548 resultados para priming


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Dissertação de Mestrado apresentada ao Instituto Superior de Psicologia Aplicada para obtenção de grau de Mestre na especialidade de Psicologia Social e das Organizações.

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The Bacillus subtilis DnaI, DnaB and DnaD proteins load the replicative ring helicase DnaC onto DNA during priming of DNA replication. Here we show that DnaI consists of a C-terminal domain (Cd) with ATPase and DNA-binding activities and an N-terminal domain (Nd) that interacts with the replicative ring helicase. A Zn2+-binding module mediates the interaction with the helicase and C67, C70 and H84 are involved in the coordination of the Zn2+. DnaI binds ATP and exhibits ATPase activity that is not stimulated by ssDNA, because the DNA-binding site on Cd is masked by Nd. The ATPase activity resides on the Cd domain and when detached from the Nd domain, it becomes sensitive to stimulation by ssDNA because its cryptic DNA-binding site is exposed. Therefore, Nd acts as a molecular 'switch' regulating access to the ssDNA binding site on Cd, in response to binding of the helicase. DnaI is sufficient to load the replicative helicase from a complex with six DnaI molecules, so there is no requirement for a dual helicase loader system.

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The essential Bacillus subtilis DnaD and DnaB proteins have been implicated in the initiation of DNA replication. Recently, DNA remodeling activities associated with both proteins were discovered that could provide a link between global or local nucleoid remodeling and initiation of replication. DnaD forms scaffolds and opens up supercoiled plasmids without nicking to form open circular complexes, while DnaB acts as a lateral compaction protein. Here we show that DnaD-mediated opening of supercoiled plasmids is accompanied by significant untwisting of DNA. The net result is the conversion of writhe (Wr) into negative twist (Tw), thus maintaining the linking number (Lk) constant. These changes in supercoiling will reduce the considerable energy required to open up closed circular plectonemic DNA and may be significant in the priming of DNA replication. By comparison, DnaB does not affect significantly the supercoiling of plasmids. Binding of the DnaD C-terminal domain (Cd) to DNA is not sufficient to convert Wr into negative Tw, implying that the formation of scaffolds is essential for duplex untwisting. Overall, our data suggest that the topological effects of the two proteins on supercoiled DNA are different; DnaD opens up, untwists and converts plectonemic DNA to a more paranemic form, whereas DnaB does not affect supercoiling significantly and condenses DNA only via its lateral compaction activity. The significance of these findings in the initiation of DNA replication is discussed.

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T-cell based vaccines against human immunodeficiency virus (HIV) generate specific responses that may limit both transmission and disease progression by controlling viral load. Broad, polyfunctional, and cytotoxic CD4+ T-cell responses have been associated with control of simian immunodeficiency virus/HIV-1 replication, supporting the inclusion of CD4+ T-cell epitopes in vaccine formulations. Plasmid-encoded granulocyte-macrophage colony-stimulating factor (pGM-CSF) co-administration has been shown to induce potent CD4+ T-cell responses and to promote accelerated priming and increased migration of antigen-specific CD4+ T-cells. However, no study has shown whether co-immunisation with pGM-CSF enhances the number of vaccine-induced polyfunctional CD4+ T-cells. Our group has previously developed a DNA vaccine encoding conserved, multiple human leukocyte antigen (HLA)-DR binding HIV-1 subtype B peptides, which elicited broad, polyfunctional and long-lived CD4+ T-cell responses. Here, we show that pGM-CSF co-immunisation improved both magnitude and quality of vaccine-induced T-cell responses, particularly by increasing proliferating CD4+ T-cells that produce simultaneously interferon-γ, tumour necrosis factor-α and interleukin-2. Thus, we believe that the use of pGM-CSF may be helpful for vaccine strategies focused on the activation of anti-HIV CD4+ T-cell immunity.

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Na sociedade moderna, constata-se que os indivíduos multilingues compreendem e falam várias línguas e que essa proficiência tem despertado, ao longo de várias décadas, a curiosidade de numerosos especialistas no assunto (neurolinguistas, psicolinguistas). Sobre este assunto, Dijkstra (2003:11) preconiza que os falantes multilingues devem ter armazenado um vasto número de palavras no seu léxico mental no qual parece ser difícil recuperar uma palavra. Neste contexto, o presente trabalho, que se enquadra no Mestrado em Português Língua Não Materna, oferece uma revisão da literatura sobre o conhecimento neurolinguístico e psicolinguístico da aquisição multilingue do léxico em sujeitos bi / multilingues ; tendo como ponto nevrálgico o controlo inibitório e o acesso lexical. No decorrer desta pesquisa, a qual pôde contar com a participação de trinta e três informantes falantes de português europeu (3 monolingues, 3 bilingues, 5 trilingues e 22 multilingues), procurou-se averiguar se os informantes bi / multilingues seriam mais rápidos e precisos do que os monolingues, aquando da realização de tarefas trilingues de nomeação oral de imagens e de decisão lexical. A investigação realizada fornece dados para uma reflexão sobre o modo como falantes com estas particularidades acedem ao (s) seu (s) reportório (s) linguístico (s), bem como sobre a forma como exercem o controlo inibitório sobre o seu vasto campo linguístico.

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Finnish youth are constantly exposed to music and lyrics in English in their free time. It is likely that this has a positive effect on vocabulary learning. Learning vocabulary while simultaneously accompanied with melodies is likely to result in better learning outcomes. The present thesis covers a study on the vocabulary learning of traditional and music class ninth graders in a south-western upper comprehensive school in Finland, mainly concentrating on vocabulary learning as a by-product of listening to pop music and learning vocabulary through semantic priming. The theoretical background presents viable linguistic arguments and theories, which provide clarity for why it would be possible to learn English vocabulary via listening to pop songs. There is conflicting evidence on the benefits of music on vocabulary learning, and this thesis sets out to shed light on the situation. Additionally, incorporating pop music in English classes could assist in decreasing the gap between real world English and school English. The thesis is a mixed method research study consisting of both quantitative and qualitative research materials. The methodology comprises vocabulary tests both before and after pop music samples and a background questionnaire filled by students. According to the results, all students reported liking listening to music and they clearly listened to English pop music the most. A statistically significant difference was found when analysing the results of the differences in pre- and post-vocabulary tests. However, the traditional class appeared to listen to mainstream pop music more than the students in the music class, and thus it seems likely that the traditional class benefited more from vocabulary learning occurring via listening to pop songs. In conclusion, it can be established that it is possible to learn English vocabulary via listening to pop songs and that students wish their English lectures would involve more music-related vocabulary exercises in the future. Thus, when it comes to school learning, pop songs should be utilised in vocabulary learning, which could also in turn result in more diverse learning and the students could, more easily than before, relate to the themes and topics of the lectures. Furthermore, with the help of pop songs it would be possible to decrease the gap between school English and real-world English.

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Potato is the most important food crop after wheat and rice. A changing climate, coupled with a heightened consumer awareness of how food is produced and legislative changes governing the usage of agrochemicals, means that alternative more integrated and sustainable approaches are needed for crop management practices. Bioprospecting in the Central Andean Highlands resulted in the isolation and in vitro screening of 600 bacterial isolates. The best performing isolates, under in vitro conditions, were field trialled in their home countries. Six of the isolates, Pseudomonas sp. R41805 (Bolivia), Pseudomonas palleroniana R43631 (Peru), Bacillus sp. R47065, R47131, Paenibacillus sp. B3a R49541, and Bacillus simplex M3-4 R49538 (Ecuador), showed significant increase in the yield of potato. Using – omic technologies (i.e. volatilomic, transcriptomic, proteomic and metabolomic), the influence of microbial isolates on plant defence responses was determined. Volatile organic compounds of bacterial isolates were identified using GC/MS. RT-qPCR analysis revealed the significant expression of Ethylene Response Factor 3 (ERF3) and the results of this study suggest that the dual inoculation of potato with Pseudomonas sp. R41805 and Rhizophagus irregularis MUCL 41833 may play a part in the activation of plant defence system via ERF3. The proteomic analysis by 2-DE study has shown that priming by Pseudomonas sp. R41805 can induce the expression of proteins related to photosynthesis and protein folding in in vitro potato plantlets. The metabolomics study has shown that the total glycoalkaloid (TGA) content of greenhouse-grown potato tubers following inoculation with Pseudomonas sp. R41805 did not exceed the acceptable safety limit (200 mg kg-1 FW). As a result of this study, a number of bacteria have been identified with commercial potential that may offer sustainable alternatives in both Andean and European agricultural settings.

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Este trabajo es una revisión de literatura que abarca una selección de artículos disponibles en bases de datos especializadas y publicados en el periodo comprendido entre los años 2006 a 2016 para artículos científicos y entre los años 2000 a 2016 para libros. En total se revisaron: 1 tesis doctoral, 1 tesis magistral, 111 artículos y 9 libros o capítulos de libros. Se presentan diversas definiciones de mindfulness y formas de conceptualizarla, sus mecanismos de acción, sus enfoques psicoterapéuticos predominantes, los efectos de su práctica estable, sus principales campos de acción y la importancia de la formación de los docentes que imparten la práctica. Finalmente se presentan algunas conclusiones acerca del diálogo entre la literatura psicológica sobre mindfulness y algunas de las concepciones de la tradición budista en torno a la meditación.