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No. of bitstreams: 1 souza_icg_dr_bot.pdf: 1921899 bytes, checksum: cce906c3ffd78242151254d344282fb0 (MD5) Previous issue date: 2016-07-29 / Este trabalho teve como objetivo propor um novo método de melhoramento para Eucalyptus visando obter materiais genéticos para plantio operacional, por meio dos seguintes estudos: a) avaliar o ganho em produtividade volumétrica de madeira na seleção aplicada numa população clonal de Eucalyptus grandis oriunda da mistura de clones selecionados em progênies de polinização controlada (irmãos completos); b) avaliar a variabilidade genética a partir de marcadores moleculares microssatélites na população clonal selecionada, quando submetida a diferentes condições ambientais e níveis de seleção. Com as sementes obtidas no cruzamento controlado foi instalado um plantio experimental na Fazenda Ribeirão Grande (município de Salesópolis/SP) de propriedade da Suzano Papel e Celulose no ano de 2001. Aos 6 anos de idade foi realizada a seleção de árvores superiores (clones) com base nos caracteres silviculturais e densidade básica da madeira. Foram selecionados 57 clones (irmãos completos), os quais foram propagados vegetativamente, misturados para a formação do minijardim clonal e posterior produção de mudas para plantio operacional na empresa. Este material genético foi recomendado para plantio em 2010. Para este trabalho foram selecionadas três áreas de plantio operacional desse material genético, denominado aqui população clonal inicial com idade de 3,3 a 3,5 anos. Em cada local foram estabelecidas 3 parcelas de 100 árvores e realizada a avaliação silvicultural das árvores. Com os dados de DAP foi feita a ordenação dos valores (maior para o menor) e selecionadas as 40 árvores com maiores valores de DAP e boa forma do tronco, e as 15 árvores com menores valores de DAP totalizando 405 árvores. O perfil genético por meio de marcadores moleculares microssatélites foi realizado para as árvores selecionadas, clones que formam a população clonal inicial e clones genitores. O Incremento Médio Anual com casca aos 7 anos de idade (IMA7; m3/ha/ano) foi estimado por parcela e níveis de seleção de 10%, 20%, 30% e 40%. Com o perfil genético das 40 árvores selecionadas por parcela obteve-se os clones presentes na seleção, como também, os clones presentes nas 15 árvores de comportamento silvicultural inferior. Os resultados mostram que o método de seleção e melhoramento em populações clonais de E. grandis é promissor, diminuindo o tempo de obtenção de materiais genéticos para plantio operacional. Os níveis de seleção de 30% e 40% são os mais indicados para conservar um bom número de clones e obter ganhos em produtividade em curto prazo. / The objective of this work was to propose a new method for breeding to Eucalyptus in order to obtain genetic material for commercial planting by the following studies: a) evaluate the gain, in wood volumetric productivity, in the selection applied to a clonal population of Eucalyptus grandis coming from a selected clonal mixture obtained in cross-pollinated progenies (full sibling); b) evaluate the genetic variability using microsatellite markers obtained in a selected clonal population when subjected to different environmental conditions and levels of selection. Using the seeds obtained in the hand pollination, an experimental plantation was established, in 2001, at the Ribeirão Grande Farm in the municipality of Salesópolis, State of São Paulo, Brazil, owned by Suzano Pulp and Paper. At the age of 6 years old, a selection of superior trees (clones) was carried out based on silvicultural traits and wood density. Fifty-seven clones (full-siblings) were selected, propagated using vegetative technique and used for mixed plantation in a clonal garden to subsequent production of seedlings for commercial planting. This genetic material (initial clonal population) was recommended for planting in 2010. For this study, three commercial areas planted with these genetic materials at ages ranging from 3.3 to 3.5 years old were selected. On each site, 3 plots of 100 trees were established and the silvicultural evaluation of trees was made. The trees were ordered using the diameter at breast height (DBH) data (highest to lowest) and the 40 trees with the highest DBH values and good shape of the trunk were selected, and the 15 trees with lower DBH values, totaling 360 trees. The genetic profile through microsatellite markers was conducted for selected trees, for clones that form the initial clonal population and for parental clones. The Annual Average Increase in shell to the 7 years old (IMA7; m3 / ha.year) was estimated per plot and 10, 20, 30 and 40% selection of levels. Throughout the genetic profile of the 40 selected trees per plot, the presented clones in the selection were obtained, but also, the clones present in 15 trees with lower silvicultural behavior. The results show that the method selection and improvement in clonal populations of E. grandis is promising, decreasing the time for obtaining genetic materials for commercial plantation. Check levels of 30 and 40% are the most suitable to preserve a good number of clones and to obtain short-term productivity gains.

Made available in DSpace on 2016-03-15T19:37:43Z (GMT). No. of bitstreams: 1 Luiz Antonio Carraro.pdf: 1226702 bytes, checksum: 3cef29694a4e26f233b0aae16da69cf0 (MD5) Previous issue date: 2012-02-16 / Universidade Presbiteriana Mackenzie / A container is a broadly used solution for the cargo storage to be transported between ports, playing a central role in international trade. Consequently, ships grew in size in order to maximize their container transportation capacity in each trip. Due to increasing demand, container terminals face the challenges of increasing their service capacity and optimizing the loading and unloading time of ships. Optimization problems, such as these, often present features that make it impossible to obtain closed analytical solutions, requiring iterative search procedures in high-dimensional spaces, or subject to a combinatorial explosion of possible solutions. This dissertation presents the proposal of a novel meta-heuristic based on the Clonal Selection Algorithm, named MRC, to minimize the number of reshuffles in operations involving piles of containers. The performance of the proposed model was evaluated through simulations and results comparison with those obtained by algorithms from the literature under the same test conditions. The results obtained show that MRC is competitive in terms of minimizing the need of reshuffles, besides presenting a reduced processing time compared with models of similar performance. / A utilização de contêineres é uma solução amplamente adotada para o armazenamento da carga a ser transportada entre portos, tornando-se de grande importância no comércio internacional e, consequentemente, navios cresceram de tamanho com o objetivo de transportar a maior quantidade possível de contêineres em cada viagem. Devido à crescente demanda, terminais de contêineres enfrentam os desafios de aumentar a sua capacidade de atendimento e otimizar os tempos de carregamento e descarregamento de navios. Problemas de otimização como estes geralmente apresentam características que inviabilizam a obtenção de soluções analíticas fechadas, requerendo processos iterativos de busca em espaços de dimensão muitas vezes elevada, ou ainda sujeitos a explosão combinatória de possíveis soluções. Esta dissertação apresenta a proposta de uma meta-heurística bioinspirada baseada no Algoritmo de Seleção Clonal para a minimização de rearranjos em operações que envolvem pilhas de contêineres, denominado MRC. O desempenho do algoritmo foi avaliado por meio de simulações e comparação dos resultados com os obtidos por algoritmos da literatura sob as mesmas condições de teste. Os resultados obtidos permitem concluir que o MRC possui resultados competitivos em termos de minimização de rearranjos, além de apresentar um tempo de processamento reduzido quando comparado aos modelos tradicionalmente empregados na solução desse tipo de problema.

Despite the two-fold advantage to asexual reproduction and its prevalence in a variety of organisms, sexual reproduction is prevalent across all taxa. The maintenance of two sexes is required to ensure genetic diversity and to prevent “evolutionary dead ends,” especially in clonal organisms. Many mechanisms have been proposed for the maintenance of two sexes, ranging from environmental variation and stochasticity, parasites and predators, and mutation rates. Spatial configuration, the size and location of populations with respect to other populations, can allow two competitors to coexist when one would normally be lost. This is especially important when the two competitors are the two sexes. In the clonal organism Marchantia inflexa, I determined that spatial configuration of populations can directly influence the maintenance of both sexes in a population and in an aggregate of populations (a metapopulation) using a combination of theoretical models and field studies. Based on field studies, population size has a significant influence on whether a subpopulation will contain both sexes, with populations smaller than 1m2 being more likely to contain only one sex while populations greater than 1m2 are more likely to contain both sexes. Based on mathematical models, the spatial arrangement of subpopulations within a metapopulation can greatly influence whether a metapopulation maintains both sexes as well as whether the metapopulation persists once one sex has been lost. Field data suggest that distance to nearest neighbor, a measurement of spatial arrangement, influences the maintenance of the sexes within subpopulations, but could affect maintenance differently depending on the metapopulation identity. In some metapopulations, both sexes are maintained when the nearest neighbor is close, while in other streams, one sex is lost when the nearest neighbor is close. When mathematical models are used to explicitly simulate natural metapopulations, the mathematical model predicts the observed sex ratios in one metapopulation, predicts the observed bias in another metapopulation, and fails to predicted observed values in two other metapopulations. Understanding spatial configuration helps us understand the maintenance and loss of sex, but other factors, such as environmental differences, may be required to accurately predict which sex will be lost.

Trade-offs in resource allocation underlie key life history traits of organisms. My dissertation focuses on the size-number trade-off in clonal broods of offspring using the polyembryonic wasp, Copidosoma bakeri parasitizing immature stages of the moth Agrotis ipsilon. I aim to characterize responses of wasp brood size and individual body mass by manipulating the environments in order to understand the allocation pattern in the size number trade-off. In reviewing the functional forms of trade-off relationships in relation to resource constraints, I distinguish among three main trade-off types based on graphical representations of the relationship between the trade-off variables: linear, convex (inverse), and concave. The size-number trade-off in C. bakeri shows convex relationship. Characteristics of the trade-off are sex specific: female broods have larger body mass but smaller brood size than do male broods. When food intake of the host was increased, the trade-off between wasp body mass and brood size for both sexes shifts toward both higher wasp brood size and higher body mass. When the host has better access to food late in development, the size-number trade-off curve moves up and to the right on the graph. However, the trait combinations shift along the same trade-off curve toward greater wasp body mass but smaller brood size when the host development time is shorter due to more resources in early in development. I also investigate temperature effects on the size-number trade-off. C. bakeri brood size significantly increases with high temperature early in host development. There is no shift in the allocation pattern of the size-number trade-off with temperature. Finally, I test effects of body mass on longevity, fecundity, and mating competitiveness of C. bakeri. Larger body size increases female longevity, and mated females produce more eggs than unmated females. There are no significant relationships between male body mass and longevity or mating competitiveness. Mating reduces male longevity independent of body mass. The different impact of body mass on fitness between male and female wasps suggests the observed sex-specific allocation patterns of the size-number trade-off. Implications of the experiments and possible follow-up work are discussed.

A new adaptive learning Artificial Immune System (AIS) based committee machine is developed in this thesis. The new proposed approach efficiently tackles the general problem of clustering high-dimensional data. In addition, it helps on deriving useful decision and results related to other application domains such classification and prediction. Artificial Immune System (AIS) is a branch of computational intelligence field inspired by the biological immune system, and has gained increasing interest among researchers in the development of immune-based models and techniques to solve diverse complex computational or engineering problems. This work presents some applications of AIS techniques to health problems, and a thorough survey of existing AIS models and algorithms. The main focus of this research is devoted to building an ensemble model integrating different AIS techniques (i.e. Artificial Immune Networks, Clonal Selection, and Negative Selection) for classification applications to achieve better classification results. A new AIS-based ensemble architecture with adaptive learning features is proposed by integrating different learning and adaptation techniques to overcome individual limitations and to achieve synergetic effects through the combination of these techniques. Various techniques related to the design and enhancements of the new adaptive learning architecture are studied, including a neuro-fuzzy based detector and an optimizer using particle swarm optimization method to achieve enhanced classification performance. An evaluation study was conducted to show the performance of the new proposed adaptive learning ensemble and to compare it to alternative combining techniques. Several experiments are presented using different medical datasets for the classification problem and findings and outcomes are discussed. The new adaptive learning architecture improves the accuracy of the ensemble. Moreover, there is an improvement over the existing aggregation techniques. The outcomes, assumptions and limitations of the proposed methods with its implications for further research in this area draw this research to its conclusion.

<p>Regeneration is a remarkable feat of developmental regrowth and patterning. The blastema is a mass of progenitor cells that enables complete regeneration of amputated salamander limbs or fish fins. Despite years of study, methodologies to identify and track blastemal cell progenies have been deficient, restricting our understanding of appendage regeneration at a cellular and molecular level. To bridge this knowledge gap, gene expression analysis, the generation of transgenic and mutant zebrafish, qualitative and quantitative analyses, morphological measurements, and chemical treatments were used to assess molecular and cellular processes involved in fin regeneration. Two main findings arose from these methods. The first provides evidence that connective tissue progenitors are rapidly organized into a scalable blueprint of lost structures, and that amputation stimulates resident cells to reset proximodistal positional information. The second identifies a fibroblast subpopulation near uninjured fin joints that contributes to the blastemal progenitor population. These findings reveal insights on cellular and molecular mechanisms of appendage regeneration and provide a basis for work exploring how cells in an adult vertebrate bone appendage coordinately rebuild a new structure.</p> / Dissertation

Canyon shaped reservoirs are characteristic by specific environmental horizontal gradients, so they enable existence of several species of the Daphnia longispina complex in one water body. Due to preference of distinct environmental conditions Daphnia species occurre in the different localities. The aim of my thesis was to analyze detail taxonomical and clonal structure of Daphnia longispina group by ten microsatellite markers on longitudinal gradient and compare it between two consecutive seasons. Simultaneously I received newly discovered divergent mitochondrial lineage from Želivka reservoir. It was confirmed, that the distribution of species and their hybrids in water reservoir was non-concidental and the taxonomic spatial distribution is in two consecutive seasons relatively constant. On the contrary the spatial and temporal distribution of clones was very heterogeneous. Clonal diversity in the interspecific hybrids was lower than in the coexisting parental species. This finding supports the hypothesis, that there exist reproductive barriers between parental genomes. Most of the clones were substantively variable, but several clones that occurred in both seasons in similar localities were found. It is possible that some clones are able of overwintering in hypolimnion and in the spring...

OBJECTIVE: Cardiovascular disease (CVD) remains the leading cause of death globally. Reducing high blood cholesterol, which is a dominant risk factor for CVD events, is an essential goal of medical treatment. Statins are known as first‐choice agents. However, clinical trials report that some statins increased the risk for type 2 diabetes (T2D). Our objective was to investigate the effect of different statins on insulin secretion and content from pancreatic β-cells after chronic and acute exposure and determine the underlying mechanisms. METHODS: The effects of simvastatin, lovastatin and pitavastatin on GSIS and content were studied in clonal pancreatic β-cells (INS-1 832/13) cultured in high glucose (12 mM). Insulin content and secretion were measured after chronic and acute incubation of statins using homogenous time-resolved fluorescence (HTRF) insulin assay kit (Cisbio). Intracellular Ca2+ was measured using fura-2 AM (Invitrogen). RESULTS: Simvastatin (25-200 nM) and lovastatin (50-200 nM) significantly inhibited GSIS and depleted insulin content in a dose-dependent manner after 72-hour exposure. When the secretion level was normalized for content, the inhibitory effect was not observed. Simvastatin (200 nM) also increased the amplitude of intracellular Ca2+ oscillations at low glucose, but this was not reflected in the amplitude of oscillatory insulin release. In contrast, pitavastatin (25-200 nM) did not affect GSIS and only decreased insulin content at the highest dose tested. CONCLUSION: Inhibition of GSIS by simvastatin and lovastatin could be due to depletion of insulin content. Decreased Ca2+ sensitivity may also contribute to inhibition of GSIS by simvastatin. Pitavastatin had less inhibitory effect on GSIS and insulin content as compared to simvastatin and lovastatin indicating that not all lipophilic statins have a detrimental impact on GSIS. We suggest that statins may have differential mechanistic effects on β-cells some of which may contribute to the risk of T2D.

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.