Dissertations / Theses on the topic '12 Mile'

The following is a design for a new Catholic church for the Sts. Peter and Paul parish in rural 12 Mile, Kentucky. The existing church building is set into a hillside, mostly underground. It was originally intended to be the basement but had to be used as the church once funding ran low during construction. Through my design I wanted to create a space that would be both welcoming for the parishioners to gather each Sunday yet feel spiritual even when mass is not taking place. To achieve this I wanted to make a space used for nothing but the functions of the Liturgy, any circulation, bathrooms, mechanical services or even confessionals should not detract from the space. By using two massive double concrete walls that would encompass all of the non-essential functions of the church, the space between the walls was freed up to be used as the body of the church where nothing but the Liturgy of the Eucharist would take place.
Master of Architecture

Doxorubicin (DOX) is a broad-spectrum and effective chemotherapeutic agent, but its use in oncologic practice is limited by dose-dependent cumulative cardiotoxicity. DOX-induced cardiotoxicity is in large part due to its ability to cause oxidative stress. Multidrug resistance associated protein 1 (MRP1/ABCC1) is a member of the ATP-binding cassette (ABC) transporter superfamily. By effluxing a wide variety of endogenous and exogenous substrates, Mrp1 plays important physiological roles in multiple tissues and also protects normal tissues against toxicants. However, the role of MRP1 in heart is largely unknown. The role of Mrp1 in DOX-induced cardiotoxicity was investigated in Mrp1 null (Mrp1-/-) and their C57BL (WT) littermates. Chronic DOX caused body weight loss and hemotoxicity, and these adverse effects were significantly exacerbated in Mrp1-/- vs WT mice. Importantly, loss of Mrp1 potentiated DOX-induced cardiotoxicity, presenting as worsened cardiac function and more cellular apoptosis in DOX treated Mrp1-/- mice. Mrp1 also protected neonatal mouse cardiomyocytes (CM) and cardiac fibroblasts (CF) culture against DOX cytotoxicity in vitro. This was demonstrated by the decreased cell survival, more apoptosis and more DNA damage in DOX treated Mrp1-/- vs WT cells. In addition, the effects of deletion of Mrp1 was studied on glutathione (GSH)/glutathione disulfide (GSSG) homeostasis, glutathione conjugate of 4-hydroxy-2-nonenal (GS-HNE) accumulation, protein oxidative damage and expression of antioxidant enzymes. Loss of Mrp1 led to significantly higher GSH and GSSG basal levels in heart. Following DOX treatment, Mrp1-/- CM and CF showed increased GSH and GSSG levels vs WT cells. Meanwhile, DOX increased expression of the GSH synthesis enzymes in Mrp1-/- but not WT cells. Thus, increased GSH synthesis may contribute to the further increase in the GSH pool in DOX-treated Mrp1-/- cells. DOX induced comparable increases of GS-HNE concentration in WT and Mrp1-/- mice hearts. Finally, expression of extracellular superoxide dismutase (ECSOD/SOD3) was significantly lower in Mrp1-/- vs. WT CM treated with either saline or DOX. In summary, this study is the first to document a protective role of Mrp1 in DOX-induced cardiotoxicity. It gives critical information regarding the potential adverse sequelae of introduction of MRP1 inhibitors as adjuncts to clinical chemotherapy of multidrug resistant tumors.

Pattern recognition receptors (PRR) detect microbial products and induce cytokines which shape the immunological response. Interleukin-12 (IL-12) is a proinflammatory cytokine important for the differentiation of T helper 1 (Th1) cells which produce IFN-γ to activate macrophages and eradicate intracellular pathogens. In contrast, interleukin-10 (IL-10) is an immunosuppressive cytokine that minimises immune-driven host pathology, but can also lead to defective pathogen clearance. The regulation of IL-10 and IL-12 is therefore of interest due to their central roles in the orchestration of an effective but regulated immune response. C57BL/6 and BALB/c mice differ significantly in their resistance to several pathogens. We observed that macrophages generated from these mice produce reciprocal levels of IL-10 and IL-12 in response to the bacterial ligands LPS and Pam3CSK4, which activate TLR4 and TLR2 respectively, and heat-killed Burkholderia pseudomallei, a Gram-negative bacterium which activates TLR2 and TLR4. We have investigated this differential cytokine production in order to further dissect the molecular mechanisms underlying the regulation of IL-10 and IL-12. Detailed analyses of protein production, signal transduction and transcriptional kinetics have identified a type I IFN dependent, but IL-27 independent mechanism for the differential production of IL-10 in LPS and heat-killed B.pseudomallei stimulated C57BL/6 and BALB/c macrophages. Microarray analysis of LPS stimulated C57BL/6 and BALB/c macrophages further revealed potential regulatory networks that may differ between these mouse strains. These findings highlight key pathways responsible for the regulation of IL-10 and IL-12, and may provide valuable information on factors contributing to the ability of C57BL/6 and BALB/c mice to clear bacterial infections.

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Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil
Leishmaniose visceral zoonótica (LV) é uma doença que afeta homens e cães e é causada por protozoários das espécies Leishmania infantum e Leishmania chagasi. O cão doméstico é o principal reservatório do agente causal. Uma vacina efetiva contra a LV canina poderá contribuir para o controle da infecção e/ou doença humana e canina. A resposta imune protetora contra a LV canina é do tipo celular (Th1). Em nosso laboratório, dentre um grupo de cinco antígenos recombinantes selecionados de uma biblioteca de cDNA de L. chagasi, usando-se uma mistura de soro de cães naturalmente infectados e que exibiam resposta imune humoral e celular específica, um fragmento da extremidade carboxila de uma cinesina com cauda de 6 histidinas (rLci2B-NH6), para permitir a purificação, foi escolhido para a avaliação do seu potencial como candidato a componente de uma vacina contra LV canina. No presente trabalho foi avaliada a imunização de camundongos BALB/c com rLci2BNH6/ saponina em associação com os plasmídeos pcDNA3.1-scmu-IL-12 e/ou pcDNA3.1- mu-IL-2. Os animais imunizados com rLci2B-NH6/saponina em associação com a IL-12 e/ou IL-2 produziram anticorpos IgG, IgG2a e IgG1 específicos. Esplenócitos desses animais exibiram proliferação, mas não produziram IFN-γ ou IL-5 após estimulação in vitro com rLci2B-NH6. Esses resultados sugerem que a adição de pcDNA3.1-mu-IL-2 no protocolo de imunização não é capaz de promover uma resposta imune predominantemente Th1. Também foi avaliada a imunização de camundongos com proteína recombinante quimérica (rLci2-NTCT- NH6) formada pelos domínios não repetitivos presentes nas extremidades amina e carboxila codificadas pelo gene da cinesina. Camundongos BALB/c imunizados com rLci2- NT-CT-NH6 ou uma proteína recombinante químerica controle, contendo 5 domínios repetitivos presentes em rLci2B-NH6 entre os domínios não repetitivos de rLci2-NT-CTNH6, apresentaram produção de IgG, IgG2a e IgG1. Esplenócitos desses animais exibiram proliferação, mas não produziram IFN-γ e apenas esplenócitos dos camundongos imunizados com rLci2-NT-5R-CT-NH6 sintetizaram IL-5, após estimulação in vitro com as respectivas proteínas. Contudo, durante a purificação de rLci2-NT-CT-NH6 e rLci2-NT-5R-CT-NH6 ocorreu intensa proteólise que pode ter resultado na destruição de epítopos capazes de induzir resposta Th1. Previamente, em nosso laboratório, foi observada a morte de alguns camundongos injetados com 100 μg saponina/dose, por isso, doses menores de saponina (50 ou 25 μg) foram avaliadas como adjuvante na indução de resposta imune a rLci2B-NH6. Os resultados da mensuração dos anticorpos foram semelhantes nos três grupos. Curiosamente, somente esplenócitos do grupo injetado com 25 μg/dose exibiram proliferação específica. O grupo injetado com 100 μg/dose produziu IFN-γ e IL-5 e o grupo injetado com 50 μg/dose sintetizou IFN-γ, mas não produziu IL-5, após estimulação in vitro com rLci2B-NH6.
Zoonotic visceral leishmaniasis (VL) is a disease that affects humans and dogs and is caused by protozoan species Leishmania infantum and Leishmania chagasi. There is evidence to suggest that the domestic dog is the main reservoir of the causative agent. An effective vaccine against canine VL may contribute to infection control and/or human and canine disease. The protective immune response against canine VL is cellular (Th1 type). In our laboratory, among a group of five recombinant antigens selected from a cDNA library of L. chagasi, using a mixture of sera from naturally infected dogs that showed humoral and cellular specific responses, one fragment of the carboxyl terminus of a kinesin with a 6 His- Tag (rLci2B-NH6) was chosen to evaluate its potential as a candidate component of a vaccine against canine VL. In the present study, we evaluated the immune response of BALB/c immunized with rLci2B-NH6/saponin in combination with pcDNA3.1-scmu-IL-12 and pcDNA3.1-mu-IL-2. The animals immunized with rLci2B-NH6 associated IL-12 and/or IL-2 produced IgG, IgG2a and IgG1 reactive to rLci2B-NH6. Splenocytes of these animals exhibited proliferation but failed to produce IFN-γ and IL-5 after in vitro stimulation with rLci2B-NH6. These results suggest that adding IL-2 was not enough to induce a predominantly Th1 immune response. We also analyzed the immunization of mice with recombinant chimeric (rLci2-NT-CT-NH6) formed by non-repetitive domains present in the amine and carboxyl ends encoded by the gene for kinesin. BALB/c mice immunized with rLci2-NT-CT-NH6 or a control chimeric recombinant protein containing 5 repeated domains present in rLci2B-NH6 between the domains of non-repetitive rLci2-NT-CT-NH6 showed production of IgG, IgG2a and IgG1. Splenocytes of these animals exhibited proliferation, but failed to produce IFN-γ and only splenocytes from mice immunized with rLci2-NT-5R-CTNH6 synthesized IL-5 after in vitro stimulation with the respective proteins. However, during the purification rLci2-NT-CT-NH6 and rLci2-NT-5R-CT-NH6 occurred intense proteolysis that may have resulted in the destruction of epitopes capable of inducing Th1. Previously, in our laboratory, it was observed the death of some mice injected with 100 μg saponin/dose, therefore, lower doses of saponin (50 or 25 μg) were evaluated as an adjuvant to induce an immune response to rLci2B-NH6. The results of measuring the antibodies were similar in the three groups. Interestingly, only splenocytes from the group injected with 25 μg/dose exhibited specific proliferation. The group injected with 100 μg/dose produced IFN-γ and IL- 5 and the group injected with 50 μg /dose synthesized IFN-γ, but did not produce IL-5 after in vitro stimulation with rLci2B-NH6.

Orientador: Jose Martins Filho
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
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Resumo: Dois temas de suma importância para a saúde foram analisados neste estudo: o aleitamento materno, base para a promoção da saúde desde o nascimento, e a cárie dentária cuja prevenção é essencial para a saúde oral e integral. Investigou-se a relação entre a cárie dentária e o aleitamento materno em 158 crianças de ambos os gêneros, de 2 a 12 anos de idade, atendidas nos consultórios da Escola de Aperfeiçoamento Profissional da Associação Paulista de Cirurgiões-Dentistas. Foi realizado um exame intraoral para obtenção dos índices de cárie e aplicado questionário para análise dos hábitos dos pacientes, em especial os relacionados à amamentação e a sua alimentação, com ênfase no primeiro ano de vida. Os dados obtidos foram analisados estatisticamente e pôde-se concluir que os índices de cárie estão acima das metas estabelecidas pela Organização Mundial de Saúde; que a prevalência da cárie dentária foi menor quando houve maior duração do período de amamentação exclusiva e do período total de aleitamento materno, misto e/ou complementado, e teve relação com a época do início da higiene oral e com as escolaridades materna e paterna
Abstract: Two subjects of utmost importance for health has been analyzed in this study: the breast feeding which is the basis for the promotion of health from birth, and cavities whose prevention is essential for both oral and integral health. It was investigated the relation between cavities and breast feeding in 158 children of both genres, from 2 to 12 years of age treated in consulting offices at School of Professional Improvement of São Paulo Association of Surgeon-Dentists. An intra-oral exam was done and a questionnaire was applied to get the indexes of cavities and analysis of the patients? habits, mainly the ones related to breast feeding and his feeding, with emphasis in the first year of life. The results obtained were statistically analyzed and it was concluded that the indexes of cavities are above the goals established by World Health Organization; that the prevalence of cavities is related to the duration of the exclusive breast feeding period and with the total breast feeding time (both mixed and/or complemented), with the time of the beginning of oral hygiene and also with the maternal and paternal educational background
Doutorado
Saude da Criança e do Adolescente
Doutor em Saude da Criança e do Adolescente

A presente pesquisa teve por objetivo analisar as influências do modelo de gestão nas estratégias de duas cooperativas mistas de leite e café da região sul de Minas Gerais. Especificamente, fez-se a análise externa do ambiente (ameaças e oportunidades) ao qual as cooperativas estão expostas e também a análise interna (pontos fortes e fracos) dessas organizações; investigou-se as articulações políticas entre as chapas envolvidas no último processo eleitoral; analisou-se o grau da influência da mudança de gestão na formulação e implementação das estratégias nas cooperativas, comparando-as com indicadores de eficiência financeira, econômica e social. Finalmente, foram levantadas as opiniões dos cooperados a respeito da validade dos princípios doutrinários que regem o cooperativismo e do modelo de gestão de suas cooperativas. Quanto à metodologia, a pesquisa classifica-se como descritiva conclusiva e, também na tipologia de estudos de casos. Adotou-se, para a coleta de dados, a triangulação de técnicas: entrevista pessoal, análise documental (parte qualitativa da pesquisa) e questionário estruturado (parte quantitativa) ao qual aplicou-se a análise discriminante para verificar as divergências nas opiniões dos cooperados considerando-se o porte de produtor rural. Optou-se por omitir os nomes das cooperativas para manter o sigilo tanto das organizações quanto dos entrevistados. Os resultados demonstraram que as principais ameaças do ambiente às cooperativas são: concorrência de produtos lácteos (principalmente com as multinacionais e o mercado informal de leite); concorrência com os distribuidores de insumos agropecuários; dependência das cooperativas em relação à Cooperativa Central de Laticínios do Estado de São Paulo (Central Paulista), a qual estão filiadas e adaptação do modelo de gestão cooperativista ao regime capitalista. As principais oportunidades detectadas são: possibilidade de ampliação do mercado regional de produtos lácteos a ser explorado com as marcas próprias; boa localização geográfica das cooperativas no país (região sudeste); duplicação da rodovia Fernão Dias, o que deve ampliar o desenvolvimento do mercado regional e, finalmente, diversificação vertical na industrialização de café. Os entrevistados têm consciência de que as cooperativas estão expostas às influências ambientais, entretanto, há consenso que a resposta dessas organizações ao ambiente tem sido lenta e burocrática devido, principalmente, ao fato das decisões estratégicas serem tomadas de forma colegiada em esporádicas assembléias que, no geral, têm apresentado baixa participação dos associados. As articulações políticas que ocorreram na última eleição em ambas cooperativas, podem ser consideradas como um momento crítico nessas organizações, provocando reflexos traumáticos em todos os envolvidos no processo (membros das chapas vencedora e perdedora, cooperados e funcionários). Após a eleição na Cooperativa MINAS-1, verificou-se que metade dos membros da nova Diretoria Executiva e Conselho de Administração havia participado em cargos diretivos nas duas gestões anteriores, o que contribuiu para a continuidade de todas as estratégias formuladas na gestão anterior que tiveram influências consideradas positivas, negativas e nulas. Na Cooperativa MINAS-2, após a eleição a chapa de oposição assumiu o poder. Nenhum dos membros da nova Diretoria Executiva e Conselho de Administração havia participado de cargos administrativos nas duas gestões anteriores. Devido a divergências ideológicas nessa transição, houve descontinuidade em algumas estratégias traçadas na gestão anterior, fato que julgou-se como influência negativa. Nas demais estratégias, as influências foram consideradas como positivas, negativas e nulas. Nesse sentido, verificou-se que houve relação entre a estabilidade dos dirigentes nos cargos administrativos com a continuidade das estratégias das cooperativas. A comparação das estratégias com índices de eficiência financeira, econômica e social demonstrou que a Cooperativa MINAS-1 apresentou-se mais eficiente em relação a Cooperativa MINAS-2 devido, principalmente, a estrutura organizacional mais enxuta e menores custos administrativos. Por outro lado, a Cooperativa MINAS-2 mostrou-se mais eficiente na prestação de serviços. Os resultados da análise discriminante, considerando-se o porte de produtor rural, demonstrou que as opiniões do grupo dos mini e pequenos produtores divergiram do grupo dos médios e grandes produtores nas seguintes variáveis da pesquisa: o princípio da adesão livre deve ser eliminado; a cooperativa deve desenvolver novos critérios para pagamento dos cooperados por volume e qualidade; a cooperação entre cooperativas deve ser feita com base na eficiência e interesses estratégicos dos parceiros, seja outra cooperativa ou até mesmo uma empresa não cooperativa; o capital social deve ser corrigido com base nos juros de mercado e não em no máximo 12% ao ano; não é bom para a cooperativa quando grupos se apropriam do poder e ficam muito tempo na diretoria e, finalmente, o cooperado que não cumpre seu papel deveria ser excluído da cooperativa. Contudo, considerando o modelo de gestão das cooperativas, verificou-se que os fatores que mais influenciaram nas estratégias das cooperativas estudadas foram: o amadorismo gerencial dos dirigentes, a ausência de planejamento estratégico e, principalmente, as divergências políticas entre os grupos de interesse envolvidos na disputa pelo poder.
The present research aimed at analyzing the influences of the management model on the strategies of two milk and coffee cooperatives in the southern region of Minas Gerais. Specifically speaking, an external analysis of the environment (threats and oportunities) in which cooperatives in general are exposed to was performed, as well as an internal analysis (positive and negative aspects) of such organizations; political articulations among the groups involved in the previous elections were investigated; the degree of influence of the change of the board, in formulation and implemention of the cooperatives strategies, was analysed, related to financial, economical and social indicators. Finally, the cooperators opinions about the validity of the principles which guide cooperativism, as well as their opinions about their own cooperatives management model were raised. As for the methodology, the research is descriptive conclusive, and also in the typology of case study. As for the collection of data, three techniques were adopted: individual interviews, documental analysis (the qualitative part of the research), and structured questionaire (the quantitative part) to which the discriminant analysis was applied aiming at verifying divergences in the cooperators opinions considering the rural producers size. The names of the cooperatives were omitted to protect both the organizations and the ones interviewed. The results showed that the main environmental threats to the cooperatives are: competition with dairy products (especially those produced by multinationals and the informal milk market); competition with agricultural input dealers; the cooperatives dependence on the Cooperativa Central de Laticínios do Estado de São Paulo (Central Paulista) to which they are affiliated, and the adaptation of the management model to the capitalist regime. The main oportunities detected were: the possibility of enlargement in the regions market of milk products to be explored through the producers own brand of products; convenient location of the cooperatives throughout the countrys southeastern region; the improvement of the Fernão Dias highway, which should enhance the regional market development, and finally, the vertical diversification in the coffee industrialization. The ones interviewed are aware that the cooperatives are exposed to enviromental influences. There is, however, a consensus that the responses from those organizations to the environment has been slow and bureaucratic, due mainly to the fact that the strategical decisions are made in a collegiate manner, in sporadic assembly meetings which, in general, have presented low participation of associates. The political articulations that occurred in the last elections, in both cooperatives, may as well be seen as having caused critical moments in those organizations, traumatically reflecting on everyone involved in the process (members of the winning and of the defeated groups, cooperators and employees). Following the elections at Cooperativa MINAS-1, it was verified that half of the members of the new Executive Board and Management Council had had directive job positions in the two previous administrations, which contributed to the continuity of the strategies formulated in the previous administrations whose influences were considered as positive, negative and nule. At Cooperativa MINAS 2, the oposition group took over. None of the members of the Executive Board and Council of Administration had had any administrative jobs at that cooperative during the two previous administration periods. Due to ideological divergences in the transition some strategies adopted by the previous administrators were not continued, which was considered as negative influence. As for the other strategies, the influences were considered as positive, negative and nule. In that sense, it was verified that there was a relationship between the directors stability in their administrative positions and the maintainance of the cooperatives strategies. The comparison of the strategies to financial, economical and social efficiency indexes has shown that Cooperativa MINAS-1 had a higher performance than that of Cooperativa MINAS-2 due to a smaller organizational structure and lower administrative costs. On the other hand, however, Cooperativa MINAS-2 proved to be more efficient as far as service rendering is concerned. The results of the discriminant analysis, considering the rural producers size, showed that the opinions given by the mini-producers and the small-size producers group diverged from those given by the medium and big producersgroup in the following reasearch variables: the principle of free adhesion should be eliminated; the cooperative should develop new criteria as for the cooperators payment based on volume and quality; the cooperation among cooperatives should be performed with basis on the partners efficiency and strategical interests, be it or not another cooperative or even a non-cooperative entity. Capital stock should be updated with basis on market interests instead of the usual maximum of 12% a year; it is not advantageous, or even good for the cooperative, when groups take over the power and stay in charge for a long period of time and, finally, cooperator who fail to play his part should be excluded from the cooperative. However, considering the management models in the cooperatives, it was verified that the most influencial factors in the cooperatives investigated were: the directors amateur managerial level, the absence of strategical planning, and, mainly, the political divergences among the interest groups involved in the dispute for power.

A paracoccidioidomicose (PCM) é uma doença sistêmica de caráter granulomatoso, causada pelo fungo termodimórfico Paracoccidioides spp. A PCM é endêmica na America Latina e aproximadamente 80% do pacientes vivem no território brasileiro. O tratamento medicamentoso é eficiente, entretanto, é longo e vários pacientes acabam abandonando e recidivas são comuns neste grupo. A utilização de uma vacina terapêutica poderia resultar na redução do tempo de tratamento assim como, recuperar a resposta imune do hospedeiro frente ao fungo. As vacinas de DNA são uma abordagem promissora na imunoterapia e podem ser injetadas por via intramuscular, intradérmica ou via mucosa. As proteínas de choque térmico (HSPs) são proteínas que estão ligadas a homeostase celular e também possuem efeitos imunológicos em diversos casos como doenças infecciosas e autoimunes. No presente trabalho, analisamos o esquema vacinal terapêutico em camundongos BALB/c previamente infectados intratraquealmente com 3x105 leveduras de P. brasiliensis Pb18, 60 dias depois, submetidos a imunização com pcDNA3 contendo sequências codificadoras de PbHSP60 e/ou IL-12 murina e/ou vetor vazio. Foi observada redução significativa no número de unidades formadoras de colônia (UFCs) nos pulmões de camundongos imunizados com PbHSP60. Os grupos que receberam PbHSP60+pcDNA3 vazio ou PbHSP60x2 apresentaram os maiores índices de redução da cargas fúngicas. A inclusão do plasmídeo contendo o inserto de mIL-12, resultou em um efeito deletério. A análise dos cortes histológicos indicou que os animais vacinados apresentavam áreas bem preservadas e com poucos ou nenhum foco de granuloma. Detectamos um perfil de citocinas típico Th1/Th2. Nossos resultados sugerem que a imunização utilizando plasmídeo contendo o inserto HSP60, tem grande potencial vacinal
The paracoccidioidomycosis (PCM) is a systemic granulomatous disease of character, caused by the thermally dimorphic fungus Paracoccidioides spp. The PCM is endemic in Latin America and approximately 80% of patients are living in Brazil. The medical treatment is effective, however, is long and many patients end up abandoning and relapses are common in this group.The use of a therapeutic vaccine could result in the reducing time of treatment as well as recover the host immune response against the fungus. DNA vaccines are a promising approach for immunotherapy and can be injected by intramuscular, intradermal, or mucosal route. The heat shock proteins (HSPs) are proteins that are linked to cellular homeostasis and also have immunological effects in many cases as infectious and autoimmune diseases. In the present study, we analyzed the therapeutic vaccine schedule in BALB/c mice previously infected intratracheally with 3x105 yeast of P. brasiliensis strain 18, and 60 days after, undergoing immunization with pcDNA3 containing coding sequences PbHSP60 and / or murine IL-12 and / or empty vector. Significant reduction was observed in the number of colony forming units (CFU) in the lungs of mice immunized with PbHSP60. The groups that received empty pcDNA3 and PbHSP60 or PbHSP60x2 have higher rates of reduced fungal loads. The inclusion of the plasmid containing the insert mIL-12 resulted in a deleterious effect. The analysis of histological sections indicated that vaccinated animals had wellpreserved, with few or no focus of granuloma areas. It was detected a profile typical Th1/Th2 cytokines. Our results suggest that immunization using plasmid containing the insert HSP60 vaccine has great potential

O Sistema Agroindustrial do Leite (SAG) foi alvo de mudanças institucionais importantes nos últimos anos. Dentre essas mudanças podemos ressaltar a IN51 em 2002 e que mais tarde foi substituída pela IN 62 no ano de 2012. O objetivo da normativa foi de criar parâmetros técnicos de produção, identidade e qualidade do leite tipo A, de regular tecnicamente a identidade e qualidade do leite cru refrigerado, pasteurizado e regular tecnicamente a coleta de leite cru refrigerado e seu transporte a granel. Esse estudo pretende discutir: Quais os impactos das mudanças institucionais nas relações entre produtores rurais e laticínios no SAG do leite no Brasil? Foram utilizados como referênciais três correntes teóricas que se integram entre si. A primeira abordagem, a Nova Economia Institucional (NEI), na vertente macro, analisa o papel das instituições sobre o desempenho econômico tendo como precursor o laureado Nobel Douglass North. Para North (1990), a instituições são as regras do jogo de uma sociedade, enquanto os jogadores são as organizações. As instituições definem os incentivos aos agentes econômicos estabelecendo as condições em que as atividades empresariais possam florescer. Ou seja, as instituições importam para definir o ambiente de negócios. A segunda abordagem, a Organização Industrial (OI) fornece instrumentos para analisar os determinantes da estrutura de mercado, tais como ganhos de escala e escopo ou outros condicionantes de criação de barreiras à entrada. A terceira, a Economia dos Custos de Transação (ECT), permite analisar as escolhas estratégicas a partir dos condicionantes institucionais e de mercado, assim como fornece suporte para responder a questão de pesquisa, qual seja: entender a lógica econômica das mudanças nas estruturas de governança decorrentes das novas regulamentações que afetam o SAG do leite. Com base nas abordagens teóricas foram desenvolvidas duas preposição concorrentes, sobre o impacto na relação entre produtores e processadores. A primeira sob a ótica da ECT propôs que a regulamentação leva à queda da especificidade dos ativos (temporal e locacional) e, portanto relações mais tênues entre os produtores rurais e laticínios deveriam emergir. A segunda propôs o oposto, que a IN leva ao aumento do mercado relevante e, sob a ótica da OI, cresce a necessidade de estratégias dos laticínios de relações mais próximas com o produtor (fidelização). Nas regiões produtoras de leite estudadas (Ponte Nova-Minas Gerais e Castro - Paraná) constatou-se aumento de estratégias de fidelização, com contratos formais de pagamento por qualidade e assistência técnica.
The Milk Agribusiness System (MAS) has been subject of important institutional changes in the past few years. Among these changes, we highlight the IN51 as of 2002, that was later replaced by IN 62 as of 2012. The objective of these change was that of creating normative technical parameters for the production, identity and quality of the A type milk, of technically regulating the identity and quality of refrigerated pasteurized and raw milk, and that of technically regulating the collection of refrigerated raw milk and its transportation in bulk. This study aims to discuss: What are the impacts of institutional changes in the relationship between dairy farmers and dairy regulators in the MAS in Brazil? We have used as benchmark three theoretical approaches that integrate together. The first approach, the New Institutional Economics (NIE), in its macro view, examines the role of institutions on economic performance, having as a precursor the Nobel laureate Douglass North. According to North (1990), institutions are the rules of a society, while the players are the organizations. Institutions define the incentives given to economic agents, by establishing the conditions in which business activities can flourish. This means that institutions matter to define the business environment. The second approach, the Industrial Organization (IO), provides tools to analyze the determinants of market structure, such as economies of scale and scope or other constraints; to create entry barriers. The third, Transaction Cost Economics (TCE) allows for the analysis of strategic choices out of the institutional and market constraints, as well as provides support to answer the research question, which is: understanding the economic logic of the governance shifts arising from the new regulations affecting the MAS. Based on these theoretical approaches, two competing hypotheses about the impact on the relationship between producers and processors have been developed. The first, in the perspective of TCE, proposed that regulation leads to lower asset specificity (temporal and locational) and therefore, more tenuous relations between farmers and dairy producers should emerge. The second proposed the opposite, that IO leads to an increase in the relevant market and, from the perspective of IO, a growth in the need for forthcoming strategies between processors and producers (loyalty relationship). In the milk-producing regions studied (Ponte Nova Minas Gerais and Castro - Paraná) we have found and increase in loyalty strategies, with formal contracts of payment in exchange for quality and service.

We analyzed gene expression during t10c12-CLA-induced body fat reduction in a polygenic obese line of mice. Adult mice (N=185) were allotted to a 2x2 factorial experiment consisting of a non-obese (ICR-control) and an obese (M16-selected) line of mice fed a 7% fat, purified diet containing either 1% linoleic acid (LA) or 1% t10c12-CLA. Body weight (BW) gain by day 14 was 12% lower in CLA compared to LA fed mice (P<0.0001). By day 14, t10c12-CLA reduced weights of epididymal, mesenteric and brown adipose tissues as a percentage of BW in both lines by 30, 27 and 58%, respectively, and increased liver weight/BW by 34% (P<0.0001). Total RNA was isolated and pooled (4-5 mice per composite) from epididymal adipose (day 5 & 14) and liver (day 14) of the obese mice to analyze gene expression profiles using Agilent mouse oligo microarray slides (4 per tissue?day) representing >20,000 genes. Numbers of genes differentially expressed by ≥ two fold in epididymal adipose (day 5 & 14) and liver (day 14) were 29, 125, and 80, respectively. Of particular interest in adipose, CLA putatively increased expression of the uncoupling proteins (1 and 2), carnitine palmitoyltransferase (L and M), and carnitine translocase, but decreased expression of PPAR-gamma, GLUT-4, perilipin, caveolin-1, adiponectin and resistin (P<0.01). In conclusion, this experiment has revealed candidate genes that will be useful in elucidating mechanisms underlying the potent anti-adipogenic effects of t10c12-CLA.

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The trans-10, cis-12 conjugated linoleic acid is known to inhibit fat synthesis in the mammary gland of many animal species. The objective of this study was to analyze the effect of PPARγ on mammary lipogenesis and gene expression, through a specific chemical agonist and its response to trans-10, cis-12 CLA. Twenty four 70 ± 3 days in milk (DIM) and body weight (BW) 60 ± 0.45 kg lactating ewes were randomly assigned to one of the four treatments for 7 days: 1) Control (100 mL/day of sterile saline solution, intravenous); 2) Thiazolidinedione (TZD) (4mg/kg of BW/day in 100 mL of sterile saline solution, intravenously); 3) CLA (27g/d orally-dosed rumen-unprotected 29.9% trans-10, cis-12 CLA); 4) TZD+CLA. Compared to Control, milk fat was 22.3% lower in CLA (P=0.05), tended to be 20.7% lower in TZD+CLA (P=0.06) and did not change in the TZD treatment (P=0.39). The lactose content and milk yield and production of components were not affected by treatments. The protein content was lower in the CLA compared to TZD (P=0.01) and tended to be higher with the TZD compared to Control (P=0.08). In the mammary gland, CLA reduced expression of PPARγ, SREBP1 and SCD1, but TZD did not stimulate the expression of these genes. In adipose tissue, PPARγ expression was not affected by treatments, whereas the SREBP1 had more expression in TZD treatment, CLA and TZD + CLA and the SCD1 had more expression with TZD+CLA, compared to the other treatments. In conclusion, the CLA negatively affected the expression of genes involved in lipid synthesis and the TZD was unable to stimulate gene expression and lipogenesis in mammary gland
O ácido linoleico conjugado trans-10, cis-12 é conhecido por inibir a síntese de gordura na glândula mamária de diversas espécies animais. O objetivo deste estudo foi analisar o efeito do PPARγ sobre a lipogênese mamária e expressão gênica, através de um agonista químico específico e sua resposta ao CLA trans-10, cis-12. Vinte e quatro ovelhas em lactação, com 70 ± 3 dias em lactação (DEL) e peso corporal (PC) de 60 ± 0,45 kg, foram distribuídas aleatoriamente em um dos quatro tratamentos, por 7 dias: 1) Controle (100 mL/dia de solução salina estéril, intravenosa); 2) Tiazolidinediona (TZD) (4mg/kg de PC/dia em 100 mL de solução salina estéril, intravenosa); 3) CLA (27g/dia de CLA desprotegido da bio-hidrogenação ruminal, com 29,9% de trans-10, cis-12, dosado oralmente); 4) TZD+CLA. Comparado ao Controle, a gordura do leite foi 22,3% menor no tratamento CLA (P=0,05), tendeu a ser 20,7% menor no tratamento TZD+CLA (P=0,06) e o TZD não afetou o teor de gordura (P=0,39). O teor de lactose e as produções de leite e dos componentes não foram afetados pelos tratamentos. O teor de proteína foi menor no CLA comparado ao TZD (P=0,01) e tendeu a ser maior com o TZD comparado ao Controle (P=0,08). Na glândula mamária, o CLA reduziu a expressão do PPARγ, SREBP1 e SCD1, porém o TZD não estimulou a expressão destes. No tecido adiposo, a expressão do PPARγ não foi afetada pelos tratamentos, enquanto que o SREBP1 teve maior expressão nos tratamentos TZD, CLA E TZD+CLA e a SCD1 teve maior expressão com TZD+CLA, comparada aos demais tratamentos. Concluindo, o CLA afetou negativamente a expressão dos genes envolvidos na síntese de lipídeos e o TZD não estimulou a expressão gênica e lipogênese na glândula mamária

Thesis (Ph.D.)--Southern Illinois University Carbondale, 2007.
"Department of Molecular, Cellular, and Systemic Physiology." Includes bibliographical references (leaves 71-82). Also available online.

Orientador: Ricardo de Lima Zollner
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas
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Resumo: O camundongo NOD é utilizado como modelo experimental por desenvolver diabetes mellitus tipo 1 (DM-1) espontâneo similar ao diabetes mellitus humano resultando na destruição das ilhotas, orquestrada pelos linfócitos T que induzem a liberação de citocinas, entre elas a IL-1, promovendo o processo inflamatório. A diacereína possui propriedades antiinflamatórias, inibindo IL-1. Entretanto, os mecanismos envolvidos na modulação imunológica ainda estão incompletamente compreendidos. No presente estudo, soro, pâncreas, ilhotas e células mononucleares esplênicas de animais tratados com diferentes doses (5, 10 e 50mg/kg/dia) de diacereína e o grupo controle o qual recebeu solução salina, foram isolados para investigar a expressão de IL-1ß, IFN-?, IL-12 e TNF-a e a relação com o início do diabetes, aspectos morfológicos e a dose-dependência da diacereína. Os resultados mostraram o aumento do RNAm de células esplênicas e ilhotas e a diminuição da concentração no soro de IL-1 em 10mg/kg/dia de diacereína, comparado com os outros grupos e aumento da concentração de IFN-? no soro do grupo de 50mg/kg/dia. Esses resultados sugerem que a diacereína interfere na instalação do diabetes e a freqüência, pela modulação de citocinas pró-inflamatórias como IL-1ß, IFN-?, IL-12 e TNF-a. Além disso, os dados sugerem regulação pós-transcricional de IL-1ß pela diacereína
Abstract: The NOD mice is used as a experimental model of spontaneously develop type1 diabetes mellitus (DM1) similar to human disease resulting in the destruction of the islet, orchestrated by T lymphocytes that induce a cytokines release, among then the IL-1, promoting inflammatory process. Diacerhein has antiinflammatory properties, inhibiting IL-1. However, the mechanisms involved in immune modulation are still incompletely understood. In the present study, serum, pancreas islets and spleen mononuclear cells from animals treated by different doses (5, 10 and 50mg/kg/day) and the group control which received solution saline, were isolated to investigate the IL-1ß, IFN-?, IL-12 and TNF-a expression and relationship with diabetes onset, morphological aspects and diacerein dose dependence. The results demonstrate upregulation of mRNA spleen cells and islets and downregulation of serum concentration of IL-1 in 10mg/kg/day diacerein treated group compared with others groups and increase of IFN-? serum concentration for 50 mg/kg/day group. These results suggest that the diacerhein interfere with the diabetes onset and frequency by modulation of pro-inflammatory cytokines as IL-1ß, TNF-a, IFN -? and IL- 12. Furthermore, the data suggest a post-transcriptional downregulation for IL-1ß by diacerhein
Mestrado
Ciencias Basicas
Mestre em Clinica Medica

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Feeding trans-10, cis-12 CLA has resulted in milk fat depression in dairy cows, ewes and goats. However, its effects in goats are less understood in comparison with other ruminant species. Therefore, the objective of the present study was to evaluate the effect of increasing doses of a rumen unprotected trans-10, cis-12 CLA supplement on milk production, secretion of milk components and milk fatty acid profile in dairy goats. Eight Toggenburg non pregnant goats (135 ± 21 DIM) were used in a duplicate 4 x 4 Latin Square design (12-d treatment periods separated by 6-d intervals) according to the order of lactation (primiparous and multiparous). The animals within each group were randomly assigned to the following dietary treatments: CLA0: 45 g/d of Megalac-E; CLA15: 30 g/d of Megalac-E+15 g/d CLA; CLA30: 15 g/d of Megalac-E+30 g/d of CLA; CLA45: 45 g/d of CLA. The lipid supplements were mixed in the concentrate and fed individually to the animals after morning and afternoon milkings. The CLA supplement (Luta-CLA 60) had 29.9% of trans-10, cis-12 CLA as methyl esters, resulting in doses of 0, 4.48, 8.97 and 13.45 g/d of this CLA isomer for CLA0, CLA15, CLA30 and CLA45 treatments, respectively. Dry matter intake, milk yield, content and secretion of milk protein and lactose, body condition score and body weight were unaffected by the dietary treatments., The comparison of milk fat content and yield observed on the last day of each treatment period (d 12) with those found on day 0 showed that CLA0 treatment increased the milk fat content. Thus, milk fat content and yield on day 0 were considered as 100%. The CLA15, CLA30 and CLA45 treatments reduced milk fat yield by 8.1, 26.1 e 32.7% and milk fat content by 4.5, 21.5 e 28.3%, respectively. The increase in dietary trans-10, cis-12 CLA dose reduced the concentration of milk fatty acids arising from de novo synthesis and increased the concentration of those derived from blood circulation. However, the milk secretions of both classes of fatty acids were reduced linearly as the CLA dose increased. The increase in dietary trans-10, cis-12 CLA also caused a linear reduction in milk fat C14:1/C14:0, C16:1/C16:0, C17:1/C17:0 and C18:1/C18:0 dessaturase indexes. Milk fat trans-10, cis-12 CLA content and secretion increased in goats fed increasing doses of trans-10, cis-12 CLA, which, corresponded to transfer efficiencies from diet to milk of 1.18, 1.17 and 1.21% for CLA15, CLA30 and CLA45 treatments, respectively. The energy balance (EB) increased linearly in goats fed increasing doses of trans-10, cis-12 CLA
O fornecimento de CLA trans-10, cis-12 através da dieta tem resultado na depressão da gordura do leite de bovinos, ovinos e caprinos. No entanto, os seus efeitos em caprinos são menos compreendidos perante as outras espécies de ruminantes. Assim, o objetivo do presente estudo foi avaliar o efeito de doses crescentes de CLA trans-10, cis-12 desprotegido da bio-hidrogenação ruminal sobre a produção, secreção dos componentes e perfil de ácidos graxos do leite de cabras leiteiras. Para isso, foram utilizados 8 animais da raça Toggenburg não prenhes com 135 ± 21 dias em lactação, separadas de acordo com o número de lactações para compor dois Quadrados Latinos 4 X 4 (12 dias de tratamento separados por 6 dias de intervalo), um formado por primíparas e o outro por multíparas. Os animais foram submetidos aleatoriamente aos seguintes tratamentos: CLA0) 45 g/d de Megalac-E; CLA15) 30 g/d de Megalac-E+15 g/d de CLA; CLA30) 15 g/d de Megalac- E+30 g/d de CLA; CLA45) 45 g/d de CLA. Os tratamentos foram inclusos no concentrado fornecido aos animais, assim, todos os tratamentos continham 45 g/d de suplemento lipídico. O suplemento de CLA (Luta-CLA 60) continha 29,9% do isômero trans-10, cis- 12 na forma de ésteres metílicos, perfazendo doses de 0, 4,48, 8,97 e 13,45 g/d deste ácido graxo para os respectivos tratamentos citados anteriormente. O consumo de matéria seca de silagem, a produção de leite, a produção e o teor de proteína e lactose do leite, o escore de condição corporal e o peso vivo não foram afetados pelas doses de CLA trans-10, cis- 12. Ao final de cada período experimental (d 12), a avaliação do teor e produção de gordura do leite em relação ao encontrado no dia 0, demonstrou que o tratamento CLA0 levou ao aumento da gordura do leite. Assim, as comparações de produção e teor de gordura do leite foram feitas considerando os valores obtidos no dia 0 como 100%. Os tratamentos CLA15, CLA30 e CLA45 reduziram, respectivamente, a produção de gordura do leite em 8,1, 26,1 e 32,7% e o teor de gordura do leite em 4,5, 21,5 e 28,3%. O uso de doses crescentes de CLA trans-10, cis-12 na dieta alterou o perfil de ácidos graxos do leite, com reduções da concentração dos ácidos graxos oriundos da síntese de novo e aumento da concentração dos provenientes da circulação sanguínea. Porém, quando a secreção destes mesmos ácidos graxos foi avaliada houve redução da secreção de ambas as classes de ácidos graxos. A suplementação de CLA trans-10, cis-12 também acarretou na redução dos índices de dessaturação C14:1/C14:0, C16:1/C16:0, C17:1/C17:0 e C18:1/C18:0. Tanto a concentração como a secreção do CLA trans-10, cis-12 aumentaram conforme o seu aumento na dieta, e correspondeu a uma eficiência de transferência da 1,18, 1,17 e 1,21% para os tratamentos CLA15, CLA30 e CLA45 (respectivamente), sendo que as mesmas não diferiram entre si. Quando o balanço energético (BE) dos animais foi avaliado, as inclusões crescentes do CLA trans-10, cis-12 na dieta aumentaram linearmente o BE

Tsang Sai Leong.
Thesis submitted in: December 2004.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2005.
Includes bibliographical references (leaves 96-104).
Abstracts in English and Chinese.
Abstract (in Chinese) --- p.(i)
Abstract --- p.(iii)
Table of contents --- p.(v)
Chapter Chapter 1 --- Introduction
Chapter 1.1 --- The interferon --- p.1
Chapter 1.1.1 --- About type I IFN --- p.1
Chapter 1.1.2 --- IFN α/β receptor and signal transduction --- p.3
Chapter 1.1.3 --- IFN induction --- p.3
Chapter 1.1.4 --- Functions --- p.4
Chapter 1.1.5 --- MuIFN α subtypes --- p.8
Chapter 1.1.6 --- Gene expression --- p.9
Chapter 1.2 --- Aim of study: Functions and gene expression --- p.9
Chapter Chapter 2 --- Materials and Methods
Chapter 2.1 --- Materials --- p.15
Chapter 2.1.1 --- "Cell line, bacterial strain, virus strain and plasmid vector" --- p.15
Chapter 2.1.2 --- Chemicals --- p.20
Chapter 2.1.3 --- "Culture media, buffer and other solutions" --- p.20
Chapter 2.1.4 --- Reagents and nucleic acids --- p.21
Chapter 2.1.5 --- Reaction kits --- p.22
Chapter 2.1.6 --- Solutions --- p.22
Chapter 2.1.7 --- Major equipments --- p.24
Chapter 2.1.8 --- Primers used --- p.24
Chapter 2.2 --- Methods --- p.26
Chapter 2.2.1 --- "Cloning of MuIFN αl2, MuIFN αl and MuIFN α4 from L929 genomic DNA and their subcloning into pEGFP-Nl mammalian expression vector" --- p.26
Chapter 2.2.1.1 --- PCR of MuIFN αl2 --- p.26
Chapter 2.2.1.2 --- Gel purification of MuIFN αl2 PCR product --- p.26
Chapter 2.2.1.3 --- Ligation of MuIFN αl2 PCR product into pGEM-T vector --- p.26
Chapter 2.2.1.4 --- Sequencing of clones which were positive in PCR screening --- p.26
Chapter 2.2.1.5 --- Subcloning of the gene from pGEM-T vector to pEGFP-Nl --- p.28
Chapter 2.2.1.6 --- Construction of expression vectors for MuIFN αl and MuIFN a4 gene --- p.28
Chapter 2.2.2 --- Preparation ofplasmid DNA --- p.29
Chapter 2.2.3 --- Preparation of cell culture medium --- p.30
Chapter 2.2.4 --- Production of recombinant MuIFN α (rMuIFN α) --- p.30
Chapter 2.2.5 --- Production of native MuIFN α by polyI：polyC induction --- p.31
Chapter 2.2.6 --- Influenza A virus strain A/NWS/33 preparation and titration --- p.31
Chapter 2.2.7 --- Virus infection in Influenza A virus challenge assay --- p.32
Chapter 2.2.8 --- Cell culture techniques --- p.32
Chapter 2.2.9 --- "MTT cell proliferation assay of JCS cell line, for measuring MuIFN α anti-proliferation activity" --- p.33
Chapter 2.2.10 --- Quantitative analysis of MuIFN α --- p.34
Chapter 2.2.11 --- Flow cytofluorometric analysis of cell cycle of MuIFN α treated JCS cells by propidium iodide staining --- p.34
Chapter 2.2.12 --- FACS study on the effect of MuIFN α on MHC-I up-regulation in JCS cells --- p.35
Chapter 2.2.13 --- FACS study on the effect of MuIFN α on MHC-I up-regulation on primary macrophages from Balb/c mice --- p.35
Chapter 2.2.14 --- Anti-viral activity by transfection of MuIFN α gene --- p.36
Chapter 2.2.15 --- Sequencing of MuIFN al2 coding region from genomic DNA of L929 and JCS cell lines --- p.37
Chapter 2.2.16 --- "RNA extraction from L929 cell lines, with or without Influenza A virus infection or polyI:polyC induction" --- p.37
Chapter 2.2.17 --- RNA extraction from tissues of Balb/c mouse --- p.38
Chapter 2.2.18 --- Reverse transcription --- p.39
Chapter 2.2.19 --- Polymerase Chain Reaction (PCR) --- p.39
Chapter Chapter 3 --- Results
Chapter 3.1 --- Overview --- p.40
Chapter 3.2 --- "Subcloning of MuIFN α 12, MuIFN αl and MuIFN α4 coding sequences into the pEGFP-Nl vector" --- p.40
Chapter 3.3 --- The growth inhibitory effect of different MuIFN α subtypes on murine myeloid leukemia cell line JCS --- p.41
Chapter 3.4 --- Quantitation of MuIFN α subtype samples --- p.50
Chapter 3.5 --- Cell cycle analysis of MuIFN α treated JCS cells --- p.50
Chapter 3.6 --- FACS analysis of the effect of different MuIFN α subtypes on MHC-I expression in JCS cell line --- p.57
Chapter 3.7 --- FACS analysis of the effect of different MuIFN α subtypes on MHC-I expression in primary macrophages in Balb/c mice --- p.65
Chapter 3.8 --- Effect of MuIFN α subtype transgenes on L929 cells challenged with Influenza A virus --- p.72
Chapter 3.9 --- Sequencing of MuIFN αl2 coding region from genomic DNA of L929 and JCS cell line --- p.78
Chapter 3.10 --- "MuIFN αl2 expression in untreated, Influenza A virus infected or polyl:polyC induced L929 cells" --- p.78
Chapter 3.11 --- Detection of MuIFN α12 transcripts in tissues of the 8-10 week untreated Balb/c mice --- p.85
Chapter Chapter 4 --- Discussion --- p.89
Chapter 4.1 --- Overview --- p.89
Chapter 4.2 --- rMuIFN α 12 has anti-proliferative and apoptotic effects on JCS cell line --- p.89
Chapter 4.3 --- "Up-regulation of MHC-I expression in JCS cells and primary macrophages by rMuIFN αl2, rMuIFN αl, rMuIFN α4 and mixed type I IFN" --- p.91
Chapter 4.4. --- Transfection of MuIFN α12 gene could induce anti-viral state in L929 cell line --- p.91
Chapter 4.5 --- Gene regulation of MuIFN al2 in L929 cells infected with Influenza A virus or induced by polyI:polyC --- p.92
Chapter 4.6 --- Gene expression of MuIFN αl2 in different tissues of Balb/c mice --- p.94
Conclusion --- p.95
Reference List --- p.96
List of figures:
Fig. 1.1 The 3D structure of recombinant human interferon alpha (HuIFN α) subtype 2B --- p.11
Fig. 1.2 Current model of lFN induction --- p.12
Fig. 1.3 Activation of RNase L --- p.13
Fig. 2.1 Graphical map of plasmid vector pEGFP-Nl --- p.17

The As- and Mo- bearing secondary mineral phases formed during the neutralization of uranium mill wastes were studied for a variety of ore blends including current and future ore sources at the Key Lake milling operation, northern Saskatchewan, Canada. A lab-scale plant model was employed to characterize secondary precipitates obtained during the mill waste neutralization process. Three scenarios of ore blends were processed through the lab-scale plant to produce mill waste solutions for neutralization before combination into final tailings. Slurry samples (n = 12) were collected from the secondary precipitates formed during the neutralization of mill wastes (raffinate) by precipitation with Ca(OH)2 (slaked lime) from pH 1.5 to 10.5. Synchrotron based X-ray absorption spectroscopy of mill and lab-scale plant precipitates showed arsenate adsorbed to ferrihydrite was the dominant As mineral phase regardless of pH or sample blend (53-77%), with fractional contributions from ferric arsenates, and adsorption to aluminum phases (AlOHSO4, As(OH)3 and hydrotalcite). Molybdate adsorbed to ferrihydrite was the dominant Mo mineral phase, regardless of pH or sample blend, with fractional contribution decreasing with increasing pH, and minor contributions from calcium molybdate, ferric molybdate and nickel molybdate. These results were used in geochemical modelling to predict the source terms for these mineral phases in tailings facilities. Sequestration of As and Mo in the model showed solubility was controlled by adsorption to both Fe and Al oxide surfaces as well as by direct precipitation with other dissolved constituents (Ni, Ca and SO4).The models developed pH profiles of mineral phase precipitation to explain the solubility of As, Mo, Fe, Al, Mg and Ni during sequestration from pH 1.5 to 10.5 that were consistent regardless of ore blend used in simulations. Since adsorption of anions to the surface of ferrihydrite has been shown to slow conversion to crystalline forms of Fe oxides (goethite and hematite) and sequestration of arsenate effectively controls As solubility at high pH (pH >10), As-bearing mineral phases are expected to be stable for thousands of years. With adsorption as well as direct precipitation considered, Mo phases though effectively sequestering below pH 8, became unstable and released Mo back into the tailings porewater (pH >10), as predicted by the thermodynamic model. Historical data obtained from as-discharged tailings as well as previously published U mill tailings studies agree with these findings.

Environmental microbiology has traditionally been performed using culture-based methods. However, in the last few decades, the emergence of molecular methods has changed the field considerably. The latest development in this area has been the introduction of next-generation sequencing, including pyrosequencing. These technologies allow the massively parallel sequencing of millions of DNA strands and represent a major development in sequencing technologies. The purpose of this study was to use both pyrosequencing and traditional culture-based techniques to investigate the diversity and dynamics of bacterial populations within milk and untreated sewage sludge samples. Pasteurized and raw milk samples were collected from grocery stores and dairies within Texas. Milk samples were analyzed by plating, pyrosequencing, and an assay for the presence of cell-cell signaling molecules. Samples were processed, stored, and then evaluated again for spoilage microflora. The results of this study showed that raw milk had a considerably higher bacterial load, more diversity between samples, and a significantly higher concentration of pathogens than pasteurized milk. Additionally, this study provided evidence for varying spoilage microflora between raw and pasteurized milk, as well as evidence for the production of cell-cell signaling molecules by bacterial organisms involved in milk spoilage. Four samplings of untreated sewage sludge were collected from wastewater treatment plants in seven different municipalities across the United States. Samples were subjected to quantification of selected bacterial organisms by culture and a pyrosequencing analysis was performed on extracted community DNA. The results of this study showed that untreated sewage sludge is inhabited by a huge diversity of microorganisms and that certain municipalities may have distinct bacterial populations that are conserved over time. Additionally, this study provided some evidence for seasonal differences in several of the major bacterial phyla. Lastly, this study emphasized the challenges of comparing results obtained by culture and pyrosequencing. In conclusion, this study showed that both milk and sewage are highly diverse, dynamic environments that can contain organisms of public health concern. The use of both culture-based methods and pyrosequencing in this study proved a complementary approach, providing a more comprehensive picture of both microbial environments.

碩士
中山醫學大學
醫學研究所
96
Angiostrongylus cantonensis, the rat lugworm, is the principal cause of eosinophilic meningitis or meningoencephalitis. In the previous study, matrix metalloproteinase-9 (MMP-9) has been proved involved in the pathogenesis of eosinophilic meningitis or meningoencephalitis. In this study, we used 5-week BALB/c male mice infected 50 A. cantonensis larvae. The results showed that MMP-12 is found in the brain tissues and cerebral spinal fruid (CSF) of infected mice and significantly increases from day 5 PI. Similarly, the expression of elastin, the substrate of MMP-12 is also increased. By immunohistochemistry, confirmed that MMP-12 distributed in leukocytes that infiltrated into subarachnoid space and mainly expressed in macrophages and eosinophils. The mice treated with albendazole (10mg/kg per day) alone, doxycycline (30mg/kg per day) alone, or a combination of albendazole (10mg/kg per day) and doxycycline (30mg/kg per day) for 7 consecutive days on day 5 and 15 post-inoculation (PI), respectively. The treatment of albendazole-doxycycline co-therapy on day 5 PI significantly reduced the infiltration of leukocytes and the expression of MMP-12 in infected mice. These results suggested that MMP-12 may contribute to the develop of eosinophilic meningitis or meningoencephalitis caused by A. cantonensis. This new approach may be beneficial to treat the parasitic meningitis or meningoencephalitis.

Within the context of contemporary politics, Christian, Muslim and Liberal traditions have been, in many instances, at odds with each other regarding how humankind‘s social political future should be ordered. Such a conflicting condition has been aggravated by the global circulation of democratic ideals, which has significantly disseminated Western liberal values and made those ideals an almost universal desirable social commodity. In support of this argument, one can observe the unprecedented and controversial assumption that liberal democracy has become the ultimate form of political governance. It is in the context of these end-times liberal aspirations, whether self desired or imposed through external pressure, that some competing and conflicting elements are introduced into the political landscape of Christian and Muslim groups. By presenting itself as the universal and final solution for humanity‘s future, liberalism appears to create uneasiness among religious people who, indeed, see its secular and religious-privatizing tendencies as a secular eschatological competitor. Despite this perceived end-times conflict, there may be hope for a constructive dialogue among these groups.

In recent years there have been two cycloterpenals, molecules consisting of a cyclohexadienal core, isolated from nature. Cyclocitral, the condensation product of citral, has been isolated from the North Sea bryozoans Flustra foliacea. In the human eyes, cycloretinal has been isolated and is a toxic by product of the vision cycle. This retinal dimer is believed to contribute to age related macular degeneration, the leading cause of blindness in the elderly. In 1992, it was discovered that if [beta]-ionylideneacetaldehyde was incubated with beta-lactoglobulin ([beta]-LG), the principal whey protein in dairy milk, that it would mediate the formation of cyclo-[beta]-ional. No follow up studies were performed on this protein mediated reaction or what biological activities these cycloterpenals may possess. This dissertation investigates the biological roles of substituted cyclohexadienes including cycloterpenals and cyclohexadiene enaminonitriles. To mimic the protein mediated reaction, we developed a synthetic procedure to produce a wide array of cycloterpenal by utilizing L-proline. Over 100 cycloterpenals were synthesized and screened for their biological activities against an array of cell based screens. The phenotypic effects of these cycloterpenals were screened against a PC12 assay where dramatic effects were observed on neurite outgrowth. During the synthesis of starting materials for the production of our cycloterpenal library, it was discovered that if excess base was added to the Horner-Wadsworth-Emmons reaction between a methyl-ketone and diethyl-(cyanomethyl)-phosphonate, conversion of the [alpha]-[beta]-unsaturated nitrile into an enaminonitrile was observed. This new synthetic procedure was optimized to generate a library of enaminonitriles as well as their quinazoline derivatives. The work within also includes the investigation of the [beta]-LG mediated reaction formation of cycloterpenals with natural and unnatural [beta]-methyl aldehydes. We were able to demonstrate that [beta]-LG could mediate the conversion of [alpha],[beta]-unsaturated aldehydes into their corresponding cycloterpenal. In vitro analysis was also performed with store bought milks and the [beta]-LG present was able to mediate the formation of cyclocitral. An in vivo experiment was also performed by utilizing New Zealand White rabbits to demonstrate the formation of cycloretinal within the blood stream by feeding a source of [beta]-LG with retinal. Interestingly, in human blood, [beta]-LG is present at concentrations of 0.7-1.2 g/dL. The protein has been identified within drusen pigments and lipofuscin granules that accumulate in the retina of macular degeneration patients. As humans do not produce beta-lactoglobulin, the source of this protein is from milk and milk products. With these experiments, we clearly demonstrate that under the appropriate conditions, cycloretinal can be produced with [beta]-LG. We have clearly established a direct link between [beta]-LG chemistry and age-related macular degeneration.

If renewable energy and biofuels are to attain success in the market place, each step of their production and the system as a whole must be optimized to increase material and energy efficiency, reduce production cost and create a competitive alternative to fossil fuels. Systems optimization techniques may be applied to product selection, process design and integration, feedstock procurement and supply chain management to improve performance. This work addresses two problems facing a biorefinery: technology selection and feedstock scheduling in the face of varying feedstock supply and cost. Also addressed is the optimization of a biorefinery supply chain with respect to distributed processing of biomass to bio-products via preprocessing hubs versus centralized processing and facility location selection. Two formulations are proposed that present a systematic approach to address each problem. Case studies are included to demonstrate model capabilities for both formulations. The scheduling model results display model sensitivity to feedstock price and transport distance penalized through carbon dioxide emissions. The distributed model shows that hubs may be used to extend the operating radius of a biorefinery and thereby increase profits.

Unique among the intracellular lipid binding proteins, acyl CoA binding protein (ACBP) exclusively binds long chain fatty acyl CoAs (LCFA-CoAs). To test if ACBP is an essential protein in mammals, the ACBP gene was ablated by homologous recombination in mice. While ACBP heterozygotes appeared phenotypically normal, intercrossing of the heterozygotes did not result in any live homozygous deficient (null) ACBP^(-/-) pups. Heterozygous and wild type embryos were detected at all postimplantation stages, but no homozygous ACBP null embryos were obtained– suggesting that an embryonic lethality occurred at a preimplantation stage of development, or that embryos never formed. While ACBP null embryos were not detected at any blastocyst stage, ACBP null embryos were detected at the morula (8- cell), cleavage (2-cell), and zygote (1-cell) preimplantation stages. Two other LCFACoA binding proteins, sterol carrier protein-2 (SCP-2) and sterol carrier protein-x (SCPx) were significantly upregulated at these stages. These findings demonstrate for the first time that ACBP is an essential protein required for embryonic development and its loss of function may be initially compensated by concomitant upregulation of two other LCFA-CoA binding proteins only at the earliest preimplantation stages. The fact that ACBP is the first known intracellular lipid binding protein whose deletion results in embryonic lethality suggests its vital importance in mammals.

This dissertation has four interrelated studies, namely (1) the characterization of milk purchase choices which included the purchase of organic milk, both organic and conventional milk and conventional milk only; (2) the estimation of a single-equation household demand function for organic and conventional milk; (3) the assessment of binary choice models for organic milk using the Brier Probability score and Yates partition, and (4) the estimation of demand systems that addresses the censoring issue through the use of econometric techniques. In the first paper, the study utilized the estimation of both multinomial logit and probit models in examining a set of causal socio-demographic variables in explaining the purchase of three outcome milk choices namely organic milk, organic and conventional milk and conventional milk only. These crucial variables include income, household size, education level and employment of household head, race, ethnicity and region. Using the 2004 Nielsen Homescan Panel, the second study used the Heckman two-step procedure in calculating the own-price, cross-price, and income elasticities by estimating the demand relationships for both organic and conventional milk. Results indicated that organic and conventional milk are substitutes. Also, an asymmetric pattern existed with regard to the substitution patterns of the respective milk types. Likewise, the third study showed that predictive outcomes from binary choice models associated with organic milk can be enhanced with the use of the Brier score method. In this case, specifications omitting important socio-demographic variables reduced the variability of predicted probabilities and therefore limited its sorting ability. The last study estimated both censored Almost Ideal Demand Systems (AIDS) and Quadratic Almost Ideal Demand System (QUAIDS) specifications in modeling nonalcoholic beverages. In this research, five estimation techniques were used which included the usage of Iterated Seemingly Unrelated Regression (ITSUR), two stage methods such as the Heien and Wessells (1990) and the Shonkwiler and Yen (1999) approaches, Generalized Maximum Entropy and the Dong, Gould and Kaiser (2004a) methods. The findings of the study showed that at various censoring techniques, price elasticity estimates were observed to have greater variability in highly censored nonalcoholic beverage items such as tea, coffee and bottled water.

碩士
臺北醫學大學
醫學科學研究所
96
The T helper type 2 cells (Th2) play a curial role in the initiation of allergic asthma. Asthma is characterized by reversible airway obstruction, eosinophilic infiltration, airway remodeling, mucus hypersecretion and airway hyperresponsiveness. In our studies, we use IL-10 or IL-12 expessing adenoviral vector to observe whether combination treatment can suppress the airway inflammation of allergic asthma. Firstly, we applied the Ad-IL-10/IL-12 to the bone marrow derived dendritic cells (DCs) and found such modified DCs expressed increased surface makers of CD 80, CD 86 and MHC class II. These modified DCs drived activated CD4+ T cells to differiate to specific T cell line. These special T cells secreted high levels of IL-10 and IFN-γ and suppressed the proliferation of effector T cells through cell-cell contact rather than the cytokine mediation. In the in vivo experiments, Ad-IL-10/IL-12 infected DCs were injected into OVA-induced asthmatic animal model to investigate whether modified DCs could prevent the development of asthma. The data showed that modified DCs suppressed the development of airway hyper-responsiveness, reduced airway inflammation and enhanced the expression of IL-10 and IFN-γ. We suggest that Ad-IL-10/IL-12 infected DCs can prevent the rise of asthma effectively. In addition, this modified DCs applied to the established asthmtic animal model and the same therapeutic effect was observed. In conclusion, we suggest that Ad-IL-10/IL-12 infected DCs induce a group of IL-10+ and IFN-γ+ T cells in vivo and such specific T cells can suppress the airway inflammation of asthmatic animal. Thus, such modified DCs may provide a more appropriate tool in future application of gene therapy for allergic diseases.

This dissertation is a comprehensive description of the grammar of Sierra Popoluca (SP, aka Soteapanec), a Mixe-Zoquean language spoken by approximately 28,000 people in Veracruz, Mexico. This grammar begins with an introduction to the language, its language family, a typological overview of the language, a brief history of my fieldwork, and the methodology undertaken in this study. The grammar continues with a description of the phonology of SP, followed by an overview of the word classes, including verbs, nouns, relational nouns/postpositions, adjectives, adverbs, and numbers, and formative types. The bulk of this grammar is devoted to the morphosyntax of Sierra Popoluca, including nouns and nominal morphology, verbs and verbal morphology, and the mechanisms for expressing tense, aspect, mood, and modality. The grammar also describes the complex predicate formation strategies and sentence-level syntax. A compilation of interlinearized texts appears in the appendix. Sierra Popoluca is an agglutinating, polysynthetic, head-marking language with a complex verbal system. It has ergative-absolutive alignment and its grammar is sensitive to animacy and saliency hierarchies, evident in the case marking and `split' plural systems. Its constituent order is verb-initial, although word order is pragmatically determined. Sierra Popoluca has a number of strategies to form complex predicates, which include verb serialization, noun incorporation, and dependent verb constructions. The data available in this grammar contributes a body of data and descriptive analysis to broad theoretical areas of linguistics as well as existing research on the Mixe-Zoquean language family, languages throughout Mesoamerica, and especially the Gulf branch of the Zoquean family.
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The objective of this study was to examine the effects of feeding extruded and non-extruded mixtures of wheat dried distillers grains with solubles with peas (WP) or canola meal (WC) on ruminal fermentation, total tract nutrient digestion and milk production in dairy cows. Eight dairy cows (712 ± 54 kg body weight; 90 ± 31 days-in-milk at the beginning of the experiment) were used in a replicated 4 × 4 Latin square design (28-d periods) with a 2 × 2 factorial arrangement of dietary treatments. Four cows in one Latin square were fitted with rumen cannulas for the measurement of ruminal fermentation characteristics. Treatment diets contained either WP or WC combinations fed in an extruded or non-extruded form (16% of DMI). Diets were isonitrogenous (17.1% crude protein; CP) and contained approximately 53% concentrate and 47% forage (DM basis). Dietary treatment had no significant effect on DMI (P > 0.10). Starch intake was higher for cows fed extruded diets compared to those fed non-extruded diets (P = 0.028) and was also higher for cows fed WP compared to those fed WC (P = 0.042). Cows fed extruded diets had higher apparent ruminal digestion of DM (P = 0.02) and a tendency (P = 0.05) for a higher OM apparently digested in the rumen compared to those fed non-extruded diets. Total tract digestibilities of organic matter (P < 0.01), CP (P < 0.01), ether extract (P < 0.01) and starch (P = 0.047) were higher for cows fed extruded diets compared to those fed non-extruded diets. Total tract digestibility of ether extract was lower (P = 0.011) but digestibility of starch was higher (P < 0.01) and CP digestibility tended to be higher (P = 0.08) for cows fed WP compared to those fed WC. Fecal N excretion was lower in cows fed extruded diets compared to those fed non-extruded diets (P < 0.01), but there was no difference in N retention, productive N, RDP or RUP between diets (P > 0.10). Ruminal pH was higher for cows fed non-extruded WC compared to those fed extruded WC, but there was no difference between WP diets (interaction; P = 0.047). Ruminal acetate displayed the opposite interaction where concentration was highest for cows fed extruded WC and lowest for those fed non-extruded WC but there was no difference between WP diets (interaction; P = 0.019). Ruminal ammonia-N concentration tended to be higher for cows fed WC compared to those fed WP (P = 0.06). Ruminal propionate concentration was higher for cows fed extruded diets compare to those fed non-extruded diets (P = 0.026). Ruminal isobutyrate concentration was higher for cows fed WC compared to those fed WP (P < 0.01). Ruminal butyrate (P < 0.01) and isovalerate (P < 0.01) concentrations were higher for cows fed extruded WC compared to those fed non-extruded WC, but concentrations decreased for cows fed extruded WP compared to those fed non-extruded WP. Plasma glucose concentration was higher for cows fed WC compared to those fed WP but concentration was highest for cows fed extruded WC but lowest for cows fed extruded WP (interaction; P < 0.01). Milk protein yield (P = 0.047) was higher and milk yield tended to be higher (P = 0.06) for cows fed WP compared to those fed WC diets. Milk protein content was not affected by diet; however, milk fat content (P = 0.04) and MUN (P = 0.011) were lower, whereas milk yield (P = 0.030), 3.5% fat corrected milk yield (P = 0.027), milk fat yield (P = 0.027), lactose content (P = 0.011) and lactose yield (P < 0.01) were higher in cows fed the extruded diets compared to those fed non-extruded diets. In summary, these results indicate that extrusion had positive effects on overall milk production and total tract nutrient digestion.

Des enfants de 9 à 12 ans peuvent-ils assumer le rôle de metteur en scène dans un groupe de création composé d'adultes? Cette question nous a amenée à nous pencher sur la mise en place d'un processus qui leur permettrait d'assumer ce rôle. Dans ce mémoire, nous ne décrirons donc pas, au jour le jour, chaque atelier, mais mettrons plutôt en évidence les éléments clefs qui ont permis, à notre avis, à nos jeunes metteurs en scène d'assumer complètement leur rôle et aux adultes de ne pas dénaturer leurs choix artistiques. De même, ce document n'a pas eu pour objectif de cerner ou d'analyser les particularités de l'imaginaire enfantin en comparant une mise en scène enfantine, à celle d'un adulte. En effet, si cette recherche a été envisagée, il nous est très vite apparu qu'elle demanderait des moyens humains, financiers et de temps dépassant largement le cadre d'une recherche de maÎtrise. Notre premier chapitre rend compte de l'évolution progressive des notions de jeunes publics et de théâtre pour enfants ainsi que de la légitimation des activités théâtrales à l'école. Nous y soulignons également de quelle manière notre recherche-création s'inscrit de façon naturelle dans ce courant qui, depuis plus de trente ans, a fait évoluer le théâtre jeunes publics. De même, y sont présentées les raisons qui nous ont poussée à choisir comme cadre théorique le rôle du metteur en scène et, décrites, les tâches qui lui incombent. Enfin, nous y précisons brièvement l'éthique de travail que les adultes ont dû respecter à l'égard des enfants. Ces balises posées, nous décrivons dans notre deuxième chapitre le processus et les stratégies qui permirent aux enfants d'écrire des textes fidèles à leur imaginaire et de se préparer à assumer leur futur rôle de metteur en scène. Abordant le coeur de notre recherche, nous présentons et analysons, dans notre troisième chapitre, le processus, les exercices et les stratégies qui ont été mis en place afin de permettre aux jeunes d'assumer pleinement leur fonction, bien que de façon peu conventionnelle. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Enfants metteurs en scène, Mise en scène, Théâtre jeunes publics, Enseignement de la mise en scène.