Thesis (Ph. D.)--University of Missouri--Columbia, 1999. / "December 1999." Typescript. Vita. Includes bibliographical references (leaves 137-154). Also available on the Internet.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 84-91).

Thesis (Ph. D)--University of Washington, 2000. / Vita. Includes bibliographical references (leaves 126-137).

Deformed epidermal autoregulatory factor 1 (DEAF1) is a transcription factor that binds to (T/C)TCG(G/T) half-sites and has been shown to be involved in human diseases of cancer, diabetes, depression and intellectual disorders. We used chromatin immunoprecipitation assays to assess endogenous levels of DEAF1 and RNA polymerase II occupancy on the promoter and 5'UTR of the DEAF1 gene. In exponentially growing HEK293 cells, low levels of DEAF1 bind to sequences between -718 and +232, with +1 marking the start of translation. Within 0.5 hr of treating the cells with 500 µM H2O2, DEAF1 occupancy is increased between 7-18 fold at B (-718/-569), -577/-444, C (-432/-299), D (-205/-112) and E(-97/17). There were no statistically significant changes in either RNA polymerase II phospho-serine 5 (RNA PolII pS5) or RNA polymerase II phospho-serine 2 (RNA PolII pS2) binding with H2O2 treatment compared to control. With media change, there is an increase in RNA PolII pS2 and pS5 occupancy at both a distal site -1462/-1326 and in the coding region at 133/232, while no significant change in DEAF1 occupancy was detected. DEAF1 occupancy at the DEAF1 promoter and 5'UTR are inversely correlated with RNA polymerase II occupancy, however, there were no measurable differences in DEAF1 RNA levels at 0.5 hr and 1 hr time points. In summary, these data indicate that there is increased occupancy of DEAF1 at its own promoter following stress, which inversely affects occupancy of RNA polymerase at proximal promoter and 5'UTR sites of the DEAF1 gene.

A flavomicina é um antimicrobiano registrado no Ministério da Agricultura do Brasil e que tem sido amplamente utilizado na avicultura e suinocultura. Seu uso foi intenso até o final dos anos 1990, sendo este reduzido posteriormente. Este produto está registrado como promotor de crescimento até a dose máxima de 2 ppm no Brasil. A definição desta dose provêm de trabalhos executados há muitas décadas e, portanto, é possível que as doses que otimizem seu uso sejam diferentes na avicultura atual, especialmente em um cenário de limitadas ferramentas para melhora de desempenho de frangos de corte. O objetivo deste trabalho foi reavaliar a utilização da flavomicina em frangos de corte a partir de um experimento dose-resposta. Foram utilizados 900 frangos de corte machos Cobb x Cobb 500 de um dia de idade, divididos em 5 tratamentos com 12 unidades experimentais cada. Os tratamentos foram constituídos de doses crescentes de flavomicina formuladas com níveis similares de EMAn e nutrientes baseados em níveis utilizados pela indústria brasileira: 0, 2, 4, 8 e 16 ppm de flavomicina. A avaliação de desempenho foi feita semanalmente até os 42 dias com avaliações de ganho de peso, conversão alimentar corrigida para mortalidade, consumo de ração e mortalidade. No período de 1 a 21 dias de idade, as dosagens ótimas calculadas pela equação de regressão foram de 16 ppm , 12,17 ppm e 14,68 ppm para consumo de ração, conversão alimentar e ganho de peso, respectivamente. De 22 a 42 dias, as dosagens foram de 10,31 e 9,83 ppm para conversão alimentar e ganho de peso. Pela equação de regressão do período acumulado de 1 a 42 dias, houve uma melhora crescente até o platô de inclusão de 10,9 e 10,14 ppm, para ganho de peso e conversão alimentar. O estudo nos mostra que, nas condições experimentais, o desempenho de modernas linhagens de frangos de corte podem ser maximizados de dosagens que variam de 9,83 até 16 ppm, acima dos padrões estabelecidos na legislação brasileira. / Flavophospholipol is an antimicrobial registered in Ministério of Agricultura of Brazil that has been widely used in poultry and swine production. Its use was intense until the end of 1990, which was later reduced. This product is registered as a growth promoter until the maximum dosis of 2 ppm in Brazil. These doses derived from research performed many decades ago and, therefore, this dose is likely to different from the one that optimizes broiler performance, especially in a scenario of limited tools for improvements of performance of broilers. The objective of this study was to reassess the use of flavophospholipol for broilers in a dose-response trial. A total of nine hundred male Cobb x Cobb 500 broilers were placed across five treatments with 12 replications each. The treatments consisted of increasing doses of flavophospholipol: 0, 2, 4, 8, 16 ppm. The diets were formulated with similar levels of metabolizable energy based on levels and nutrients used by the Brazilian industry. Performance evaluation was done weekly until the day 42. The evaluated parameters were: body weight gain, feed conversion ratio corrected for mortality, feed intake and percentage of mortality. In the period equation were 16 ppm, 12.17 ppm and 14.68 ppm for feed intake, feed conversion ratio and body weight gain, respectively. From 22 to 42 days, the estimated doses were 10.31 and 9.83 ppm for feed conversion and body weight gain. In the cumulative period from 1 to 42 days it was found an increased improvement until the plateau which was 10.9 and 10.14 ppm, for body weight gain and feed conversion ratio. The study showed that under the experimental conditions used, the performance of modern broilers could be optimized from 9.83 to 16 ppm, above the standards established by Brazilian legislation.

Master of Agribusiness / Department of Agricultural Economics / Kevin Dhuyvetter / Many companies use surveys to establish customer satisfaction metrics. This OEM has been using surveys to analyze customer satisfaction with their products, services, and distribution channel for several decades. Satisfaction metrics are established for the brand, product, and channel partners. The product metric is derived from a question on the survey asking customers how satisfied they are with the product. There are subsequent questions thereafter inquiring about satisfaction with specific functional areas of the product. It is common practice to use Partial Least Squares (PLS) regression analysis to evaluate what impacts the functional area questions have on the overall satisfaction question. The model results are used to understand what areas of the machine should be focused on to improve customers’ experiences with the machine. These results are compared to other data sources such as warranty, field reports, customer focus groups, etc. The results from these models are sometimes questioned based on what common intuition would suggest. Typically the top three drivers to the product metric are understandable, but there are often one or two key areas that do not make logical sense. The objective of this thesis was to understand whether PLS modeling is appropriate given the nature of customer survey data. Models were estimated using existing survey data on a specific model in the tractor product line. PLS models assume data are linear with no bounds. This in itself likely makes this type of model inappropriate for analyzing customer survey data. Responses are bounded on an 11 point scale from 0-10, however, the PLS model being non-bounded assumes there can be a score under 0 or over 10. The model also assumes a linear slope that would indicate each covariate answer 0-10 has the same level of effect on the response variable. This research has found that each covariate answer is in fact non-linear. For example, a customer answering a 2 to quality of manufacturing workmanship has a different impact on the overall satisfaction score than a customer who answers 8. Finally, this research discovered that the PLS models produce negative coefficients of significant value that are not reported to the enterprise. Binary and ordered logistic (logit) models were estimated as an alternative to PLS. Logistic models are non-linear and are commonly used to evaluate bounded data. Response data were separated into two groups based on Net Promoter Score (NPS) Methodology (Reicheld 2006). Using the NPS methodology, 0-6 scores are considered detractors, 7-8 scores are considered passives, and 9-10 scores are considered promoters. The logistic models demonstrate that the top two drivers to customer satisfaction scores are still quality of manufacturing workmanship and reliability/operational availability (similar to results of the PLS model). The unresolved problems question on the survey was included in the models and demonstrated that the predicted probability of a customer being a promoter is much higher in both binary and ordered logit models if no unresolved problems exist. Finally, the model found engine oil consumption remained negative and is statistically significant suggesting that even with the alternative modeling approach there still may be data issues related to the survey. It is recommended that the OEM implement logistic modeling for analyzing customer survey data. It is also recommended that a new survey design be constructed to eliminate issues with correlated data that can lead to spurious and unexplainable results.

A flavomicina é um antimicrobiano registrado no Ministério da Agricultura do Brasil e que tem sido amplamente utilizado na avicultura e suinocultura. Seu uso foi intenso até o final dos anos 1990, sendo este reduzido posteriormente. Este produto está registrado como promotor de crescimento até a dose máxima de 2 ppm no Brasil. A definição desta dose provêm de trabalhos executados há muitas décadas e, portanto, é possível que as doses que otimizem seu uso sejam diferentes na avicultura atual, especialmente em um cenário de limitadas ferramentas para melhora de desempenho de frangos de corte. O objetivo deste trabalho foi reavaliar a utilização da flavomicina em frangos de corte a partir de um experimento dose-resposta. Foram utilizados 900 frangos de corte machos Cobb x Cobb 500 de um dia de idade, divididos em 5 tratamentos com 12 unidades experimentais cada. Os tratamentos foram constituídos de doses crescentes de flavomicina formuladas com níveis similares de EMAn e nutrientes baseados em níveis utilizados pela indústria brasileira: 0, 2, 4, 8 e 16 ppm de flavomicina. A avaliação de desempenho foi feita semanalmente até os 42 dias com avaliações de ganho de peso, conversão alimentar corrigida para mortalidade, consumo de ração e mortalidade. No período de 1 a 21 dias de idade, as dosagens ótimas calculadas pela equação de regressão foram de 16 ppm , 12,17 ppm e 14,68 ppm para consumo de ração, conversão alimentar e ganho de peso, respectivamente. De 22 a 42 dias, as dosagens foram de 10,31 e 9,83 ppm para conversão alimentar e ganho de peso. Pela equação de regressão do período acumulado de 1 a 42 dias, houve uma melhora crescente até o platô de inclusão de 10,9 e 10,14 ppm, para ganho de peso e conversão alimentar. O estudo nos mostra que, nas condições experimentais, o desempenho de modernas linhagens de frangos de corte podem ser maximizados de dosagens que variam de 9,83 até 16 ppm, acima dos padrões estabelecidos na legislação brasileira. / Flavophospholipol is an antimicrobial registered in Ministério of Agricultura of Brazil that has been widely used in poultry and swine production. Its use was intense until the end of 1990, which was later reduced. This product is registered as a growth promoter until the maximum dosis of 2 ppm in Brazil. These doses derived from research performed many decades ago and, therefore, this dose is likely to different from the one that optimizes broiler performance, especially in a scenario of limited tools for improvements of performance of broilers. The objective of this study was to reassess the use of flavophospholipol for broilers in a dose-response trial. A total of nine hundred male Cobb x Cobb 500 broilers were placed across five treatments with 12 replications each. The treatments consisted of increasing doses of flavophospholipol: 0, 2, 4, 8, 16 ppm. The diets were formulated with similar levels of metabolizable energy based on levels and nutrients used by the Brazilian industry. Performance evaluation was done weekly until the day 42. The evaluated parameters were: body weight gain, feed conversion ratio corrected for mortality, feed intake and percentage of mortality. In the period equation were 16 ppm, 12.17 ppm and 14.68 ppm for feed intake, feed conversion ratio and body weight gain, respectively. From 22 to 42 days, the estimated doses were 10.31 and 9.83 ppm for feed conversion and body weight gain. In the cumulative period from 1 to 42 days it was found an increased improvement until the plateau which was 10.9 and 10.14 ppm, for body weight gain and feed conversion ratio. The study showed that under the experimental conditions used, the performance of modern broilers could be optimized from 9.83 to 16 ppm, above the standards established by Brazilian legislation.

A flavomicina é um antimicrobiano registrado no Ministério da Agricultura do Brasil e que tem sido amplamente utilizado na avicultura e suinocultura. Seu uso foi intenso até o final dos anos 1990, sendo este reduzido posteriormente. Este produto está registrado como promotor de crescimento até a dose máxima de 2 ppm no Brasil. A definição desta dose provêm de trabalhos executados há muitas décadas e, portanto, é possível que as doses que otimizem seu uso sejam diferentes na avicultura atual, especialmente em um cenário de limitadas ferramentas para melhora de desempenho de frangos de corte. O objetivo deste trabalho foi reavaliar a utilização da flavomicina em frangos de corte a partir de um experimento dose-resposta. Foram utilizados 900 frangos de corte machos Cobb x Cobb 500 de um dia de idade, divididos em 5 tratamentos com 12 unidades experimentais cada. Os tratamentos foram constituídos de doses crescentes de flavomicina formuladas com níveis similares de EMAn e nutrientes baseados em níveis utilizados pela indústria brasileira: 0, 2, 4, 8 e 16 ppm de flavomicina. A avaliação de desempenho foi feita semanalmente até os 42 dias com avaliações de ganho de peso, conversão alimentar corrigida para mortalidade, consumo de ração e mortalidade. No período de 1 a 21 dias de idade, as dosagens ótimas calculadas pela equação de regressão foram de 16 ppm , 12,17 ppm e 14,68 ppm para consumo de ração, conversão alimentar e ganho de peso, respectivamente. De 22 a 42 dias, as dosagens foram de 10,31 e 9,83 ppm para conversão alimentar e ganho de peso. Pela equação de regressão do período acumulado de 1 a 42 dias, houve uma melhora crescente até o platô de inclusão de 10,9 e 10,14 ppm, para ganho de peso e conversão alimentar. O estudo nos mostra que, nas condições experimentais, o desempenho de modernas linhagens de frangos de corte podem ser maximizados de dosagens que variam de 9,83 até 16 ppm, acima dos padrões estabelecidos na legislação brasileira. / Flavophospholipol is an antimicrobial registered in Ministério of Agricultura of Brazil that has been widely used in poultry and swine production. Its use was intense until the end of 1990, which was later reduced. This product is registered as a growth promoter until the maximum dosis of 2 ppm in Brazil. These doses derived from research performed many decades ago and, therefore, this dose is likely to different from the one that optimizes broiler performance, especially in a scenario of limited tools for improvements of performance of broilers. The objective of this study was to reassess the use of flavophospholipol for broilers in a dose-response trial. A total of nine hundred male Cobb x Cobb 500 broilers were placed across five treatments with 12 replications each. The treatments consisted of increasing doses of flavophospholipol: 0, 2, 4, 8, 16 ppm. The diets were formulated with similar levels of metabolizable energy based on levels and nutrients used by the Brazilian industry. Performance evaluation was done weekly until the day 42. The evaluated parameters were: body weight gain, feed conversion ratio corrected for mortality, feed intake and percentage of mortality. In the period equation were 16 ppm, 12.17 ppm and 14.68 ppm for feed intake, feed conversion ratio and body weight gain, respectively. From 22 to 42 days, the estimated doses were 10.31 and 9.83 ppm for feed conversion and body weight gain. In the cumulative period from 1 to 42 days it was found an increased improvement until the plateau which was 10.9 and 10.14 ppm, for body weight gain and feed conversion ratio. The study showed that under the experimental conditions used, the performance of modern broilers could be optimized from 9.83 to 16 ppm, above the standards established by Brazilian legislation.

Atualmente o Brasil ocupa lugar de destaque entre os produtores de citros sendo o maior produtor de laranja doce do mundo. Entretanto, essa produção vem sendo gravemente afetada, por doenças como o huanglongbing (HLB), que tem causado perdas significativas para toda cadeia citrícola. O HLB está associado a bactérias Gram-negativas, restritas ao floema das plantas, denominadas Candidatus Liberibacter spp., que além de reduzir a produção de frutos, podem, em casos mais severos da doença, ocasionar a morte da planta. Uma importante estratégia para controle desta doença é a produção de plantas transgênicas expressando genes, especificamente no local de colonização do patógeno. O objetivo deste trabalho foi a obtenção de plantas transgênicas de Citrus sinensis cvs. \'Pera\' e \'Natal\', contendo o gene atacina A (attA) que codifica um peptídeo antibacteriano, dirigido por promotores preferencialmente expressos no floema: AtSuc2 (transportador de sacarose) ou AtPP2 (proteína de floema 2), clonados de Arabidopsis thaliana, ou CsPP2 (proteína de floema 2) clonado de Citrus sinensis. A identificação de 65 plantas transgênicas foi realizada, por meio da análise de PCR. Foi verificado um número menor de eventos transgênicos, utilizando-se a construção gênica pCAtSuc2/attA em relação ao número de eventos transgênicos obtidos com as construções gênicas pCCsPP2/attA e pCAtPP2/attA. Plantas identificadas como transgênicas pela análise de PCR, foram aclimatizadas e transferidas para casa-devegetação certificada para o cultivo de plantas transgênicas. Análises de Southern blot foram realizadas em plantas aclimatizadas que apresentaram desenvolvimento suficiente, confirmando-se a integração do gene attA. A expressão do gene attA foi confirmada pela análise de RT-qPCR. As plantas transgênicas obtidas neste trabalho, contendo o gene attA dirigido por promotores preferencialmente expressos no floema, serão avaliadas em uma etapa futura para resistência a Candidatus Liberibacter spp. / Currently, Brazil is a major citrus producer and the world\'s largest producer of sweet oranges. However, diseases, such as huanglongbing (HLB) have seriously affected this production, causing significant losses in citrus production chain. HLB is associated with Gram-negative bacterias, restricted to the phloem of plants, called Candidatus Liberibacter spp., which besides reducing fruit production, can lead to plant death. An important strategy to control this disease is the production of transgenic plants expressing genes, specifically at the region of pathogen colonization. The aim of this study was to obtain transgenic plants of Citrus sinensis cv. \'Natal\' and \'Pera\', containing the gene attacin A (attA) that encodes an antibacterial peptide, driven by preferentially expressed in phloem promoters: AtSuc2 (sucrose transporter) or AtPP2 (phloem protein 2), cloned from Arabidopsis thaliana, or CsPP2 (phloem protein 2) cloned from Citrus sinensis. The identification of 65 transgenic plants was performed by PCR analysis. A lower number of transgenic events were verified using the gene construct pCAtSuc2/attA in relation events obtained with the gene constructs pCCsPP2/attA and pCAtPP2/attA. Plants identified as transgenic by PCR analysis were acclimatized and transferred to a greenhouse certified for growing transgenic plants. The Southern blot analyses were performed in acclimatized plants, which had sufficiently developed, confirming the integration of attA gene. The expression of attA gene was confirmed by RT-qPCR analysis. The transgenic plants obtained in this work, containing the gene attA directed by preferentially expressed in phloem promoters, will be further evaluated for resistance to Candidatus Liberibacter spp.

Zona pellucida glycoproteins (ZP) are important proteins for maturation of the oocytes in eukaryotes, these proteins are encoded by cluster of zp genes. zp2.3 and zp3.5 genes are expressed during the developing and maturation of the oocytes in zebrafish ovaries. Both of the gens have a CCAAT box in their promoter regions, playing a big role in the expression of the both genes in zebrafish oocytes. The transcription of the genes in the eukaryotes requires transcription factors to initiate and promote the transcription, the transcription factors can bind to the promoter region and initiate the transcription process. The nuclear factor y (NFY) regulates the genes by binding to the CCAAT boxes in their promoter regions, it consist from many subunit such as NF-YA and NF-YB. Here in this study we characterize the expression pattern of NF-YA and NF-YB by screening these genes expression in several organs and tissues, also to determine its roles in the expression of the zp2.3 and zp3.5 genes in the adultzebrafish ovary.