No description available.

Peptídeos de sinalização ou hormonais desempenham papéis importantes nas plantas por serem determinantes no crescimento, desenvolvimento e defesa. RALF é um peptídeo ubíquo no reino vegetal e na planta modelo Arabidopsis thaliana os peptídeos RALF formam uma família multigênica de 37 membros alguns com expressão gênica tecido-específica e outros com expressão em toda a planta. A isoforma AtRALF1 é a mais estudada e é expressa principalmente na raiz e no hipocótilo. Uma das funções deste peptídeo é a regulação da expansão celular, um processo que também envolve auxina, giberelina, etileno, brassinosteróides e citocininas. O objetivo deste trabalho foi estudar a relação entre o AtRALF1 e o hormônio etileno, principalmente na expansão celular e alcalinização celular que são efeitos característicos do peptídeo. AtRALF1 inibe o crescimento da raiz primária porém plantas de arabidopsis tratadas simultaneamente com o peptídeo e inibidores da biossíntese ou percepção de etileno não tiveram o crescimento de suas raízes inibido. Etileno induz a deposição de calose e plantas selvagens expostas ao AtRALF1 e plantas que super-expressam AtRALF1 exibiram depósito de calose nas células da ponta da raiz. Curiosamente, quando a alcalinização do meio de células em suspensão induzida por AtRALF1 foi avaliada perante o aumento da produção de etileno ou perante o bloqueio de sua síntese ou percepção, não foi observada alteração na resposta. Os resultados aqui apresentados demonstram que a resposta de inibição da expansão celular ocasionada por AtRALF1 é dependente de etileno e sugere que os efeitos de alcalinização do meio extracelular e da inibição do crescimento da raiz primária estão dissociados. / Peptide hormones or signaling peptides play important roles that determine growth, development and defense in plants. RALF is a ubiquitous peptide in the plant kingdom and in model plant Arabidopsis thaliana comprises a multigene family of 37 members, some of them with tissue-specific gene expression and others that are expressed throughout the plant. AtRALF1 isoform is the most studied and is expressed in roots and hypocotyls. One of the peptide functions is the negative regulation of cell expansion, a process also controled by auxin, gibberelin, ethylene, brassinosteroids, cytokinin. The aim of this work was to study ethylene and AtRALF1 peptide relation, mainly in the cell alkalinization and inhibition of cell expansion responses that characteristic of the peptide. AtRALF1 inhibits root growth but simultaneous treatment with both AtRALF1 and inhibitors of the ethylene perception or biosynthesis show no inhibition on root growth. Ethylene increases callose deposition and wild-type plants treated with AtRALF1 or plants overexpressing the AtRALF1 gene show increased callose plate formation on root tips. Curiously, when the extracellular alkalinization induced by AtRALF1 was evaluated against ethylene production or against ethylene inhibitors, no alteration was observed. The data presented here reveal that the cell expansion inhibition caused by AtRALF1 is ethylene dependent and suggest that the extracellular alkalinization response and root growth inhibition are dissociated.

Growing evidence from animal and clinical studies suggests that cardiac cell therapy can restore perfusion and improve function in the ischemic/infarcted myocardium. However, cell therapy is hindered by insufficient cell numbers, inefficient cell homing and engraftment, and inadequate cellular interactions. Furthermore, the biological mechanisms and local effects of transplanted cells have not been well-elucidated. The research presented herein attempts to address some of these issues. In manuscript #1, a new subpopulation of circulating progenitor cells (CPCs), termed derived CD133+ cells, was generated from the CD133- fraction of human peripheral blood. The derived CD133+ progenitors appeared to have superior vasculogenic potential in vitro, which may prove to be beneficial in inducing vasculogenesis in ischemic tissues. Positron emission tomography (PET) with direct cell labeling and reporter gene techniques were employed to assess the fate of transplanted human CPCs in vivo at different subjects of investigation, and different stages of cell transplantation. In manuscript #2, PET imaging with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) direct cell labeling was used to demonstrate that collagen-based matrices improve the early homing and retention of delivered CPCs in a rat ischemic hindlimb model. This mechanism conferred by the matrix may have implications on cell therapy at the early stages after transplantation. In manuscript #3, a more efficient, stable and accurate labeling method, hexadecyl-4-[18F]fluorobenzoate (18F-HFB) direct cell labeling, was developed to quantify cell distribution of transplanted CPCs in a rat myocardial infarction model. PET imaging of 18F-HFB-CPCs revealed significant cell washout from the myocardium immediately after intramyocardial injection, with only a small proportion of transplanted CPCs remaining in the target area in the first 4 hours after delivery. In manuscript #4, human CPCs transduced with lentiviral vectors showed stable expression of PET reporter genes. This reporter gene based-cell labeling technique can be developed for noninvasive tracking cells within a bioengineered matrix by PET, while preserving cell phenotype, viability and function. These studies contribute important insights into the biology and physiology of transplanted stem cells and the ability of delivery matrices to improve transplanted cell engraftment, survival, and function. I believe with further refinement, cell expansion, tissue engineering and PET imaging could facilitate the clinical applications of cell therapies in years to come.

Growing evidence from animal and clinical studies suggests that cardiac cell therapy can restore perfusion and improve function in the ischemic/infarcted myocardium. However, cell therapy is hindered by insufficient cell numbers, inefficient cell homing and engraftment, and inadequate cellular interactions. Furthermore, the biological mechanisms and local effects of transplanted cells have not been well-elucidated. The research presented herein attempts to address some of these issues. In manuscript #1, a new subpopulation of circulating progenitor cells (CPCs), termed derived CD133+ cells, was generated from the CD133- fraction of human peripheral blood. The derived CD133+ progenitors appeared to have superior vasculogenic potential in vitro, which may prove to be beneficial in inducing vasculogenesis in ischemic tissues. Positron emission tomography (PET) with direct cell labeling and reporter gene techniques were employed to assess the fate of transplanted human CPCs in vivo at different subjects of investigation, and different stages of cell transplantation. In manuscript #2, PET imaging with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) direct cell labeling was used to demonstrate that collagen-based matrices improve the early homing and retention of delivered CPCs in a rat ischemic hindlimb model. This mechanism conferred by the matrix may have implications on cell therapy at the early stages after transplantation. In manuscript #3, a more efficient, stable and accurate labeling method, hexadecyl-4-[18F]fluorobenzoate (18F-HFB) direct cell labeling, was developed to quantify cell distribution of transplanted CPCs in a rat myocardial infarction model. PET imaging of 18F-HFB-CPCs revealed significant cell washout from the myocardium immediately after intramyocardial injection, with only a small proportion of transplanted CPCs remaining in the target area in the first 4 hours after delivery. In manuscript #4, human CPCs transduced with lentiviral vectors showed stable expression of PET reporter genes. This reporter gene based-cell labeling technique can be developed for noninvasive tracking cells within a bioengineered matrix by PET, while preserving cell phenotype, viability and function. These studies contribute important insights into the biology and physiology of transplanted stem cells and the ability of delivery matrices to improve transplanted cell engraftment, survival, and function. I believe with further refinement, cell expansion, tissue engineering and PET imaging could facilitate the clinical applications of cell therapies in years to come.

Growing evidence from animal and clinical studies suggests that cardiac cell therapy can restore perfusion and improve function in the ischemic/infarcted myocardium. However, cell therapy is hindered by insufficient cell numbers, inefficient cell homing and engraftment, and inadequate cellular interactions. Furthermore, the biological mechanisms and local effects of transplanted cells have not been well-elucidated. The research presented herein attempts to address some of these issues. In manuscript #1, a new subpopulation of circulating progenitor cells (CPCs), termed derived CD133+ cells, was generated from the CD133- fraction of human peripheral blood. The derived CD133+ progenitors appeared to have superior vasculogenic potential in vitro, which may prove to be beneficial in inducing vasculogenesis in ischemic tissues. Positron emission tomography (PET) with direct cell labeling and reporter gene techniques were employed to assess the fate of transplanted human CPCs in vivo at different subjects of investigation, and different stages of cell transplantation. In manuscript #2, PET imaging with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) direct cell labeling was used to demonstrate that collagen-based matrices improve the early homing and retention of delivered CPCs in a rat ischemic hindlimb model. This mechanism conferred by the matrix may have implications on cell therapy at the early stages after transplantation. In manuscript #3, a more efficient, stable and accurate labeling method, hexadecyl-4-[18F]fluorobenzoate (18F-HFB) direct cell labeling, was developed to quantify cell distribution of transplanted CPCs in a rat myocardial infarction model. PET imaging of 18F-HFB-CPCs revealed significant cell washout from the myocardium immediately after intramyocardial injection, with only a small proportion of transplanted CPCs remaining in the target area in the first 4 hours after delivery. In manuscript #4, human CPCs transduced with lentiviral vectors showed stable expression of PET reporter genes. This reporter gene based-cell labeling technique can be developed for noninvasive tracking cells within a bioengineered matrix by PET, while preserving cell phenotype, viability and function. These studies contribute important insights into the biology and physiology of transplanted stem cells and the ability of delivery matrices to improve transplanted cell engraftment, survival, and function. I believe with further refinement, cell expansion, tissue engineering and PET imaging could facilitate the clinical applications of cell therapies in years to come.

Peptídeos de sinalização ou hormonais desempenham papéis importantes nas plantas por serem determinantes no crescimento, desenvolvimento e defesa. RALF é um peptídeo ubíquo no reino vegetal e na planta modelo Arabidopsis thaliana os peptídeos RALF formam uma família multigênica de 37 membros alguns com expressão gênica tecido-específica e outros com expressão em toda a planta. A isoforma AtRALF1 é a mais estudada e é expressa principalmente na raiz e no hipocótilo. Uma das funções deste peptídeo é a regulação da expansão celular, um processo que também envolve auxina, giberelina, etileno, brassinosteróides e citocininas. O objetivo deste trabalho foi estudar a relação entre o AtRALF1 e o hormônio etileno, principalmente na expansão celular e alcalinização celular que são efeitos característicos do peptídeo. AtRALF1 inibe o crescimento da raiz primária porém plantas de arabidopsis tratadas simultaneamente com o peptídeo e inibidores da biossíntese ou percepção de etileno não tiveram o crescimento de suas raízes inibido. Etileno induz a deposição de calose e plantas selvagens expostas ao AtRALF1 e plantas que super-expressam AtRALF1 exibiram depósito de calose nas células da ponta da raiz. Curiosamente, quando a alcalinização do meio de células em suspensão induzida por AtRALF1 foi avaliada perante o aumento da produção de etileno ou perante o bloqueio de sua síntese ou percepção, não foi observada alteração na resposta. Os resultados aqui apresentados demonstram que a resposta de inibição da expansão celular ocasionada por AtRALF1 é dependente de etileno e sugere que os efeitos de alcalinização do meio extracelular e da inibição do crescimento da raiz primária estão dissociados. / Peptide hormones or signaling peptides play important roles that determine growth, development and defense in plants. RALF is a ubiquitous peptide in the plant kingdom and in model plant Arabidopsis thaliana comprises a multigene family of 37 members, some of them with tissue-specific gene expression and others that are expressed throughout the plant. AtRALF1 isoform is the most studied and is expressed in roots and hypocotyls. One of the peptide functions is the negative regulation of cell expansion, a process also controled by auxin, gibberelin, ethylene, brassinosteroids, cytokinin. The aim of this work was to study ethylene and AtRALF1 peptide relation, mainly in the cell alkalinization and inhibition of cell expansion responses that characteristic of the peptide. AtRALF1 inhibits root growth but simultaneous treatment with both AtRALF1 and inhibitors of the ethylene perception or biosynthesis show no inhibition on root growth. Ethylene increases callose deposition and wild-type plants treated with AtRALF1 or plants overexpressing the AtRALF1 gene show increased callose plate formation on root tips. Curiously, when the extracellular alkalinization induced by AtRALF1 was evaluated against ethylene production or against ethylene inhibitors, no alteration was observed. The data presented here reveal that the cell expansion inhibition caused by AtRALF1 is ethylene dependent and suggest that the extracellular alkalinization response and root growth inhibition are dissociated.

Growing evidence from animal and clinical studies suggests that cardiac cell therapy can restore perfusion and improve function in the ischemic/infarcted myocardium. However, cell therapy is hindered by insufficient cell numbers, inefficient cell homing and engraftment, and inadequate cellular interactions. Furthermore, the biological mechanisms and local effects of transplanted cells have not been well-elucidated. The research presented herein attempts to address some of these issues. In manuscript #1, a new subpopulation of circulating progenitor cells (CPCs), termed derived CD133+ cells, was generated from the CD133- fraction of human peripheral blood. The derived CD133+ progenitors appeared to have superior vasculogenic potential in vitro, which may prove to be beneficial in inducing vasculogenesis in ischemic tissues. Positron emission tomography (PET) with direct cell labeling and reporter gene techniques were employed to assess the fate of transplanted human CPCs in vivo at different subjects of investigation, and different stages of cell transplantation. In manuscript #2, PET imaging with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) direct cell labeling was used to demonstrate that collagen-based matrices improve the early homing and retention of delivered CPCs in a rat ischemic hindlimb model. This mechanism conferred by the matrix may have implications on cell therapy at the early stages after transplantation. In manuscript #3, a more efficient, stable and accurate labeling method, hexadecyl-4-[18F]fluorobenzoate (18F-HFB) direct cell labeling, was developed to quantify cell distribution of transplanted CPCs in a rat myocardial infarction model. PET imaging of 18F-HFB-CPCs revealed significant cell washout from the myocardium immediately after intramyocardial injection, with only a small proportion of transplanted CPCs remaining in the target area in the first 4 hours after delivery. In manuscript #4, human CPCs transduced with lentiviral vectors showed stable expression of PET reporter genes. This reporter gene based-cell labeling technique can be developed for noninvasive tracking cells within a bioengineered matrix by PET, while preserving cell phenotype, viability and function. These studies contribute important insights into the biology and physiology of transplanted stem cells and the ability of delivery matrices to improve transplanted cell engraftment, survival, and function. I believe with further refinement, cell expansion, tissue engineering and PET imaging could facilitate the clinical applications of cell therapies in years to come.

No description available.

Successful treatment of cancer with adoptive immunotherapy (AIT) is dependent on the ability to produce large numbers of tumor-specific, functional T cells. The purpose of this thesis is to explore ways in which T cell expansion could be augmented. We have focused on exploring alternate gamma chain cytokines as stimulators of T cell proliferation and differentiation in addition to investigating the potential usefulness of gemcitabine (GEM) in abrogating the immunosuppressive effects of myeloid derived suppressor cells (MDSCs). B16 melanoma sensitized draining lymph node cells that have been activated in vitro with bryostatin-1 and ionomycin (B/I) were expanded in either IL-7/15 or in IL-2. We found that IL-7/15 was superior to IL-2 in expanding T cells for AIT of pulmonary metastases. Expansion of antitumor T cells was also improved by suppressing accumulation of MDSCs in mice bearing 4T1 mammary carcinoma using GEM. GEM directly inhibits both 4T1 mammary carcinoma cells and MDSCs. Its inhibition of MDSCs rescued tolerant T cells, augmenting both expansion and response to tumor antigen.

La transformation de la paroi de l’ovaire en un péricarpe charnu implique une coordination entre les divisions cellulaires et l’expansion cellulaire. Des données considérables sur le développement et la maturation du fruit de tomate ont été établies, mais la coordination des divisions cellulaires, de l’expansion cellulaire et de l’endoréduplication durant la mise à fruit ainsi que durant la croissance du fruit de tomate reste grossièrement caractérisée au sein du péricarpe et de nombreuses questions ne sont pas résolues : comment ces deux processus sont-ils régulés et coordonnés durant le développement du fruit d’un point de vue cellulaire? Quand commence l’endoréduplication dans les tissus du fruit et quelle est sa fonction? La première partie de ce mémoire concerne la coordination des divisions cellulaires et de l’expansion cellulaire durant la fin du développement de l’ovaire et le début du développement du fruit. Une différenciation précoce des assises cellulaires composant la paroi de l’ovaire puis le péricarpe a été démontrée. Les divisions cellulaires se font principalement au sein de l’épiderme externe et montrent une synchronisation partielle tandis que l’expansion cellulaire se fait principalement dans le mésocarpe. L’endoréduplication semble être initiée avant l’anthèse. La deuxième partie est consacrée à l’analyse du RNA-seq nucléaire en fonction de quatre niveaux de ploïdie (4, 8, 16 et 32C). La majorité des gènes montrent une augmentation proportionnelle de leurs expressions en fonction des niveaux de ploïdie. Cependant, certains gènes révèlent une surexpression ou une sous-expression en fonction des niveaux de ploïdies. / The transformation of the ovary wall into a fleshy pericarp involves a coordinated pattern of cell division and cell expansion. Considerable data have been reported on tomato fruit development and ripening, but the pattern of cell division, cell expansion and endoreduplication at the tomato fruit set and during fruit growth remains grossly appreciated at the whole pericarp level and many questions are not yet resolved: How are cell division and cell expansion coordinated in tomato fruit a cellular level and according to developmental time? When does endoreduplication begin in fruit tissues and what is its function? The first part of this deals with the coordination of cell division and cell expansion during the end of tomato ovary development and the beginning of fruit growth. Evidence for early differentiation of cell layers in the ovary wall and then in fruit pericarp are presented. Cell division happens mainly in the external epidermis and shows partial synchronization, whereas cell expansion happens mostly in mesocarp cell layers. Endoreduplication is initiated as soon as before anthesis. The second part of this work is devoted to RNA-seq based transcriptome profiling of pericarp nuclei which have been sorted according to four ploidy levels (4, 8, 16 and 32C). We demonstrate that the expression of most of the pericarp-expressed genes shows a proportional increase according to ploidy level, on a nuclear basis. However, a significant amount of genes has been identified as over-expressed or under-expressed according to ploidy level.