Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (p. 118-122).

Thesis (Ph. D.)--University of Washington, 2004. / Vita. Includes bibliographical references (p. 126-132).

Thesis (Ph. D.)--University of Chicago, Department of Physics, December 1999. / Includes bibliographical references. Also available on the Internet.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (p. 252-274).

RNA interference (RNAi) is a post-transcriptional gene silencing process elicited by double-stranded RNA, such as micro-RNA (miRNA) and small interfering RNA (siRNA). They are 18-25 nucleotide long, small non-coding RNAs acting as critical regulators in eukaryotic genome expression. They play an important role in regulating a wide range of biological processes such as cell cycle control, differentiation, aging and apoptosis. However, their role in supporting skeletal development and bone homeostasis is still poorly understood. Osteoporotic fractures constitute a tremendous and growing problem in our ageing populations, with an annual incidence of approximately 60000 osteoporotic fractures in Sweden. Osteoporosis is referred as the “Silent epidemic” because bone loss is gradual and a basically symptomless development until a fracture occurs. Results presented in this thesis provide a novel insight into crucial roles of   miRNAs in regulating bone homeostasis. The initial aim for the thesis was to perform global miRNA expression profiling in human bone cells, and to correlate these levels to global mRNA levels. We identified and functionally characterized several miRNAs that were differentially expressed and acted in important bone signaling pathways such as the Wnt and BMP pathways. These miRNAs included hsa-miR-29b, hsa-miR-30c2 and hsa-miR-125b, which we found targeting genes highly relevant to bone metabolism e.g. COL1A1, SPARC, RUNX2, BGLAP and FRZB. Thereafter, the effect on the microRNAome upon external stimuli (e.g., Dexamethasone and Parathyroid hormone) was assessed by SOLiD sequencing. We observed a substantial difference in the expression of miRNAs between PTH and DEX treated cells. Understanding the changes in miRNAome in human bone cells under different conditions could provide new insight in bone remodeling, specifically differentiation and functional properties of osteoblasts. Based on these studies, we furthermore identified Dlx5 as potential common target of miR-203 and miR-320b and these miRNAs negatively regulate BMP-2-induced osteoblast differentiation. To activate the RNAi pathway, siRNA or miRNA molecules must be conveyed into the cytoplasm of target cells. Since challenges in cellular delivery of these small silencing RNA molecules so far have limited their clinical utility, we developed a new siRNA design that demonstrates a novel carrier-free cellular delivery. This development could potentially have a major impact in RNAi therapeutics. In conclusion, this thesis provides novel insight of miRNAs that play a major role in the regulation of bone remodeling and differentiation and functional properties of osteoblasts. Our findings may have diagnostic and/or therapeutic implications in disorders of bone metabolism.

The T2K experiment is a long-baseline neutrino oscillation experiment, in which a muon neutrino beam is directed over a 295 km baseline from the J-PARC facility to the Super-Kamiokande detector. This allows neutrino oscillation to be studied in two channels: disappearance of &nu;_&mu; and appearance of &nu;_e. The T2K beam can be run either in neutrino or antineutrino configuration, for a neutrino beam which is predominantly composed of &nu;_&mu; or anti -&nu;_&mu; respectively. This thesis presents the first oscillation analysis to combine neutrino-mode and antineutrino-mode data sets, giving the first ever sensitivity to the CP-violating phase &delta;_CP from T2K data alone, as well as the most precise T2K measurement of the other neutrino oscillation parameters. The analysis uses a Markov Chain Monte Carlo method to construct a sample from the Bayesian posterior distribution, from which the oscillation parameters of interest and their uncertainties are estimated. Data samples from the T2K near detector, ND280, are fit simultaneously with data from the far detector in order to reduce the uncertainty in the far-detector prediction. When fitting the T2K data alone, the best-fit oscillation parameter values are sin² &theta;₂₃= 0.519^+0.031<sub style='position: relative; left: -2.8em;'>-0.059</sub>, sin² &theta;₁₃= 0.0257^+0.0106<sub style='position: relative; left: -3.2em;'>-0.0440</sub>, &Delta;m²<sub style='position: relative; left: -0.4em;'>32</sub>= 2.54^+0.12<sub style='position: relative; left: -2em;'>-0.10</sub> x 10^-3eV²,and &delta;_CP=-1.91 rad. The 90&percnt; credible interval for &delta;_CP excludes values around &pi;/2: &delta;_CP &notin; [0.38, 2.60] rad. When fitting the T2K data with a constraint on sin² &theta;₁₃ from measurements by reactor experiments, the best-fit oscillation parameter values are sin² &theta;₂₃= 0.519^+0.061<sub style='position: relative; left: -2.8em;'>-0.029</sub>, sin² &theta;₁₃= ^+0.0014<sub style='position: relative; left: -3.2em;'>-0.0014</sub>, Dm²<sub style='position: relative; left: -0.4em;'>32</sub>= 2.54^+0.12<sub style='position: relative; left: -2em;'>-0.12</sub> x 10^-3eV², and &delta;_CP=-1.72 rad. The 90&percnt; credible interval for &delta;_CP contains values &delta;_CP &isin; [-3.10,-0.17] rad, excluding the CP-conserving values 0 and ± &pi; at 90&percnt; probability.

Using 3 fbˉ1 of proton-proton collisions, collected at centre-of-mass energies of √s = 7 and 8 TeV by the LHCb detector, several measurements of charmless B⁰s meson decays are made. A search is also performed for a highly suppressed B⁰ decay. First, the branching fraction of the B0⁰s→ ØØ decay is measured to be B(B⁰s→ ØØ) = (1.84 ± 0.05(stat) ± 0.07(syst) ± 0.11(fs=fd) ± 0.12(norm))X10ˉ⁵; where the third and fourth uncertainties arise from the fragmentation fraction fs/fd and the branching fraction of the normalisation mode. This represents a factor of five reduction in the statistical uncertainty compared to the previous best measurement. An upper limit on the branching fraction of the mode B⁰→ ØØ is set at B(B⁰→ ØØ) < 2.8 X 10ˉ⁸ (90% CL). This is a factor of seven improvement over the previous best measurement. An amplitude analysis of the B⁰s→ ØK+Kˉ decay is performed, wherein first observations of the decay modes B⁰s→ Øf´2(1525) and B⁰s→ ØØ(1680) are made. The branching fraction of the B⁰s→ Øf´2(1525) decay is measured to be B(B⁰s→ Øf´2 (1525)) = (1.63 ± 0.18(stat) ± 0.12(syst) ± 0.29(model) ± 0.17(norm)) X 10ˉ⁶; where the 'model' uncertainty arises from the choice of amplitude model. The longitudinal polarisation fraction of the decay B⁰s→ Øf´2(1525) is measured to be F0 = (86.6 ± 3.4 ± 0.8 ± 2.0 (model))%.

Made available in DSpace on 2015-03-26T13:54:53Z (GMT). No. of bitstreams: 1 texto completo.pdf: 443172 bytes, checksum: 25810212202085897d024a4831c5557d (MD5) Previous issue date: 2008-07-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The present study involved two experiments that aimed to evaluate nutrient intake and total digestibility, pH and ruminal ammonia concentration (first experiment) as well as the productive performance of beef cattle (second experiment) that were fed with diets containing soybean (SS) 0; 25; 50; 75; 100%. The rate forage:concentrate was 60:40, based on the dry matter. In the first experiment, five HxZ crossbred animals fistulated in the rumen and abomasum that had an initial weight of about 390kg were used. The animals were distributed in a 5x5 Latin square. Chromc oxide was used to estimate the fecal excretion. The intakes of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF), nonfiber carbohydrates (NFC), total carbohydrates (TC) and total digestible nutrients (TDN), expressed as kg.dia-1, showeda decreasing linear behavior while the intakes of EE and CP adjusted to the linear response plateau (LRP) model, thus estimating maximum values for the rates of 84 and 57% of corn silage in the forage, respectively. Diet effects were not detected (P>0.05) on the total apparent digestibility of DM, OG, CP, NDF and TC they revealed and average of 66.96; 69.49; 63.49; 57.60; 82.34 and 71.10%, respectively. On the other hand, EE aparent digestibility increased linearly following the corn silage rates. Quadratic effect of time collection for the ammonia concentration and the ruminal pH was observed where the maximum and minimal values 17.42 mg/dL and 6.23 mg/dL at 2,54and 4,27 hours were respectively estimated after feeding. In the second experiment, 35 anerolados and emasculated animals initially weighing 370 kg were used.The animals were distributed into a randomized block design with five treatments and seven repetitions. The experiment, which lasted 99 days long, was divided into a 15-day adaptation period and three 28-day periods. Five animals that were used as references were killed after the adaptation period in order to estimate the carcass gain. For estimating fecal excretion, ingestible neutral detergent fiber (iNDF) was used as an indicator, after the 240 hours "in situ" incubation. The intakes of NFC and CHO decreased linearly (P<0.05) according to the soybean silage rates in the diets, while the EE intake adjusted to a LRP model, with maximum value at the 89.55% rate of soybean silage in the forage. Lower intakes (P<0.05) of NCF and TC were observed for the diets that had higher rates of soybean silage than corn silage. Intakes for the other nutrients, expressed as kg/day, were not influenced by the diets. Weight and carcass gain, carcass yield and food conversion were not influenced by the diets either, they showed average values of 1.12 kg.day-1, 0.67 kg.day-1, 53.00% and 8.05, respectively. Soybean silage may be used as the single forage source in diets for anerolados cattle since it does not compromise the cattle productive performance or the ruminal fermentation patterns. However, its use may become and interesting alternative when associated to corn silage. / O presente estudo envolveu dois experimentos e foi realizado objetivando-se avaliar o consumo e a digestibilidade total dos nutrientes, o pH e a concentração de amônia ruminal (experimento1), bem como o desempenho produtivo de bovinos de corte (experimento 2) alimentados com dietas contendo 0; 25; 50; 75; 100% de silagens de soja (SS). A relação volumoso:concentrado foi de 60:40, com base na matéria seca. No primeiro experimento, foram utilizados cinco animais mestiços Holandês × Zebu, fistulados no rúmen e no abomaso, com peso inicial médio de 390 kg, distribuídos em um quadrado latino 5 × 5. Usou-se o óxido crômico para estimar a excreção fecal. Os consumos de matéria seca (MS), matéria orgânica (MO), proteína bruta (PB), fibra em detergente neutro (FDN), carboidratos nãofibrosos (CNF), carboidratos totais (CHOT) e nutrientes digestíveis totais (NDT), expressos em kg.dia-1, apresentaram comportamento linear decrescente, enquanto os consumos de EE e PB ajustaram-se ao modelo linear com resposta platô (LRP), estimando-se valores máximos para os níveis de 84 e 57% de silagem de soja no volumoso, respectivamente. Não foi detectado efeito (P>0,05) das dietas sobre as digestibilidades aparentes totais de MS, MO, PB, FDN, CNF, CHOT, cujas médias foram de 66,96; 69,49; 63,49; 57,60; 82,34; 71,10%, respectivamente. Por sua vez, a digestibilidade aparente do EE aumentou linearmente com os níveis de silagem de soja. O tempo decoleta teve efeito quadrático sobre a concentração de amônia e o pH ruminal, que apresentaram valores máximo e mínimo de 17,42 mg/dL e 6,23 mg/dL às 2,54 e 4,27 horas após a alimentação, respectivamente. No experimento 2, foram utilizados 35 animais anerolados, castrados, com peso inicial de 370 kg, distribuídos em delineamento de blocos casualizados, com cinco tratamentos e sete repetições. O experimento teve duração de 99 dias, divididos em três períodos de 28 dias, após 15 dias de adaptação. Cinco animais- referência foram abatidos após o período de adaptação, para estimativa do ganho de carcaça. A estimativa da excreção fecal foi feita utilizando-se a fibra em detergente neutro indigestível (FDNi) como indicador, após incubação in situ por 240 horas. Os consumos de CNF e carboidratos totais (CHOT) decresceram linearmente (P<0,05) com os níveis de silagem de soja nas dietas, enquanto o consumo de EE ajustou-se ao modelo LRP, com valor máximo para o nível de 89,55% de silagem de soja no volumoso. Os menores consumos de CNF e CHOT (P<0,05) foram observados nos animais mantidos com as dietas com maiores proporções de silagem de soja. Os consumos dos demais nutrientes, em kg/dia, não foram influenciados pelas dietas. O ganho de peso, ganho de carcaça, rendimento de carcaça e a conversão alimentar também não foram influenciados pelas dietas e apresentaram médias de 1,12 kg.dia-1, 0,67 kg.dia-1, 53,00% e 8,05, respectivamente. A silagem de soja pode ser utilizada como fonte única de volumoso em dietas para bovinos anelorados, uma vez que não compromete o desempenho produtivo e o padrão de fermentação ruminal desses animais. No entanto, proporciona melhores resultados se utilizada em associação à silagem de milho.

Orientador: Vicente Pleitez / Banca: Ricardo Jorge González Felipe / Banca: Orlando Luis Goulart Peres / Banca: José Antonio Martins Simões / Banca: Dionizio Bazeia Filho / Resumo: A construção de modelos de unificação continua sendo de grande interesse fenomenológico. As teorias baseadas no grupo de simetrias SU (5), apesar de terem suas versões mínimas, com apenas um multipleto da representação fundamental e um decupleto fermiônicos, praticamente excluídas pelos dados experimentais, continuam sendo de grande interesse desse ponto de vista. Modelos experimen- talmente plausíveis, onde o decaimento do próton é evitado pela separação dos setores de léptons e quarks do Modelo Padrão (MP) em diferentes multipletos, em conjunto com a implementação de si- metrias cíclicas discretas, 'dzeta IND. n', poderiam explicar o porquê do próton ter se mantido estável até a idade atual do universo. De fato, ao mesmo tempo que as simetrias cíclicas protegem o próton de operadores que induzem seu decaimento, elas são naturalmente implementadas quando o problema de CP forte é resolvido via áxion invisível, mantendo, ao mesmo tempo, o áxion livre de possíveis efeitos gravitacio- nais para uma simetria cíclica 'dzeta IND. 13'. Entretanto, a fenomenologia completa para os modelos SU (5) deve ser cuidadosamente formulada, uma vez que deseja-se que qualquer nova teoria que venha comple- mentar o MP implemente, por exemplo, massa para todas as partículas conhecidas. Todavia, a simples estruturação da teoria pode não ser suficiente para a solução das massas dos neutrinos. De fato, os modelos mais simples, enquanto explicam os dados de oscilação dos neutrinos, geram problemas para o decaimento do próton, uma vez que introduzem fases da simetria 'dzeta IND. n' que geram operadores perigosos. A solução para esse problema é a inclusão de massas apenas pela quebra espontânea de uma simetria adicional. O termo de Yukawa direto na Lagrangiana é acoplado à uma U{1) quiral adicional, que é espontaneamente quebrada pelo singleto acoplado aos neutrinos ativos. Mostra-se que... / Abstract: The construction of unification models remain of great phenomenological interesting. The theories based on SU(5) symmetry group, despite having their minimalist versions, with only a fermionic mul- tiplet of fundamental representation an one decuplet, almost excluded from experimental data, remain of great interest from this point of view. Experimentally plausible models, where the proton decay is avoided by separation of Standard Model (SM) lepton and quark sectors in different multiplets, together with the implementation of cyclic symmetries, 'dzeta IND. n', could explain why the proton has remained stable until the current age of the universe. Indeed, while protecting the proton from operators that induces their decay, discrete symmetries are naturally implemented when the strong CP problem is solved by invisible axion, keeping, at the same time, the axion protected against possible semi-classical gravitational effects for a cyclical 'dzeta IND. 13' symmetry. However, the complete phenomenology for SU{5) models should be carefully formulated, once one want to have that any new theory, which comple- ment the SM, implement, for example, mass for all known particles. However, the simple structure of the theory may not be sufficient for the solution of the neutrino masses. In fact, the simplest models, while explaining the data of neutrino oscillation, create problems for proton decay, since introduces 'dzeta IND. n' symmetry charges that generate dangerous operators. The solution to this problem is the inclusion of mass term only by additional symmetry. The direct Yukawa term in the Lagrangian is coupled to an additional chiral U(1) symmetry, which is spontaneously broken by the singlet coupled to the active neutrinos. It shows that when these symmetries are implemented locally, complementing the gauge structure of the SM, the seesaw mechanism of types I and III can not be simultaneously... / Doutor

Made available in DSpace on 2014-06-12T15:54:00Z (GMT). No. of bitstreams: 2 arquivo5189_1.pdf: 2087717 bytes, checksum: 1718a5a73d49c7021a82fe893f78dbfe (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2006 / A Ressonância Magnética Nuclear (RMN) e a Cromatografia em fase gasosa acoplada à espectrometria de massas (GC/MS) fornecem informações sobre a estrutura química de substâncias, bem como, sobre a constituição qualitativa e quantitativa de amostras sem a necessidade de padrões de análise ou pré-tratamento de amostras. Uma jovem estudante de 19 anos morreu de forma súbita após inalar um aerossol cujo componente principal, o 1-1-dicloro-1-fluoretano (HCFC-141b), foi identificado por meio de RMN e GC/MS. Nas vísceras e no sangue da vítima não foram detectados venenos, álcool ou outras drogas. Os resultados dos exames necroscópico, toxicológico e microscópico não excluem a hipótese de morte por inalação do HCFC-141b e estabelecem dois mecanismos de morte como viáveis: asfixia e arritmia cardíaca. As análises realizadas e os dados do inquérito policial suportam probabilisticamente a hipótese de morte por arritmia cardíaca fatal que pode ter ocorrido de forma associada com asfixia. Algumas técnicas de RMN como ¹H, ¹³C, Distortionless Enhancement by Polarization Transfer (DEPT), Cross-Polarization and Magic Angle Spinning (CP/MAS) and Dipolar Dephasing (DD) foram usadas para caracterizar amostras de cocaína. Os deslocamentos químicos dos espectros obtidos foram atribuídos por meio de Heteronuclear Correlation (HETCOR) e Heteronuclear Multiple Bond Correlation (HMBC). As amostras foram primariamente caracterizadas por espectrometria de massas cujos espectros foram comparados com dados obtidos na literatura. Os resultados obtidos indicam que ¹H, ¹³C e DEPT são técnicas bastante eficientes para caracterização de amostras ilícitas de cocaína no estado líquido e que por meio de CP/MAS e CP/MAS/DD RMN é possível distinguir o crack do cloridrato de cocaína de uma maneira não destrutiva. Portanto, a RMN é uma excelente ferramenta para caracterização de amostras ilícitas de cocaína