Um dos objetivos permanentes da Computação Gráfica é a representação fiel, e por consequência realista, da aparência de objetos e cenas sintetizadas. Para objetos e cenas do Reino Vegetal pode-se dizer que houve um grande avanço nos métodos para modelagem da forma e propriedades de reflexão de elementos individuais, como folhas, e de coleções de elementos, como árvores, vegetação em geral e florestas. Por outro lado, as técnicas para modelagem de padrões visuais presentes em muitos objetos naturais do Reino Vegetal, como listras de melancias ou padrões em folhas e flores, não tem recebido a mesma atenção. O presente trabalho tem por objetivos principais o estudo de técnicas de Computação Gráfica aplicáveis ao Reino Vegetal, principalmente explorar o modelo procedural Mosaico de Clones (MClone), desenvolvido em pesquisas anteriores, para a síntese de padrões visuais familiares do Reino Vegetal. Nossos resultados mostram que, neste contexto, um modelo procedural tem vantagens sobre outras técnicas, como, por exemplo, mapeamento de texturas, pois geralmente esses padrões são necessários em grande quantidade, e, ao menos para algumas plantas, com muitas variações geométricas, a atribuição de coordenadas de textura consistentes é um desafio. Nós mostramos resultados para frutos, cactus e pequenas folhagens decorativas, que confirmam a aplicabilidade do MClone para uma ampla gama de padrões do Reino Vegetal. / There has been a lot of progress in modeling and rendering elements of our Natural World for computer graphics tasks. In the Plant Kingdom, techniques for modeling the visual patterns presented in many natural objects (such as stripes on a watermelon or spots on flowers) have advanced far less than methods for modeling the shape and reflectance properties of individual or large collections of elements (such as leaves and trees). In this work we want to explore the procedural Clonal Mosaic Model (MClone), developed in previous research, for synthesis of many familiar visual patterns from the Plant Kingdom. Our results show that in this context a procedural model has advantages over other techniques such as texture mapping, since these patterns are often needed in large quantities, and at least for some plants, with many geometric variation, assigning consistent texture coordinates is a challenge. We show results for fruits, cactus, and small decorative plants, which confirm the applicability of the MClone model for synthesis of a large group of visual patterns in the Plant Kingdom.

O produto do gene thi1 de Arabidopsis thaliana está provavelmente envolvido na biossíntese de tiamina (vitamina B1) e na proteção do DNA organelar contra danos. Estudos sobre a biossíntese da tiamina em plantas sugerem uma localização plastidial para este mecanismo, o que está de acordo com a existência de um peptídeo de trânsito cloroplástico (TP) na região N-terminal de THI1. Por outro lado, em leveduras a tiamina é sintetizada em mitocôndrias. Interessantemente, o cDNA de thi1 de A. thaliana complementa uma cepa de levedura com disrupção no gene homólogo thi4. A análise da seqüência de aminoácidos de THI1 revelou a presença de uma região capaz de formar uma estrutura do tipo a-hélice anfifílica, freqüentemente encontrada em preseqüências mitocondriais, localizada logo após o peptídeo de trânsito cloroplástico. O papel desta região na localização da proteína THI1 nas mitocôndrias foi comprovado a partir de ensaios envolvendo construções de genes quiméricos (contendo ou não a putativa seqüência de direcionamento mitocondrial) e um gene repórter (uidA). Estas construções foram introduzidas em plantas de tabaco e a localização da atividade GUS foi determinada nas frações subcelulares das plantas transgênicas. Análise direta da presença de THI1 nas mitocôndrias e cloroplastos de Arabidopsis foi realizada via imunolocalização. Também foram fornecidas evidências que as duas isoformas organelares são codificadas por um único transcrito nuclear. Experimentos de transcrição/tradução in vitro indicaram a ocorrência de dois produtos da tradução a partir de códons de iníciação em fase de leitura. Mutações sítio-específicas na seqüência de thi1 acoplados à experimentos usando a proteína fluorescente verde (GFP) mostraram que a tradução no primeiro AUG determina a localização da proteína nos cloroplastos, enquanto que a tradução no segundo AUG é responsável pelo endereçamento da proteína às mitocôndrias. A análise do contexto para início da tradução revelou que a região em torno do primeiro AUG é mais favorável para a tradução do mRNA de thi1. Além disso, observou-se a presença de uma forte estrutura em "grampo de cabelo" próximo ao segundo códon AUG, indicando um contexto subótimo capaz de interferir na tradução. Estas observações confirmaram os dados obtidos a partir da tradução in vitro na qual a iniciação se dá preferencialmente no primeiro AUG o que pode sugerir uma maior necessidade da proteína nos plastídeos. / Arabidopsis thaliana thi1 gene product is probably involved in both thiamine biosynthesis as well as protection of organellar DNA from damage. Studies of thiamine biosynthesis in plants suggest a plastid location for the pathway, which is in agreement with the predicted THI1 N-terminal chloroplastic transit peptide (TP). On the other hand, thiamine is synthesized in mitochondria in yeast cells. Interestingly, A. thaliana thi1 cDNA complements a yeast strain disrupted for the homologous thi4 gene. Analysis of THI1 amino acid sequence revealed the presence of a putative amphiphilic a-helix, which is typical for mitochondrial presequences, located downstream of the chloroplast transit peptide. The role of this sequence on mitochondrial import has been shown by chimeric gene constructs (carrying or not the putative mitochondrial presequence) and the uidA reporter gene. These constructions have been introduced into tobacco plants and the GUS activity has been measured in subcellular fractions of transgenic plants. Direct analysis of THIp in mitochondria and chloroplasts has been done via ImmunoGold labelling experiments. Additional evidence suggested that the two organellar isoforms were encoded by a single nuclear transcript. In vitro transcription/translation experiments revealed the presence of two translational products by a differential usage of two in-frame translational start codons. Coupling site-specific mutations on THI1 encoding sequence with green fluorescent protein (GFP) gene fusions showed that translation initiation in the first AUG directs translocation of THI1 to plastids. However, when translation initiates from the second AUG THI1 is addressed to mitochondria. Analysis of the translation efficiency of thi1 mRNA revealed that the best context for translation initiation is present at the first AUG. In addition, it has been shown a suboptimal context at the second AUG and a strong stem-and-loop structure which is likely to slow translation. These observation confirm the in vitro translation data in which translation occurs preferentially in the first AUG, what could suggest a higher requirement of the protein in plastids.

O objetivo deste trabalho foi avaliar o efeito da enxertia seriada e micropropagação no rejuvenescimento de matrizes adultas de Tectona grandis. A experimentação foi realizada nas empresas Floresteca S.A e Bioteca Ltda, Mato Grosso, utilizando três materiais genéticos diferente, sendo dois clones, com 35 anos idade e mudas de semente. Como técnica de macropropagação optou-se pela enxertia seriada com delineamento inteiramente aleatorizado com três tratamentos representados pelos materiais genéticos com sete repetições. Para micropropagação, utilizou-se meio MS modificado. O experimento in vitro foi dividido em etapas: resgate do meristema com delineamento inteiramente aleatorizado, com os tratamentos sendo três materiais genéticos com trinta repetições; estabelecimento da cultura com os tratamentos arranjados em fatorial 3x5 sendo três materiais genéticos com cinco concentrações de BAP, totalizando quinze tratamentos com cinco repetições; multiplicação/rejuvenescimento com delineamento inteiramente aleatorizado, com os tratamentos arranjados em fatorial 3x5 sendo três materiais genéticos com cinco subcultivos, totalizando quinze tratamentos com três repetições e enraizamento das microestacas. Para macroprogação, o parâmetro avaliado foi a percentagem de pega dos enxertos. Para micropropagação, na fase de resgate, avaliou-se o número de meristemas viáveis; na fase de estabelecimento da cultura, avaliou-se o tamanho, número de nós e número de folhas dos explantes; e na fase de multiplicação/rejuvenescimento, avaliou-se o desenvolvimento de brotos nos sucessivos subcultivos. Os enxertos das mudas obtidas por semente apresentaram brotação aos sete dias de enxertados comparados aos clones que iniciaram as brotações aos quinze dias de enxertados. A sobrevivência dos meristemas apicais introduzidos que não apresentaram oxidação ou contaminação foi superior a 84%. A concentração de 1,5 mg.L-1 de BAP foi a mais significativa na fase de estabelecimento da cultura. O número de brotação e o rejuvenescimento do material adulto aumentaram à medida que aumentou o número de subcultivos, bem como a percentagem de enraizamento. O estabelecimento de minijardim clonal de Tectona grandis é possível mediante uso de brotações rejuvenescidas in vitro. / The aim of this study was to evaluate the effect of serial grafting and micropropagation in rejuvenation of Tectona grandis. The experiments were carried out in Floresteca SA and Bioteca Ltda. companies, Mato Grosso, using two clones and seedlings. The clones were 35 years old and the seedlings were six months old. Serial grafting was chosen for macropropagation technique on a completely randomized statistical design, with three treatments corresponding to genetic material with seven replications. For micropropagation, solid modified MS medium was used. The in vitro experiment was divided into stages: meristem rescue on a completely randomized design with the treatments corresponding to three genetic material with seven replications; establishment in vitro culture on completely randomized design arranged in 3x5 factorial scheme, corresponding to three genetic material with five concentrations of BAP resulting in fifteen treatments with five replications; multiplication/rejuvenation on completely randomized design arranged in 3x5 factorial scheme, corresponding to three genetic material with five subcultures concentrations of resulting in fifteen treatments with three replications on a completely randomized 3x5 and microcuttings rooting. For macropropagation, the parameter evaluated was the percentage of grafting takes. For micropropagation, in the rescue phase, it was evaluated the number of viable meristems. In the establishment phase of culture, the parameters evaluated were the size, number of nodes and number of leaves of the explants; and in the phase of multiplication/rejuvenation it was evaluated the development of the shoots in successive subcultures. The shoots sprouting from seedlings grafts at seventh day compared with grafted clones that started sprouting on the fifteenth day after grafting. The survival of apical meristems without oxidation or contamination was above 84%. The concentration of BAP (1.5 mg.L-1) that was the most significant in the establishment phase of culture. The number of shoots produced and the rejuvenation process increases with increasing number of subcultures, as well as the percentage of rooting. The establishment of clonal minigarden of Tectona grandis coud be possible through rejuvenation of trees in vitro.

The heart arises from two sources of mesoderm progenitors, the first (FHF) and the second heart field (SHF) progenitors. Mesp1 has been proposed to mark the most primitive multipotent cardiac progenitors (MCPs) common for both heart fields. However, it remains unclear whether at the single cell level, Mesp1 progenitors represent a common progenitor for the FHF and SHF. Using mosaic tracing and inducible clonal analysis with a multicolor reporter strategy, we investigated the contribution of Mesp1 cardiovascular progenitors in a temporally controlled manner during the early gastrulation. Our data indicated that the myocardium derives from ~250 Mesp1 expressing cardiac progenitors born during gastrulation. Temporal analysis of clonally labeled Mesp1 cells revealed the existence of temporally distinct populations of Mesp1 progenitors that are restricted to either the FHF or the SHF. FHF progenitors were unipotent, while SHF progenitors, were either uni- or bipotent. Microarray and single cell RT-PCR analysis of Mesp1 progenitors revealed the existence of molecularly distinct populations of Mesp1 progenitors, consistent with their lineage and regional contribution. Moreover biophysical analysis of clonal data revealed that, despite arising at different time points and contributing to different heart regions, the temporally distinct cardiac progenitors present very similar clonal dynamics. Altogether, these results provide insights into the number of cardiac progenitors and their mode of growth. Moreover they provide evidence that heart development arises from distinct populations of unipotent and bipotent cardiac progenitors expressing Mesp1 independently at different time points during gastrulation. Our data reveal that the regional segregation and lineage restriction of cardiac progenitors occurs very early during embryonic development. / Doctorat en Sciences biomédicales et pharmaceutiques (Médecine) / info:eu-repo/semantics/nonPublished

Palms provide valuable commercial resources in the tropics and are dominant species in tropical lowland forests. While general biology of palms is well studied, there are gaps in the literature on palm growth through life stages and in response to environmental conditions. Literature gaps on palm growth could be caused by the slow growth of palms; it is difficult to monitor morphology and architecture for the periods of time necessary to capture changes. Acoelorrhaphe wrightii is a threatened palm native to southern Florida with an unusual adult architecture. The purpose of this dissertation was to study growth A. wrightii throughout its life stages and in response to changes in environmental conditions. In order to do study growth, I first had to understand the evolutionary history and types of vegetative branching in palms to identify vegetative branching possibilities in A. wrightii. I described branching types for 1903 species from all 181 genera using literature reviews and hands-on analysis. I then studied adult morphology and architecture in a common garden setting by monitoring leaf morphology, ramet growth and architecture of A. wrightii in two gardens in Miami, FL, over a two year period. I tested the effects of water and light on germination and growth of juvenile plants in a mesocosm where water and light were manipulated, following growth for a year. Finally, I compared leaf morphology and architecture of adult individuals in four populations in Belize and Florida. I found five branching types were present in the palms: lateral axillary branching, shoot apical division, false vivipary, abaxial branching and leaf-opposed branching. In the garden, Acoelorrhaphe wrightii displayed two types of lateral axillary branching: basal suckering and rhizomatous branching. The two branching types produced tiers in adult clones, which were used to model architecture. Ramets had an establishment period and growth varied seasonally in establishing and established phases. Low water levels and full sun yielded greater germination of A. wrighti and produced juveniles with a greater number of leaves, more root mass and more branches. Variability between populations and environmental conditions was observed in adult individuals in the field but differences were minimal.

Most clonal plants exhibit a modular structure at multiple levels. At the level of the organs, they are characterized by functional modules, such as, internodes, leaves, branches. At the level of the genetic individual (clone or genet), they possess independent evolutionary and physiological units (ramets). These evolutionary units arise through the widespread phenomenon of clonal reproduction, achieved in a variety of ways including rhizomes, stolons, bulbils, or lateral roots. The focus of this study was Populus tremuloides, trembling aspen, a dioecious tree that reproduces sexually by seed and asexually through lateral roots. Local forest patches in western populations of Populus tremuloides consisted largely of multiple genotypes. Multi-clonal patches were dominated by a single genotype, and in one population (Riske Creek) we found several patches (five out of 17) consisting of a single genotype. A second consequence of modularity is that during the repeated cycle of ramet birth, development and death, somatic mutations have the opportunity to occur. Eventually, the clone becomes a mosaic of mutant and non-mutant cell lineages. We found that neutral somatic mutations accumulated across 14 microsatellite loci at a rate of between 10^-6 and 10^-5 per locus per year. We suggest that neutral genetic divergence, under a star phylogeny model of clonal growth, is an alternative way to estimate clone age. Previous estimates of clone age couple the mean growth rate per year of shoots with the area covered by the clone. This assumes a positive linear relationship between clone age and clone size. We found, however, no repeatable pattern across our populations in terms of the relationship of either shape or size to the number of somatic changes. A final consequence of modularity is that during clonal growth, natural selection is relaxed for traits involving sexual function. This means that mutations deleterious to sexual function can accumulate, reducing the overall sexual fitness of a clone. We coupled neutral genetic divergence within clones with pollen fitness data to infer the rate and effect of mildly deleterious mutations. Mutations reduced relative sexual fitness in clonal aspen populations by about 0.12x10^-3 to 1.01x10^-3 per year. Furthermore, the decline in sexual function with clone age is evidence that clonal organisms are vulnerable to the effects of senescence. / Medicine, Faculty of / Medical Genetics, Department of / Graduate

Macrophage-conditioned medium (MacCM) inhibits the differentiation of rodent and human preadipocytes. Previous studies report that murine J774A.1-MacCM inhibits clonal expansion (early required phase of adipogenesis), including Rb phosphorylation. I hypothesized that MacCM induced alterations in cyclins and/or cyclin-dependent kinases (CDKs) were responsible for impairing Rb phosphorylation. My first objective was to assess the effect of J774A.1-MacCM on CDK4, CDK2, and their regulatory cyclins. Murine 3T3-L1 preadipocytes were differentiated with control medium or J774A.1-MacCM. Expression of cyclin D and A was inhibited by J774A.1-MacCM. Inhibition of cyclin A expression was associated with reduced differentiation-induced CDK2 activity. My second objective was to assess the expression patterns of cell cycle proteins in differentiating human abdominal subcutaneous preadipocytes, which do not undergo clonal expansion in culture. Cyclin E expression increased with differentiation. THP-1-MacCM (a human macrophage cell line) further enhanced this increase. My studies suggest MacCM leads to alterations in cyclin/CDK regulation during adipogenesis in murine and human preadipocyte models.

Les lignées clonales sont un outil puissant pour une meilleure caractérisation génétique des organismes utilisés en recherche. En utilisant les techniques de reproduction uniparentale, de telles lignées peuvent être développées chez les poissons en seulement deux générations. L'androgenèse avait déjà été réalisée chez plusieurs espèces d'eau douce et a été tentée chez le loup, une espèce d'intérêt économique et scientifique. Le faible nombre d'individus contenant effectivement uniquement le patrimoine génétique du père, après irradiation des œufs aux UV, a soulevé des questions quant à la présence de composés photoprotecteurs dans les œufs pélagiques. Les acides aminés de type mycosporines et le gadusol ont été recensés dans de nombreux organismes marins et d'eau douce, mais leur présence n'a pu être reliée au comportement flottant ou coulant des œufs. Tandis que le gadusol apparaissait en plus grande quantité dans les œufs marins flottants, ce résultat était beaucoup moins clair en eau douce et le type de composés photoprotecteurs semble lié à la phylogénie. D'autres études sur les mécanismes de photoréparation de l'ADN pourraient éclairer les résultats obtenus dans les expériences d'androgenèse. La gynogenèse avait déjà été reportée chez le loup mais le grand nombre d'individus méiotiques contaminant les descendances homozygotes produites a conduit à valider des marqueurs microsatellites efficaces à distinguer les individus mitotiques. De plus, un retard de développement des gonades a été observé, augmentant les difficultés à obtenir les lignées clonales. La grande variabilité observée entre les individus quant au succès des reproductions uniparentales a montré que la caractérisation et la qualité des gamètes sont un préalable indispensable. / Clonal lines are a powerful scientific tool for improved genetic characterization of organisms used in research. Inbred fish lines can be produced in only two generations using uniparental reproduction techniques. Androgenesis, achieved with variable success in several freshwater species, has been attempted in the European sea bass (Dicentrarchus labrax L), a marine fish of commercial and scientific interest. The low yields of progenies inheriting only the paternal genome after UV-irradiation of eggs led to considerations on the occurrence of UV screening compounds in pelagic eggs. Mycosporine-like amino acids and gadusol were found in many marine and freshwater organisms, but their occurrence in fish eggs was not clearly related to a behavioral pattern and while gadusol appeared in higher proportions in pelagic marine eggs compared to benthic species, this statement did not apply in freshwater, and moreover the kind of compounds was related to phylogeny. Further studies on DNA photorepair could enlighten hypotheses to understand the mechanisms underlying the disparate results obtained in inducing androgenesis in different fish species. Gynogenesis was reported successful to produce clonal founders in the sea bass, but high numbers of meiotic individuals contaminating fully homozygous progenies highlighted the need for efficient DNA markers to distinguish mitotic gynogenetic individuals. Furthermore, gonad development was highly delayed in gynogenetic progenies enhancing the difficulties to produce clonal lines. A high variability between individuals in the success of uniparental reproduction brought out gamete characterization and quality as a prerequisite.

Multiple Myeloma (MM) is the second most common hematological malignancy, and although patient outcomes have significantly improved since the introduction of autologous stem cell transplantation (ASCT) and novel pharmacological agents such as immunomodulators (IMID), proteasome inhibitors (PI), and monoclonal antibodies (mAb), the disease remains incurable. The pathological complexity of MM results from accumulating mutations in clonal populations of malignant B-cells, which are cytogenetically heterogenous and selectively sensitive to different therapeutic agents. Drug regimens therefore include a diverse combination of therapeutics designed to target specific pathways to inhibit cell proliferation. Recent advances in genomic analytics and novel pharmacological agents potentially allow for more targeted treatments which improve patient outcomes and frequency of remission with minimal adverse effects. Only recently have studies began to correlate an increased understanding of the many subtypes of MM with optimal treatment regimens, and practices such as “Direct to Drug” screening can give clinicians a look at a patient’s likely response to a combination of drugs. By incorporating emerging pharmaceutical agents into studies based on patient characteristics, the management of MM is making incremental strides towards a more targeted treatment paradigm.

京都大学 / 0048 / 新制・課程博士 / 博士(理学) / 甲第22286号 / 理博第4600号 / 新制||理||1660(附属図書館) / 京都大学大学院理学研究科生物科学専攻 / (主査)教授 工藤 洋, 教授 田村 実, 准教授 高山 浩司 / 学位規則第4条第1項該当 / Doctor of Science / Kyoto University / DGAM