br />CHs résulte parfois de changements dans l'activité magnétique voisine.<br />Les mesures d'incertitudes sur les reconstructions par stéréovision n'ont pas permis de valider une méthode générale pour visualiser la couronne globalement en 3D. Par contre, le principe a pu être utilisé avec succès pour la reconstruction en 3 dimensions de boucles coronales. Ainsi après analyse des paramètres physiques de 30 boucles EUV de températures intermédiaires d'une région active, il est apparu que contrairement aux boucles chaudes en X, elles étaient en équilibre hydrostatique.<br />Les boucles d'une région active émergentes sont apparues comme se détorsadant au fur et à mesure de son grandissement ce qui correspond à un transfert de l'hélicité.<br />Des corrélations entre des brusques détorsadages et des éruptions ont aussi été établis dans les cas où la torsion initiale est trop importante et permet le développement<br />d'instabilités.<br />Ces études 3D vont pouvoir permettre de mieux contraindre les bilans énergétiques pour le chauffage coronal gràce à la conservation de l'hélicité et d'améliorer la prévision de la météorologie spatiale. L'utilisation de techniques d'imageries adaptées (comme<br />le dégrillage ou le Modèle de Vision Multi-échelle) a permis de faire ressortir certains détails dans la formation de CMEs par exemple.

L’immunothérapie anticancéreuse est basée sur la capacité du système immunitaire à reconnaître les cellules tumorales comme étrangères et à les éliminer. Les stratégies immunothérapeutiques abordées dans ce travail, incluent l’activation du système immunitaire par l’expression de facteurs immunomodulateurs (l’interleukine-2) via l’utilisation d’un vecteur dérivé du parvovirus MVM, ou par présentation des antigènes tumoraux par la machinerie des cellules dendritiques (DC), via la génération d’hybrides entre DC et cellules tumorales (TC).<p>L’intérêt majeur du parvovirus autonome MVM en tant que vecteur pour la thérapie génique du cancer vient de son expression préférentielle dans les cellules transformées (oncotropisme) et de son aptitude à lyser celles-ci (oncolyse). Les vecteurs générés au laboratoire conservent l’unité de transcription NS et expriment l’IL2 humaine sous contrôle du promoteur P38, à la place des protéines de capside. Malgré les améliorations apportées à la production de vecteurs recombinants, la faible concentration des stocks reste un problème. Il a été montré que, de nombreux virus sont mieux produits en conditions de faible tension en oxygène (hypoxie). Nous avons tenté d’améliorer les titres des vecteurs en les produisant sous faible tension d’oxygène mais sans y parvenir (annexe 1). Dans un modèle in vivo utilisant la lignée de mélanome K-1735 dans des souris immunocompétentes, des cellules tumorales infectées in vitro avant leur implantation en sous-cutané ont montré un effet anti-tumoral du vecteur MVM/IL2 (annexe 2). Afin de mettre en évidence l’apport de l’oncolyse parvovirale dans l’activité anti-tumorale, nous avons mis au point des expériences, dans le même modèle de tumeur, visant à comparer l’efficacité du vecteur MVM/IL2 à celle d’autres vecteurs, Ad/IL2 et Rétrovirus/IL2, ne possédant pas d’activité oncolytique. Dans le but de mettre en évidence une éventuelle réponse immune in vivo, nous avons utilisé le modèle de tumeur TC-1 mais ce modèle s’est montré moins sensible à l’effet du vecteur MVM/IL2 et nous n’avons pas pu démontrer d’activation de cellules cytotoxiques spécifiques de la tumeur.<p>Il a été proposé d’utiliser des hybrides entre DC/TC pour la vaccination anti-tumorale pour optimaliser la présentation des antigènes tumoraux. Une lignée cellulaire exprimant la protéine fusogène du virus de la leucémie du Gibbon (GaLV-FMG, Gibbon ape leukemia virus) a été dérivée de la lignée cellulaire CHO (cellules ovariennes de hamster chinois) au laboratoire. Cette lignée CHO-FMG, utilisée comme partenaire intermédiaire, a permis la fusion entre cellules tumorales et dendritiques (annexe 3). Nous avons montré que l’expression transitoire après infection par un vecteur AAV-FMG ou après transfection transitoire ne génère pas un pourcentage significatif d’hybrides. En effet, le niveau d’expression ainsi que le pourcentage de cellules transduites exprimant FMG s’est révélé trop faible. Ceci a mis en valeur l’efficacité de la lignée stable CHO-FMG comme intermédiaire de la fusion. De plus, nous avons intégré dans la lignée fusogène, le gène de l’interleukine-2, qui devrait permettre d’augmenter l’efficacité de l’induction de la réponse immune. <p>\ / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Cancer is a worldwide health problem and despite advances in traditional treatments i.e. surgery, chemotherapy and radiotherapy, the cure rate remains disappointing for some cancers. Different novel therapeutic strategies are being developed. In this thesis two nontraditional cancer therapy approaches are studied: gene therapy using viral vectors and antitumour vaccination with dendritic cell - tumour cell (DC/TC) hybrids.<p>We have developed a novel ELISPOT titration method for viral vectors that is based on the actual expression of the transgene in target cells. This method was developed with recombinant parvovirus MVM-IL2, but it should be adaptable for other vectors carrying expression cassettes for secreted transgene products for which antibodies are available. The ELISPOT titration method allows for faster and better quantification of transducing units present in vector stocks as opposed to titration by in situ hybridisation (annexe I). The MVMIL2 vector has shown an anti-tumour effect against melanoma in an immunocompetent mouse model (annexe IV). Previous work concerns photodynamic inactivation of adenoviral vectors for biosafety and an in vivo study in which a synergistic effect of antiangiogenesis gene therapy combined with radiotherapy could be shown (annexes V and VI).<p>DC/TC hybrids have been proposed as cancer vaccines for their simultaneous expression of antigen presentation machinery and tumour associated antigens. Hybrids are classically generated by polyethylene glycol (PEG) or electrofusion. These methods however require special skills and equipment and cause rather high cell lethality. Fusion via the expression of viral fusogenic membrane glycoproteins (FMG), such as the vesicular stomatitis virus-G (VSV-G) (annexe III) or the Gibbon ape Leukemia Virus (GaLV) FMG, have recently been described. We have mainly focussed on the latter. Transduction of cells with GaLV-FMG proved to be a limiting step for an efficient generation of hybrids. On the other hand, constitutive expression of GaLV-FMG leads to lethal syncytia formation in human cells. Therefore we developed a novel fusion strategy for the generation of DC/TC cell hybrids that involves the use of a non-human fusogenic cell line that constitutively expresses the GaLV-FMG. With this method we were able to generate reproducible yields of DC/TC triparental hybrids. The formation of tri-parental hybrids via the fusogenic cell line is an interesting alternative to existing DC/TC fusion methods because of its simplicity and its flexibility in the choice of fusion partners, i.e. autologous or allogeneic DCs and tumour cells.<p>Moreover, the tri-parent hybrid system offers the possibility to further enhance the immune response by the addition of transgenes that code for immuno-modulating factors to the fusogenic cell line (annexe II). / Doctorat en sciences biomédicales / info:eu-repo/semantics/nonPublished