Les écosystèmes marins côtiers sont soumis à des pressions anthropiques et naturelles qui placent les organismes aquatiques dans des situations de multi-stress.Ce travail doctoral avait pour objectif d’évaluer l’impact et de mieux comprendre les mécanismes d’adaptation des premiers stades de vie de moule exposés à deux facteurs environnementaux majeurs : la pollution métallique et l’accroissement des températures des eaux marines côtières. Les effets induits par l’exposition à deux polluants métalliques modèles (Cu et Ag) et à un stress thermique modéré seuls ou en combinaison ont été évalués sur les stades précoces de développement de deux espèces de moule : Mytilus galloprovincialis et Mytilus edulis ainsi que leurs hybrides. Nos résultats montrent une augmentation significative du pourcentage de larves D ma lformées avec l’augmentation de la température. Par ailleurs, l’Ag apparaît significativement plus toxique que le Cu pour les larves. De plus,la toxicité des métaux s’accroit avec l’augmentation de la température. La coexposition aux métaux et à une température modérée augmente les activités enzymatiques antioxydantes de la catalase (CAT), de la superoxyde dismutase (SOD)et de la glutathion-S-transférase (GST) et accroît le contenu cellulaire en métallothionéines et la peroxydation lipidique. A une température plus élevée de 22 °C,une diminution significative des activités des enzymes antioxydantes est observée.Les dommages à l’ADN chez les larves-D de moule M. gallo provincialis ont été évalués à l’aide du test des comètes avec et sans traitement par la Formamidopyrimidine ADN glycosylase. L’exposition aux métaux et/ou à la température aaugmenté de manière significative les lésions de l’ADN sur les larves de moule,avec un effet plus accentué sur les dommages oxydatifs. Il a été montré également que le Cu et l’Ag sont accumulés différentiellement dans les larves en fonction de la température d’exposition. L’étude de l’expression par RT-qPCR de 18 gènes impliqués dans les défenses antioxydantes, la réparation de l’ADN, l’apoptose,la protéolyse, la transcription, les réponses au stress thermique et la détoxification métallique a été conduite chez les larves-D de moule M. galloprovincialis.En cas d’un stress thermique modéré, une tendance à la surexpression des gènes impliqués dans les défenses cellulaires est observée. Toutefois, en cas d’une coexposition métallique et thermique, les voies d’apoptose et d’altération cellulaires sont activées.Un plugin et une macro ont été développés pour le logiciel ImageJ afin d’évaluer et de caractériser le comportement de nage des larves D de M. galloprovincialis.Une augmentation significative de la vitesse maximale des larves-Davec l’augmentation de la température est observée sans que la vitesse moyenne soit affectée. En revanche, l’exposition au Cu et à l’Ag à 22 °C augmente significativement les vitesses moyenne et maximale des larves. En condition contrôle à18 °C, les larves suivent principalement des trajectoires rectilignes (88 %). Avec l’augmentation de la température et/ou l’exposition aux métaux, les trajectoires circulaires augmentent considérablement au détriment des trajectoires rectilignes.Ces modifications de comportement pourraient être liées d’une part à l’induction de malformations mais aussi à la modulation de l’activité neuronale. Ainsi, une augmentation significative de l’activité acétylcholine estérase (AChE) des larves a été observée suite à l’exposition à un gradient de températures et/ou aux métaux.Des larves D viables et normalement développées ont été obtenues par hybridation de M. galloprovincialis et M. edulis. Les larves de M. edulis apparaissent plus sensibles à la température que les larves de M. galloprovincialis. Aucune différence significative de sensibilité n’a été observée entre les moules hybrides et pures quelque soit le métal testé. (...) / Coastal marine ecosystems are subject to anthropogenic and natural pressures that place aquatic organisms in multi-stress situations. The aim of this doctoral work was to assess the impact and better understand the adaptation mechanisms of mussel early life stages exposed to two major environmental factors : metallic pollution and increasing temperatures in coastal marine waters. The effects induced by exposure to two model metal pollutants (Cu and Ag) and moderatethermal stress alone or in combination were evaluated on the early life stages of development of two mussel species : Mytilus galloprovincialis and Mytilus edulisas well as their hybrids. Our results show a significant increase in the percentageof malformed D-larvae with increasing temperature. Moreover, Ag is significantly more toxic than Cu for larvae. In addition, the toxicity of metals increases with increasing temperature. Co-exposure to metals and a moderate temperature increases the antioxidant activity of catalase (CAT), superoxide dismutase (SOD)and glutathione-S-transferase (GST), and increases cellular content of metallothioneinsand lipid peroxidation. At a higher temperature of 22 °C, a significant decrease in the activities of the antioxidant enzymes is observed. DNA damage in M. galloprovincialis larvae was evaluated using the comet assay with and without Formamidopyrimidine DNA glycosylase treatment. Co-exposure to metals and/ortemperature increase significantly increased DNA lesions on mussel larvae, witha more pronounced effect on oxidative damage. It has also been shown that Cuand Ag are accumulated differently in the larvae as a function of the exposure temperature. The study of the gene expression by RT-qPCR of 18 genes involvedin antioxidant defenses, DNA repair, apoptosis, proteolysis, transcription, thermal stress and metal detoxification was conducted in D-larvae of M. galloprovincialis.Under moderate thermal stress, a tendency of over expression of the cell defense pathways is observed. However, in the case of metallic and thermal co-expositions, the pathways of apoptosis and cellular alteration are rather induced. A pluginand macro were developed for the evaluation and characterization of behavior oflarvae of M. galloprovincialis. A significant increase in the maximum speed of D larvaewith increasing temperature is observed without affecting the mean velocity. In contrast, exposure to Cu and Ag at 22 °C significantly increases average and maximum larval speed. Under controlled conditions at 18 °C, larvae mainly follow rectilinear trajectories (88 %). With increasing temperature and/or exposure to metals, circular trajectories increase considerably at the expense of rectilinear trajectories. These changes in behavior may be related to the induction of malformations but also to the modulation of neuronal activity. Thus, a significant increase in the acetylcholinesterase (AChE) activity of the larvae is observed following exposure to a temperature gradient and/or metals. Viable and normally developed D-larvae were obtained by hybridization of M. galloprovincialis and M.edulis. Larvae of M. edulis are more sensitive to temperature than larvae of M. galloprovincialis.No significant sensitivity differences between species are observed whatever the metal used. Moreover, the hybrid larvae from female M. galloprovincialis are more tolerant to the increase of temperature compared to the hybrid larvae issues from rom female of M. edulis.

The diploma thesis analyses the commercial use of internet in the Czech Republic. It briefly describes the development of the internet towards the development of electronic commerce in the world and in the Czech Republic and defines the various stages of e-commerce. It refers to the various advantages and disadvantages of participation in online trading and analyses development of selected quantitative and qualitative indicators in the past approximately five years. From the consumer perspective it focuses on trends in the number of internet users, their shopping behaviour and their relationship to payments on the internet. Approaching business it examines corporate internet access, website presentation and development of e-shops in CR. The conclusion of the thesis is dedicated to the selected barriers to the further development of online business at both national and international level within the European Union.

As características relacionadas à frutificação do cafeeiro são altamente influenciadas pelo ambiente. Portanto, a associação de análises agronômicas, fenológicas e genômicas são necessárias para o entendimento desse processo. Esses estudos fazem parte do programa de melhoramento genético do café, com o objetivo de obter plantas com maior uniformidade de maturação, duração de ciclo e bebidas específicas. Nesse contexto, os objetivos desse trabalho foram estabelecer padrões agronômicos e fenológicos comparativos entre diferentes cultivares e safras de café, caracterizar genes diferencialmente expressos durante o desenvolvimento dos frutos de cafeeiro arábica e correlacionar genes diferencialmente expressos com atributos fenológicos e agronômicos. Os estudos foram realizados no Centro de Café do IAC em Campinas, SP. As cultivares de Coffea arabica utilizadas foram: Mundo Novo, Catuaí Vermelho, Icatu Vermelho, Obatã e Icatu Precoce, nas safras de 2004/2005 e 2005/2006. Os atributos fenológicos foram avaliados com base no desenvolvimento do ciclo fenológico e na porcentagem de frutos maduros na colheita. As avaliações agronômicas foram avaliadas baseadas nas características tecnológicas do produto como, produção e rendimento, tipos de sementes e tamanho dos grãos. A expressão gênica diferencial foi avaliada por método semi-quantitativo e quantitativo RT-PCR em todos os estádios de desenvolvimento dos frutos. Seqüências de genes específicos foram identificadas por buscas em bancos de dados do Programa Genoma Café e no Genbank, possibilitando a construção de oligonucleotídeos específicos para cada gene. Em geral, os resultados das análises confirmaram a influência ambiental no desenvolvimento dos frutos do cafeeiro, especialmente as variações hídricas e de temperatura. Diferenças significativas foram identificadas entre as cultivares, em relação aos atributos fenológicos e agronômicos, principalmente na safra 2005/2006 quando foi possível discriminar as cultivares quanto à duração do ciclo fenológico. A expressão gênica durante o desenvolvimento dos frutos, seguiu padrão similar entre as cultivares, mesmo comparando diferentes anos agrícolas. Conseqüentemente, essas análises permitiram a identificação de genes candidatos a serem usados como marcadores genéticos das fases de frutificação de C. arabica, como: crescimento (PAL, CHS e manB), transição do crescimento para o início da maturação (csp1, GLA e ER5), maturação e amadurecimento (CS, AAT, ACS, ACO, ETR e ICL) e transição da maturação para o amadurecimento (PAL). Esses genes marcadores em associação com atributos agronômicos e fenológicos, podem ser utilizados como parâmetros moleculares para a definição de estádios adequados à colheita, assegurando composição final específica dos frutos e da qualidade da bebida. / Coffee fruit development and ripening are biological processes largely influenced by environmental conditions. Therefore, the association of agronomic, phenologic and genomic analyses is necessary for a broad comprehension of these processes. This type of approach is already part of coffee breeding programs, which aimed the development of coffee cultivars bearing maturation uniformity, controlled life cycle and specific cup qualities. In this context, the main objectives of this dissertation are to establish comparative patterns of fruit ripening among different coffee cultivars, to characterize gene expression during fruit development, and to correlate possible differential gene expression with agronomic and phenologic traits. All investigations were performed at the experimental field of the Coffee Center/IAC, Campinas, SP. The Coffea arabica cultivars Mundo Novo, Catuaí Vermelho, Icatu Vermelho, Obatã and Icatu Precoce, years 2004/2005 and 2005/2006, were evaluated regarding phenologic cycle and percentage of cherry fruits at harvesting time. Agronomic traits evaluated included productivity and outturn, type of seeds and grain size. Differential gene expression was evaluated through semi-quantitative and quantitative RT-PCR, in all stages of fruit development. Sequences of selected genes were identified through blast searches in the database of the Coffee Genome EST Project and the Genbank, and used to design gene-specific primers. In general, analyzed results confirmed the environmental influence over fruit development, especially temperature variations during evaluated seasons. Significant differences were identified among cultivars regarding phenologic and agronomic traits, especially in the year 2005/2006 where life cycle duration allowed cultivar discrimination. Gene expression during fruit development followed a similar pattern among evaluated cultivars, even when comparing different harvesting years. Therefore, these patterns allowed the identification of candidate genes for use as genetic markers of C. arabica fruit development phases, such as growing (PAL, CHS and manB), transition from growing to maturation (csp1, GLA, ER), maturation and ripening (CS, AAT, ACS, ACO, ETR, ICL) and transition from maturation to ripening (PAL). These gene-markers, in association with agronomic and phenological traits, may be used as molecular parameters for the definition of best colleting stage, ensuring specific final coffee fruit composition and cup quality.

A cultura do porongo ou cabaça [Lagenaria siceraria (Mol.) Standl] é importante para a agricultura familiar em várias regiões do Rio Grande do Sul, Brasil, porque sua rentabilidade, que viabiliza a pequena propriedade, e propicia empregos no campo, no processamento e na comercialização da cuia e artesanatos derivados. A pesquisa foi realizada com a finalidade de estudar a morfologia, fenologia e biologia floral do porongueiro, como base para outras pesquisas, e, também, para subsidiar o manejo da cultura. No experimento sobre densidade de plantas (6.666, 3.333, 2.222 e 1.666 plantas ha-1) avaliou-se a produtividade e a qualidade da cuia, assim como realizou os estudos sobre a fenologia e morfologia. A biologia floral foi estudada em casa de vegetação. Identificou as fases fenológicas e a duração das mesmas em graus-dia acumulados. A relação área foliar por fruto foi muito alta, e a taxa de fecundação baixa; e a cor pardo-escura do pedicelo do fruto serviu como indicador do ponto de colheita. A polinização das flores dependeu de agente biótico. A produtividade aumentou com a densidade de plantas, sem, no entanto, ficar estabelecido a população máxima. Os atributos morfológicos de qualidade da cuia não variaram com a densidade de plantas, no intervalo da pesquisa. / The cultivation of the bottle gourd [Lagenaria siceraria (Mol.) Standl] is an important crop for the familiar farming of several regions of Rio Grande does Sul, states Brazil. Due to profitability enables the small property, and also provides jobs in the field, in the processing and marketing of the gourd and derivative handicrafts. The research was conducted in order to study the morphology, phenology and floral biology of the bottle gourd plant, as a basis for further researches, and also to subsidize the crop management. The experiment of the plants density (6.666, 3.333, 2.222 and 1.666 plants ha-1), in which was evaluated productivity and the quality of the gourd, as well as studies conducted on the phenology and morphology. The floral biology was studied in a greenhouse. Identified phenological phases and the duration of the same in degree-days. The relative leaf area per fruit was very high, and low fertilization rate, and the dark-brown color of the fruit pedicel served as an indicator of harvest time. The pollination of flowers depended on biotic agent. The productivity of the gourd increased with the density of the plants, without establishing the maximum population. The morphological attributes of quality of the gourd did not varied with the plants density, in the interval of the research.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Fontes ricas em compostos antioxidantes sÃo importantes para a manutenÃÃo da saÃde humana, e nos Ãltimos anos, uma maior atenÃÃo tem sido dada aos antioxidantes encontrados nos frutos, isso se deve a estudos epidemiolÃgicos que demonstraram que uma alta ingestÃo de frutos està estritamente associada à reduÃÃo da mortalidade por doenÃas vasculares e cÃnceres, sendo justificado pela atividade antioxidante presente nesses produtos. Com isso, esse experimento objetivou a caracterizaÃÃo fÃsica, qualidade e atividade antioxidante total de quatro clones de cajueiro anÃo precoce: CCP 09, CCP 76, BRS 189 e BRS 265, oriundos do Campo Experimental de Pacajus-CE, selecionando entre esses clones os que apresentam os melhores resultados quando analisados em sete diferentes estÃdios de maturaÃÃo. Sendo caracterizados quanto Ã: tamanho, peso, firmeza, coloraÃÃo, pH, sÃlidos solÃveis (SS), acidez titulÃvel (AT), SS/AT, vitamina C, carotenÃides totais (CT), antocianinas totais (ANT), flavonÃides amarelos (FA), polifenÃis extraÃveis (PE), atividade antioxidante total (AAT), atividade da catalase (CAT), da dismutase do superÃxido (SOD) e da peroxidase do ascorbato (APX). A atividade antioxidante dos pedÃnculos de caju analisados deve-se, sobretudo ao conteÃdo de polifenÃis extraÃveis, e sÃo, portanto, considerados fontes relevantes de compostos antioxidantes, que sÃo necessÃrios para a manutenÃÃo da saÃde humana. CorrelaÃÃes positivas e significativas foram encontradas entre polifenÃis extrable e atividade antioxidante. Os clones que tiveram destaque foram o BRS 265 e o CCP 09. Os maiores teores de Ãcido ascÃrbico foram encontrados no clone BRS 265, durante todo o crescimento e maturaÃÃo dos pedÃnculos. Os mais elevados nÃveis de polifenÃis extraÃveis e capacidade antioxidante total foram encontrados em CCP 09 nos primeiros cinco estÃdios de maturaÃÃo. Os pedÃnculos sÃo consumidos frescos, o que significa que um potencial de mercado, inexplorado para eles existe. Portanto, os clones representam um potencial real para o desenvolvimento de novos produtos com propriedades funcionais, podendo ser explorados pela indÃstria de suplementos antioxidantes ou proveitosamente estudados pela indÃstria alimentar para o desenvolvimento de diversos produtos. / Rich sources of antioxidants are important for the maintenance of human health, and in recent years, increasing attention has been given to the antioxidants found in fruits, this is due to epidemiological studies that have shown that high intake of fruit is associated with reduced mortality due to vascular diseases and cancers, as justified by the antioxidant activity in these products. This work aimed at to physical characterization, quality and total antioxidant activity of four early dwarf cashew clones: CCP 09, CCP 76, BRS 189 and BRS 265 were harvest from the Experimental Farm of Pacajus-CE, selecting among these clones that presenting the best results when analysed in seven different stages of maturation. The clones were characterized to results: size, weight, firmness, color, pH, soluble solids (SS), titrable acidity (AT), SS/TA ratio, vitamin C, total carotenoids (TC), total anthocyanins (TAN), yellow flavonoids (YF), extractable polyphenols (EP), total antioxidant capacity (TAC), catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX). Activity antioxidant the cashew apples analysed are mainly attributed contents of extrable polyphenols, and this therefore considered as relevant sources of antioxidant compounds, which are necessary for the maintenance of human health. Positive and significant correlations were found between extrable polyphenols and antioxidant activity. The clones that had prominence were the BRS 265 and the CCP 09. The highest ascorbic acid contents were found in the BRS 265 clone, throughout growth and maturation of the cashew apples. The highest levels extrable polyphenols and antioxidant capacity were found in CCP 09 in first five maturity stages. These cashew apples are consumed fresh, which means a potential, and untapped, market for them exists. Therefore, these cashew apples represent a real potential for the development of new products with functional properties, may be explored for antioxidant supplements industry or usefully studied by the industry food for the development of diverse products.

Made available in DSpace on 2016-04-27T14:30:55Z (GMT). No. of bitstreams: 1 Cristiane Tavares Casimiro de Oliveira.pdf: 6754831 bytes, checksum: a7161c08ce444d0c247ed69a2263b95f (MD5) Previous issue date: 2012-03-16 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The present research is part of the Programa de Pós-Graduação em Educação: Currículo PUCSP, alligned with the area of research Novas Tecnologias na Educação. The study was conducted within the context of the course in Technical Marketing, offered in the distance learning (DL) mode by the Instituto Federal de Educação Ciência e Tecnologia do Tocantins (IFTO) and derived from the following problem: what is the relationship between the internship on technological appropriation of teachers and pedagogical use of TDIC during the course? The field research was conducted with former teachers, distance tutors, mentors and managers involved in the course. Accordingly, the qualitative and quantitative approach was selected, using direct observation, application of two questionnaires with closed and open questions, semi-structured interviews, records in the logbook, photographic records, and focus group data collected in the AVA. Based on data analyzed within the theoretical framework the following aspects were observed: the teachers under survey are in the second stage of technological appropriation, according to a study Information Commmunication Technology in Educacion: a curriculum for schools and programe of teacher development published by UNESCO (2002) known as Application of TDIC in the area of specialization. However, considering the study by Sandholtz et.al. (1997), we can infer that teachers are in transition between stages of Adaptation and Appropriation. There are indications that the group has an understanding of the pedagogical use of TDIC which is close to a more advanced phase of technological appropriation than those shown by the analysis of AVA records. According to AVA, the use of TDIC did not meet the expectations of a collaborative and interactive work, as shown by the studies of Almeida (2000, 2003, 2005, 2011). Nevertheless, it presented a character content inconsistent with the characteristics of a work based on a constructionist approach. Although the course is based on the constructionist approach, the lack of teacher training for pedagogical work with TDIC was an important factor, hampering the development of activities based on this approach. We concluded that aspects extrinsic to individuals but that permeate the pedagogical work with the TDIC as well as advancement of the stage of technological appropriation of teacher may often overlap and determine the intrinsic ones, insofar as the structural aspects ( therefore non-pedagogical ones ), determine evasion, rotation and the very existence of a faculty. Thus, while reflecting upon the teacher´s appropriation of the technology and pedagogical use of TDIC one must study the context, the questions posed by the environment in which not only appropriation but also the teacher´s action takes place / Esta pesquisa insere-se no Programa de Pós-graduação em Educação: Currículo, da PUC-SP, na linha de pesquisa Novas Tecnologias em Educação. A investigação foi realizada no contexto do curso Técnico em Marketing, oferecido na modalidade à distância (EaD) pelo Instituto Federal de Educação Ciência e Tecnologia do Tocantins (IFTO), a partir do seguinte problema: qual a relação entre o estagio de apropriação tecnológica dos professores e a utilização pedagógica das TDIC no decorrer do curso? A pesquisa de campo foi realizada com professores formadores, tutores a distância, tutores presenciais e gestores do curso. Para tanto, valeu-se da abordagem qualiquantitativa, utilizando observação direta, aplicação de dois questionários com questões fechadas e abertas, entrevistas semiestruturadas, registros em diário de bordo, registro fotográfico, grupo focal e dados levantados no AVA. Com base nos dados analisados à luz do referencial teórico, foi possível perceber que os professores pesquisados encontram-se no segundo estágio de apropriação tecnológica, conforme estudo Padrões de competências em TIC para professores divulgado pela UNESCO (2002) como Aplicação das TDIC na área de especialização. Porém, considerando o estudo publicado por Sandholtz et.al. (1997), pode se inferir que os professores se encontram em fase de transição entre os estágios de Adaptação e de Apropriação. Há indícios de que o grupo tem um entendimento acerca da utilização pedagógica das TDIC que se aproxima de uma fase de apropriação tecnológica mais avançada do que indicou a análise dos registros no AVA. De acordo esses registros, a utilização das TDIC não se aproximou das expectativas de um trabalho colaborativo e interativo, como apontam os estudos de Almeida (2000, 2003, 2005, 2011); no entanto, apresentou um caráter conteudista, incongruente com as características de um trabalho amparado em uma abordagem construcionista. Embora a oferta do curso esteja fundamentada na abordagem construcionista, a falta de formação do professor para o trabalho pedagógico com as TDIC mostrou-se importante fator dificultador do desenvolvimento das atividades nessa abordagem. Conclui-se que os aspectos extrínsecos aos indivíduos, mas que permeiam o trabalho pedagógico com as TDIC, e o avanço do estágio de apropriação tecnológica do professor, muitas vezes, podem se sobrepor e determinar os intrínsecos, na medida em que os aspectos estruturais - e portanto não-pedagógicos- determinam a evasão, rotatividade e a própria existência de um corpo docente. Dessa forma, ao se refletir sobre apropriação tecnológica do professor e uso pedagógico das TDIC, há que se analisar o contexto, as questões postas pelo ambiente em que se dá a apropriação e no qual a atuação do professor se insere

The impact of change facilitator styles (CFS) on elementary teachers' stages of concerns (SoC) about adopting outcome-based education (OBE) in their schools was studied. The group studied was 266 teachers from the Texas Network for Outcome-Based Education. Principal styles are based on the Concerns-Based Adoption Model (CBAM. Styles were determined by the Change Facilitator Style Questionnaire, and teachers' concerns profiles were measured by the Stages of Concern Questionnaire. ANOVA and t tests were conducted to assess the effects of CFS at each of the seven stages of concern. ANOVA assessed teachers' educational level, experience with teaching and OBE, principal gender and type of community related to SoC. Chi-square addressed the relationship among the demographic variables and CFS. With schools as the unit of analysis, significant differences at stages 0,1,2 were found. When teachers were the unit of analysis, significant differences were found at stages 0,1,2, and 3. Concerns of teachers with Initiator style principals were significantly lower at these stages. All teachers demonstrated concerns typical of nonusers, indicating resistance to OBE. Concerns were significantly lower for teachers with master's degree than for bachelor's at stages 0 to 3. Teachers with the least experience with OBE had significantly higher concerns. Chi-square compared change facilitator styles with the demographic variables. The only significant results were more males at the management style than expected. These findings support the CBAMtheory that the initiator style is more effective at impacting SoC and improving success in adopting an innovation. Teacher demographic variables do not affect SoC or CFS. The study indicates problems implementing OBE but suggests effective leadership could impact teachers' concerns.

O parasita Schistosoma mansoni é um dos principais causadores da esquistossomose, doença que acomete 200 milhões de indivíduos no mundo. O parasita possui um complexo ciclo de vida composto de seis estágios evolutivos em dois hospedeiros. S. mansoni possui um sofisticado sistema de interação com seus hospedeiros de modo a escapar da resposta imune e interagir com moléculas produzidas por eles. Alguns trabalhos na literatura descreveram o efeito do TNF-α humano sobre o processo de ovoposição do parasita adulto. Nosso trabalho teve por objetivo analisar o perfil de expressão gênica do S. mansoni ao longo de seus estágios de desenvolvimento, avaliar o efeito do TNF-α humano sobre o perfil de expressão gênica do parasita em dois estágios de desenvolvimento e descrever o gene homólogo ao receptor de TNF-α humano em S. mansoni. Para isso, duas plataformas de microarrays distintas foram utilizadas: uma composta por 4000 sondas de cDNA dupla fita produzida pelo nosso grupo de pesquisa e a outra, composta por 44000 sondas de oligonucleotídeos desenhadas pelo nosso grupo e produzida pela empresa Agilent Technologies. Com estas plataformas foi detectada a expressão de 5798 genes em vermes adultos, sendo que 156 genes apresentavam a transcrição nas fitas senso e anti-senso; 6 destes tiveram confirmadas a transcrição nas duas fitas por transcrição reversa fita específica seguida de PCR em Tempo Real. Adicionalmente foram identificados 229 genes diferencialmente expressos entre vermes adultos machos e fêmeas. A análise de expressão gênica entre 5 estágios de desenvolvimento do parasita mostrou dois tipos de conjuntos de dados: (i) 1423 genes diferencialmente expressos entre dois estágios subseqüentes de desenvolvimento e (ii) 342 genes com expressão enriquecida em um estágio exclusivamente. 68 destes são transcritos intrônicos que não possuem potencial codificador de proteinas. Um gene ortólogo ao receptor de TNF-α humano (SmTNFR) foi clonado e sequenciado. O transcrito SmTNFR possui 1967pb e codifica uma proteína de 599 aminoácidos. Outros 9 genes codificando elementos conservados da via de sinalização de TNF-α também foram identificados no conjunto de ESTs público, revelando uma via completa de sinalização de TNF-α no parasita. Por fim, identificamos com microarrays o conjunto de genes com expressão alterada pela adição de TNF-α humano em esquistossômulos e vermes adultos. 340 genes tiveram expressão alterada em vermes adultos utilizando a plataforma de cDNA. 411 genes sofreram mudanças de expressão em esquistossômulos e 1093 genes em vermes adultos detectados na plataforma de oligonucleotídeos. Este trabalho representa uma importante contribuição no entendimento da relação parasita-hospedeiro em nível molecular. / The parasite Schistosoma mansoni is one of major causative agents of schistosomiasis, a disease which affects 200 million people in the world. The parasite has a complex life cycle with six developmental stages in two hosts. S. mansoni has a sofisticated system of interaction with the hosts, permitting it to escape the immune response and to interact with molecules produced by the hosts. The effect of human TNF-α on adult parasite egg-laying has been described in the literature. The present work intended to analyse the gene expression profile of S. mansoni among its developmental stages, to evaluate the effect of human TNF-α on gene expression profile in two different developmental stages and to describe a homologous gene to human TNF-α receptor in S. mansoni. Two microarrays platfoms were used: one comprised by 4000 cDNA probes and printed by our research group and another, comprised by 44000 oligonucleotide probes designed by our group and printed by Agilent Technologies Company. With these platforms, we detected the expression of 5798 genes in adult worms, of which 156 showed transcription in sense and anti-sense strands; 6 of them had their expression levels confirmed by strand specific Real Time PCR. 229 genes were identified as differentially expressed between male and female adult worms. Gene expression analysis among 5 parasite developmental stages identified two data sets: (i) 1423 differentially expressed genes between two subsequent developmental stages and (ii) 342 expressed genes enriched in one exclusive stage. 68 of them are intronic transcripts with no protein-coding potential. An ortologue to human TNF-α receptor (SmTNFR) was cloned and sequenced. SmTNFR transcritpt has 1967bp and encodes a 599-amino acid protein. Other 9 genes encoding conserved elements of the TNF-α signaling pathway were identified among the public S. mansoni ESTs dataset, thus revealing a complete TNF-α signaling pathway in the parasite. Finally, we identified with microarrays the set of genes that have an altered gene expression upon exposure of schistosomula and adult worms to human TNF-α. 340 genes were identified with altered expression in adult worms using the cDNA platform. 411 genes changed their expression pattern in schistosomula and 1093 genes in adult worms using the oligonucleotide platform. This work represents an important contribution to the understanding of host-parasite interaction at the molecular level.

The shift in health care and nursing philosophy and practice from a holistic approach to a highly technological, cure-oriented approach has been attributed to effective pharmaceuticals made to prolong life. Recently medical professionals have shifted their focus to a combination of spiritual healing and medicine. Hospice care in particular have taken a key interest in integrating spirituality within their health care. The problem is that due to the complications in defining spirituality and appropriate training and education of spirituality within nursing curriculum, assessing patients' spiritual distress may be difficult for many hospice nurses which may be at a loss when attempting to integrate spirituality within their practice. This study used a phenomenological approach to explore the infusion of spirituality in nursing practice and the hospice nurses perceptions of assessing spiritual distress needs of terminally ill patients. Frankl's existential theory and Kubler- Ross's stages of grief theory framed the study. Participants included 8 hospice nurses working in a Pacific Northwestern state. Face-to-face interviews were conducted to explore the essence of the experience of integrating spirituality as well as their views and concerns regarding assessment instruments used to assess spiritual distress. Data was analyzed for content themes. The study found that spiritual courses were merged into hospice nursing as a teaching unity making it difficult for hospice nurses in a Pacific Northwestern State to fully grasp the concept of spirituality. Further findings suggested that only a handful of schools had spiritual nursing as an independent course. The study may impact social change by informing the advancement of hospice nurses and hospice administrators in the practice of including spirituality within healthcare and integrating extensive existential support training within nurses' curriculum.

本研究以三階段資料包絡分析探討在95到97學年度三年間國內近年升格的私立技職院校的經營績效。研究藉由三階段的調整，期望觀察出各技職院校在摒除環境和產出品質的影響下，真實的學校經營效率表現。結果顯示，當學校屬性為中部學校、南部學校、中型規模學校、科技大學等與各項投入差額之間，具有正向關係，而學校屬性為商科重點或其他科的學校、大型學校等與各項投入差額之間，具有負向關係。而在排除外在因素與統計干擾的影響後，本研究發現各高等技職院校的技術效率、純技術效率與規模效率的差異會減少，多數高等技職院校管理效率仍有改善空間，且亦有部分無效率來自於規模無效率，除部分技職院校處於固定規模報酬階段外，其餘多數技職院校皆處於規模報酬遞增階段。 / This paper uses the three-stage data envelopment analysis (DEA) to measure performance efficiency of the private vocational and technological colleges which are elevated recently in Taiwan during the 2006-2008 periods. With the three-stage adjustment, the researcher presented the relationship between the outside environmental factors among the considered colleges and the input slacks. After excluding the impacts from outside environmental factors and statistical noises, the study finds that the differences among the technical efficiency, the pure technical efficiency, and scale efficiency of the considered colleges would be decreased. In addition, it is also found that most considered colleges have opportunity to improve their efficiency and are in the phase of increasing returns to scale.